HUMREP35 Suppl1 ESHRE 2020 - V4 PDF

Abstracts
36th Virtual Annual Meeting of the

European Society of
Human Reproduction and Embryology
5 to 8 July 2020
BK-OUP-ESHRE2020_ABSTRACTS-200150-Prelims.indd 1 22/06/20 5:11 PM

Abstracts
36th Virtual Annual Meeting of the
European Society of
Human Reproduction and Embryology,
5 to 8 July 2020
The abstracts are available on-line to all Human Reproduction/Update/Molecular Human Reproduction
subscribers and are also freely available to all visitors to the following website www.humrep.oxfordjournals.
org, and on the ESHRE website: www.eshre.eu
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human
human
reproduction
reproduction Editor-in-Chief
C.B. Lambalk (The Netherlands)
Deputy Editors
Editor-in-Chief
C. De Geyter (Switzerland) K. Kirkegaard (Denmark) M. van Wely (The Netherlands)
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Professor Cornelis (Nils) Lambalk
Associate Editors
Deputy Editors
Jason Abbott (Australia) Lucia De Santis (Italy) Maris Laan (Estonia) Peter Ruane (United Kingdom)
Samir Babayev (USA) C. DeZaira
Geyter K. KirkegaardSusan MM.van
Donarelli (Italy) Wely
Laird (United Kingdom) Geetanjali Sachdeva (India)
Elisabetta Baldi (Italy) Hakan Duran (USA) William Ledger (Australia) Laurel Stadtmauer (USA)
Emily Barrett (USA) Silke Dyer (SouthAssociate
Africa) Editors Jung Ryeol Lee (South Korea) Anne Steiner (USA)
J. Abbott,
Valerie Australia
Baker (USA) I. Demeestere, Belgium(France) J. James,
Arnaud Fauconnier NewLevine
Hagai Zealand(Israel) V. Provoost, Belgium
Judy Stern (USA)
S. Babayev,
Gurkan BozdagUSA(Turkey) L. De Santis,
Elpida Italy (United
Fragouli S.G. Kristensen, Denmark
Artur Ludwin (Poland) G. Quinn, Jan-Bernd
USA Stukenborg (Sweden)
E. Baldi,
Paula Italy(USA)
Brady Z. Donarelli,
Kingdom)Italy M. Laan, EstoniaMain (Denmark) B. Reed, USA
Katharina Katja Teerds (The Netherlands)
E. Barrett, USA
Pablo Bermejo-Alvarez (USA) H. Duran,
ThomasUSAFréour (France) S. M. Laird, UKMatsuzaki (France)P. Ruane, Kristen
Sachiko UK Upson (USA)
V. Baker, USA S. Dyer, South Africa W. Ledger, Australia G. Sachdeva, India
Stephen Brown (USA) Sonia Goedeke (New Zealand) Carmen Messerlian (USA) Asli Uyar (USA)
G. Bozdag, Turkey A. Fauconnier, France J.R. Lee, South Korea L. Stadtmauer, USA
Astrid Cantineau
P. Brady, USA (The David Handelsman
E. Fragouli, UK (Australia) H. Levine,
DeanIsrael
Morbeck (New Zealand) A. Steiner,Aafke
USA van Montfoort (The
Netherlands)
P. Bermejo-Alvarez, USA Elizabeth
T. Fréour, Hatch (USA)
France Mariana
A. Ludwin, Moura-Ramos
Poland Netherlands)
J. Stern, USA
Juan CarlosUSA
S. Brown, Castillo (Spain) Trine BeritNew
S. Goedecke, Haugen (Norway) K. Main,
Zealand (Portugal)
Denmark CorrineSweden
J-B. Stukenborg, Welt (USA)
Judit Castillo (Spain)
A. Cantineau, Netherlands Patrick Henriet
D Handelsman, (Belgium)
Australia MonicaFrance
S. Matsuzaki, Muratori (Italy) K. Teerds,Carrie Williams (United
Netherlands
J.C. Castillo,
Nicholas Spain(USA)
Cataldo E. Hatch, USA
Anat Hershko-Klement (Israel)C. Messerlian, USA(Israel)
Galia Oron K. Upson,Kingdom)
USA
J. Castillo,Chambers
Georgina Spain (Australia) T.B.Karla
Haugen,
HuttNorway
(Australia) D. Morbeck,
JulianaNew (Portugal) A. Uyar, USA
Zealand
Pedro Christine Wyns (Belgium)
N. Cataldo,
Jorge ChavarroUSA(USA) P. Henriet,
Vasanti Belgium
Jadva (United Kingdom) M. Moura-Ramos, (Belgium) A. van Montfoort, Netherlands
Portugal
Veerle Provoost
G. Chambers,
Anne DelbaereAustralia
(Belgium) A. Hershko-Klement, Zealand) M. Muratori,
Joanna James (NewIsrael Italy Quinn (USA) C. Welt, USA
Gwendolyn
J. Chavarro, USA K. Hutt, Australia G. Oron, Israel C. Williams, UK
Isabelle Demeestere (Belgium) Stine Gry Kristensen (Denmark) Beverley Reed (USA)
A. Delbaere, Belgium V. Jadva, UK J. Pedro, Portugal C. Wyns, Belgium
Statistical Advisory
Advisory Board
Board
Olga Basso (Canada) Stephen RobertsS.(United
O. Basso (Canada) RobertsKingdom)
(UK) Stacey Missmer(USA)
S. Missmer (USA) Christos Venetis
C. Venetis (Greece) L. Lauren
(Greece) Wise (USA)
Wise (USA)
Founding Editor
R.G. Edwards
R.G. Edwards
Editors Emeriti
Editors Emeriti
D.H. Barlow
D.H. Barlow
A.Van
A. Van Steirteghem
Steirteghem
J.L.H. Evers
J.L.H. Evers
Managing Editor
Managing Editor
A.C. Williams (United Kingdom)
A.C. Williams
AssistantManaging
Assistant ManagingEditors
Editor
J.M. Hastings
J.M. (United
HastingsKingdom)
K.R. Watkins
K.R.(United
WatkinsKingdom)
Editorial Administrator
Editorial Administrator
Emma J Andrew
K.E. (United
Parks Kingdom)
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Published for the

European Society of Human Reproduction and Embryology
by Oxford University Press,
Oxford, UK

ESHRE COMMITTEES
Executive Committee (2019 – 2021) Committee of National Nebojsa Radunovic (Serbia)

Chair Representatives (2017-2020) Milan Reljic (Slovenia)
Cristina Magli (Italy) Petya Andreeva (Bulgaria) Catherine Rongieres (France)
Christiana Antoniadou (Cyprus) Jesper M.J. Smeenk (The Netherlands)
Chair-elect Tamar Barbakadze (Georgia) Robert Spaczynski (Poland)
Carlos Calhaz-Jorge (Portugal) Raminta Baušytė (Lithuania) Patrik Stanic (Croatia)
Members Ursula Bentin – Ley (Denmark) Oliver Sterthaus (Switzerland)
Richard Anderson (United Kingdom) Wolfgang Biasio (Austria) Martin Stimpfel (Slovenia)
Baris Ata (Turkey) Virginia N. Bolton (United Kingdom) Isabelle Streuli (Switzerland)
Basak Balaban (Turkey) Pierre Boyer (France) Lela Surlan (Serbia)
Valerie Blanchet De Mouzon (France) Jean Calleja-Agius (Malta) Mátyás Szabolcs (Hungary)
Edith Coonen (The Netherlands) Lia Chkonia (Georgia) Greta Verheyen (Belgium)
Thomas Ebner (Austria) Susana M. Chuva de Sousa Lopes Kjell Wånggren (Sweden)
Anja Pinborg (Denmark) (The Netherlands) Mary Wingfield (Ireland)
Karen Sermon (Belgium) Monica Marina Dascalescu (Romania) Christine Wyns (Belgium)
Ioana Adina Rugescu (Romania) Lucia De Santis (Italy) Hakan Yarali (Turkey)
Thomas Strowitzki (Germany) Francisco Dominguez (Spain)
Isabel Doria Reis (Portugal) Current International Scientific
Snežana Vidakovic (Serbia) Committee
Giovanni Coticchio (Italy) – Petros Drakakis (Greece)
Sozos J. Fasouliotis (Cyprus) María Isabel Acien (Spain)
(Ex officio SIG Chair) Giuliana Baccino (Spain)
Gianluca Gennarelli (Italy)
Immediate Past Chair Gareth Greggains (Norway) Carlos Calhaz-Jorge (Portugal)
Roy Farquharson (United Kingdom) Marie Louise Groendahl (Denmark) Susana M. Chuva de Sousa Lopes
Mykola Gryshchenko (Ukraine) (The Netherlands)
Special Interest Groups Chair Giovanni Coticchio (Italy)
Andrew Horne (United Kingdom)
Giovanni Coticchio (Italy) Roy Farquharson (United Kingdom)
Anna Janicka (Poland)
Lale Karakoc Sokmensuer (Turkey) Francesco Fiorentino (Italy)
Central Office
Tatyana Kodyleva (Russia C.I.S.) Nicolás Garrido Puchalt (Spain)
Christine Bauquis
Péter Kovács (Hungary) Annick Geril (Belgium)
Lieve Buggenhout
Markus S. Kupka (Germany) Georgios Lainas (Greece)
Andres De Nutte
Joaquin Llacer (Spain) Nicholas Macklon (Denmark)
Veerle De Rijbel
Ana Luisa M.S.Teixeira De Sousa Ramos Cristina Magli (Italy)
Veerle Goossens
(Portugal) Heidi Mertes (Belgium)
Nathalie Le Clef
Sirpa Makinen (Finland) Rebecca Nakalema (Belgium)
Karen Maris
Alice Malenovska (Czech Republic) Anja Pinborg (Denmark)
Saria Mcheik
Corina Manolea (Romania) Heidi Roijemans (Belgium)
Rebecca Nakalema
Ieva Masliukaite (The Netherlands) Andrea Romano (The Netherlands)
Catherine Plas
Laure C. Morin – Papunen (Finland) Virginie Rozée (France)
Erika Mar Rodriguez Raes
Sergei Nikitin (Russia C.I.S.) Ioannis Sfontouris (Greece)
Heidi Roijemans
Georgi Nikolov (Bulgaria) Kelly Tilleman (Belgium)
Anne-Julie Van Bever
Verena Nordhoff (Germany) Bettina Toth (Austria)
Bruno Van den Eede
Kazem Nouri (Austria) Kirsten Louise Tryde Macklon (Denmark)
Sarah Vandersteen
Øyvind Nytun (Norway) Bruno Van den Eede (Belgium)
Titia Van Roy
Ine Van Wassenhove Dinka Pavicic Baldani (Croatia)
Nathalie Vermeulen Michael Pelekanos (Greece)

human
reproduction
Volume 35, Suppl July 2020 https://academic.oup.com/humrep
ORAL PRESENTATIONS
Monday, 06 July 2020

08:30 - 09:30 Session 01: Keynote session . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 1
10:00 - 11:30 Session 02: Blastocyst transfer and freezing��Parallel 1
10:00 - 11:30 Session 03: Strategies to improve the outcomes of ovarian stimulation 1��Parallel 2
10:00 - 11:30 Session 04: Male fertility related predictors and their use . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 3
10:00 - 11:30 Session 05: Endometriosis and uterine disorders. New clinical insights. . . . . . . . . . . . . . . . . . . . . . . . . Parallel 4
10:00 - 11:30 Session 06: Frozen versus fresh embryo transfer. An ongoing challenge on
children’s health��Parallel 5
09:50 - 11:40 Session 07: Male and female fertility preservation - clinical aspects. . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 6
11:45 - 12:45 Session 08: Novel oocyte and embryo biomarkers. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 2
11:45 - 12:45 Session 09: Data reporting session: the European perspective (EIM and PGT). . . . . . . . . . . . . . . . . . . Parallel 3
11:45 - 12:45 Session 10: Updated terminology for early pregnancy assessment. . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 4
11:45 - 12:55 Session 11: Patient priorities. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 5
14:00 - 15:00 Session 12: ASRM exchange session - controversies in ART. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 2
14:00 - 15:00 Session 13: Challenging scenarios in IVF patients. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 3
14:00 - 15:00 Session 14: The way forward for fertility preservation. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 4
14:00 - 15:00 Session 15: In the name of the father. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 5
14:00 - 15:00 Session 16: Breaking news in current practice . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 6
15:15 - 16:30 Session 17: Cellular characteristics of embryo development . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 1
15:15 - 16:35 Session 18: Cellular and molecular markers of ovarian ageing. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 2
15:15 - 16:30 Session 19: RIF and endometrial factors: does it matter?. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 3
15:15 - 16:30 Session 20: Reproductive (EPI)genetics 1. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 4
15:15 - 16:30 Session 21: Impact of new technologies on human reproduction. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 5
15:15 - 16:35 Session 22: Updates on ART outcomes, barriers and predictions: an
international overview ��Parallel 6
17:00 - 18:00 Session 23: Recent advances in endometriosis. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 2
17:00 - 18:00 Session 24: Promoting fertility awareness in your own backyard. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 3
17:00 - 18:00 Session 25: The future of andrology. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 4
(continued overleaf )

17:00 - 18:00 Session 26: The day after. Fertility preservation and embryo transfer in patients
with cancer diagnosis. ��Parallel 5
17:00 - 18:00 Session 27: Frontiers in developmental biology. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 6
Tuesday 07 July 2020

08:30 - 09:30 Session 28: Revisiting early embryo development. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 1
08:30 - 09:30 Session 29: Building bridges towards harmonisation. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 2
08:30 - 09:30 Session 30: Nurse or midwife led e-health care interventions. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 3
10:00 - 11:30 Session 31: Predictive algorithms in clinical embryology. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 1
10:00 - 11:30 Session 32: Which are the optimal ovarian stimulation protocol? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 2
10:00 - 11:30 Session 33: Predictors. Technology and processes improving outcomes in andrology. . . . . . . . . . . . . . Parallel 3
10:00 - 11:30 Session 34: Endometriosis - pathogenesis and diagnosis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 4
10:00 - 11:30 Session 35: Impact of ART on health outcomes of children. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 5
10:00 - 11:30 Session 36: Covid-19 session . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 6
11:45 - 12:45 Session 37: MHR symposium - fundamentals on making oocytes. . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 2
11:45 - 12:45 Session 38: Laboratory session - time-lapse in 2020. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 3
11:45 - 12:55 Session 39: Strategies to improve the outcomes of ovarian stimulation 2. . . . . . . . . . . . . . . . . . . . . . . . Parallel 4
14:00 - 14:45 Session 40: Global ART monitoring. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 2
14:00 - 15:00 Session 41: ALMER exchange session - IVF laboratory automation. . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 3
14:00 - 15:00 Session 42: Stress and infertility - the chicken or the egg. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 4
15:15 - 16:30 Session 43: ICSI in 2020. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 1
15:15 - 16:30 Session 44: What are the optimal regimes for frozen embryo transfer?. . . . . . . . . . . . . . . . . . . . . . . . . Parallel 2
15:15 - 16:30 Session 45: Spermatogenesis subtle regulatory effectors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 3
15:15 - 16:30 Session 46: Reproductive (EPI)genetics 2. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 4
15:15 - 16:35 Session 47: Does emotional balance before being parents and after exist?. . . . . . . . . . . . . . . . . . . . . . Parallel 5
15:15 - 16:30 Session 48: Relating the relevance of biomarkers to infertility. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 6
17:00 - 18:00 Session 49: Embryo metabolism and development. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 1
17:00 - 18:00 Session 50: Androgen treatment in fertility management. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 2
17:00 - 18:10 Session 51: RM: new diagnostic and therapeutic aspects. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 3
17:00 - 18:00 Session 52: AI. A new tool to assess art outcomes and help patients?. . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 4
17:00 - 18:00 Session 53: Controversies in ART. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 5
17:00 - 18:00 Session 54: Modern techniques promote variety in fertility nursing research . . . . . . . . . . . . . . . . . . . . Parallel 6
Wednesday 08 July 2020

08:30 - 09:30 Session 55: Cochrane session - better evidence, better policies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 1
08:30 - 09:30 Session 56: Frontiers in andrology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 2
08:30 - 09:30 Session 57: Evidence-based surgical interventions. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 3
08:30 - 09:30 Session 58: Improving sperm crysopreservation outcomes. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 4

10:00 - 11:45 Session 59: New morphokinetic insights of embryo development. . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 1
10:00 - 11:45 Session 60: Long term health, obstetrics and neonatal outcomes relating to infertility treatment. . . . . Parallel 2
09:50 - 11:55 Session 61: Understanding spermatogenesis beyond histology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 3
10:00 - 11:45 Session 62: Pathophysiologic aspects of implantation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 4
10:00 - 11:45 Session 63: Protecting gamete quality. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 5
10:00 - 11:45 Session 64: Prospective carrier screening of ART couples. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 6
12:00 - 13:00 Session 65: Biomarkers of failed pregnancy. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 1
12:00 - 13:00 Session 66: Synthetic embryology: myth or reality?. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 2
12:00 - 13:00 Session 67: COVID-19 - Psychosocial impact of delayed treatment . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 3
12:00 - 13:00 Session 68: Genetic determinants of embryo quality . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 4
14:00 - 15:15 Session 69: Biomarkers of developmental competence. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 1
14:00 - 15:15 Session 70: Ovarian stimulation strategies in IVF and IUI. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 2
14:00 - 15:15 Session 71: About how sperm quality and male infertility relate to genetics. . . . . . . . . . . . . . . . . . . . . Parallel 3
14:00 - 15:15 Session 72: Pregnancy loss: what to consider. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 4
14:00 - 15:15 Session 73: Endometriosis and ART. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 5
14:00 - 15:15 Session 74: Oocyte and embryo evaluation. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parallel 6
• INVITED SESSIONS
• SELECTED ORAL COMMUNICATION SESSIONS

Human Reproduction, Vol.35, No.Suppl, pp. i1–i513, 2020
human
reproduction
ESHRE 2020 / Oral presentations

only one association – of earlier pubic hair development with propyl paraben –
with peripubertal concentrations.
INVITED SESSION Conclusions: We found associations of altered pubertal timing with early life
exposure to chemicals in personal care products in girls, but little evidence in
SESSION 01: KEYNOTE SESSION
boys. This study contributes to a growing literature that suggests that exposure
06 July 2020 Parallel 1 08:30 - 09:30 to certain endocrine disrupting chemicals may impact timing of puberty in
children.
O-001 Harnessing the power of data: Application of data science

in reproductive medicine SELECTED ORAL COMMUNICATIONS
S. Nelson1 SESSION 02: BLASTOCYST TRANSFER AND FREEZING
1
University of Glasgow, Royal Fertility Clinic, Glasgow, United Kingdom 06 July 2020 Parallel 1 10:00 - 11:30
O-002 Association of phthalates, parabens and phenols found in
personal care products with pubertal timing in girls and boys
K.G. Harley1, K. Berger1, K. Kogut1, K. Parra2, R. Lustig3,
L. Greenspan4, A. Calafat5, B. Eskenazi1 O-003 The effect of laser assisted hatching on vitrified-warmed
1
UC Berkeley, School of Public Health, Berkeley- CA, U.S.A. blastocysts: an interim analysis of a multicentric randomized
2
University of Arizona, Mel and Enid Zuckerman College of Public Health, controlled trial
Tucson- AZ, U.S.A. A. Alteri1, C. Guarneri2, L. Corti1, L. Restelli2, M. Reschini2,
3
University of California- San Francisco, Department of Pediatrics, San Francisco, N. Barberis1, E. Papaleo1, A. Paffoni3, P. Viganò4, E. Somigliana2
U.S.A. 1
IRCCS San Raffaele Scientific Institute, Obstetrics and Gynecology Department,
4
Kaiser Permanente, Department of Pediatrics, San Francisco, U.S.A. Milan, Italy ;
5
Centers for Disease Control and Prevention, National Center for Environmental 2
Fondazione IRCCS Ca’ Granda Ospedale Maggiore Policlinico, Obstetrics and
Health, Atlanta- GA, U.S.A. Gynaecology Department, Milan, Italy ;
3
ASST Lariana, Infertility Unit, Cantù, Italy ;
Background: Certain chemicals in cosmetics and personal care products, 4
IRCCS San Raffaele Scientific Institute, Division of Genetics and Cell Biology, Milan, Italy
including low molecular weight phthalates, parabens, and phenols, have
demonstrated endocrine disrupting properties. These chemicals have been Study question: Does assisted hatching of vitrified/warmed blastocysts
associated with altered pubertal timing in animal studies, but few human improve clinical outcomes?
studies exist. Summary answer: No significant difference was found in clinical pregnancy
Methods: Data were from the Center for the Health Assessment of Mothers rate (CPR).
and Children of Salinas (CHAMACOS) longitudinal cohort study that followed What is known already: Recently, the Human Fertilization and Embryology
338 children in the Salinas Valley, California from before birth to adolescence. (HFEA) declared that the assisted hatching (AH) is an unproven add-on inter-
Pregnant women were enrolled in 1999-2000. Mothers were mostly Latina, living vention and the guidelines of the National Institute for Clinical Excellence (NICE)
below the federal poverty threshold, and had not completed high school. We do not recommend the use of AH to improve pregnancy rates. Of note, available
measured concentrations of three phthalate metabolites (monoethyl phthalate data are mainly focused on the effect of AH in fresh-cleavage stage embryo
[MEP], mono-n-butyl phthalate, and mono-isobutyl phthalate), methyl and propyl transfers while data regarding the effect of laser AH (LAH) on blastocysts, espe-
paraben, and four other phenols (triclosan, benzophenone-3, and 2,4- and cially after cryopreservation, are inconclusive since they are based on studies
2,5-dichlorophenol) in urine collected from mothers during pregnancy and from with small sample sizes and may hide a type II error.
children when they were 9 years of age. Pubertal timing was assessed among 179 Study design, size, duration: A multicentric prospective comparative study
girls and 159 boys every 9 months between ages 9 and 13 using clinical Tanner with a parallel randomized controlled design. The sample size (700 blastocysts)
staging. Accelerated failure time models were used to obtain mean shifts of was calculated considering a 10% difference in live birth rate (LBR) between
pubertal timing associated with concentrations of prenatal and peripubertal LAH-group and no LAH group. The primary outcome was live birth rate and
biomarkers. the secondary outcomes were implantation, biochemical pregnancy, clinical
Results: In girls, we observed associations of earlier onset of pubic hair devel- pregnancy, miscarriage, multiple pregnancies, obstetrical and neonatal compli-
opment with increased prenatal urinary MEP concentrations and earlier men- cations rates. An interim analysis based on CPR was performed including cycles
arche with increased prenatal triclosan and 2,4-dichlorophenol concentrations. between September 2018 and October 2019.
With peripubertal biomarkers, we observed earlier breast development, pubic Participants/materials, setting, methods: Women ≤ 40 years and with
hair development, and menarche with increased methyl paraben urinary con- ≤ 2 previous oocyte retrievals undergoing IVF cycles and scheduled for elective
centrations; earlier menarche with propyl paraben concentrations; and later single embryo transfer (eSET) with vitrified/warmed blastocysts were considered
pubic hair development with 2,5-dichlorophenol concentrations. In boys, we for enrollment in each of the two units. Warmed blastocysts were randomized
observed no associations with prenatal urinary biomarker concentrations, and immediately after warming to the study group (LAH) or to the control group
© The Author(s) 2020. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved.
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BK-OUP-ESHRE2020_ABSTRACTS-200150-Monday.indd 1 22/06/20 8:43 PM

i2 Abstracts of the 36th Annual Meeting of the ESHRE, 5 to 8 July 2020
(without LAH). The allocation sequence was stratified according to woman’s 293 first transfers were included (139 FPS- and 154 LPS-derived euploid blas-
age and blinded to the physicians and participants. tocysts). All transfers were performed in an artificial cycle.
Main results and the role of chance: Since the beginning of the study, 291 Main results and the role of chance: To achieve 80% power (α=0.05) to
women were recruited (n=145 in LAH-group and n= 146 in the control group). rule-out a 15%-difference in LBR between FPS- and LPS-derived euploid blasto-
At the time of transfer, hatching and hatched blastocyst rates were significantly cysts ~300 first SETs were required. The blastocysts in the two groups were
higher in the LAH group compared to the control group (respectively 55% vs equally distributed according to morphology and day of development. The study
16%, p<0.001). The clinical pregnancy rate in the whole cohort was similar in arms were similar for age, sperm factor, kind and cause of infertility and indication
LAH and control group, being 45% and 43%, respectively (p=0.81). The clinical to PGT. The positive-pregnancy-tests-rates were 59.7% (n=83/139) and 63.6%
pregnancy rate in the age group <38 years was 45% in the LAH group compared (n=98/154) from FPS- and LPS-derived euploid blastocysts, respectively. The
to 39% in the control group (p=0.37). In addition, in the age group ≥ 38 years, BPL rates were 10.8% (n=9/83) and 9.2% (n=9/98). The MRs were 10.8%
the clinical pregnancy rates were 44% in the LAH group and 56% in the control (n=8/74) and 11.2% (n=10/89). The LBRs were 47.5% (n=66/139) and 51.3%
group, respectively (p=0.30). (n=79/154;p=0.6). Among patients with euploid blastocysts produced after both
Limitations, reasons for caution: Being an interim analysis, the sample size FPS and LPS, the LBRs were also similar (51.9% [n=28/54] and 54.3% [n=25/46];
is not powered enough to detect differences <10% between groups. At this step, p=0.8). Four out of 66 (6.0%) FPS-derived pregnancies showed gestational issues
it is not possible to evaluate LBR which is the main outcome of the study. (one gestational diabetes, one icterus, one polyhydramnios, one embryo twin-
Wider implications of the findings: This study could clarify the real potential ning). Five out of 79 (6.3%) LPS-derived pregnancies showed gestational diabetes.
of AH on vitrified-warmed blastocysts in terms of live birth leading to evi- Gestational age (38.1±1.3weeks versus 38.2±1.7weeks;p=0.6), birthweight
dence-based changes in current clinical practice. (3309±438g versus 3232±471g;p=0.4) and length (47.9±10.2cm versus
Trial registration number: NCT03623659 47.6±11.4cm;p=0.9) were similar. Four out of 67 (6.0%) FPS-derived new-borns
showed perinatal issues (one neonatal respiratory distress involving 7 days in the
O-004 The euploid blastocysts produced after either follicular- neonatal-intensive-care-unit, one diaphragmatic hernia, one single kidney, one
phase or luteal-phase-stimulation show similar live-birth rates: hydrocephalous).No neonatal issue has been reported among LPS-derived
a prospective multicenter study including 293 vitrified-warmed new-borns.
single-embryo-transfers Limitations, reasons for caution: LPS-derived embryos were produced only
A. Vaiarelli1, D. Cimadomo1, S. Colamaria1, E. Trabucco2, in a DuoStim approach. The reproductive competence of poor-quality blastocysts
A. Sansone2, E. Alviggi2, A. Della Ragione2, F. Golia2, F. Fiorini3, was not assessed. The study was powered to investigate the primary outcome,
N. Barnocchi3, L. Papini3, A. Conforti4, C. Alviggi4, L. Rienzi1, hence the analysis of secondary outcomes should be considered observational.
F.M. Ubaldi1 Wider implications of the findings: The adoption of unconventional ovarian
1
Clinica Valle Giulia, Genera centers for reproductive medicine, roma, Italy ; stimulation protocols for the treatment of peculiar populations of poor-prog-
2
Clinica Ruesch, Genera centers for reproductive medicine, Naples, Italy ; nosis infertile and oncologic patients is increasing worldwide. This study further
3
Genera Umbria, Genera centers for reproductive medicine, Umbertide, Italy ; reinforces the hypothesis that FPS- and LPS-derived embryos have comparable
4
University of Naples- Federico II, Department of Neuroscience- Reproductive competence. Therefore, if required, LPS might be adopted.
Science and Odontostomatology, Naples, Italy Trial registration number: none
Study question: Is there any difference in the reproductive competence of O-005 Cleavage stage embryo transfer impairs cumulative
euploid blastocysts produced after follicular-phase-stimulation (FPS) or lute- live birth rates and time to livebirth as compared to blastocyst
al-phase-stimulation (LPS) during vitrified-warmed single-embryo-transfers? transfer in oocyte recipients. A randomized controlled trial
Summary answer: LPS-derived vitrified-warmed euploid blastocysts showed E. Clua Obradó1, I. Rodríguez1, G. Arroyo1, F. Martínez1,
similar live-birth-rate (LBR), obstetrical and perinatal outcomes as FPS- L. Latre1, B. Coroleu1, N.P. Polyzos1
derived ones. 1
USP Instituto Universitario Dexeus, Medicina de la Reproducción, Barcelona, Spain
What is known already: Multiple follicular waves arise during a single
ovarian cycle in humans, thereby highlighting a novel folliculogenesis pattern Study question: Does embryo transfer day (D5 vs D3) affect cumulative
overtaking the classic theory. Several studies conducted by numerous groups pregnancy (CPR), cumulative live birth rates (CLBR) and time to livebirth (TTL)
worldwide consistently showed a similar developmental competence between in oocyte donation programs?
FPS- and LPS-derived cohorts of oocytes in terms of fertilization, blastulation, Summary answer: Embryo transfer at cleavage stage(D3) results in ~20%
and euploidy rates. These observations supported the further implementation relative reduction in CPR and 15% in CLBR, while increases the TTL as compared
of unconventional protocols for ovarian stimulation in poor-prognosis with blastocyst(D5) transfer.
patients, like patients fulfilling the Bologna criteria, advanced-maternal-age What is known already: Blastocyst embryo transfer has been traditionally
women, oncologic patients: random-start,LPS-only and FPS+LPS in the same associated with very good pregnancy outcomes due to embryo self-selection
ovarian cycle (DuoStim). Nevertheless, it still needs to be outlined whether after the embryonic genome activation on day 3.
LPS-derived blastocysts show similar reproductive competence as FPS- In IVF/ICSI cycles among infertile women using their own oocytes, evidence
derived ones. from RCTs suggest that blastocyst transfer results in significantly higher live birth
Study design, size, duration: Multicenter study conducted between rates; still no difference has been identified between D3 and D5 in terms of
October2015–March2019 including all vitrified-warmed euploid single blastocyst cumulative live birth rates. However, it is unclear whether D3 or D5 embryo
transfers after DuoStim with preimplantation-genetic-testing (PGT). Only first transfer may result in different CPR and CLBR in women included in an oocyte
transfers of good quality blastocysts (≥BB according to Gardner and Schoolcraft’s donation program.
classification) were included. In the presence of euploid blastocysts produced Study design, size, duration: In a single-center randomized trial conducted
after both FPS and LPS (n=100/293,34%), the embryo to transfer was chosen between March 2017-August 2018, 134 oocyte recipients were randomized
in a blinded fashion (random.org). The primary outcome was the LBR per trans- to cleavage stage (D3-group) or to blastocyst stage (D5-group) embryo
fer. Biochemical-Pregnancy-Loss (BPL), Miscarriage-Rate (MR) and obstetrical/ transfer.
perinatal outcomes were also monitored. Participants/materials, setting, methods: Eligible women were recipients
Participants/materials, setting, methods: DuoStim was proposed to between 18-50 years, in their 1st/2nd synchronous cycle (excluding: PGT-A,
poor-prognosis patients respecting two of these criteria: AMH≤1.5ng/ml, implantation failure). At first consultation and prior to oocyte donation cycle,
antral-follicle-count≤6, ≤5 oocytes from previous cycle(s), advanced-mater- recipients were randomly allocated to D3 or D5 transfer.
nal-age. Ovarian stimulations were performed with recombinant-gonadotrophins Co-primary outcomes were, CPR (first pregnancy) and CLBR (first live birth)
in an antagonist protocol. LPS was started five days after the first retrieval. per patient, within 12 months from the first embryo transfer, considering fresh
Embryos were cultured to blastocyst, underwent trophectoderm biopsy and and subsequent frozen embryo transferred. In addition, we analyzed TTL (time
vitrification. Only full-chromosome meiotic aneuploidies were reported. Overall, from 1st embryo transfer until 1st livebirth).

Abstracts of the 36th Annual Meeting of the ESHRE, 5 to 8 July 2020 i3
Main results and the role of chance: Whereas 250 recipients were planned 13.7% (19/139), p=0.73), 2 (18.4% (16/87) vs. 22.9% (30/131), p=0.42), 3
to be enrolled, the study was prematurely terminated, with an unplanned interim (34.8% (31/89) vs. 31.8% (28/88), p=0.67), 4 (26.8% (22/82) vs. 39.1%
analysis, when 134 women completed treatment, for ethical reasons, due to the (27/69), p=0.11), 5 (36.4% (24/66) vs. 36.4% (16/44), p=1) and ≤5 zygotes
clinically and statistically significant inferior outcomes in D3 arm. (26.1% (100/383) vs. 25.5% (120/471), p=0.83) were all comparable. Patients
Although the number of supernumerary frozen embryos was significantly having ≤5 zygotes were also analyzed by logistic regression and transfer day did
higher in D3-group (5.3±2.7) than in D5-group (3.6±2.3), pregnancy outcomes not matter at all. Outcome of embryos with a known implantation status were
were substantially inferior in patients allocated to D3-group. compared between d5 and d3 groups and implantation rates were comparable
CPR were significantly lower in the D3 vs D5 group (67.2% vs. 85.9%) with (9.1% (13/143) vs. 12.95% (32/247) respectively, p=0.12). Although implanta-
a proportion difference of -18.7% 95%CI [-32.8% to -4.3%]. Similarly, CLBR tion rate per number of transferred embryos of d5 group was significantly higher
were lower in the D3 vs D5 group (55.2% vs. 70.3%) with a proportion difference (31.28% (183/585) vs. 20.52% (157/765), p<0.0001), the number of transfer-
of -15.1% 95%CI [-30.9% to 1.6%], although results did not reach statistical able embryos (embryos transferred plus cryopreserved) were significantly higher
significance for this outcome. in d3 group (29.14% (681/2337) vs. 46% (961/2088), p<0.0001).
Finally, TTB was significantly longer in D3 group as compared with D5 log Limitations, reasons for caution: The retrospective nature of this study
rank test p=0.027. In order to reach a 50% cumulative live birth rate, D3 group may not eliminate potential bias. On the contrary, the strength of our study is
required 6 months more than D5 group and always D5 group maintained a that patients were allocated to the d5 and d3 groups regardless of patient and
higher live birth rate. cycle characteristics. More research is needed to prove our findings.
Limitations, reasons for caution: This randomized trial was prematurely Wider implications of the findings: Developmental potential of embryos
terminated, with an unplanned interim analysis, due to the clinically and statisti- was neither compromised nor supported by transferring earlier or later. Embryo-
cally significant inferior results for the D3 group following evaluation by the endometrium synchronization and in vitro versus in vivo embryo development
institutional review board. does not appear to make any significant contribution to treatment outcome.
Wider implications of the findings: Transfer of embryos in cleavage stage Embryo selection in extended culture makes sense in patients with a larger
(D3) should be discouraged in oocyte recipients with good prognosis since it zygote cohort.
significantly impairs cumulative pregnancy rates, appears to reduce cumulative Trial registration number: not applicalble
live birth rates, while significantly increases time to live birth and costs due to
higher number of cryopreserved embryos. O-007 High grade trophectoderm is associated with monozygotic
Trial registration number: NCT03088735 twinning in frozen-thawed single blastocyst embryo transfer
Q. Zheng1, S. Xu1, M. Mo1, F. Xiong1, X. Hu1, Y. Zeng1
O-006 Blastocyst versus cleavage stage transfers: who benefits? 1
Shenzhen Zhongshan Urology Hospital, Fertility center, Shenzhen, China
E.K. Dirican, Ph.D1, Ş. Olgan1, M. Çağlar1, M. Sakıncı1,
O. Ertürk, Kadir1, Z. Kasabalı1, U. Çakmaklı1 Study question: What specific risk factors make monozygotic splitting at the
1
Akdeniz University- Faculty of Medicine, Obstetrics & Gynecology- Reproductive blastocyst embryo stage?
Endocrinology and Infertility, Antalya, Turkey Summary answer: High grade trophectoderm (TE), but not inner cell mass
(ICM) or blastocyst expansion, is associated with monozygotic splitting after
Study question: What is the outcome of cleavage and blastocyst transfers frozen-thawed single blastocyst ET.
with regards to transfer cancellation, embryo selection and embryo-endome- What is known already: Elective single blastocyst embryo transfer (eSET)
trium synchronization? contributes to high live-birth rate per embryo transfer cycle and low multi-preg-
Summary answer: Transfer cancellations due to extended culture did not nancy rate, and is well perceived in clinical practice. However, blastocyst transfer
affect the pregnancy outcomes and patients with 6 or more zygotes benefited is considered as the major risk factor of monozygotic splitting.
from embryo selection. Study design, size, duration: This was a retrospective observational study
What is known already: Despite improvements in embryo culture, there is still including a cohort of 2863 single blastocyst-transferred pregnancies between
a concern whether the in vitro extended culture supports embryo development January 2011 and June 2019 in a single fertility center in South China. The study
to the same extent as in vivo or not. In addition, transfer cancellations often result was approved by the hospital’s Ethics Committee.
in an unpleasant patient counseling. As a precaution for transfer cancellation, some Participants/materials, setting, methods: Monozygotic splitting pregnancy
clinics offer extended culture only when abundant embryos are available. However, was identified as the number of foetuses exceeded the number of gestational sacs
the relative contributions of transfer cancellation against embryo selection during (GSs). The incidence of splitting regarding the oocyte age, paternal age, ovarian
extended culture to the cycle outcome have not been fully documented. On the stimulation protocol, insemination method, type of frozen cycle, Gardner grading
other hand, the effects of the proposed asynchrony between the cleavage stage of expansion, ICM and TE were calculated. The serum E2, P level and endometrium
embryo and the endometrium are also contradictory. thickness on hCG/progesterone day, and serum HCG levels on day 11 after ET
Study design, size, duration: This was a single center retrospective com- were compared between the splitting and non-splitting pregnancies
parative cohort study in a university based medical assisted reproduction (MAR) Main results and the role of chance: There were 51 splitting cases in total
center. 1710 entries from January 2018 to December 2019 were evaluated. In among the 2863 single blastocyst-transferred pregnancies, the incidence of split-
this cohort, embryos were not transferred on weekends. Regardless of patient ting was 1.78%. High TE grade was associated with monozygotic splitting
and cycle characteristics, Monday and Tuesday egg retrievals were called back (P=0.022), aOR for grade A and B TE were 3.96 (95% CI: 1.20-13.06) and 4.24
for transfer on day 3 (d3) (n=584), Wednesday and Thursday retrievals were (95% CI: 1.24-14.46) compared to that of grade C TE respectively. While the
on day 5 (d5) (n=532). Friday retrievals were excluded. ICM grade and blastocyst expansion were not statistically significant. There were
Participants/materials, setting, methods: Fresh cycles were included and no statistically significant differences in oocyte age, ovarian stimulation protocol,
primary outcomes were clinical pregnancy and implantation rate. Cycles were insemination method, type of frozen cycle, serum E2, P and endometrium thick-
stratified on the basis of zygote numbers and analyzed using the intention to ness between the splitting and non-splitting pregnancies. Serum hCG levels on
treat principle. Outcome of embryos with a known implantation status were 11 d after ET were significantly higher in the splitting cases than those in the
used to demonstrate the effects of transferring earlier or later on embryo devel- non-splitting pregnancies (P=0.020). We hypothesized that increased secretion
opment. Statistical analyses were performed with T or chi-squared tests. Logistic of HCG from the high grade TE may widen the implantation window and support
regression analysis was carried out to calculate the effects of confounders. the monozygotic splitting.
Main results and the role of chance: For the patients having 6 or more Limitations, reasons for caution: The primary limitation of this study was
zygotes, clinical pregnancy rates were significantly higher in the d5 group com- its retrospective nature and small sample size. The second limitation was that
pared with d3 (48.3% (72/149) vs. 25.7% (29/113), p<0.0001). A logistic regres- MZT pregnancy was mainly diagnosed by ultrasound evaluation but not con-
sion was performed to account for most relevant confounders and transfer day formed by the DNA profiling for the zygosity.
was the only significant factor (OR (95% CI) = 3.12 (1.8 – 5.43), p<0.0001). Wider implications of the findings: Clinicians should consider whether to
The clinical pregnancy outcome of d5 versus d3 groups for 1 (11.9% (7/59) vs. counsel couples about the slightly increased risk of monozygotic splitting

associated with high grade TE. The transfer of an embryo with an optimal TE Wider implications of the findings: The results of this original meta-analysis
reduces monozygotic splitting. confirm that delayed blastulation is associated with a poorer prognosis in terms
Trial registration number: SZSM201502035; 2018YFC1003904 of euploid rate and pregnancy outcomes following frozen transfers. On the other
hand, the results presented do not support the discharge of embryos deriving
O-008 Day7 blastocyst: is it worth it? A systematic review and from a delayed blastulation.
meta-analysis Trial registration number: PROSPERO Registration Number CRD42017067270
G.C. Cermisoni1, L. Corti1, A. Alteri1, P. Vigano’2, E. Giacomini3,
M. Noventa4
1
San Raffaele Scientific Institute - Milan - Italy, U.O. Obstetrics and Gynecology, SELECTED ORAL COMMUNICATIONS
Milan, Italy ;
2
San Raffaele Scientific Institute - Milan - Italy, Reproductive Sciences Laboratory, SESSION 03: STRATEGIES TO IMPROVE THE OUTCOMES
Milan, Italy ; OF OVARIAN STIMULATION 1
3
San Raffaele Scientific Institute - Milan - Italy, Reproductive Sciences Lab - Division 06 July 2020 Parallel 2 10:00 - 11:30
of Genetics and Cell Biology, Milan, Italy ;
4
University of Padua, Department of Woman and Child Health-, Padua, Italy
Study question: What are the differences in pregnancy outcomes and euploid
rate between blastocysts developed on Day7 (D7) and faster blastocysts devel- O-009 Progestins as an alternative to Gonadotropin-Releasing
oped on Day5 (D5) or Day6 (D6)? hormone analogues: A retrospective study comparing in
Summary answer: Frozen D7 blastocyst transfers are associated with lower vitro fertilization outcomes during follicular and luteal phase
clinical pregnancy and live birth rates than D5/6 blastocysts. Euploid rate of stimulation.
screened D7 blastocysts similarly lower. L. Yamakami1, M. Nakano1, F. Ramos1, L. Matsumoto1,
What is known already: Human embryos optimally reach the blastocyst R. Tomioka1, O. Duarte Filho1, V. Heinrich-Oliveira1
stage after five days of culture but some have a slower development. Albeit 1
VidaBemVinda, Medical department, Sao Paulo, Brazil
blastocysts developing on Day 7 can be viable and result in a healthy live birth,
slowly developing embryos after D6 are routinely discarded. Recent studies Study question: May progestins instead Gonadotropin-Releasing Hormone
provide evidence supporting the idea that D6 blastocysts have a decreased analogues (GnRH-analogues), for suppressing premature Luteinizing Hormone
reproductive potential compared to D5 ones. There is however scanty evidence (LH) surge, affects embryo viability and in vitro fertilization (IVF) outcomes?
regarding the clinical outcomes of Day 7 blastocysts compared to blastocysts Summary answer: Ovarian stimulation using Progestins does not affect chro-
developing on D5 or D6. mosomal rearrangements and pregnancy outcomes; however, it increases the
Study design, size, duration: Systematic review and meta-analysis of pub- fertilization rate.
lished studies that have evaluated the effect of delayed blastulation in frozen What is known already: The use of progestin during ovarian stimulation is
ART cycle. The intervention group consisted of patients with blastocysts cryo- effective in preventing LH surge. Development of this protocol is supported by
preserved on D7. Searches were conducted on 28/08/2019 using the following the freezing all strategy and by multiple follicular recruitment waves, respectively,
search terms: blastocyst, Day 5, Day 6, pregnancy, implantation, live birth and preventing detrimental effects of progesterone on embryo implantation odds,
embryo transfer. The primary outcome was the clinical pregnancy rate (CPR). and enabling greater flexibility to physicians and patients. Nonetheless, although
Secondary outcomes were euploid and survival rates after thawing and live birth it has been demonstrated that progestin does not influence neither the number
rate (LBR). or the quality of the oocytes, greater depth about the embryo viability and
Participants/materials, setting, methods: The systematic review was pregnancy achievement is required.
written following the PRISMA guidelines. Sixty-three full-text articles were pre- Study design, size, duration: This retrospective study included data from
selected based on title and abstract. Study selection and data extraction were 140 “freeze-all” cycles conducted between August/2018 and December/2019
performed according to Cochrane methods. The pooled results of euploid rate, and split according to pituitary suppression regimen: Gonadotropin-releasing
CPR and LBR were compared according to the day of blastocyst development. hormone antagonists at follicular phase (GnRH-antagonists, n=53), Progestin at
Odds ratios (OR) with 95% confidence intervals (CI) were calculated for out- follicular phase (FP, n=38), and Progestin at luteal phase (LP, n=49). We consider
come measures. Random-effect meta-analysis was performed and a p-value less follicular phase as day cycle 2 or 3 and luteal phase was defined based on features
than 0.05 was considered statistically significant. of transvaginal ultrasound performed after presumed ovulation.
Main results and the role of chance: Data from 11 relevant articles were Participants/materials, setting, methods: Women received gonadotropins
extracted. The meta-analysis of 7 studies showed a lower proportion of euploid for follicular recruitment and their respective method for pituitary suppression.
embryos comparing D7 vs. D5 (OR 0.47, 95%CI 0.39-0.57; p<0.0001) with Laboratorial procedures were conducted according to standard practice. When
moderate heterogeneity (I2=69%) and comparing D7 vs. D6 (OR 0.68, 95%CI indicated, a blastocyst biopsy was performed for 24-chromosome analysis by
0.61-0.75, p<0.0001) with low heterogeneity (I2=19%). Next-Generation Sequencing (NGS). The effects of stimulation protocols on
A lower proportion of CPR (evaluated in 7 studies) was evident when trans- IVF outcomes and embryo impairments in terms of chromosomal rearrange-
fers of blastocysts frozen in D7 were compared to those of blastocysts frozen ments were evaluated using adjusted general linear models. SPSS Statistics 20
in D5 (OR 0.37, 95%CI 0.23- 0.59; p<0.0001) with moderate heterogeneity was used for data analysis and an α of 5% was adopted.
(I2=66%). Similarly, CPR was reduced comparing D7 vs. D6 (OR 0.49, 95%CI Main results and the role of chance: There were no differences between
0.35- 0.69, p<0.0001) with moderate heterogeneity (I2=38%). A lower propor- groups regarding to female age, body mass index, basal follicle stimulation hor-
tion of LBR (evaluated in 11 studies) was found comparing transfers of blasto- mone (FSH), anti-müllerian hormone (AMH) and antral follicle count. Total
cysts frozen in D7 to those of blastocysts frozen in D5 (OR 0.21, 95%CI gonadotropin doses were different between GnRH-antagonist and progestin
0.16-0.27; p<0.0001) with low heterogeneity (I2=0%) and comparing D7 vs. D6 groups (GnRH-antagonist: 2199±719 vs FP: 2512± 543; p=0.04 and LP:
(OR 0.34, 95%CI 0.26-0.45, p<0.0001) with low heterogeneity (I2=0%). These 2695±438; p<0.01). Twenty-nine cycles (54%) from GnRH-antagonist group,
findings were confirmed in a subgroup of PGT-A screened embryos. A lower twenty-seven (71%) from FP group and twenty-six (53%) from LP group had
survival rate (evaluated in 5 studies) was demonstrated for blastocysts frozen in their embryos undergoing to NGS. The euploidy rate was similar among the
D7 vs. D5 (OR 0.31, 95%CI 0.11-0.82; p=0.02) with moderate heterogeneity tree protocols, as well as the biochemical and clinical pregnancy rates. However,
(I2=72%), while the comparison of D7 vs. D6 was not statistically different. fertilization rate was significantly higher in the FP (86.0%) and LP (90.5%) groups
Limitations, reasons for caution: The validity of meta-analysis results depends when compared to GnRH-antagonists (86.0% vs 90.5% vs 72.9%, respectively,
mainly on the number and the quality and of the included studies: this meta-analysis p<0.01) with Pearson’ correlation (r=0.43). There was no difference among the
included retrospective studies only. The low number of D7 embryo transfers and three groups regarding endometrial priming protocol and mean endometrial
the heterogeneity of laboratory strategies are limitations of this paper. thickness for frozen embryo transfers.

Limitations, reasons for caution: The present study has limitations inherent cycle with antagonists level of satisfaction has been very high / high with respect
to this retrospective design, as a wide range of patient ages, as so different drugs to the previous cycle in 92.74%.
for ovarian stimulation and triggering. Moreover, it was a single center study with Limitations, reasons for caution: This is a non-inferiority study with number
a relatively small cohort. of retrieved oocytes as the primary outcome. The limitations of this RCT include
Wider implications of the findings: Progestin use for pituitary suppression, that treatment could not be blinded, because of the different administration
either in follicular and luteal phases, does not affect euploid blastocyst or clinical route of the medication in study. Another limitation to take into account is that
pregnancy rates, which go against the belief that high levels of progesterone oocyte recipients were not randomized.
during ovarian stimulation impair oocyte and embryo quality. In addition, we Wider implications of the findings: We observed comparable oocyte retrieval,
highlighted that progestins provides better oocyte competence for endocrine profile, viable embryo numbers and similar pregnancy outcomes in the two
fertilization. groups. Therefore, MPA is useful for OS in OD and provides a more friendly approach.
Trial registration number: not applicable Trial registration number: NCT03300960
O-010 Can serve Medroxiprogesterone acetate (MPA) as O-011 Optimising follicular development, down regulation,
pituitary suppressor instead of GnRH antagonist during ovarian triggering and luteal phase support during in vitro fertilisation
stimulation (OS) in oocyte donation (OD) cycles trigger with (IVF): a Delphi consensus
GnRH agonist? C. Alviggi1, C. Venetis2, H. Fatemi3, T. D’Hooghe4, E. Gravotta5,
J. Giles1, P. Alama1, P. Gamiz2, C. Vidal1, P. Badia1, E. Bosch1 M. Horton6, M. Grynberg7, R. Fischer8, S. Longobardi9,
1
IVIRMA Valencia, Human Reproduction, Valencia, Spain ; S. Esteves10, S.K. Sunkara11, Y. Li12, Y. Koloda13, R. Orvieto14
2 1
IVIRMA Valencia, IVF Laboratory, Valencia, Spain Universita Federico II - University of Naples, Department of Neuroscience
Reproductive Science and Odontostomatology, Napoli, Italy ;
Study question: Is ovarian response of oocytes donors when pituitary is 2
UNSW Australia, Centre for Big Data Research in Health, Sydney, Australia ;
suppressed with MPA comparable to the conventional treatment with GnRH 3
IVI Middle-East Fertility Clinic, Abu Dhabi & Dubai and Muscat Royal Marina
antagonist cycles?. Village, Abu Dhabi, United Arab Emirates ;
Summary answer: MPA serve as pituitary suppressor during OS in OD since 4
Merck KGaA, Global Medical Affairs Fertility, Darmstadt, Germany ;
it does not present lower number of MII or worse reproductive outcome com- 5
Merck Serono S.p.A, Medical Affairs Fertility- Endocrinology and General Medicine,
pared to antagonist. Rome, Italy ;
What is known already: Administering progestins orally in the follicular phase 6
Pregna Medicina Reproductiva, Head- Assisted Reproduction Lab and Quality
since the beginning of OS is an efficient alternative to prevent LH from peaking, Control, Buenos Aires, Argentina ;
related to its effect on LH pulse frequency and amplitude, with similar results 7
Hôpital Antoine Béclère, Department of Reproductive Medicine & Fertility
to conventional protocols. Preservation, Clamart, France ;
Progestins have been successfully used in normo-ovulating patients, Polycistic 8
MVZ Fertility Center Hamburg GmbH, Medical Director of the Fertility Center,
Ovarian Syndrome, endometriosis and low-responders. Hamburg, Germany ;
A randomized controlled trial (RCT) in OD reported no differences with 9
Merck KGaA, Global Clinical Development Fertility, Darmstadt, Germany ;
GnRH antagonist in OS parameters, mature oocytes and top quality embryos. 10
ANDROFERT- Andrology and Human Reproduction Clinic, Referral Center for
However, pregnancy outcomes were lower. Male Reproduction, Campinas, Brazil ;
In contrast, no differences were observed between the two groups in terms 11
King’s College London- UK, Division of Women’s Health- Faculty of Life Sciences
of reproductive outcomes in other recently published retrospective studies. and Medicine, Barking, United Kingdom ;
Study design, size, duration: University-affiliated infertility clinic. Prospective 12
Beijing Chaoyang Hospital affiliate to the Capital Medical University,
RCT study, from October 2017 to June 2019, to evaluate ovarian response in Reproductive Medical Center for Human, Beijing, China ;
terms of number of oocytes. We randomized 318 donors in two groups in a 13
Russian Medical Academy of Continuous Professional Education, Department of
1:1 ratio. A difference of ± 3 oocytes respect a mean of 21 in the reference Obstetrics and Gynecology, Moscow, Russia C.I.S. ;
group was considered as an equal response (NCT03300960). 14
Sheba Academic Medical Centre Hospital, Director- Infertility and IVF unit,
Cycle outcome of the recipients were later analysed retrospectively. Oocytes Ramat Gan, Israel
obtained were assigned to 364 recipients (1910-VLC-091-JG).
Participants/materials, setting, methods: In MPA group 161 participants Study question: How can outcomes of in vitro fertilisation (IVF) be improved
received intervention (10 mg daily administered orally during OS) and 156 were using the evidence-based opinion of clinical experts?
treatad with antagonist (started once the leading follicle reached 13 mm). Summary answer: Eighteen statements were developed concerning improving
Transvaginal ultrasound and serum estradiol (E2), LH, and progesterone (P) outcomes of IVF; 17 statements reached consensus during the first vote and
were performed during monitoring controls. one reached consensus after a second vote.
Other parameters that were analyzed: endocrine profile (in serum and follic- What is known already: The ESHRE 2019 Guidelines provide clinicians with
ular fluid), number of MII, pregnancy outcomes. For the latest, a questionnaire valuable evidence-based recommendations to optimise ovarian stimulation and IVF.
was offered to all participants after the oocyte retrieval. However, data from such guidelines are primarily based on randomised controlled
Main results and the role of chance: No significant differences were trials (RCTs) with highly selected populations that are conducted under very con-
observed in donor demographic characteristics. The number of oocytes trolled conditions. Furthermore, these trials are limited by the fact that only ~35%
retrieved were 21.41±11.71in the MPA group vs. 21.26±9.27 in the antagonist of the general patient population has been reported to meet their inclusion criteria.
group (P=0.949) (Mean difference 0.14 [95%CI= -2.233, 2.517]). Study design, size, duration: A Delphi consensus was conducted to formu-
The total dose of rFSH, length of OS and endocrine profile in follicular fluid late expert opinion on how IVF outcomes could be improved. Step 1: state-
in the oocyte pick-up procedure (FSH, estradiol, LH, progesterone) were comments/supporting references were discussed/amended by 11 experts. Step 2:
parable between groups. LH values on the day of trigger were significantly lower 35 experts voted on their level of agreement/disagreement with each statement.
in study group (1.8±2.0 vs 0.9 ±1.1, p=<0.001), while no early luteinization was Consensus was reached if the proportion of participants agreeing/disagreeing
observed in either group. with a statement was >66%. If consensus was not achieved, the statement was
No differences between groups were observed for implantation rate (78% vs. revised and re-voted until consensus was reached. Step 3: consensus results
73.9% p=0.441), clinical pregnancy rate (78.3% vs. 73.3, p=0.383), ongoing communicated to participating experts.
pregnancy rate (70.9 vs. 67%, p=0.592) and early pregnancy loss (9.7% vs. 8.0%, Participants/materials, setting, methods: Step 1 involved the Scientific
p=0.669). Live birth rate would be presented at the congress since there are Board, comprising the Scientific Coordinator, who developed the initial statements
still gestations in progress and supporting references (which included RCTs, meta-analyses, systematic reviews,
There is a significant difference in favor of the MPA group in questions related as well as retrospective studies and review articles), and 10 additional experts. The
to ease of administration and number of injections. In donors with previous Scientific Board discussed and refined the final statements and references. Step 2

involved 35 experts who rated their level of agreement or disagreement with each Participants/materials, setting, methods: Oocyte maturation was trig-
statement and were asked to provide reasons for their rating. gered with human chorionic gonadotrophin except for those with OHSS risk
Main results and the role of chance: Consensus was achieved for 18 where GnRH agonist was substituted and fresh transfer cancelled. The primary
statements, the most relevant of which are summarised below: endpoint was ongoing pregnancy rate (presence of at least 1 intrauterine preg-
nancy with fetal heartbeat 8-9 weeks after fresh transfer). Planned sensitivity
• Follicular development/gonadotropins (n=9 statements): analyses in the modified intent-to-treat population (all randomized subjects who
• Oocyte number and live birth rate (LBR) are strongly correlated; there is a received at least 1 dose of gonadotrophin) included assessment of primary
positive linear correlation with cumulative LBR endpoint rate by infertility diagnoses.
• Exogenous FSH alone is sufficient for follicular development in normogo- Main results and the role of chance: The non-inferiority objective for the
nadotropic patients aged <35 years primary endpoint of ongoing pregnancy was met. HP-hMG was associated with
• Different FSH preparations have identical polypeptide chains but different numerically higher ongoing pregnancy rates vs rFSH (35.5% vs 30.7%, P>0.05).
glycosylation patterns, affecting the biospecific activity of r-hFSH The average number of oocytes per patient (±SD) in the rFSH arm (22.2±11.54)
• rLH supplementation demonstrates improved pregnancy rates and cost effi- was higher than in the hMG arm (15.1±10.12), a difference in ovarian response
cacy versus hMG in patients with severe FSH/LH deficiency that was accompanied by statistically significant increases in rates of OHSS
• Pituitary suppression (n=2): (21.4% vs 9.7%; p<0.05). There were no significant differences in ongoing preg-
• GnRH antagonists are associated with lower rates of any grade OHSS and nancy rate between treatment groups in those diagnosed with endometriosis,
cycle cancellation versus agonists male factor, tubal infertility, idiopathic, or other. However, among those diag-
• Final oocyte maturation triggering (n=4): nosed with oligoovulation, HP-hMG treatment (N=50) was associated with a
• hCG represents the gold standard in fresh cycles 19.2% higher ongoing pregnancy rate (95% confidence interval 1.2%-37.3%) than
• GnRH agonist trigger, in GnRH antagonist protocol, is recommended for rFSH (N=56). Relative to the rest of the trial population, those with oligoovu-
final oocyte maturation in women at risk of OHSS lation had higher mean AMH (52.10 vs. 60.95 pmol/L, p<0.001), luteinizing
• Current evidence supports significantly higher pregnancy rates with hCG + hormone (6.45 vs 7.55 U/L, p=0.007), and testosterone (1.00 vs. 1.14 nmol/L,
GnRH agonist versus hCG alone, but further evidence is needed p=0.006) although FSH and BMI were similar. Comparisons between populations
• The efficacy of hCG triggering for frozen transfers in modified natural cycles with and without oligoovulation were made using either t-tests (continuous
is controversial compared with LH peak monitoring parameters) or Fisher’s exact test (categorical parameters). HP-hMG differs
• Luteal-phase support (n=3): from rFSH in its FSH isoforms and presence of LH activity; either or both of
• Vaginal progesterone therapy represents the gold standard for luteal-phase which could account for differences observed.
support Limitations, reasons for caution: The present trial was not powered to
detect differences in pregnancy outcomes based upon infertility diagnoses.
Additional comparative trials are required to confirm this finding.
Limitations, reasons for caution: The statements only represent the col-
Wider implications of the findings: The current study highlights a possible
lective opinion of the experts included. Furthermore, not all statements reached
opportunity for optimization of stimulation protocols for predicted high-re-
100% agreement, with some statements reaching consensus even though some
sponders with oligoovulation through the use of HP-hMG.
participants disagreed with them.
Trial registration number: NCT02554279
Wider implications of the findings: This Delphi consensus provides a real-
world clinical perspective from a diverse international group of experts.
Additional guidance from clinicians on IVF strategies could complement guidelines O-013 A drop in serum progesterone levels on day of fresh
and policies, and may help to further improve treatment outcomes. blastocyst transfer, using standard luteal phase support is
Trial registration number: not applicable associated with significantly lower ongoing pregnancy rates
E. Uyanık1, M. Polat2, S. Mumusoglu1, I. Yarali Ozbek3, G. Bozdag1,
O-012 Predicted high-responder women diagnosed with P. Humaidan4, H. Yarali1,2
oligoovulation may benefit from stimulation with highly purified 1
Hacettepe University, Obstetrics and Gynecology, Ankara, Turkey ;
human menopausal gonadotrophin (HP-hMG) 2
Anatolia IVF and Women Health Center, Obstetrics and Gynecology, Ankara,
P. Heiser1, E. Foster1, A. Sinha1, O. Elci1, G. Daftary2 Turkey ;
1 3
Ferring Pharmaceuticals- Inc., Clinical Development, Parsippany, U.S.A. ; Anatolia IVF and Women Health Center, Histology and Embryology, Ankara, Turkey ;
2 4
Ferring Pharmaceuticals- Inc., Medical Affairs, Parsippany, U.S.A. Aarhus University- Skive Hospital, Obstetrics and Gynecology, Skive, Denmark
Study question: Do gonadotrophin-related differences in efficacy exist in pre- Study question: Do early- and mid-luteal serum progesterone (P4) levels
dicted high-responder women undergoing assisted reproductive technology impact ongoing pregnancy rates (OPRs) in fresh blastocyst transfer cycles, using
based upon underlying infertility diagnosis? a standard luteal phase support (LPS)?
Summary answer: Ongoing pregnancy rates after fresh blastocyst transfer Summary answer: A drop in serum P4 level from oocyte-pick up (OPU)+3
are significantly higher with HP-hMG versus recombinant follicle stimulating hor- to OPU+5, using a standard LPS, is associated with a 2.8 fold decrease in OPR.
mone (rFSH) in predicted high-responder women with oligoovulation. What is known already: In fresh embryo transfer cycles, significant inter-per-
What is known already: Women predicted to be high-responders are a sonal variation occurs in serum P4 levels during the luteal phase, possibly due to
heterogenous cohort with diverse infertility etiologies and unique treatment differences in endogenous P4 production from corpora lutea after hCG trigger
challenges. Post-hoc analyses of previous trials suggest that these patients exhibit and/or differences in bioavailability of exogenously administered P4 via different
differential treatment response based on type of gonadotrophin administered. routes. Although exogenous P4 may ameliorate this drop in serum P4 in fresh
Use of HP-hMG is associated with fewer oocytes retrieved, higher pregnancy transfer cycles, there is paucity of data exploring the possible impact on repro-
rates, and fewer interventions for ovarian hyperstimulation syndrome (OHSS) ductive outcomes of a reduction in P4 levels.
versus rFSH (Anckaert 2012, Arce 2012, Arce 2013). These findings required Study design, size, duration: Using a prospective cohort study design, a
further evaluation through prospective clinical trials. total of 101 patients were consecutively enrolled between February 2019 to
Study design, size, duration: Planned analysis of a multicenter, randomised, November 2019. The inclusion criteria were; female age <40 yr-old, retrieval
assessor-blind, controlled non-inferiority trial in 620 women, 21-35 years, with of ≥3 oocytes irrespective of ovarian reserve, number of previous failed cycles
BMI 18-30 kg/m2 and serum anti-Mullerian hormone (AMH) ≥35.7 pmol/L ≤2, body-mass index (BMI) < 35 kg/m2, single or double fresh blastocyst transfer.
undergoing intracytoplasmic sperm injection and single blastocyst transfer. Each patient was included only once. The primary outcome measure was OPR,
Patients were randomised to a 150IU dose of rFSH (N=309; GONAL-F, Merck) as defined by pregnancy ≥12 weeks of gestational age.
or HP-hMG (N=311; MENOPUR, Ferring), receiving 150 IU daily for the first Participants/materials, setting, methods: A GnRH-agonist (n=23) or
five days with 75 IU adjustments permitted thereafter in a gonadotrophin releas- GnRH-antagonist (n=78) protocol was used. rhCG was used for trigger in the
ing hormone (GnRH) antagonist protocol. majority of cycles (n=95). Vaginal progesterone gel (Crinone 8%, Merck) once

daily was used for LPS. Serum P4 levels were measured in patients on five cancellation rate, embryo quality, clinical pregnancy rate. Statistical test: Poisson
occasions; on the day of ovulation trigger, day of OPU, OPU+3, OPU+5 and regression.
OPU+14 days; the timing of blood sampling was standardized to be 4-5 hours Main results and the role of chance: Forty-nine patients (group long-testos-
after the morning administration of vaginal gel. terone= 17, short-testosterone= 17, control= 14) completed the study as 14 out
Main results and the role of chance: Female age, BMI, number of previous of the 63 randomised abandoned or were excluded due to several reasons. Basal
cycles, number of oocytes, number and quality of blastocysts transferred were characteristics of the patients were as follows: age (36.51+/-2.99), BMI (23.21+/-
comparable among patients with (n=48) or without (n=53) OP. Similarly, mean 3.6), AMH (4.37 pmol/L+/- 2.54) and days of stimulation (10.15 +/- 2.26). There
P4 level on the day of trigger, day of OPU, and OPU+3 was comparable between were no differences between groups. Testosterone levels and free androgen index
two groups. However, patients with OP had significantly higher P4 levels on at the time of starting stimulation were significantly higher in groups receiving
OPU+5 (103.6±34.0 vs 86.9±37.5 ng/ml, p=0.021). More importantly, a drop testosterone compared to controls. There were no differences between groups
in P4 level from OPU+3 to OPU+5 was seen in 35% of patients (negative-Δ= for androstendione, SHBG or DHEA. Mean number of oocytes retrieved was 3.8
OPU+5 – OPU+3), and was associated with a significantly lower OPR when +/- 3.17 and mean number of MII was 2.56 +/- 2.68. For the primary outcome
compared with positive-Δ counterparts (31.4% vs 56.3%;OR= 2.80 (95% CI; there were no differences between groups (long-testosterone=2.12+/-2.66,
1.17-6.68; p=0.02); this decrease in OPR was due to lower initial pregnancy short-testosterone 2.71+/-2.95, control 2.92+/-2.43, p=0.98). The rest of results
rates rather than increased early pregnancy loss rates. For negative-Δ patients, are still under analysis so we can only report on the primary outcome at this
magnitude of negative-Δ was a significant predictor of OP (AUC=0.80; 95% CI; moment. Shortly we will have the remainder of results regarding cycle parameters
0.65-0.96); with an optimum threshold of -18.1 ng/ml, sensitivity and specificity and outcomes and we will update this abstract accordingly.
were 54.2% and 90.9%, respectively. Of interest, for positive-Δ patients, mag- Limitations, reasons for caution: The drop-out rate was higher than
nitude of Δ was not a predictor of OP (AUC=0.51; 95% CI; 0.37-0.66). When expected (22%, sample size calculated for 15%) which could affect the power
logistic regression analysis was performed, only negative–ΔP4, but not serum to detect differences. We present only partial results regarding mainly the pri-
P4 level on OPU+5 , was noted to be an independent predictor of ongoing mary objective.
pregnancy (OR= 0.34;95% CI; 0.11-0.99;p=0.047). Wider implications of the findings: Based on these preliminary results the
Limitations, reasons for caution: The physiological circadian pulsatile secre- use of testosterone in POR, either in a short or long course does not appear to
tion of P4 during the mid-luteal phase is a limitation when analyses are based on increase the number of MII retrieved and therefore should not be considered
one blood sample, only. as a priming strategy.
Wider implications of the findings: Two measurements (OPU+3 and Trial registration number: NCT03378713
OPU+5days) of serum P4 may delineate those patients with a drop in P4 (~ 35%
of whole cohort), associated with 2.8 fold lower OPRs. Rescuing these IVF
cycles with additional supplementation of P4 or adopting a freeze-all policy should
be tested in future randomized controlled trials. SELECTED ORAL COMMUNICATIONS
Trial registration number: NCT04128436 SESSION 04: MALE FERTILITY RELATED PREDICTORS AND
THEIR USE
O-014 Testosterone priming (short or long course) before IVF 06 July 2020 Parallel 3 10:00 - 11:30
does not improve the number of oocytes retrieved in poor ovarian
responders: a randomized controlled trial.
J. Subira1,2, A. Algaba1, S. Vazquez1, R. Taroncher Dasi1,
S. Monzó Fabuel1, V. Baydal1, A. Ruiz Herreros1, N. Garcia
Camuñas1, J.M. Rubio Rubio1 O-015 Paternal contribution to embryo morphokinetics in a time-
1 lapse incubator system
University Hospital La Fe, Assisted Reproduction, Valencia, Spain ;
2
IVIRMA Castellon, Assisted Reproduction, Castellon, Spain A. Setti1,2, D. Braga1,2, R. Provenza3, A. Iaconelli Jr.4, E. Borges Jr.2,4
1
Fertility Medical Group, Scientific research, São Paulo, Brazil ;
Study question: Does follicular preparation with testosterone increase the 2
Instituto Sapientiae- Centro de Estudos e Pesquisa em Reprodução Assistida,
number of metaphase II oocytes retrieved in poor ovarian responders (POR) Scientific research, São Paulo, Brazil ;
according to Bologna criteria? 3
Fertility Medical Group, Andrology laboratory, São Paulo, Brazil ;
Summary answer: The use of testosterone either in a short or long course 4
Fertility Medical Group, Clinical department, São Paulo, Brazil
before IVF does not increase the number of MII oocytes retrieved.
What is known already: POR is characterized by an androgen-depleted Study question: Can paternal age and semen quality influence embryo mor-
follicular environment. Follicular preparation with testosterone has been used phokinetic events in a time-lapse incubator system (TLS)?
in several studies showing an increase in oocyte recovery in some of them while Summary answer: Embryo morphokinetic parameters are negatively influ-
in others no benefit was shown. Recently, some have advocated that the possible enced by paternal age and positively influenced by the seminal quality.
effect of testosterone would only be achieved when given for several weeks What is known already: Although male-factor infertility is known to play a
prior to IVF, given the duration of folliculogenesis in women. Most of the studies role in 50% of the cases of infertility, the impact of male partner characteristics
published have limited the use of testosterone to the previous luteal phase before on IVF is often ignored. The sperm contribution to the success of IVF shouldn’t
starting ovarian stimulation and/or were not randomized. Thus, follicular tes- be underestimated, however, few studies have focused on the influence of male
tosterone preparation in POR remains a controversial intervention. factors on IVF, with conflicting results. The TLS provides the opportunity of
Study design, size, duration: Randomized controlled trial comparing three multiple observations of the embryo developmental changes, while optimal
groups: long-testosterone (testosterone transdermal gel 12.5 mg/day during culture conditions are maintained. Therefore, the identification of morphokinetic
previous two cycles), short-testosterone (testosterone transdermal gel 12.5 events affected by paternal factors may contribute to a better understanding of
mg/day during previous luteal phase) and control group (no testosterone). morphologic mechanisms in fertilization and behavior of early human embryos.
Single-blinded for physicians involved. Sample size powered to detect a difference Study design, size, duration: Kinetic data were analyzed in 139 patients and
of at least 2 MII: 21 patients per group (N=63). Serum androgen determination 1220 zygotes cultured until day five in a TLS between March/2019 and
at randomization, before starting stimulation and on the day of trigger. Follow-up November/2019. Timing of specific events from the point of insemination was
until 12th week of gestation. determined using time-lapse imaging. Abnormal cleavage patterns, such as
Participants/materials, setting, methods: University hospital La Fe. POR reverse cleavage and direct uneven cleavage, and the presence of multinucleation
patients according to Bologna criteria. Short-antagonist protocol, fixed dose 300 were also recorded. Multivariate linear regression analyzes were used to evaluate
IU hMG throughout stimulation. Primary outcome: number of MII retrieved. the influence of paternal factors on embryo morphokinetic events.
Secondary outcomes: serum androgen levels at the start of stimulation, antral Participants/materials, setting, methods: This study was performed in a
follicles at the start of stimulation, number of follicles on the day of trigger, private university–affiliated IVF center. Recorded kinetic markers were: pronuclei

appearance (tPNa), timing to pronuclei fading (tPNf ), timing to two (t2), three Limitations, reasons for caution: This is a large observational study based
(t3), four (t4), five (t5), six (t6), seven (t7), and eight cells (t8), and timing to on retrospective data collection. Despite our robust methodological approach,
blastulation (tB). Durations of the second (t3-t2) and third (t5-t3) cell cycles the presence of biases related to retrospective design cannot be excluded
(cc2 and cc3, respectively) and timing to complete synchronous divisions s1 Wider implications of the findings: Our findings underscore the importance
(t2-tPNf ), s2 (t4-t3), and s3 (t8-t5) were also calculated. of a complete endocrine evaluation of the subfertile male. Efforts to evaluate
Main results and the role of chance: The paternal age was directly cor- and maybe optimize even asymptomatic low TT in male partners should be
related with longer t2 (B: 0.043, p: 0.024), t3 (B: 0.056, p: 0.044), t4 (B: 0.066, considered. Future research might need to identify a new cut-off in the definition
p: 0.012), t6 (B: 0.080, p: 0.042), tB (B: 0.120, p: 0.041), and the presence of of “normal” TT, in an infertility context
multinucleation (Exp(B): 1.027, p: 0.004), while the implantation rate (B: -1.933, Trial registration number: N/A
p<0.001) and the odds of pregnancy rate (Exp(B): 0.784, p<0.001) were neg-
atively affected by paternal age. The progressive sperm motility was negatively
correlated with t4 (B: -0.016, p: 0.037), t6 (B: -0.028, p: 0.027), t7 (B: -0.044, O-017 Negative impact of elevated DNA fragmentation index
p: 0.003), t8 (B: -0.054, p: 0.002), s3 (B: -0.033, p: 0.019), and tB (B: -0.033, p: (DFI) and human Papillomavirus (HPV) presence in sperm on the
0.019). In addition, the total motile sperm count was inversely correlated with outcome of intra-uterine insemination (IUI)
t8 (B: -0.009, p: 0.021), S3 (B: -0.009, p: 0.005), and tB (B: -0.011, p:0.019). C. Depuydt1, G. Donders2, L. Verstraete1, J. Beert1, G. Salembier1,
Limitations, reasons for caution: Retrospective nature of this study and E. Bosmans1, D. Nathalie3, W. Ombelet3
the small sample size may be a reason for caution. 1
AML- Sonic Healthcare, Department of Hormonology and Reproductive
Wider implications of the findings: The association between embryonic Health- AML, Antwerp, Belgium ;
growth rate and embryo quality is well recognized. Implantation and live-birth 2
University Hospital Antwerpen, Department of Obstetrics and Gynecology,
rates are lower with slow-growing embryos. We demonstrated that increasing Antwerp, Belgium ;
paternal age and poor seminal quality correlates with delayed cell cleavage and 3
Genk Institute for Fertility Technology- ZOL Hospitals, Zorgprogramma
blastulation. This finding highlights the importance of paternal contribution for Reproductieve Geneeskunde B ZRGB, Genk, Belgium
the IVF success.
Trial registration number: Not Applicable Study question: To determine the impact of DFI and HPV-DNA positivity on
fertility outcome (biochemical and clinical pregnancy rate) in sperm before its
O-016 Low male testosterone results in a substantial decrease use in IUI.
of fresh live birth rates in couples with non-male factor infertility Summary answer: DFI and HPV status in sperm from male partners are
undergoing IVF independent predictors of clinical pregnancies (CP) in women undergoing IUI.
P. Drakopoulos1, F. Di Guardo1, C. Blockeel1, M. De Vos1, What is known already: Recent evidence identified HPV infections as a
E. Anckaert1, G. Verheyen1, S. Santos-Ribeiro2, A. Racca1, possible cause of male and couple infertility in IUI. While the pathogenesis of
L. Boudry1, S. Mackens1, H. Tournaye1, V. Vloebergs1 HPV infection in the cervix can be divided in two pathways, (an infectious,
1
UZ Brussel, Center for Reproductive Medicine, Jette- Brussels, Belgium ; virion-producing and a non-infectious, cell transforming, cancer-inducing path-
2
IVI-RMA, Department of reproductive medicine, Lisbon, Portugal way), in sperm, the HPV DNA always originates from infectious virons only
and is limited in time. HPV virions can bind syndecan-1 present at two distinct
Study question: Are male testosterone levels associated with reproductive sites along the equator of the spermatozoon’s head, not only causing detri-
outcomes in couples with non-male factor infertility undergoing IVF? mental effects on sperm parameters but also damages the DNA of the sper-
Summary answer: Low male serum testosterone levels in couples with non- matozoids, impacting on gamete interaction and causing temporal
male factor infertility are associated with a decrease in fresh live birth rates after subfertility.
fresh embryo transfer Study design, size, duration: Non-interventional prospective multi-center
What is known already: Low serum testosterone is found in approximately study (12-month study period) in which we measured both the DFI and HPV
15% of subfertile men. Although testosterone is essential in spermatogenesis, it DNA in sperm before its use in IUI in a cohort of 161 consecutive infertile
is unclear whether low testosterone levels may have a negative impact on repro- couples (209 IUI cycles).
ductive outcomes in couples with non-male factor infertility. Furthermore it is Participants/materials, setting, methods: DFI was measured with the
debatable whether the initial evaluation of the subfertile male should include an sperm chromatin structure assay and HPV DNA was detected with type specific
endocrine assessment quantitative PCRs (HPV 6,11,16,18,31,33,35,39,45,51,52,53,56,58,59,66 and
Study design, size, duration: This was a 7 years (2011 to 2018) retrospec- 68) in sperm before its use in IUI. We analyzed the impact of DFI and HPV
tive, single-center cohort study conducted at a tertiary fertility clinic. positivity on fertility outcome (biochemical and clinical pregnancy rate) and cal-
Participants/materials, setting, methods: All couples with non-male factor culated the clinical cutoff value for DFI.
infertility who underwent their first IVF cycle in a GnRH antagonist protocol Main results and the role of chance: A DFI criterion value of 25.7% was
were included. All asymptomatic men provided morning blood samples (before calculated by ROC curve analysis. Couples with DFI >26% had significantly less
10 am) and none had been on exogenous testosterone or other relevant med- CP than couples with DFI ≤26% (OR 31.8, 1.8-510.9, p = 0.017). In total, 31
ication. Low total testosterone (TT) was defined as TT<264 ng/dL in line with sperm samples from 169 different men were HPV positive, resulting in an HPV
the recently published guidelines of the endocrine society. Testosterone assay prevalence of 14.8% per IUI cycle and a high-risk HPV prevalence of 9.6% per
used was the same for all patients. IUI cycle (20/209). HPV positive sperm samples had a significantly higher DFI
Main results and the role of chance: In total 1026 couples were included compared to HPV negative sperm samples (29.8% vs 20.9%; p=0.011). None
in the analysis. Among them, 136 (13.3 %) had low TT. Semen concentration, of the 31 inseminations in which the sperm tested positive for HPV led to
progressive (A+B) motility (WHO 2010) and morphology using Kruger’s strict pregnancy, even when DFI% was below 26%.
criteria were lower, but not significantly different between patients with low and Limitations, reasons for caution: HPV positivity (virions) in female partners
normal TT [median (IQR): 52.8 x 106 (28-109) vs. 54.3 x 106 (27-93), 58% (41- could also lead to DNA damage of spermatozoa and influence the clinical preg-
68) vs. 59 (45-69) and 5% (4-8) vs. 6% (4-10), p value=0.6, 0.3 and 0.3, respec- nancy rate per cycle even more. Therefore, to have a complete picture of the
tively]. However, fresh live birth rates (LBR) after fresh embryo transfer were prediction model both partners should be tested for HPV and ideally each time
significantly lower in patients with low TT (13.2% vs. 23.2%, p value=0.009). an IUI cycle is performed.
Multivariate regression analysis allowing adjustment for relevant confounders Wider implications of the findings: Because men can easily infect women
revealed that TT status was significantly associated with fresh LBR. The odds of and these HPV infected women have longer duration of transient infections
fresh LB decreased by 65% in couples whose male partner had low TT (adjusted resulting in (longer) infertility period. Considering the low IUI success rates IVF
OR =0.35, 95% CI=0.15-0.79,p value=0.01). The results were also replicated or ICSI might be indicated in HPV-positive couples and in couples with a DFI
when TT was considered as a continuous variable and when free calculated above 26% to increase the pregnancy rate.
testosterone was used in the regression model Trial registration number: EudraCT number: 2017-004791-56

O-018 Proteomic characterization of spermatozoa from patients Summary answer: Idiopathic azoospermia cases with low AMH level (<1.0ng/
with idiopathic infertility. ml) would have more opportunity to present heterogeneous seminiferous
I. Urizar Arenaza1, B. Navarro2, B. Gómez-Giménez1, tubules when micro-TESE was performed and had higher SRR.
A. Odriozola1, S. Martín-González1, I. Muñoa-Hoyos1, M. Gianzo1, What is known already: For infertile patients with non-obstructive azoosper-
T. Ganzabal2, N. Subiran1 mia (NOA), micro-TESE is considered to have higher SRR than traditional meth-
1 ods. However, Serum inhibin B, follicle-stimulating hormone (FSH) and various
University of the Basque Country UPV/EHU, Physiology, Leioa, Spain ;
2 clinical parameters are not reliable predictors for the presence of focal sper-
Quirón Bilbao, Assisted Reproduction Unit., Bilbao, Spain
matogenesis and SRR. SRR of micro-TESE between NOA patients with different
Study question: Which is the proteomic profile of spermatozoa belonging to etiology varies widely. Idiopathic azoospermia would have much lower SRR than
patients with idiopathic infertility? orchitis, cryptorchidism, AZFc deletion, Klinefelter syndrome and other cases
Summary answer: Compared to normozoospermic samples, we identified when micro-TESE is performed. Therefore, it is still controversial that whether
a total of 385 differentially expressed proteins that are involved in human sperm micro-TESE have an advantage in patients with idiopathic azoospermia.
metabolism. Study design, size, duration: This is a retrospective study. From Sept.
What is known already: Infertility has become a medical and social prob- 2014 to Jan. 2020, 163 idiopathic azoospermia patients treated with micro-
lem that affects over 186 million people worldwide and male factors repre- TESE by single surgeon were divided into three groups according to their
sent approximately 45% of clinical cases. Among other factors, anatomical serum AMH level: group A <1.0ng/ml (n=41), group B 1.0-5.0ng/ml (n=68),
or genetic abnormalities, problems in spermatogenesis or environmental group C >5.0ng/ml (n=54). Patient’s age, testicular volume, serum FSH and
factors are responsible for abnormal sperm parameters which contribute to AMH level, SRR and heterogeneous seminiferous tubules presenting rate were
male infertility. However, approximately 20-30% of men with normal sperm evaluated.
parameters have impaired fertility caused by unknown deficiencies. Thereby, Participants/materials, setting, methods: Micro-TESE was performed
the molecular mechanisms underlying idiopathic infertility are poorly at x10 to x20 magnification. An attempt was made to identify seminiferous
understood. tubules that were larger and more opaque than others. The procedure was
Study design, size, duration: We used 80 normozoospermic samples and terminated when sperm were retrieved. If all tubules were seen to have an
80 pathological samples belonging to men with idiopathic infertility. Male idio- identical appearance, at least three samples (upper, middle, and lower) were
pathic infertility was considered when spermatozoa presented normal sperm extracted. Venous blood samples were drawn from each patient (7–10 AM)
parameters but there was a repeated failure in the Assisted Reproduction after an overnight fast. FSH and AMH were measured by electrochemilumi-
Techniques. Samples were obtained from the Quirón Bilbao Clinic and were nescence immunoassay.
isolated and capacitated by swim up. Main results and the role of chance: Testicular sperms were successfully
Participants/materials, setting, methods: We adopted a Tandem Mass- retrieved in 55 cases (SRR=33.7%). There was no statistical difference in age,
Tag (TMT) 6-plex isotopic labeling strategy and generated samples for testicular volume, FSH level between the patients who succeeded and failed to
LC-MS/MS. We extracted proteins from normozoospermic samples and obtain sperm (33.3±5.1 years, 5.4±3.0 ml, 26.7±15.7 IU/L vs. 32.5±5.7 years,
samples with idiopathic infertility and after proteomic procedures we run the 6.0±2.8 ml, 26.1±12.6 IU/L, p>0. 05 respectively). The patients who obtained
generated peptides in a Q Exactive mass spectrometer. Protein searches sperms had lower serum AMH level than those without sperm (2.87±3.79ng/
were performed using the MaxQuant Software. Perseus software (v.1.6.0.7) ml vs. 4.99±4.64ng/ml, p<0. 05). SRR and heterogeneous seminiferous tubules
was employed for the calculation of the statistical significance and fold presenting rate were 56.1% (23/41), 70.7% (29/41) in group A vs. 30.9%
changes between samples with idiopathic infertility and normozoospermic (21/68), 27.9% (19/68) in group B vs. 20.4% (11/54), 13.0% (7/54) in group
samples (p<0.05). C (p<0. 05 respectively). In group A (AMH=0.36±0.29ng/ml), SRR and het-
Main results and the role of chance: In the present study we identified a erogeneous seminiferous tubules presenting rate were highest, meanwhile they
total of 1722 human sperm proteins. Differential proteomic studies showed an were lowest in group C (AMH=9.26±4.37ng/ml).
increase in the relative abundances of 176 proteins and a decrease in the relative Limitations, reasons for caution: Seminal AMH level should be included in
abundances of 209 proteins. According to the proteins that are upregulated, the following study to confirm our conclusion. Randomized controlled trial com-
there are involved in metabolical processes, being the most enriched ones the paring micro-TESE and traditional TESE would demonstrate that whether idio-
acetyl-CoA biosynthetic process, ADP transport and 2-oxoglutarate metabolic pathic azoospermia patients with serum AMH level higher than 1.0ng/ml could
process. Regarding proteins that are downregulated, there are involved in ATP not have benefit by microsurgery.
metabolic processes, protein targeting and nucleotide processes. According to Wider implications of the findings: Recently, in our pathologic research,
our results, human sperm metabolism may be key in the finding of targets that NOA patients with extremely lower serum AMH level are observed to have more
may cause idiopathic infertility. This finding suggests that altered metabolic param- opportunity to present severe hyalinization in their seminiferous tubules. Tubules
eters may lead human sperm to idiopathic infertility. with severe hyalinization have less Sertoli cells and seem very thin. Therefore,
Limitations, reasons for caution: We need further studies to identify and tubules with spermatogenesis would be easy to identify during micro-TESE.
validate specific protein targets that may alter specific metabolic processes and Trial registration number: Not Applicable
consequently cause idiopathic infertility.
Wider implications of the findings: The study of human sperm metabolism O-020 MACS Vs TESA for raised sperm DNA fragmentation
may be essential to understand the causes of many idiopathic infertility cases. index – A RCT
This supports the idea that proteins involved in metabolic processes could be K.C. Mantravadi1, D.R. Gedela1, S. Thakur1, P. Shetty1
used as therapeutic targets in the treatment of idiopathic infertility. 1
Oasis Fertility, Fertility, Hyderabad, India
Trial registration number: CEISH/61/2011.
Study question: In Individuals with raised Sperm DNA Fragmentation Index
(DFI), sperm selection by magnetic activated cell sorting (MACS) or surgical
O-019 extremely low serum AMH level correlates with a high retrieval of testicular sperms (TESA) optimize the reproductive outcomes?
sperm retrieval rate of micro-TESE in men with idiopathic Summary answer: TESA seems like a beneficial intervention to optimize sperm
azoospermia selection and reproductive outcomes for Individuals with raised sperm DFI.
J. Zhang1, X. Liang1 What is known already: It is evident that raised sperm DFI negatively affects
1
the Sixth Affiliated Hospital- Sun Yat-sen University, reproductive center, the reproductive outcomes. Management for raised sperm DFI to optimize
Guangzhou- Guangdong, China reproductive outcomes is still elusive.
Study design, size, duration: This is an ongoing Randomised Control Trial
Study question: To investigate the value of serum anti-Müllerian hormone (RCT) with prior approval from institutional Ethical Committee (IEC). This is
(AMH) level in predicting the sperm retrieval rate (SRR) of micro-TESE for preliminary data of the pilot study. Study duration 6months (March – October
patients with idiopathic azoospermia. 2019). Couples undergoing IVF stimulation with raised sperm DFI were

randomised into MACS group (n=30) and TESA group (n=30) for sperm selec- Participants/materials, setting, methods: Participants: Women with endo-
tion. Couples with history of one failed IVF cycle were offered testing for sperm metriosis confirmed with a visual diagnosis at diagnostic or operative laparoscopy.
DFI. Individuals with sperm DFI>30% were included in the study. Intervention: Laparoscopic intervention. Comparison: Another laparoscopic or
Participants/materials, setting, methods: Sperm DFI testing was done robotic intervention, holistic or medical intervention or diagnostic laparoscopy.
with SCSA method and randomized using software. Intra Cytoplasmic Sperm Risk ratios (RR) were calculated for dichotomous data and mean differences
Injection (ICSI) was the method of insemination in all cases. Extended embryo (MD) for continuous data, with 95% confidence intervals (CI). Heterogeneity
culture till blastocyst stage was done and a freeze all policy was opted. Two was examined via the I2 statistic. Primary analysis was conducted on data per
Blastocysts that showed 100% survivals were transferred in a Frozen Embryo woman randomised. Primary outcomes: overall pain and live birth.
transfer (FET) cycle. Implantation rates (IR) and Clinical Preganacy Rates (CPR) Main results and the role of chance: For this update, we screened 812 titles
were compared between both groups. and abstracts. We identified 5 new studies (including one ongoing study). We
Main results and the role of chance: MACS group showed 38% blastocyst included a total of 14 studies with 1102 women in the review. Laparoscopic surgery
formation rates with a CPR of 50% and IR of 43% (ablation or excision) was associated with decreased overall pain (measured as ‘pain
TESA group showed 31% blastocyst formation rates with a CPR of 83% and better or improved’) at 6 months (RR6.58, 95%CI3.31to13.10, three RCTs, n=171,
IR of 72% I2=0%, moderate-quality evidence) and 12 months postoperatively (RR10.00,
Though MACS group had slightly more percentage of blastocyst formation; (95%CI3.21to31.17, one RCT, n=69, low-quality evidence) compared to diagnostic
TESA group had significantly higher CPR and IR. laparoscopy only. Laparoscopic surgery was associated with an increased live birth
Individuals with history of failed implantation and raised sperm DFI, TESA or ongoing pregnancy rate (RR1.94, 95%CI1.20to3.16, two RCTs, n=382, I2=0%,
seems to be beneficial intervention to optimize reproductive outcomes. moderate-quality evidence).Laparoscopic surgery was associated with decreased
Limitations, reasons for caution: Small sample size. TESA is a surgical overall pain at 12 months (measured as ‘pain free at 12 months’) postoperatively
intervention. compared to diagnostic laparoscopy and medical treatment (RR5.63,
Wider implications of the findings: Testicular sperm seem to have better 95%CI1.18to26.85, one RCT, n=35, low-quality evidence).On comparing laparo-
DNA quality than ejaculated sperm. In couples with failed IVF attempts and scopic ablation with laparoscopic excision, there was no evidence of a difference in
raised sperm DFI we can offer TESA as an active intervention to optimize repro- overall pain (12 months) (MDon0to10VAS0.00, 95%CI-1.22to1.22, one RCT,
ductive outcomes. n=103, low-quality evidence). Live birth was not reported in any of the included
Trial registration number: REF/2019/07/026887 trials. We included three studies in this comparison, while one study is awaiting
classification.Due to the limited available evidence it is unclear if a certain surgical
technique is superior. We included 2 studies, both are awaiting classification.
Limitations, reasons for caution: The quality of some of the included
SELECTED ORAL COMMUNICATIONS
studies, the heterogeneity of the disease among the different studies and limited
SESSION 05: ENDOMETRIOSIS AND UTERINE DISORDERS. long-term follow-up are the major limitations of this systematic review.
NEW CLINICAL INSIGHTS Furthermore, there were few studies for each comparison and meta-analysis
06 July 2020 Parallel 4 10:00 - 11:30 could rarely be performed.
Wider implications of the findings: Surgical (laparoscopic) treatment of
endometriosis appears to beneficially influence pain and pregnancy outcomes.
Further high-quality RCT are required to further differentiate between various
O-021 Laparoscopic surgery for endometriosis: a Cochrane treatment options (whether or not surgical) and their corresponding effect on
systematic review (Cochrane update). the different outcomes of the core outcome set for endometriosis.
C. Bafort1,2, Y. Beebeejaun3, C. Tomassetti1,2, J. Bosteels2,4, Trial registration number: NA
J. Duffy5
1 O-022 Long-term treatment with norethindrone acetate
UZ Leuven, OBGYN/Leuven University Fertility Center, Leuven, Belgium ;
2 decreases the postoperative recurrence of deep endometriosis at
KULeuven, Dept of Development and Regeneration, Leuven, Belgium ;
3 long-term follow-up
Guy’s and St. Thomas’ Hospital NHS Foundation Trust, Assisted Conception Unit,
London, United Kingdom ; S. Ferrero1, C. Scala2, G. Evangelisti1, F. Barra1
4 1
Imelda Hospital, Department of obstetrics and gynaecology, Bonheiden, Belgium ; IRCCS Ospedale Policlinico San Martino - University of Genoa, Academic Unit of
5
University of Oxford, Nuffield Department of Primary Care Health Sciences, Obstetrics and Gynecology, Genoa, Italy ;
2
Oxford, United Kingdom Gaslini Hospital, Unit of Obstetrics and Gynecology, Genoa, Italy
Study question: Does laparoscopy improve pain and/or infertility associated Study question: Does postoperative treatment with norethindrone acetate
with endometriosis compared to any other laparoscopic or robotic intervention, (NETA) decrease the risk of endometriosis recurrence following surgical treat-
holistic or medical treatment or diagnostic laparoscopy only? ment of endometriosis?
Summary answer: This updated review shows that laparoscopic surgery Summary answer: Postoperative administration of NETA significantly
reduces pain and improves live birth rates. When comparing different surgical decreases the risk of recurrence of both endometriosis related symptoms and
techniques, no difference in outcomes was shown. ultrasonographically diagnosed endometriotic lesions at long-term follow-up.
What is known already: Endometriosis is defined by the presence of endo- What is known already: Progestins are widely used to treat endometrio-
metrial glands and stroma in ectopic locations such as the pelvic peritoneum, sis-related symptoms. Postoperative administration of hormonal therapies may
ovaries and rectovaginal septum. Symptoms include pain and/or infertility. decrease the recurrence of endometriosis-related pain and ovarian endometri-
Treatment options for endometriosis include medical therapy (hormonal therapy omas. Limited information is available on the role of postoperative treatment
and non-steroidal anti-inflammatory drugs) and surgery. Laparoscopic surgery with NETA in preventing recurrence of endometriosis.
may benefit in treating overall pain and infertility associated with mild to mod- Study design, size, duration: The study was performed between June 2014
erate endometriosis. Although the laparoscopic management of endometriosis and December 2019. Four hundred thirty-seven women were enrolled. The
is widely accepted, the ideal surgical techniques are still being debated. patients, after being undergone surgical excision of deep endometriosis, either
Study design, size, duration: For the update of this Cochrane systematic received continuous postoperative treatment with NETA or did not receive any
review we conducted electronic searches of the Cochrane Gynaecology and hormonal therapy, were enrolled. Patients exit the study when they interrupted
Fertility Specialised Register of Controlled Trials, CENTRAL, MEDLINE, the protocol started immediately after surgery.
EMBASE, PsycINFO and CINAHL from inception to July 2019 to identify relevant Participants/materials, setting, methods: The choice of receiving NETA
randomized controlled trials (RCTs). Three independent review authors (CB, was based on the preference of the patients (including previous experiences with
YB, CT) independently selected trials and extracted data for meta-analysis. Any hormonal therapies and their adverse effect, contraindications to hormonal ther-
disagreements were resolved by discussion or by a third review author (JD). apies and/or desire to conceive). Patients underwent a follow-up consultation

(including transvaginal ultrasonography) every six months. Presence and intensity who had a 2 g/dL hemoglobin increase from baseline and an absolute percentage
of pain symptoms were assessed. Quantity of life was evaluated using the EHP30 change from baseline hemoglobin was assessed.
and sexual function was evaluated using the Female Sexual Function Index (FSFI). Main results and the role of chance: The proportion of women with anemia
Main results and the role of chance: Demographic characteristics, severity was 33% (n=83) in the Relugolix-CT group and 32% (n=83) in the placebo group.
of endometriosis (assessed by the rASRM classification), prevalence of endome- The anemic women were notably different than the women in the overall study
triomas and length of follow-up were similar in patients who received postoper- populations of LIBERTY 1 and 2. They were more likely to be Black/African
ative NETA (n = 309) and in those who did not receive postoperative therapy (n American women (65.2% vs 51.2%) and from North America (85.9% vs 75.5%]).
= 128). At a median length of follow-up was 46 months (range, 12-73 months), Their baseline MBL volume was greater (mean: 285.5 mL vs 228.8 mL) with
the rate of ultrasonographic diagnosis of endometriosis recurrence was significantly larger proportion having baseline MBL volumes ≥ 225 ml (52.0% vs 34.8%) and
lower in patients treated with NETA (16/309; 5.2%) than in those who did not they had a larger baseline uterine volume (mean: 488 cm3 vs 408 cm3).
receive hormonal therapy (22/128; 17.2%; p < 0.001). NETA significantly Nonetheless, a significantly greater proportion of women had a clinically mean-
decreased the risk of recurrence of endometriomas (p <0.001) and of deep ingful increase in hemoglobin levels of ≥ 2 g/dL in the pooled Relugolix-CT arms
endometriotic nodules (p < 0.01). NETA decreased the occurrence of de novo (55.7%) compared with the placebo arms (11.7%; p < 0.0001). Similarly, the
endometrioma and deep endometriotic lesions during the follow-up (p < 0.05). mean percentage increase in hemoglobin concentration was greater in the pooled
Similarly, the recurrence of endometriosis-related pain symptoms was significantly Relugolix-CT arms (23.0%) compared with the placebo arms (6.4%; p <0.0001).
lower in patients treated with NETA (31/309; 10.0%) than in those who did not Limitations, reasons for caution: Patients with evidence of iron deficiency
receive hormonal therapy (46/128; 35.9%; p < 0.001). Quality of life and sexual anemia, at any time during the trial, were required to receive parenteral or oral
function were better in patients treated with NETA than in those who did not iron supplementation. However, iron therapy was not standardized. Therefore,
receive postoperative hormonal therapy (p < 0.001 and p < 0.001, respectively). outcomes may have been impacted by differences in methods of supplementa-
Limitations, reasons for caution: The study was not randomized. Only tion (oral versus parenteral) and patient adherence to therapy.
NETA was investigated in this study; it remains to be evaluated if other hormonal Wider implications of the findings: Overall, these significant improvements
therapies are more efficacious than NETA in preventing postoperative endome- in hemoglobin concentrations from baseline with Relugolix-CT treatment relative
triosis recurrence. to placebo reflect the impact that HMB had on study participants and how
Wider implications of the findings: Patients undergoing surgery for endo- reductions in MBL volume translate into clinically meaningful improvements in
metriosis who do not desire to conceive should be advised to use postoperative hemoglobin levels in patients with UF.
hormonal therapy to decrease the risk of endometriosis recurrence. Trial registration number: NCT03049735 and NCT03103087
Trial registration number: Not applicable
O-024 Relugolix combination therapy reduced patient-reported
O-023 Relugolix combination therapy improves hemoglobin levels distress from bleeding and pelvic symptoms and improved daily
in anemic women with heavy menstrual bleeding due to uterine activities in patients with uterine fibroids in the LIBERTY
fibroids: results from the LIBERTY Phase 3 program program
R. Venturella1, E.A. Stewart2, A. Al-Hendy3, J. Kang4, L. McKain4, E. Stewart1, A. Al-Hendy2, R. Venturella3, A. Poindexter III4,
A. Lukes5 C. Villarroel5, Y. Li6, R.B. Wagman6, E. Hunsche6, A. Lukes7
1 1
University Magna Graecia, Obstetrics and Gynecology, Catanzaro, Italy ; Mayo Clinic, Ob Gyn, Rochester MN, U.S.A. ;
2 2
Mayo Clinic, Department of Obstetrics & Gynecology, Rochester- Minnesota, University of Illinois/Chicago, Department of Obstetrics and Gynecology,
U.S.A. ; Chicago- Illinois, U.S.A. ;
3 3
University of Illinois/Chicago, Department of Obstetrics and Gynecology, Magna Graecia University of Catanzaro, Department of Obstetrics and
Chicago- Illinois, U.S.A. ; Gynecology, Catanzaro, Italy ;
4 4
Myovant Sciences Inc., Myovant Sciences Inc., Brisbane- California, U.S.A. ; Baylor College of Medicine and St. Luke’s Episcopal Hospital, Obstetrics &
5
Carolina Woman’s Wellness Center, Obstetrics and Gynecology, Durham- North Gynecology, Houston- Texas, U.S.A. ;
5
Carolina, U.S.A. Instituto de Investigaciones Materno Infantil IDIMI- University of Chile,
Reproductive Medicine Unit, Santiago, Chile ;
Study question: What is the impact of 24 weeks of relugolix combination 6
Myovant Sciences Inc., Myovant Sciences Inc., Brisbane- California, U.S.A. ;
therapy (Relugolix-CT) on hemoglobin in women with uterine fibroids (UF) and 7
Carolina Woman’s Wellness Center, Obstetrics and Gynecology, Durham- North
anemia at baseline? Carolina, U.S.A.
Summary answer: In women with UF and anemia at baseline, Relugolix-CT,
an oral gonadotropin-releasing hormone with estradiol/norethindrone, signifi- Study question: What is the effect of relugolix combination therapy
cantly improved hemoglobin concentrations at Week 24 (p < 0.0001). (Relugolix-CT) on patient-reported distress from common uterine fibroids (UF)
What is known already: In the LIBERTY 1 and 2 Phase 3 studies, Relugolix-CT symptoms and on activities in women with UF?
(once-daily relugolix 40 mg, estradiol [E2] 1.0 mg, and norethindrone acetate Summary answer: Relugolix-CT treatment for 24 weeks significantly reduced
[NETA] 0.5 mg) significantly reduced menstrual blood loss (MBL) in women with patient-reported distress from common UF symptoms and improved physical
UF-associated heavy menstrual bleeding (HMB) and was well tolerated. In the and social activities vs placebo (nominal p < 0.001).
pooled analysis of Phase 3 data, the proportion of responders was 72.3% in the What is known already: The efficacy and safety of Relugolix-CT treatment
Relugolix-CT group (n=254) and 16.8% in the placebo group (n=257); p < (once-daily relugolix 40 mg, estradiol [E2] 1.0 mg, and norethindrone acetate
0.0001. Additional benefits included a clinically meaningful reduction of UF-related [NETA] 0.5 mg for 24 weeks), in women with UF-associated heavy menstrual
pain and a high rate of amenorrhea. Coadministration of E2/NETA maintained bleeding (HMB) was demonstrated in the LIBERTY 1 and 2 Phase 3 trials.
bone mineral density and minimized incidence of vasomotor symptoms. Relugolix-CT achieved a statistically significant and clinically meaningful reduction
Study design, size, duration: LIBERTY 1 and 2 were randomized, dou- in menstrual blood loss and in pain among patients with moderate to severe pain
ble-blind, placebo-controlled, international, Phase 3 trials to assess the efficacy at baseline, a high rate of amenorrhea, and a clinically meaningful hemoglobin
and safety of Relugolix-CT for 24 weeks of treatment. Women were randomized increase in patients with anemia. Furthermore, bone mineral density was main-
1:1:1 to one of three arms: Relugolix-CT for 24 weeks, Delayed Relugolix-CT tained, and vasomotor symptoms minimized, as in the placebo group.
(relugolix 40 mg alone for 12 weeks followed by Relugolix-CT for 12 weeks), or Study design, size, duration: 388 and 382 premenopausal women with clin-
placebo for 24 weeks. Data were pooled from LIBERTY 1 and 2 for this analysis. ically significant uterine fibroids and alkaline-hematin documented HMB, respec-
Participants/materials, setting, methods: Premenopausal women (aged tively, were randomized 1:1:1 to: Relugolix-CT for 24 weeks, relugolix 40 mg for
18–50 years) with an MBL volume of ≥ 80 mL/cycle (assessed using the alkaline 12 weeks followed by Relugolix-CT for 12 weeks, or placebo for 24 weeks. Pooled
hematin method) and ultrasound-confirmed UF were eligible. The results of an data from both studies were used for analyses of distress from common UF
analysis of a subset of women with hemoglobin ≤ 10.5 g/dL at baseline who symptoms, measured using a new, validated patient-reported outcome tool, and
had a hemoglobin value at Week 24 are reported. The proportion of patients of activities assessed with the previously validated UFS-QoL revised activities scale.

Participants/materials, setting, methods: The Bleeding and Pelvic Participants/materials, setting, methods: EH was diagnosed on hystero-
Discomfort (BPD) scale consists of three UFS-QoL items assessing distress due scopic endometrial biopsies. First-line treatment was either oral MPA 10 mg
to HMB, passing blood clots, and pelvic pressure/tightness. The Revised daily (n=193) or LNG-IUS (n=28). In case of EH persistence, patients receiving
Activities (RA) scale contains five items relating to physical and social activities. oral MPA either switched to LNG-IUS (n=63) or continued daily MPA (n=14).
Both scales range from 0 to 100; least squares means (95% confidence interval Follow up biopsies were scheduled after 90 days of treatment, both for first and
[CI]) for change from baseline were obtained using mixed-effects model repeated second line therapy. If LNG-IUS was used, it was kept in place during biopsy and
measures, and percentage of responders were analyzed for the Relugolix-CT removed after confirmation of negative histology.
and the placebo groups. Main results and the role of chance: Baseline characteristics and possible
Main results and the role of chance: The reduction from baseline to week confounders such as age at diagnosis, BMI, parity, smoking status and duration of
24 in distress due to HMB, passing blood clots, and pelvic pressure, measured infertility did not differ between patients receiving LNG-IUS versus MPA as first-line
by the BPD scale, was significantly greater with Relugolix-CT (-45.0) than with treatment for EH (data not shown). EH regression rate at first follow up (3 months)
placebo (-16.1) (difference -29.9; p<0.001). With Relugolix-CT, mean BPD scale was higher in the LNG-IUS group compared to the oral progestins group (28/28
scores improved from 67.5 (CI: 63.4, 71.5) at baseline to 22.7 (CI: 17.8, 27.6) and 116/193 respectively, 1000% vs 60.1%, p< 0.001). EH regression after sec-
at Week 24, while the placebo group changed from 71.9 (CI: 67.8, 75.9) to 55.8 ond-line treatment was 61/63 in the group switching from MPA to LNG-IUS and
(CI: 50.9, 60.6). In addition, Relugolix-CT led to an improvement in RA scale 12/14 in the group continuing MPA (96.8 vs 85.7%, p=0.15). Overall, n=91
scores from 33.8 (CI: 29.3, 38.3) at baseline to 78.4 (CI: 73.2, 83.5) at Week patients used LNG-IUS as either first or second-line treatment for EH, while n=130
24, while the placebo group changed from 29.4 (CI: 24.9, 33.9) to 43.8 (CI: 38.8, only used MPA. The cumulative live-birth rate following ART in patients ever
48.9); the difference between Relugolix-CT and placebo (30.7) was statistically receiving LNG-IUS (21/91, 25.4%) was similar to the cumulative live-birth rate
significant (p < 0.0001). Percentages of treatment responders, defined as those patients observed in patients only receiving MPA (33/130, 23.1%, p=0.75).
with a clinically meaningful change from baseline to Week 24 of ≥ 20 points, Confounders including age at diagnosis (36.8 ± 1.8 vs 37.9 ± 0.3, Mean ± SD),
were significantly greater with Relugolix-CT than placebo. On the BPD scale, BMI (23.1 ± 0.4 vs 22.1 ± 0.2, Mean ± SD) and number of previous failed ART
61.7% vs 27.6% on Relugolix-CT vs placebo were responders, with a significant cycles (1.3 ± 0.2 vs 1.8 ± 0.2, Mean ± SD) did not differ between patients ever
difference of 34.2% (p<0.0001). Regarding the RA scale, 60.9% and 35.4% of receiving LNG-IUS versus patients only receiving MPA respectively.
patients on Relugolix-CT and placebo, respectively, were responders, with a Limitations, reasons for caution: Limitations of this study are the relatively
statistically significant difference of 25.5% (p<0.0001). small number of patients included and the fact that treatment’s choice was not
Limitations, reasons for caution: The BPD is a new scale that has not been based on randomization but rather on patient’s preference after thorough coun-
used previously. Hence, results cannot be compared with other studies. selling. Another relevant information that is currently missing is a cost-analysis
Wider implications of the findings: Relugolix-CT not only reduced HMB comparing the two groups.
in women with UF, but also improved patient-reported outcomes, by reducing Wider implications of the findings: LNG-IUS should be the preferred
the distress caused by frequent UF symptoms and improving daily activities. therapy for EH without atypia in infertile patients, as it provides very high curing
Responder rates for both patient-reported outcomes provide meaningful results rates already after 3 months of treatment and does not impair endometrial
that are easy to interpret and communicate to patients. receptivity in the following ART cycles. Further prospective data collection and
Trial registration number: NCT03049735 and NCT03103087 studies confirming our results will be of interest.
Trial registration number: na
O-025 Levonorgestrel-releasing intrauterine system (LNG-IUS)
versus oral medroxyprogesterone acetate (MPA) in infertile O-026 Endometrial cancer risk in women with histological proven
women with endometrial hyperplasia without atypia: prospective endometriosis or adenomyosis; a retrospective nationwide cohort
evaluation of regression rates and live-birth rates study.
V.S. Vanni1, G. Campo1, E. Papaleo2, F. Turba1, D. Casiero1, M. Hermens1, I. Velthuis2, R. Bekkers1, H. Van Vliet1, S. Bert3,
M. Sajdak2, R. Villanacci1, L. Quaranta2, F. Pasi2, M. Scarrone3, A. Van Altena2
S. Bordoli3, V. Canti3, M. Candiani1, P. Rovere-Querini3, P. Giardina2 1
Catharina Hospital, Obstetrics and Gynaecology, Eindhoven, The Netherlands ;
1 2
Università Vita-Salute San Raffaele, Obstetrics and Gynecology Dept., Milano, Italy ; Radboudumc, Obstetrics and gynaecology, Nijmegen, The Netherlands ;
2 3
IRCCS Ospedale San Raffaele, Obstetrics and Gynecology Dept., Milan, Italy ; Radboudumc, Pathology, Nijmegen, The Netherlands
3
Università Vita-Salute San Raffaele, Internal and Specialistic Medicine Dept.,
Milano, Italy Study question: What is the association between histological proven endo-
metriosis/adenomyosis and endometrial cancer?
Study question: What is the role of Levonorgestrel-releasing intrauterine Summary answer: Women with histological proven endometriosis/adeno-
system (LNG-IUS) in the treatment of infertile women with endometrial hyper- myosis have an increased risk of endometrial cancer.
plasia without atypia (EH)? What is known already: Women with histological proven endometriosis/
Summary answer: EH regression rates are higher in patients receiving LNG- adenomyosis have an increased risk of ovarian cancer. Smaller studies show
IUS compared to oral MPA. Live-birth rates following assisted reproduction conflicting results on the endometrial cancer risk in women with histological
treatments are comparable between the two groups. proven endometriosis/adenomyosis.
What is known already: LNG-IUS is considered superior to continuous oral Study design, size, duration: A population-based retrospective cohort study
progestin therapy for treatment of EH in the general population (El Behery et of 131,646 women with histologically proven endometriosis/adenomyosis,
al., 2015). Treatment of EH in infertile patients undergoing assisted reproduction matched with 132,700 women with a benign dermal nevus selected from the
treatments (ART) however requires special considerations, due to the need to Dutch nationwide registry of histopathology and cytopathology (PALGA)
preserve short-term endometrial receptivity. LNG-IUS causes downregulation between January 1990 and July 2017. In the endometriosis/adenomyosis group
of endometrial receptors to an extent that is greater than that observed after 1,820 (1.4%) women had histological reports on endometrial cancer and in the
oral progesterone administration (Vereide et al., 2005). Thus, a detrimental effect nevus group 771 (0.6%) women. Crude and age-adjusted incidence rate ratios
of LNG-IUS on fertility outcomes could be hypothesized and should be excluded (IRR) for endometrial cancer and its subtypes were estimated.
before widely adopting the use of LNG-IUS in the treatment of EH in infertile Participants/materials, setting, methods: We selected all women with
patients. histological proven endometriosis/adenomyosis and women with a benign der-
Study design, size, duration: This prospective cohort study was designed mal nevus diagnosed between 1990 and 2015 from the Dutch nationwide registry
to evaluate the role of LNG-IUS as a first or second-line treatment for EH of histopathology and cytopathology.
occurring in infertile patients undergoing ART. N=221 infertile women with a Main results and the role of chance: The age-adjusted IRR for endometrial
diagnosis of EH treated between December 2014 and April 2018 at the cancer overall was 29.60 (95%CI 26.40-33.18). Age at endometrial cancer diag-
Reproductive Medicine Unit of the San Raffaele Hospital (Milan, Italy) were nosis was similar for the endometriosis/adenomyosis and nevus group, 61 (IQR
included. After EH regression, patients were admitted to ART. 55-69) and 62 (IQR 56-68), p=0.28, respectively.

After excluding the first year of follow-up the age-adjusted IRR was 1.19 compared to 11.8% for placebo (p<0.0125). All linzagolix treatments significantly
(95%CI 0.94-1.52), indicating that endometrial cancer is most often found at (p<0.0125) reduced pain and uterine volume, and significant (p<0.0125) reduction
time of histological diagnosis of endometriosis/adenomyosis. In 25.2% of the in fibroid volume was observed for 200 mg linzagolix with and without ABT. Median
endometrial cancer cases in the endometriosis/adenomyosis group, the endo- serum estradiol levels were suppressed below 20 pg/mL in the 200 mg linzagolix
metrial cancer diagnosis was not made until after hysterectomy. These women without ABT group and were maintained from 20 to 60 pg/mL in the other groups.
had not had prior (micro)curettage or biopsy. Mean percent (95% CI) lumbar spine BMD changes from baseline to week 24
Limitations, reasons for caution: These results are limited to women with in the placebo, 100 mg, 100 mg + ABT, 200 mg and 200 mg + ABT dose groups
histological proven endometriosis/adenomyosis, and prone to detection bias. were: 0.514% (-0.019;1.048), -2.068% (-2.630;-1.506), -0.992% (-1.572;-0.412),
Furthermore, no clinical information is available on possible confounders. -4.019% (-4.668;-3.371), -1.305 (-1.923;-0.687), respectively. Changes in femoral
Wider implications of the findings: This study shows an association neck and total hip BMD showed a similar pattern but were generally smaller.
between endometriosis/adenomyosis and endometrial cancer. In most women, Limitations, reasons for caution: This Phase 3 trial of linzagolix for
diagnosis of endometrial cancer and endometriosis/adenomyosis is at time of UF-associated HMB included a relatively narrow population of primarily Caucasian
hysterectomy. To develop preventive strategies future studies should focus on European patients treated for 52 weeks. Post-treatment follow-up will provide
the detection of women at risk for endometrial cancer in the group of women more information on symptom recurrence and BMD recovery. Effects on BMD
suspect for endometriosis/adenomyosis. of calcium/vitamin D supplementation with linzagolix in UF are not known.
Trial registration number: not applicable Wider implications of the findings: Two linzagolix doses were identified
for treatment of UF-associated HMB: 200 mg with ABT, with a responder rate
O-027 Efficacy and safety of linzagolix on heavy menstrual of 94%, and 100 mg without ABT, which has the potential for long-term treat-
bleeding (HMB) due to uterine fibroids (UF): Results from a ment without ABT. Data from a similar ongoing US trial should provide infor-
placebo-controlled, randomized, Phase 3 trial mation on linzagolix in a broader population.
H. Taylor1, J. Donnez2, R. Taylor3, E. Bestel4, J.P. Gotteland5,
A. Humberstone4, E. Garner6
1
Yale University- School of Medicine, Dept. of Reproductive Endocrinology and
Infertility, New Haven- CT, U.S.A. ;
2
Catholic University of Louvain, Société de recherche pour l’infertilité, Brussels,
Belgium ; SESSION 06: FROZEN VERSUS FRESH EMBRYO TRANSFER.
3
University of Utah Health, Department of Obstetrics and Gynecology, Salt Lake AN ONGOING CHALLENGE ON CHILDREN’S HEALTH
City, U.S.A. ; 06 July 2020 Parallel 5 10:00 - 11:30
4
ObsEva SA, Clinical, Geneva, Switzerland ;
5
ObsEva SA, Research and Development, Geneva, Switzerland ;
6
ObsEva SA, Clinical, Boston, U.S.A.
Study question: Do doses of 100 or 200 mg linzagolix with or without estra- O-028 Cardiac remodeling in fetuses conceived by assisted
diol 1 mg/NETA 0.5 mg add-back therapy (ABT) reduce HMB in women reproductive technologies following fresh versus frozen embryo
with UF? transfer
Summary answer: Once daily oral doses of linzagolix 100 and 200 mg with M.L. Boutet1, G. Casals1, B. Valenzuela-Alcaraz1,
and without ABT significantly reduced HMB compared to placebo after 24 weeks L. García-Otero1, F. Crovetto1, E. Vidal1, D. Manau1,
of treatment. E. Gratacos1, F. Crispi1
What is known already: Suppression of estradiol (E2) using GnRH ana- 1
Barcelona Center for Maternal-Fetal and Neonatal Medicine Hospital Clínic and
logues has been shown to be an effective treatment for UF-associated HMB. Hospital Sant Joan de Déu- ICGON- IDIBAPS- Universitat de Barcelona- Barcelona-
Full E2 suppression to postmenopausal levels is highly efficacious but requires Spain, Institut Clínic de Ginecologia- Obstetrícia i Neonatologia, Barcelona, Spain
hormonal ABT to prevent hypoestrogenic symptoms and bone mineral density
(BMD) loss. Linzagolix is an investigational, oral GnRH receptor antagonist, Study question: Do fetuses conceived after frozen embryo transfer (FET)
which dose-dependently reduces E2 levels, providing full suppression (serum present signs of cardiac remodeling and dysfunction similar to those observed
E2 < 20 pg/mL) and partial suppression with once daily oral dosing of 200 in fresh embryo-transfer (ET)?
mg and 100 mg, respectively. We hypothesized that partial E2 suppression Summary answer: Both fresh ET and FET present signs of fetal cardiac remod-
could reduce HMB without the marked BMD loss observed with full eling and dysfunction, with more pronounced changes in the fresh ET as com-
suppression. pared to FET.
Study design, size, duration: PRIMROSE 2 is a Phase 3, double-blind, ran- What is known already: We and others have previously demonstrated that
domized, placebo-controlled, multicenter trial evaluating 100 or 200 mg linzagolix fetuses and children conceived by assisted reproductive technologies present
with or without ABT for 52 weeks. 535 patients were randomized and equally cardiac remodeling and dysfunction (Scherrer et al. Circulation 2012, Valenzuela-
distributed among treatment groups. Subjects randomized to placebo were Alcaraz et al. Circulation 2013, Valenzuela-Alcaraz et al. BJOG 2018). These
crossed-over to 200 mg linzagolix + ABT after 24 weeks. Individual active vs fetuses show more globular hearts, dilated atria, reduced longitudinal motion
placebo efficacy comparisons were conducted at 24 weeks using a 0.0125 sig- and impaired relaxation. Cardiac changes were already present in utero and
nificance level to account for multiplicity of four active treatment groups. persisted after birth. However, 90% of the fetuses included in these studies were
Participants/materials, setting, methods: Participants had HMB (> 80 mL conceived by in vitro fertilization (IVF) after fresh ET. It is unknown whether fetal
menstrual blood loss (MBL)/cycle). The primary efficacy endpoint was HMB reduc- cardiac remodeling is also present in FET.
tion at 24 weeks; responders were defined as having MBL (alkaline hematin method) Study design, size, duration: Prospective cohort study of 397 singleton
of ≤ 80 mL and ≥ 50% reduction from baseline. Other assessments included amen- pregnancies recruited from December 2010 to December 2019, including 132
orrhea, pain, uterine and fibroid volume, haemoglobin levels, and quality of life. E2 spontaneously conceived (SC) pregnancies, 116 conceived by IVF following FET
levels were measured. BMD was assessed centrally using Dual Energy X-ray and 149 IVF after fresh ET. Fetal structural and functional echocardiography was
Absorptiometry. Calcium/vitamin D were not provided or recommended. performed in all pregnancies.
Main results and the role of chance: Subjects had a mean age of 43 years Participants/materials, setting, methods: Pregnancies conceived by IVF
and 218 mL baseline MBL; 5% were Black. At 24 weeks, responder rates were were recruited from a single Assisted Reproductive center, ensuring high homo-
significantly higher (p<0.001) in all linzagolix groups compared to placebo (29.4%); geneity in IVF stimulation protocols, endometrial preparation for FET, laboratory
93.9%, 77.7%. 77.2% and 56.7% in the 200 mg with ABT, 200 mg without ABT, procedures and embryo culture conditions. SC pregnancies from fertile couples
100 mg with ABT and 100 mg without ABT groups, respectively. Amenorrhea (obtained after a period no longer than 12 months) were randomly selected
rates were 80.6%, 70.9%, 63.4%, and 34.0% in the corresponding groups, from low-risk population and paired to IVF pregnancies by maternal age. Fetal

echocardiography was performed at 29-33 weeks of pregnancy to assess cardiac conceived children (SC). It is unknown to what extent these adverse outcomes
structure and function in all pregnancies. are caused by parental or treatment factors.
Main results and the role of chance: Parental age, ethnicity, body mass Sibling comparisons help disentangle the parental and treatment contributions
index and smoking were similar among study groups. Median gestational age at to perinatal health in ART children. Previous sibling studies show conflicting
echocardiography (29-33 weeks) and estimated fetal weight (1400-2000g) were results, and most could not separate children born after fresh-ET and cryo-ET.
similar in all study groups. Both fresh ET and FET groups showed significant signs Study design, size, duration: Registry-based cohort study including liveborn
of fetal cardiac remodeling and dysfunction, with more pronounced changes in singletons in the Committee of Nordic ART and Safety (CoNARTaS) cohort
the fresh ET as compared to FET. IVF fetuses showed larger atria (right atria-to- from Denmark (1994-2014), Norway (1988-2015) and Sweden (1988-2015).
heart ratio: fresh ET median 18.1% [SD 18.1-18.2] vs FET 17.7% [17.6-17.8] vs The population level analyses included 53,345 children born after fresh-ET,
SC 17.1% [17.1-17.2]; linear tendency P-value <0.001), more globular ventricles 14,405 after cryo-ET, and 2,563,837 spontaneously conceived (SC) children. In
(right ventricular sphericity index: fresh ET 1.60 [1.58-1.62] vs FET 1.62 [1.60- total, 27,041 maternal sibling groups with at least two different conception modes
1.64] vs SC 1.68 [1.66-1.70]; <0.001) and thicker myocardial walls (relative wall were identified.
thickness: fresh ET 0.80 [0.78-0.81] vs FET 0.76 [0.73-0.77] vs SC 0.65 [0.63- Outcomes were birthweight, SGA, LGA, preterm birth (<37 weeks) and very
0.66]; <0.001) as compared to SC pregnancies. Both fresh ET and FET groups preterm birth (VPTB, <32 weeks).
had also signs of systolic and diastolic dysfunction with reduced left shortening Participants/materials, setting, methods: Data from nationwide medical
fraction (fresh ET 36% [35.9-36] vs FET 37.1% [37-37.2] vs SC 37.3 [37.1-37.3]; birth registries and ART registries/databases, were linked using unique national
<0.001) and increased left myocardial performance index (fresh ET 1.58 zscores identity numbers. We compared perinatal outcomes according to treatment
[1.54-1.61] vs FET 1.54 [1.49-1.57] vs SC 1.30 [1.27-1.32]; <0.001) as compared modes in multilevel linear and logistic models, where fixed effects (within sibling
to SC pregnancies. All differences remained statistically significant after adjust- groups) were attributed to treatment. Fixed effects models included adjustment
ment by birthweight centile, preeclampsia and prematurity. for birth year and maternal age, parity, smoking, and body mass index. For
Limitations, reasons for caution: The cardiac changes reported here are comparison, random effects models provided population estimates, additionally
subclinical, with most cardiovascular indexes lying within normal ranges. adjusted for country and maternal height.
These milder features in FET fetuses cannot condition the technique´s choice Main results and the role of chance: Children born after cryo-ET had higher
and must be considered together with the global perinatal results related to birthweights (mean difference 73 grams, 95% confidence interval [CI] 59 to 87)
these gestations. and increased risk of LGA (odds ratio [OR] 1.75, 95% CI 1.42-2.2) compared
Wider implications of the findings: Although these features are recognized as to their SC siblings. In contrast, children born after fresh-ET had lower birth-
potential cardiovascular risk factors, their association with adult cardiovascular disease weights (mean difference -56 grams, 95% CI -65 to -47) and increased risk of
remains to be proven. Identification of cardiac remodeling in these fetuses represent SGA (OR 1.31, 95% CI 1.13-1.52), compared to their SC siblings. These esti-
an opportunity to recognize them as potential high-risk population that might benefit mates were comparable to associations at the population level (random effects).
from early preventive measures to improve their future cardiovascular health. Fresh-ET showed the highest risk of PTB at the population level (OR 1.66,
Trial registration number: not applicable 95% CI 1.59-1.73), but risk was increased also for cryo-ET (OR 1.44, 95% CI
1.32-1.57). Comparing siblings, the association attenuated to a modest risk
increase, similar for fresh-ET (OR 1.26, 95% CI 1.13-1.41) and cryo-ET (OR
O-029 Maternal and treatment contributions to perinatal 1.23, 95% CI 1.03-1.48). VPTB showed stronger associations at the population
outcomes after transfer of fresh and cryopreserved embryos in level for both fresh-ET (OR 2.07, 95% CI 1.89-2.26) and cryo-ET (OR 1.70, 95%
assisted reproduction: A Nordic sibling study CI 1.40-2.04), which attenuated completely in the sibling comparisons (fresh-ET
K. Westvik-Johari1, L.B. Romundstad2, C. Bergh3, M. Gissler4, OR 1.04, 95% CI 0.81-1.34; cryo-ET OR 0.95, 95% CI 0.63-1.44).
A.K.A. Henningsen5, S.E. Håberg6, U.B. Wennerholm3, Results were independent of which conception method was the first in each
A. Tiitinen7, A.B. Pinborg8, S. Opdahl9 sibling group and remained similar when restricted to full siblings.
1
Department of Public Health and Nursing-, Norwegian University of Science and Limitations, reasons for caution: Although we could adjust for unmeasured,
Technology- Trondheim- Norway, Trondheim, Norway ; constant parental factors, we cannot exclude confounding from unmeasured,
2
Norwegian Institute of Public Health- Oslo- Norway, Center for Fertility and non-constant parental factors. Information on causes of infertility was limited.
Health, Oslo, Norway ; Consequently, we could not investigate if parental contribution depends on
3
Institute of Clinical Sciences- Sahlgrenska Academy- University of Gothenburg- infertility causes. Further, our study cannot identify specific mechanisms under-
Sahlgrenska University Hospital, Department of Obstetrics and Gynaecology, lying treatment and parental contributions.
Gothenburg, Sweden ; Wider implications of the findings: Adverse perinatal health after ART can
4
Statistics and Registers Unit, THL Finnish Institute for Health and Welfare, be attributed to a combination of parental and treatment factors. Risk of PTB
Helsinki, Finland ; is primarily caused by parental factors whearas fetal growth deviation (SGA in
5
Copenhagen University Hospital- Rigshospitalet, The Fertility Clinic, Copenhagen, fresh-ET and LGA in cryo-ET) is primarily caused by treatment factors. Further
Denmark ; investigations of responsible treatment factors are warranted.
6
Norwegian Institute of Public Health, Center for Fertility and Health, Oslo, Trial registration number: Not applicable
Norway ;
7
Helsinki University Hospital & University of Helsinki, Department of Obstetrics
and Gynaecology, Helsinki, Finland ; O-030 Birth weight and large-for-gestational-age in singletons
8
Copenhagen University Hospital- Rigshospitalet, The Fertility Clinic, , Denmark ; born after frozen compared to fresh embryo transfer by
9
Norwegian University of Science and Technology, Department of Public Health gestational week at birth: a Nordic cohort study
and Nursing, Trondheim, Norway A. Terho1, S. Pelkonen1, S. Opdahl2, L.B. Romundstad3, C. Bergh4,
U.B. Wennerholm4, A.K.A. Henningsen5, A. Pinborg5, M. Gissler6,
Study question: Are the adverse perinatal outcomes seen after transfer of A. Tiitinen7
1
fresh and cryopreserved embryos in assisted reproductive technology (ART) a Oulu University Hospital- University of Oulu, Department of Obstetrics and
consequence of maternal or treatment factors? Gynaecology, Oulu, Finland ;
2
Summary answer: Fetal growth is more strongly influenced by treatment, Norwegian University of Science and Technology, Department of Public Health
whereas risk of preterm birth is more strongly influenced by maternal factors. and Nursing, Trondheim, Norway ;
3
What is known already: Perinatal outcomes differ between children born Norwegian Institute of Public Health, Center for Fertility and Health, Oslo,
after transfer of fresh (fresh-ET) and cryopreserved (cryo-ET) embryos. Fresh-ET Norway ;
4
children have lower birthweights, higher risk of small-for-gestational age (SGA) Sahlgrenska Academy- University of Gothenburg- Sahlgrenska University Hospital,
and preterm birth (PTB), while cryo-ET children have higher birthweights, risk Department of Obstetrics and Gynaecology- Institute of Clinical Sciences,
of large-for-gestational age (LGA) and PTB, compared to spontaneously Gothenburg, Sweden ;

5 1
Copenhagen University Hospital- Rigshospitalet, The Fertility Clinic, Copenhagen, University of Medicine and Pharmacy at Ho Chi Minh City, Department of
Denmark ; Obstetrics and Gynecology, Ho Chi Minh City, Vietnam ;
6 2
THL- Finnish Institute for Health and Welfare, Statistics and Registers Unit, My Duc Hospital, IVFMD- HOPE Research Center, Ho Chi Minh City, Vietnam ;
3
Helsinki, Finland ; University of Medicine and Pharmacy at Ho Chi Minh City, Department of
7
University of Helsinki, Department of Obstetrics and Gynaecology, Helsinki, Finland Pediatrics, Ho Chi Minh City, Vietnam ;
4
My Duc Hospital, HOPE Research Center, Ho Chi Minh City, Vietnam ;
Study question: When does the intrauterine growth difference become appar- 5
University of Adelaide, Fertility SA, Adelaide, Australia ;
ent between singletons born after frozen embryo transfer (FET) and fresh 6
School of Medicine- Monash University, Department of Obstetrics and Gynecoloy,
embryo transfer (fresh ET)? Melbourne, Australia
Summary answer: Mean birth weights and proportion of large-for-gestation-
al-age (LGA) become significantly higher among FET singletons starting from Study question: What are the long-term development outcomes in children
gestational week (GW) 33 and 36, respectively. born after freeze-only versus fresh embryo transfer (ET) in women without
What is known already: In recent years there has been a steep rise in polycystic ovary syndrome (PCOS)?
recorded FET treatments, enabling widespread use of elective single embryo Summary answer: Children born from non-PCOS women undergoing frozen
transfer, thus reducing the risks associated with multiple gestations. However, ET had better problem solving and possibly better fine motor skills than children
FET singletons are heavier and there is a higher risk of LGA (birth weight > 90 born from fresh ET.
percentiles) compared to fresh ET singletons. In turn, risk of small-for-gestation- What is known already: A recent randomized controlled trial (RCT) showed
al-age (SGA, birth weight < 10 percentiles) is lower in FET singletons compared comparable ongoing pregnancy and live birth rates after use of a freeze-only
to fresh ET singletons. The reasons, timing and consequences of these differences versus fresh ET strategy in non-PCOS women undergoing IVF/ICSI
remain largely unclear. There is limited evidence that this difference in growth (NCT02471573; Vuong LN, et al. NEJM 2018;378:137-147). Birth weight was
develops by the last trimester of pregnancy. significantly higher in children born from frozen ET compared with fresh ET.
Study design, size, duration: A retrospective Nordic register-based cohort Here, we report on the development of babies born after frozen versus fresh ET.
comparing singletons born after FET (n = 17 500) to singletons born after fresh Study design, size, duration: This was a long-term follow-up study
ET (n = 69 510) and spontaneous conception (SC) (n = 3 311 588). All live born (NCT04099784) of babies born alive after the first ET in the original RCT. Of 391
singletons between the years 2000 and 2015 in Denmark, Norway and Sweden couples randomised, live birth occurred in 132 (97 singleton/35 twins) and 123
with gestational age ≥ 22 weeks at birth were included from the population-based (95 singleton/28 twins) women in the freeze-only and fresh ET group, respectively;
Committee of Nordic ART (assisted reproductive technology) and Safety 113 parents (86%) from the freeze-only group (147 babies) and 99 (80%) from
(CoNARTaS) study population. the fresh ET group (120 babies) returned questionnaires for this follow-up study.
Participants/materials, setting, methods: FET children were compared Participants/materials, setting, methods: At least 2 years after birth,
to fresh ET and SC for mean birth weight and proportion of LGA and SGA for parents of all babies born alive from the first transfer after freeze-only or fresh
each GW at birth. χ² test and test for relative proportions were used to compare ET in the RCT were contacted via postal mail and e-mail and asked to complete
categorical and Student’s t-test to compare continuous variables. Adjusted odds and return the Developmental Red Flags and Ages & Stages Third Edition (ASQ-
ratios for LGA and SGA were calculated using logistic regressions, adjusting for 3) Questionnaires. Parents received training and completed questionnaires at
year of birth, maternal age, parity, BMI, chronic hypertension, diabetes, smoking home, and returned questionnaires via postal mail, e-mail or instant messenger.
and offspring sex. Main results and the role of chance: The mean age of children at the end
Main results and the role of chance: Mean birth weights were significantly of follow-up was 37 months. Height (95.0±6.1 vs 95.7±5.6 cm; p=0.382) and
higher for FET boys compared to fresh ET boys starting from GW 33 and for weight (14.9±2.6 vs 14.8±2.6 kg; p=0.903) were comparable in the freeze-only
FET girls starting from GW 34 (range from p<0.001 to p=0.046 by week). FET and fresh ET groups (results were also similar when singletons and twins were
boys had a significantly higher proportion of LGA (11.0-15.1%) at birth between analysed separately). ASQ-3 scores for problem solving were significantly better
GW 36-42 compared to fresh ET (7.1-9.4%) (range from p<0.001 to p=0.048 in the freeze-only versus fresh ET group (overall: 53.6±8.4 vs 51.1±10.2,
by week). For FET girls this was seen between GW 37-42 (10.6-13.4%) com- p=0.041; singletons: 52.3±10.1 vs 51.0±9.9, p=0.776; twins: 55.0±5.5 vs
pared to fresh ET (6.6-8.0%) (range from p<0.001 to p=0.009 by week). 51.4±11.1; p=0.202), and there was at trend towards higher fine motor skills
The proportion of SGA was lower among FET boys (7.6-8.7%) compared to scores in the freeze-only versus fresh ET group (overall: 47.8±11.6 vs 44.9±12.6;
fresh ET (11.9-13.6%) between GW 36-42 (range from p<0.001 to p=0.016 by p=0.056; singletons: 46.0±13.4 vs 45.4±12.6, p=0.992; twins: 49.85 ± 8.72 vs
week). For FET girls this was seen between GW 38-42 (7.0-9.3%), compared 43.93 ± 12.71, p=0.060). The overall proportion of children with abnormal
to fresh ET (13.0-14.6%) (p<0.001). ASQ-3 (6.8% vs 8.3%) or abnormal Red Flags (5.4% vs 6.7%) findings in the
The rate of LGA was significantly higher for FET boys between GW 38-41 freeze-only and fresh ET groups was low and did not differ significantly between
(p<0.001) and for FET girls between GW 37-40 (range from p<0.001 to p=0.018 groups. In a quartile-based analysis including predictive variables for ASQ-3,
by week) compared to SC children. there was no significant association between estradiol or progesterone concen-
All singletons born after FET had a higher risk of LGA compared to singletons trations and ASQ-3 overall or problem solving scores.
born after fresh ET (adjusted odds ratio (aOR) 1.87, 95% confidence interval (95% Limitations, reasons for caution: This analysis includes only babies born
CI) 1.76 - 1.98) and singletons born after SC (aOR 1.28, 95% CI 1.22 - 1.35). alive after IVF/ICSI, and not all parents from the original study returned the study
Limitations, reasons for caution: There may be residual confounding factors questionnaires.
that we were not able to control for, most importantly the causes of preterm Wider implications of the findings: Use of a freeze-only strategy resulted
birth, which may also have an influence on the fetal growth. Also, the number in better problem solving and possibly better fine motor skills scores than fresh
of children born extremely preterm or post-term is limited even in a large study embryo transfer, with comparable results in other domains. This highlights the
design. importance of evaluation of normal development in children born after
Wider implications of the findings: This is, to date, the largest study on fresh compared with frozen embryo transfer.
birth weights among preterm and term ART singletons with a population-based Trial registration number: NCT04099784
design and SC control group. Growth differences are an important aspect of the
safety profile of ART. More research is needed on the long-term outcome of O-032 Health in childhood following assisted reproductive
these children. technology (HiCART) - an ongoing study of 600 children born after
Trial registration number: not applicable assisted reproductive technology (ART) in Denmark
L.L. Asserhøj1,2, A.S.L. Kjaer1,2, I. Mizrak1, G.F. Heldarskard1,
O-031 Development of children born from freeze-only versus T.D. Clausen3, G. Greisen4, K.M. Main2, P.L. Madsen5,6, R.B. Jensen2,
fresh embryo transfer: follow-up of a randomized controlled trial A. Pinborg1
T.N.L. Vuong1, T. Ly2, N. Nguyen3, L. Nguyen4, X. Le2, T. Le2, 1
Rigshospitalet, Fertility Clinic, Copenhagen, Denmark ;
K. Le2, T. Le2, M. Nguyen4, V. Dang2, R. Norman5, B. Mol6, T. Ho4 2
Rigshospitalet, Department of Growth and Reproduction, Copenhagen, Denmark ;

3 1
Nordsjællands Hospital- Hillerød, Department of Obstetrics and Gynecology, Epidemiology, Infertility, Paris, France ;
2
Hillerød, Denmark ; Université Bourgogne Franche-Comté - INSERM UMR1231, Biologie de la
4
Rigshospitalet, Department of Neonatology, Copenhagen, Denmark ; Reproduction, Dijon, France ;
5 3
Herlev-Gentofte Hospital, Department of Cardiology, Copenhagen, Denmark ; Clinique de La Muette, Unité de Génétique, Paris, France ;
6 4
Copenhagen University, Department of Clinical Medicine, Copenhagen, Denmark CHU d’Amiens, Centre d’assistance médicale à la procréation- biologie de la
reproduction, Amiens, France ;
Study question: Is body composition of children born after frozen embryo 5
Agence de la biomédecine, Pole Evaluation Biostatistique, La Plaine Saint Denis,
transfer (FET) different compared to children born after fresh embryo transfer France ;
and spontaneously conceived (SC) children? 6
Agence de la Biomedecine, Working group ART women and children health, La
Summary answer: Inclusion is ongoing, but preliminary results revealed an Plaine Saint Denis, France
increased fat-percentage in the cohort despite a BMI close to mean according
to a normal reference material. Study question: Do some specific assisted reproductive treatments (ART)
What is known already: Children conceived after FET are at increased risk present a higher risk of congenital anomalies in singletons?
of being born large for gestational age (LGA) compared with children conceived Summary answer: After multivariate adjustments, including female infertilities,
after fresh embryo transfer and SC children, whereas children conceived after the increased risks of congenital defects associated with intra-uterine insemina-
fresh embryo transfer have an increased risk of being born preterm and small tion were no longer significant, they only subsisted in IVF-group.
for gestational age (SGA) compared with SC children. Studies suggest that ART What is known already: Many epidemiological studies suggest that singletons
may induce epigenetic variations around implantation, fertilization and at the born from assisted reproductive technologies (ART) have a higher risk of birth
early embryonic stages, but the potential long-term effects of ART on the health defects, specifically for musculoskeletal, cardiovascular and urogenital disorders.
of the offspring and the underlying mechanisms are scarcely explored. However, most of these studies were established on data at birth or in neonatal
Study design, size, duration: This cohort study consists of 600 pre-pubertal period and from relatively small population or from several registries. Moreover,
singletons born after FET (n=200), fresh embryo transfer (n=200) or sponta- to our knowledge, the female infertility, a major potential confounder, was not
neous conception (n=200) in Denmark from 2009-2012. All children are iden- included into the risk calculation.
tified by their mother’s personal identification number in the national ART and Study design, size, duration: Using data from the French National System
medical birth registry. The recruitment and examination started January 2019 database (SNDS), we conducted a comparative cohort study on all singleton
and is expected to be completed in September 2020. Currently, 289 children births (deliveries ≥22 weeks of gestation and/or >500g of birthweight) that
(134 boys, 155 girls) have been examined. occurred in France over a 5-year period (2013-2017) resulting from fresh
Participants/materials, setting, methods: The children (age 6-9 years) embryo or frozen embryo transfers (fresh-ET or FET from IVF/ICSI cycles),
undergo a clinical examination with anthropometric measurements including intrauterine insemination (IUI) and natural conceptions (NC). Data was
body mass index (BMI), whole body DXA-scan, blood pressure, pubertal staging available for this cohort of children at least up to early childhood (2.5
and blood sampling (e.g. glucose profile, lipid profile, calcium homeostasis, hor- years old).
monal levels (e.g. IGF-I, IGFBP-3, androgen status, AMH) genetic and epigenetic Participants/materials, setting, methods: A total of 3,501,496 singleton
analyses). Both parents fill in a questionnaire regarding the pregnancy, medical births were included (including 20,218 from IUI, 45,303 from fresh-ET; 18,885
history and current health.Maternal, obstetric and neonatal medical data are from FET). Data were extracted from national health databases. We monitored
obtained from the national ART and birth registry. the major birth defects. Malformations were classified according to the
Main results and the role of chance: Children born after FET had higher International Classification of Disease (ICD-10). To analyse the effect of ART
birth weight (SDS) compared to children born after fresh embryo transfer and conception, multivariate analyses were performed with multiple logistic regres-
SC children (FET vs. fresh embryo transfer: 0.31 [SD 0.96] vs. -0.20 [SD 1.05], sion models adjusted for maternal age, primiparity, obesity, smoking, history of
p=0.001, FET vs. SC: 0.31 [SD 0.96] vs. -0.11 [SD 0.97], p=0.006). Mothers high blood pressure or diabetes and female infertility.
conceiving after ART were older (FET vs. SC: 35.1 vs. 32.2 years, p<0.001, fresh Main results and the role of chance: In our cohort of children, the whole
embryo transfer vs SC: 34.2 vs. 32.2 years, p=0.002) and of lower parity com- prevalence of congenital malformations was 3.8%: 3.8% after NC, 4.5% after
pared to mothers conceiving spontaneously (FET vs SC: 1.50 vs. 1.76, p=0.02, fresh-ET, 4.4% after FET, and 3.9% after IUI. Compared with infants conceived
fresh embryo transfer vs. SC 1.28 vs. 1.76, p<0.001). There were no differences naturally, children born after fresh-ET and after FET had a significantly higher
regarding pregestational BMI and gestational age.To preserve blinding and avoid prevalence of malformations, with an aOR of 1.15 [95%CI 1.10–1.21,
false preliminary conclusions results are presented only for girls and boys respec- p<0.0001] and aOR of 1.12 [95%CI 1.05–1.21, p=0.001], respectively. Among
tively. Height (SDS) was 0.19 [SD 1.04] in boys and 0.16 [SD 1.02] in girls. Weight the fifteen relevant subgroup of malformations studied, we observed a signif-
(SDS) was 0.09 [SD 1.07] in boys and 0.10 [SD 0.99] in girls. BMI (SDS) was icant increased risk for eight of them in the fresh-ET group compared with the
0.01 [SD 1.11] in boys and 0.04 [SD 1.02] in girls. These results are in accordance NC group (i.e. cleft lip and/or palate: aOR 1.39 [95%CI 1.11–1.74, p=0.004],
with Danish sex-matched references. DXA-scans showed a fat-percentage (SDS) respiratory: aOR 1.38 [95%CI 1.08–1.78, p=0.012], musculoskeletal: aOR 1.19
of 1.70 [SD 0.78] in boys and 1.06 [SD 0.69] in girls which was markedly higher [95%CI 1.09–1.30, p<0.0001], nervous: aOR 1.26 [95%CI 1.09–1.47, p=0.002],
for boys compared to reference material. digestive: aOR 1.25 [95%CI 1.09–1.44, p=0.002], urinary: aOR 1.14 [95%CI
Limitations, reasons for caution: The study is powered to detect a differ- 1.01–1.28, p=0.029], cardiovascular systems with namely heart defects: aOR
ence of 0.3 SD in BMI (primary endpoint). Thus, smaller differences may be 1.14 [95%CI 1.03–1.26, p=0.013]). In the FET group, this increased risk was
overlooked. Other outcomes may be relevant, but analyses are exploratory by observed for face (aOR 2.83 [95%CI 1.39–5.75, p=0.004]) and digestive sys-
definition. Mothers included in the two ART groups may have different infertility tems (aOR 1.27 [95%CI 1.03–1.57, p=0.002]. The overall risk of congenital
background which may bias the results. malformations was similar in the IUI group and the NC group, as well as for
Wider implications of the findings: This study will provide important infor- specific anomalies.
mation on the health of children born after ART that may influence the choices Limitations, reasons for caution: For some extremely rare malformations,
made early in the planning of a fertility treatment. The study has the potential the sample size may be too small to reliably conclude that there was no difference
to reduce some of the uncertainties associated with the long-term conse- between groups. Male infertility, the in vitro fertilization method and embryo
quences of ART. stage at transfer could not be taken into account in the analyses. Furthermore,
Trial registration number: ClinicalTrials.gov identifier: NCT03719703 residual confounding cannot be excluded.
Wider implications of the findings: In this large study, after multivariate
O-033 Do assisted reproductive treatments impact the risk of maternal adjustments the increased risks of children defects subsisted after IVF
congenital anomalies in singletons? A longitudinal national French but relatively moderate, and those associated with IUI were no longer significant.
study These findings highlight that underlying parental infertility could contribute to
J. De Mouzon1, P. Fauque2, G. Viot3, A. Devaux4, F. Pessione5, the increased risk of children defects associated with ART.
A.W.G. ART women and children health6 Trial registration number: Not applicable

findings may therefore not be applicable to biopsied tissue. These results reflect
follicle health and not follicle number.
SELECTED ORAL COMMUNICATIONS Wider implications of the findings: These results demonstrate that ovaries
SESSION 07: MALE AND FEMALE FERTILITY PRESERVATION can be stored for up to 48 h prior to cryopreservation with no adverse effect
- CLINICAL ASPECTS on primordial follicles after transplantation and in vivo development of the tissue.
This may have implications on the way ovarian tissue cryopreservation is organ-
06 July 2020 Parallel 6 09:50 - 11:40
ised on a local and national level.
Trial registration number: N/A
O-035 Ovarian function recovery in newly diagnosed advanced

O-034 Delaying cryopreservation for up to 48 hours does not Hodgkin lymphoma patients treated with PET-adapted regimen:
adversely affect ovarian tissue used for fertility preservation prospective analysis of a multicentric randomized phase 3 trial
(AHL2011).
B. Bjarkadottir1, M. Zemyarska1, X. Wei1, C. Walker1, S. Lane2,
J. Davies3, S. Williams1 I. Demeestere1, J. Dechene1, J. Racape2, R. Bouabdallah3, P. Brice4,
1
H. Ghesquieres5, A. Stamatoulas6, J. Dupuis7, A.C. Gac8,
University of Oxford, Nuffield Department of Women’s and Reproductive Health,
T. Gastinne9, B. Joly10, K. Bouabdallah11, O. Casasnovas12
Oxford, United Kingdom ; 1
2
Children’s Hospital Oxford. Oxford University Hospitals NHS Foundation Trust, Université Libre de Bruxelles, Research Laboratory on Human reproduction,
Department of Paediatric Oncology and Haematology, Oxford, United Kingdom ; Brussels, Belgium ;
2
3
Children’s Hospital Oxford- Oxford University Hospitals NHS Foundation Trust, Université Libre de Bruxelles, Research Center in Epidemiology- School of public
Oxford Cell and Tissue Biobank, Oxford, United Kingdom Health- and Biomedical Research Department of CUB-Erasme, Brussels, Belgium ;
3
Institut P. Calmette, Department of hematology-, Marseille, France ;
Study question: Does delaying processing and cryopreservation of ovarian 4
APHP- Hopital Saint Louis, Department of Hematology, Paris, France ;
tissue by 24 or 48 hours adversely affect primordial follicles? 5
Hospices Civils de Lyon- Centre Hospitalier Lyon-Sud- et Université Claude
Summary answer: A 24 or 48 hour delay before cryopreservation of ovarian Bernard Lyon-1, Department of hematology, Pierre Bénite, France ;
tissue does not negatively impact primordial follicle morphology nor follicle 6
Centre H. Becquerel, Department of hematology, Rouen, France ;
growth after 10-day xenotransplantation. 7
H. Mondor Hospital, Department of hematology, Creteil, France ;
What is known already: Ovarian tissue cryopreservation is becoming an 8
Institut d’hématologie de basse normandie, Department of hematology, Caen,
increasingly common method of fertility preservation. While in most cases the France ;
tissue is processed and cryopreserved immediately upon procurement, delaying 9
University Hospital of Nantes, Department of hematology, Nantes, France ;
processing for up to 24 or even 48 hours could help with workload management. 10
Sud Francilien Hospital, Department of hematology, Corbeil-Essonnes, France ;
In addition, ovaries may be procured at a satellite site and transported to a 11
CHU de Bordeaux, Department of Hematology, Bordeaux, France ;
cryopreservation facility, which necessitates a delay in processing. Little is known 12
University Hospital F Mitterrand and Inserm UMR1231, Department of
about the effects delayed processing may have on primordial follicle health and hematology, Dijon, France
the functional potential of the tissue.
Study design, size, duration: Lamb ovaries (n=6 pairs of ovaries) were Study question: Does de-escalated treatment strategy by switching regimen
used to model human ovarian tissue. One ovary per pair was either processed (BEACCOPP to ABVD) in early responder reduce the risk of ovarian damage
into cortical strips immediately or stored whole at 4°C for 24 or 48 h before and infertility compared to standard escalated BEACOPP regimen?
processing. Cortical strips were fixed fresh or after cryopreservation and thaw- Summary answer: Although both treatments affected the ovarian reserve,
ing. Cryopreserved-thawed tissue (immediate processing, 24 or 48 h delay) was de-escalated regimen decreased the risk of premature ovarian insufficiency com-
xenografted subcutaneously into immunocompromised mice (n=3) for 10 days pared to standard regimen in Hodgkin lymphoma patients.
to allow follicle development. Fixed samples were embedded and sectioned. What is known already: Chemotherapy-induced ovarian damage is a major
Participants/materials, setting, methods: Up to 50 primordial follicles from concern for young patients treated for advanced Hodgkin lymphoma. Six cycles
each cortical strip (one per condition/animal) stained with haematoxylin and eosin of bleomycin, etoposide, doxorubicin, cyclophosphamide, vincristine, procarba-
were classified by morphology as healthy or unhealthy. A proportional odds model zine, and prednisone (BEACOPP escalated) showed an improved progression-free
was used to determine whether a delay in processing with or without cryopres- survival rate compared to doxorubicin, bleomycin, vinblastine, and dacarbazine
ervation affected follicle health. A logistic regression model was used to determine (ABVD) in advanced stage diseases but induced high risk of immediate and long-
the odds of follicles being classified as growing after xenotransplantation. Data is term toxicities. AHL2011 trial recently demonstrated that the use of ABVD after
presented as odds ratio (OR) with 95% confidence intervals (CI). two cycles of induction with BEACOPP in early responder based on PET-driven
Main results and the role of chance: A processing delay of 24 h did not strategy was safe while minimizing toxicities (Casasnovas et al, Lancet Oncol 2019).
impact the health of primordial follicles in fresh (OR=0.78, 95%CI 0.38-1.60, Here we investigated its benefit on ovarian function in young patients.
p>0.05) and cryopreserved-thawed tissue (OR=0.93, 95% CI 0.61-1.41, p>0.05; Study design, size, duration: AHL2011 is a prospective randomized non
no delay n=6; 299 fresh follicles, 287 cryopreserved follicles; 24 h delay n=3; inferiority trial enrolling 823 patients from May 19, 2011, to April 29, 2014 in 90
149 fresh follicles, 132 cryopreserved follicles). Conversely, a processing delay centers across Belgium and France. All female patients under 45-year old par-
of 48 h (fresh tissue, n=3; 149 follicles) resulted in primordial follicles being more ticipating to AHL2011 trial were recruited for this prospective sub-analysis of
likely to be graded as unhealthy compared to immediately-processed tissue the ovarian function and fertility at baseline, end of the treatment, and every
(OR=2.47, 95%CI 1.29-4.71, p<0.01). Interestingly however, tissue cryopre- year during 5 years of follow-up.
served after a 48 h processing delay (n=3, 149 follicles) was less likely to contain Participants/materials, setting, methods: Ovarian function was assessed
unhealthy follicles compared to the immediately-processed cryopreserved con- by use of serum levels of FSH, estradiol and anti-mullerian hormone (AMH). All
trol (OR=0.56, 95% CI 0.36-0.87, p<0.01). Xenografting of cryopreserved AMH measurements were centralized. Data on menstruation, use of oral con-
ovarian tissue after 0, 24 or 48 h processing delay demonstrated that grafts from traceptive, fertility preservation, pregnancies and oncological outcomes were
all groups were functional, with no difference in the proportion of growing also collected. Acute premature ovarian insufficiency (POI) was defined as FSH
follicles between groups (immediate processing: 17.3±30.0%; 24 h delay: value above 24 IU/L twice during 5-years follow-up when available, with estradiol
46.0±2.8%; 48 h delay: 45.3±10.8%). Furthermore there was no difference in below 50pg/ml, and low ovarian reserve by AMH value below 0.16ng/ml.
the health of primordial follicles in xenografted tissue after a 24 (OR=1.30, 95% Statistics were performed using STATA15.
CI 0.59-3.22, p>0.05) or 48 h delay (OR=1.26, 95% CI 0.55-3.96, p>0.05). Main results and the role of chance: A total of 146 patients were eligible
Limitations, reasons for caution: These findings should be validated using for ovarian function assessment, 70 in the standard (BEACOPP escaladed) and
human ovarian tissue with a larger sample size to account for the natural vari- 76 in the study (PET-driven strategy) groups. Mean age was similar between
ability between subjects. Ovaries were left intact prior to processing and the both groups (26.7±6.6 and 26.9±6.6 years old, respectively) as well as

oncological characteristics. As expected, the patients in the study group received (8/61). The overall mean time to follow-up is 21.2 ± 19 months (range 1-132
significantly less cumulative doses of alkylating agents within a shorter timeframe. months), with 66% of returning patients doing so within 2 years after cancer
Ovarian function was similar between groups before treatment but became diagnosis.
significantly different during follow-up. FSH values were significantly higher in the Patients with breast cancer were more likely to return to use their gametes
standard group while AMH values were lower. Thirty-two patients faced POI (27/61: 44.3%) and had significantly higher LBR (19/27: 70.3%) in comparison
(46.1% versus 14.5% in standard and study groups, respectively). The risk of to patients with lymphoma (3/8: 37.5%) (p-value <0.001).
POI was significantly associated with age, total dose of alkylating agents, and Limitations, reasons for caution: Although we are certain of capturing the
groups with a lower risk in the study group (OR 0.20, 95% CI 0.08-0.50; outcome of those who had fertility treatment, we can’t be as certain of capturing
p<0.001). Interestingly, low ovarian reserve was not associated with groups (OR all births resulting from natural conception.
0.7, 95% CI 0.31-1.69; p=0.46). Although OR was always lower in the study A proportion of patients may need a longer time to be able to attempt preg-
group, association between low ovarian reserve and chemotherapy doses does nancy, thus the calculated LBR can be underestimated.
not reach significance except for etoposide (OR 0.36, 95% CI 0.14-0.96; p<0.04 Wider implications of the findings: This is a demonstration of how FP can
for doses <5000mg). A total of 29 and 30 patients reported pregnancies after be effective. Over nearly two decades of follow-up, 1 in 6 patients who under-
treatment in standard and study groups, respectively (p=0.81). went FP utilized their stored gametes/embryos with a good outcome. This is
Limitations, reasons for caution: Although this is the larger prospective the first published account of the utilization rate after FP and the longest reported
study analyzing ovarian function in young advanced Hodgkin lymphoma patients period.
considering a new therapeutic strategy, the number of patients remains limited Trial registration number: Not applicable
and hormonal levels were not available at all time-points for all patients during
the 5 years of follow-up. O-037 Long-term fertility and pregnancy outcomes in men
Wider implications of the findings: The PET-driven treatment in young and women with a history of childhood cancer: a nationwide
advanced Hodgkin lymphoma patients allows to reduce the risk of POI by 5 population-based linkage analysis
times compared to standard regimen. Although the ovarian reserve is also better M. Das1, C. Wang2, S. Oberg3
preserved using PET-driven strategy, around 60% of the patients had a low AMH 1
Chelsea and Westminster Hospital NHS Foundation Trust, Reproductive
levels during the follow-up in both groups.
Medicine- Obstetrics and Gynaecology, London, United Kingdom ;
Trial registration number: NCT01358747 2
Karolinska Institutet, Medical Epidemiology and Biostatistics, Stockholm, Sweden ;
3
Karolinska Institutet- Harvard T.H.Chan School of Public Health, Medical
O-036 Live birth rate and utilization rate of eggs and embryos
Epidemiology and Biostatistics, Stockholm, Sweden
following fertility preservation (FP) in 879 female cancer patients
over 19 years Study question: What are the long-term fertility and pregnancy outcomes in men
This abstract
D. Khalife , S. Alihas
, Y. been
Khalaf1chosen for, J.the
, N. Reddy press programme
Kopeika
1 1 1 1
and women with a history of childhood cancer as compared to the general
1
Guy’s and St Thomas’ NHS Foundation Trust, Assisted Conception Unit, London, population?
and will not be made public until the abstract is
United Kingdom Summary answer: Childhood cancer survivors are less likely to have had at
presented. least one biological child and are more likely to need assisted reproductive
Study question: We aim to investigate the rate of women proceeding to FP techniques to conceive.
at the time of cancer diagnosis, the return, utilization and live birth rates after What is known already: Childhood cancer survivors may face reproductive
cancer treatment. challenges due to the disease itself or the necessary cancer treatments. There is
Summary answer: Nearly half of newly diagnosed cancer patients proceed however a paucity of data from large population-based studies on the actual
to FP and return for follow-up within 21 months post-cancer treatment. The estimates of fertility, need for assisted reproduction techniques (ART) and preg-
livebirth rate (LBR) is 72.1%. nancy outcomes in young men and women childhood cancer survivors.
What is known already: FP is an established part of cancer services in many Study design, size, duration: We identified all individuals born in Sweden
countries. It is used now more and more frequently. However, very little is from 1958-1975 (n=2,703,888) and used linkage of national registers to follow
published about long term utilization of gametes/embryos after FP. In male their reproductive outcomes prospectively. Men and women with a history of
population of cancer patients concern is being raised that in spite of “routine” childhood cancer before the age of 18 (n= 6,770) were compared with the
storage of sperm, less than 10% of patients return to use the stored gametes. general population without a history of childhood cancer (n=2,019,645). We
Not much information is found for female cancer survivors. assessed the occurrence of childbearing in the entire population and of trouble
Study design, size, duration: A prospective cohort study was conducted conceiving, use of ART and pregnancy outcomes for the first birth.
on 879 young women diagnosed with cancer who sought FP counseling at the Participants/materials, setting, methods: Cancer in childhood was iden-
Assisted Conception Unit (ACU) at Guy’s and St Thomas’ Hospital (GSTT), tified from the Cancer Register, childbearing from the Multi-generation Register
London, United Kingdom between January 2000 and December 2019. and pregnancy outcomes from the Medical Birth Register. First live birth was
Participants/materials, setting, methods: Data on 879 cancer patients were modelled in Kaplan Meier and Cox regression, whereas risks related to the first
analyzed. Baseline characteristics include age, AMH, AFC, and cancer type. The birth were modelled with logistic regression. We performed a sibling comparison
primary outcome measure was total LBR. The secondary outcomes were return to account for unmeasured confounding by matching each individual with cancer
and utilization rates which were calculated as the number of patients who returned to the nearest unaffected sibling.
for follow-up and those who undergone embryo transfer. Means and frequencies Main results and the role of chance: Childhood cancer survivors were less
were used to describe continuous and categorical variables respectively. Student likely to have had at least one biological child by the age of 37; hazard ratio
t-test analysis was used with p <0.05 considered being statistically significant. adjusted for birth year and sex was 0.83 (95% CI: 0.80,0.86). Moreover, births
Main results and the role of chance: A total of 879 cancer patients received were more likely to have been preceded by trouble conceiving and use of assisted
FP counseling at GSTT with breast cancer being the most common malignant reproductive techniques (ART) in both men and women surviving childhood
disease accounting for 63.1% of the cases. The mean age, AMH, and BMI were cancer, compared to the general population. The differences were more pro-
33.8 ± 7.8 years, 18.8 ± 20.5 pmol/L and 23.7 ± 4.2 kg/m2 respectively. A total nounced in males than in females [males: infertility: OR, 95% CI: 1.29
of 373 patients (42.4%) underwent FP of whom 40.7 % opted for embryo (1.02,1.63); use of ART: 2.17 (1.67, 2.81); females: infertility: OR, 95% CI:1.15
cryopreservation, 53.4% for oocyte cryopreservation, 5.1% had both and 0.76% (1.00, 1.32); use of ART: 1.29 (1.02, 1.61)]. Female cancer survivors had a sig-
opted for ovarian tissue cryopreservation in a different facility. nificantly higher risk of preterm birth (<37 weeks gestation) compared to the
As for the return rate, 33.8% (297/879) of cancer patients returned for fol- general population (OR, 95% CI: 1.56 (1.36, 1.79), and they were also at higher
low-up for assessment of ovarian function, menopausal symptoms, Hormone risk of caesarean section. In sibling comparison, survivors of childhood cancer
Replacement Therapy, and fertility treatment. Until today, utilization rate among remained at greater risk of infertility and use of ART, and the risk of preterm
those who had frozen gametes is16.4 % (61/373) and the overall LBR is 72.1% birth was largely unchanged when female cancer survivors were compared to
(44/61) of which 9.1% (4/44) are twin births. The miscarriage rate is 12.2% their unaffected sisters [OR, 95% CI: 1.52 (1.09, 2.10)].

Limitations, reasons for caution: Detailed information on individual cancer results, further experiments with primary human tumour cells would be required
treatments was not available, so the effects of specific treatment regimens on to validate the current data.
fertility could not be assessed. Wider implications of the findings: This study provides evidence on the
Wider implications of the findings: Both male and female childhood cancer safety of COS with Letrozole for BC patients. These results could be essential
survivors had poorer reproductive outcomes. These findings will help to counsel in reassuring current indications for FP techniques and counselling to patients
young cancer survivors and their families about the effects of cancer and cancer and health-care professionals. Thus, patients with BC could safely underwent
therapies on their future fertility and pregnancy outcomes and will help to provide the gold standard FP technique, oocyte vitrification.
individual risk estimates to guide management. Trial registration number: Not applicable.
O-039 Thyroid cancer treatment and subsequent infertility
O-038 A new strategy to assess safety of controlled ovarian diagnosis in female adolescents and young adults: a population-
stimulation protocols for oocyte vitrification as a fertility based cohort study
preservation technique in breast cancer patients. M.P. Velez1, H. Imsirovic2, H. Richardson2
M.J. Soriano1, J. Martínez1, R. Lopez1, S. Herraiz1, C. Díaz-García1,2 1
Queen’s University, Obstetrics and Gynecology, Kingston, Canada ;
1 2
IVI Foundation- IIS La Fe, Reproductive Medicine, Valencia, Spain ; Queen’s University, Public Health Sciences, Kingston, Canada
2
IVI-RMA Global- IVI London, Reproductive Medicine, London, United Kingdom
Study question: Is thyroid cancer treatment associated with subsequent infer-
Study question: Could controlled ovarian stimulation (COS) protocols affect tility diagnosis?
the proliferative and metastatic potential of breast cancer (BC) cells? Summary answer: Thyroid cancer treatment is not associated with an
Summary answer: Ovarian stimulation with Letrozole for oocyte vitrification increased risk of infertility diagnosis
might be safely used as a fertility preservation (FP) technique in hormone-de- What is known already: Thyroid cancer has undergone the most rapidly
pendent breast cancer women. increasing incidence rate among all major cancers, most likely due to the increase
What is known already: BC is the most common malignancy in women at of surveillance and the use of diagnostic technologies. Normal thyroid function
reproductive age. Treatment with high-dose chemotherapy in female patients is important to maintain normal reproduction. Previous limited literature suggests
may impose deleterious effects on the ovary. COS protocols to obtain oocytes that there is little to no adverse effects of thyroid cancer treatment on gonadal
for vitrification are frequently used within FP patients but associate a rise in function (serum gonadotrophins or sex steroids), fertility (live birth rates) or
supra-physiological estradiol (E2) levels. This effect could increase the prolifer- pregnancy outcomes in women treated for thyroid cancer. However, studies
ation of tumour cells, being probably detrimental for BC women. Nevertheless, with larger sample sizes are needed to better elucidate the risk, if any, of different
standard COS has been recently adapted by using aromatase inhibitors such as types of thyroid cancer treatment on subsequent infertility diagnosis.
Letrozole, leading a maximum E2 peak similar to that found in a natural cycle. Study design, size, duration: A population-based cohort study using uni-
Unfortunately, there is still little evidence regarding safety of such approaches. versal health care databases in the province of Ontario, Canada. All women
Study design, size, duration: Experimental in vivo study. Forty 5-week old 15-39 years of age who received thyroid cancer treatment from 1992-2011 were
Nude-nu female mice were allocated to the following experimental groups: BC identified through the Ontario Cancer Registry (N=4,926) and linked to several
(n=10), BC and FSH stimulation (BC-FSH, n=10), BC and Letrozole stimulation health care datasets. Women were followed up until December 31, 2016
(BC-LTZ, n=10), Control FSH stimulation (CT-FSH, n=5) and Control Letrozole Participants/materials, setting, methods: Thyroid cancer treatment was
stimulation (CT-LTZ, n=5). BC was induced in the three first groups while controls categorized as: less than total thyroidectomy-LTT, total thyroidectomy-TOT, or
received a saline solution injection. Animals were maintained for 5 months and total thyroidectomy plus radioactive iodine therapy-TOT+RAI. Women with
then sacrificed to collect tissue and blood samples for further analysis. infertility, tubal ligation, oophorectomy, or hysterectomy previous to cancer diag-
Participants/materials, setting, methods: One million of human MCF-7 nosis were excluded. Infertility diagnosis was identified using physician billed claims
BC cells, previously transfected with the mCherry fluorescent protein, were (ICD-9 628). Modified Poisson regression models were used to calculate the risk
injected into the left renal capsule of BC, BC-FSH and BC-LTZ mice. Two days of infertility diagnosis adjusted for sociodemographic factors. Models were further
after xenograft, COS was induced by 10IU FSH or 1mg/ml Letrozole + 10IU stratified by parity at the time of cancer diagnosis (nulliparous and parous).
FSH, followed by ovarian triggering with 10IU hCG at 48h. Cell proliferation was Main results and the role of chance: Treatment distribution was: 849 (17%)
biweekly monitored by a non-invasive in vivo imagen system (IVIS) to record LTT, 2457 (50%) TOT, 1620 (33%) TOT + RAI. The mean age at cancer diagnosis
fluorescence signal and also assessed by Ki-67 immunostaining. was 31.04 years (SD=5.99). The median follow-up time for cancer survivors was
Main results and the role of chance: When tumour growth was assessed 10.78 years (IQR=6.96-15.77). A total of 563 (11%) had a subsequent diagnosis
by means of total radiant efficiency signal ([p/s]/[µW/cm2), BC and BC-LTZ of infertility. The frequency of infertility diagnosis was similar among treatments
mice presented a statistically significant lower expression when compared to (p=0.99), with a similar mean time to infertility diagnosis (p=0.69). Mean age at
BC-FSH group (6.1x1010±2.0x1010, 9.6x1010±3.2x1010 and 1.6x1011±4.5x1010; infertility diagnosis was 35.26 years (SD=5.48). Compared to women with LTT,
p<0.01 and p<0.05, respectively), five months after xenograft. Metastasis was women who received TOT or TOT+RAI did not have a higher risk of subsequent
not detected in the BC and BC-LTZ groups, nevertheless, metastatic lesions infertility diagnosis (RR=0.98, 95% CI: 0.78, 1.24, p=0.86; RR=0.93, 95% CI: 0.72,
were observed in BC-FSH mice. The in vivo monitoring results by IVIS were 1.20, p=0.55 respectively). Parity did not modify the estimates.
concordant with the histological assessment of tumour lesions after sacrifice. Limitations, reasons for caution: The accuracy of infertility diagnosis using
Tumour size was slightly lower in BC group than in BC-LTZ (0.3±0.2 cm2 vs ICD-9 codes in administrative datasets has not been validated. Non-biologic
0.5±0.2 cm2). However, lesions in BC-FSH group were considerably increased factors that may influence the likelihood of seeking a fertility assessment may
(1.2±0.3 cm2, p<0.01). Cell proliferation, by Ki-67 immunostaining, was also not be captured in administrative databases.
performed in kidney samples to validate these data. Similar proliferation levels Wider implications of the findings: Thyroid cancer treatment in female
were found in the BC and BC-LTZ groups (10.3±0.9% and 11.5±0.6%). However, adolescents and young adults is not associated with an increased risk of infertility
BC-FSH revealed a significant increase in tumour cell proliferation (28.8±1.6%, diagnosis.
p<0.01). Lastly, mean serum E2 levels of BC and both Letrozole stimulated Trial registration number: not applicable
groups were comparable (BC: 186.9±56.0 pg/ml, BC-LTZ: 193.9±78.2 pg/ml
and CT-LTZ: 119.8±50.7 pg/ml) whereas FSH-treated animals registered sig-
O-040 Impact of In Vitro Fertilization treatments on the risk
nificantly higher E2 concentrations (BC-FSH: 431.2±56.2 pg/ml and CT-FSH:
of recurrence in fertility-sparing management of endometrial
330.9±57.0 pg/ml, p<0.05). All these results confirmed that COS with Letrozole
atypical hyperplasia and grade 1 adenocarcinoma
did not induce the tumour development.
Limitations, reasons for caution: This is the first experimental study eval- M. Vaugon1, M. Peigné2, E. Larouzée1, J. Phelippeau1, C. Gonthier3,
uating the effect of COS protocols over a human BC tumour cell line using a M. Koskas1
1
non-invasive in vivo system to monitor cell growth. Although the promising Bichat Hospital, Gynecology and Obstetrics Department, Paris, France ;

2
Jean Verdier Hospital, Assisted Reproductive Technology, Bondy, France ; O-042 Revisited and novel fertilization biomarkers for embryo
3
Pitié Salpetriere Hospital, Gynecology and Obstetrics Department, Paris, quality
France I.Sfontouris1
1
Eugonia, Assisted Reproduction Unit, Athens, Greece
Study question: What is the risk of recurrence after In Vitro Fertilization (IVF)
treatment for patient with endometrial atypical hyperplasia and grade 1 adeno-
carcinoma (AH/EC) who received conservative treatment with progestins? O-043 Prediction of oocyte and embryo competence by advanced
Summary answer: There is no significant difference of the rate of recurrence image analysis and AI
at 24 months between patient who received IVF treatment or not. M. Zuccotti1, G. Fiorentino1, G. Nicora2, R. Sciajno3, F. Cavalera1,
What is known already: About 5% of endometrial cancer affect young R. Bellazzi2, A. Borini3, S. Garagna1, G. Coticchio3
women in reproductive age. In this particular case of AH/EC, conservative 1
University of Pavia, Department of Biology and Biotechnology ‘Lazzaro Spallanzani’,
treatment, such as chlormadinone acetate, can be offered to safeguard their Pavia, Italy
fertility. A quick pregnancy is recommended after progestin treatment and there- 2
University of Pavia,
fore IVF is often started. Preliminary results are encouraging: in 2013, Park’s Department of Electrical- Computer and Biomedical Engineering, Pavia, Italy
study[1] concluded that the use of fertility drugs was not associated with a higher 3
9.baby Fertility and Family Center, 9.baby Fertility and Family Center, Bologna, Italy
incidence of recurrence (5-year disease free survival rate: 73% p=0,335); in 2019
Kim’s study[2], the rate of recurrence was 27,3% with fertility drugs. However, Abstract text
they are retrospectives and not homogeneous studies. Our work follows that of four earlier studies. Adjuk et al. (2011) and Swann et
Study design, size, duration: Multicentric prospective cohort study al. (2012) first explored the potential of particle image velocimetry (PIV) (an
conducted from January 2008 to July 2019. The study is homogeneous: all optical method to measure fluid and particles dynamics) for the study of cyto-
the 60 patients had an AH/EC and received progestins treatment by chlorma- plasmic dynamics occurring during mouse fertilisation. Bui et al. (2017) and
dinone acetate at least 3 months.31 patients received IVF treatment and Cavalera et al. (2018), by complementing the detection of cytoplasmic flows
29 did not with artificial neural networks (ANN), further perfected this imaging approach,
Participants/materials, setting, methods: All the patients received expert succeeding in the prediction of the developmental competence of mouse
advice from the National Observatory of the Gynecology and Obstetrics oocytes. After the presentation of these results, we will describe our latest work
Department at Bichat Hospital (PREFERE center) and then enter the cohort. on human preimplantation embryos.
The choice between IVF treatment or not was made by the gynecologist and The ANN approach was undertaken to assess retrospectively the ability of
the patient, after advice from PREFERE center. Survival rates were calculated human embryos to develop to the blastocyst stage. The analysis focused on 113
using the Kaplan-Meier method, and differences in the survival rates between embryos generated in 32 IVF cycles, carried out between October 2015 and
groups were compared using the log-rank test. May 2018. Female age was 36.3±4.9 years. To minimize possible patient-based
Main results and the role of chance: The mean follow-up was 19 months bias, cycles were recruited ensuring to have in the same cohort both embryos
in the IVF group and 11 months in no IVF group. The end of the follow-up was able to develop to the blastocyst stage and arresting at earlier stages. Embryos
the time of the last news or the time of the surgery in the case of hysterectomy. were subject to time-lapse assessment to monitor development and perform
The probability of 2 years recurrence was 37,7% (+/- 10,41%) in the IVF group trophectoderm biopsy for preimplantation genetic testing of aneuploidy.
and 55,7% (+/- 14,02%) in the no IVF group. The difference was not significant Fertilisation was achieved by ICSI. Time-lapse monitoring started immediately
(p=0,13). We find no additional risk of recurrence according to obesity, nulli- after ICSI, with a 15 min interval between consecutive observations. Of 230
parity, PCOS, age and tumor characteristics. Twenty-three patients had a preg- embryos analysed, 112 reached the blastocyst stage (BL-group) and 118 arrested
nancy (15 in the group IVF and 8 in the group no IVF). Among all the patients sometime after the 2-cell stage (NoBL-group). ANN analysis was performed,
who had a pregnancy, 7 had a recurrence of the disease in the first 24 months at this stage, only during the first two cell divisions (44 hr, 175 frames).
(30%). Whereas among the 37 patients who didn’t have a pregnancy, 17 patients Based on morphological criteria, embryos were first blindly classified by an
had a recurrence in 24 months (45%). The difference was significant (p<0.01, expert operator as belonging to the BL or NoBL group, attaining a 75.4% accuracy,
IC 0.06-0.61). Whether the number of cycles, the E2 maximum concentration, 76.5% sensitivity and 74.3% specificity. Then, the time-lapse images corresponding
the type of protocol chosen, the dose of gonadotrophin or the thinness of the to the first 175 frames (44 hr) were anlaysed with PIV to extract their cytoplasmic
endometrium, there were no significant differences on the risk of recurrence in movement profiles, and subsequently with the AI models (k-NN and LSTM-NN).
the subgroup IVF. The results showed a classification accuracy comparable to that reported by the
Limitations, reasons for caution: The main inconvenient is the size of our operator (75.4%). Next, the results of the operator were integrated to those
groups but we chose to include only patient treated with chlormadinone acetate obtained with the AI models, and this led to a further classificaton attaining a 82.6%
in order to have a homogenous population. accuracy, 79.4% sensitivity and 85.7% specificity. Taken as a whole, these data
Wider implications of the findings: IVF treatment after fertility sparing man- indicate the possibility of predicting blastocyst development as early as the 4-cell
agement of HA/EC does not seem to increase the risk of recurrence. Therefore, stage; also, these results highlight the strenght of combining the huge experience
it is an acceptable strategy to decrease the time to pregnancy. Overall, the rate owned by an expert clinical embryologist with that obtained thanks to AI tools.
of recurrence is high in both groups which implies a close monitoring of these The study represents a proof-of-principle for the chosen embryo assessment
patients. approach. Further observations are required to trenghten these data and to
Trial registration number: not applicable assess the impact of possible confounding variables such as patient typologies,
culture conditions and time-lapse equipment.
O-041 Fertility preservation in male oncology patients - To the best of our knowledge this study represents the first attempt to classify
emergency sperm and spermatogonial stem cell retrieval very early human preimplantation embryos using AI. Further refinement of the
“Abstract withdrawn by the authors” approach is expected to impact embryo assessment ability and improve efficiency
in assisted reproduction treatments.
INVITED SESSION INVITED SESSION

SESSION 08: NOVEL OOCYTE AND EMBRYO SESSION 09: DATA REPORTING SESSION: THE EUROPEAN
BIOMARKERS PERSPECTIVE (EIM AND PGT)
06 July 2020 Parallel 2 11:45 - 12:45 06 July 2020 Parallel 3 11:45 - 12:45

O-044 Assisted Reproductive Technology (ART) in Europe (in 2016: 7.7% and 0.4%). The majority of FP interventions included the cryo-
2016 and development of a strategy of vigilance – Preliminary preservation of ejaculated sperm (n=10837 from 11 countries) and of oocytes
results generated from European registers by the ESHRE EIM (n=6551 from 11 countries).
Consortium Limitations, reasons for caution: As the methods of data collection and
C. Wyns1, C. Bergh2, C. Calhaz-Jorge3, Ch. De Geyter4, levels of completeness of reported data vary among European countries, the
M.S. Kupka5, T. Motrenko6, A. Rugescu7, J. Smeenk8, A Tandler- results should be interpreted with caution. Due to the COVID-19 pandemic,
Schneider9, S. Vidakovic10, V. Goossens11 less countries than usual have sent in data by the abstract deadline.
1
Cliniques universitaires Saint-Luc, Université Catholique de Louvain, Brussels,
Belgium
2 O-045 Data from the ESHRE PGT Consortium – year 2018
Dept of Obstetrics and Gynecology, Inst of Clinical Sciences, Göteborg University,
Göteborg, Sweden A. Van Montfoort1, M. De Rycke2, F. Carvalho3, C. Rubio4,
3
Faculdade de Medicina da Universidade de Lisboa, Portugal F. Bronet5, F. Spinella6, V. Goossens7
1
4
Reproductive Medicine and Gynecological Endocrinology (RME), Maastricht University Medical Center, Dept. of Ob/Gyn, Maastricht,
University Hospital, University of Basel, Switzerland The Netherlands
2
5
Fertility Center - Gynaekologicum, Hamburg, Germany UZ Brussels, Centre for Medical Genetics, Brussels, Belgium
3
6
Human Reproduction Center Budva, Montenegro University of Porto Faculty of Medicine, Genetics Faculty of Medicine, Porto,
7
National Transplant Agency, Romania Portugal
4
8
Elisabeth Twee Steden Ziekenhuis, Tilburg, the Netherlands Igenomix, R&D Department, Valencia, Spain
5
9
Fertility Center Berlin, Berlin, Germany IVI, IVF and PGD lab, Madrid, Spain
6
10
Institute of Obstetrics and Gynecology, Clinical Center Serbia «GAK», Serbia Genoma, Molecular Genetics Laboratories, Rome, Italy
7
11
ESHRE Central Office, Meerstraat 60, Grimbergen, Belgium ESHRE, Central Office, Grimbergen, Belgium
Study question: What are the reported data on ART cycles, IUI and fertility Study question: Which trends are shown in data collection XXI of the
preservation (FP) interventions in 2017 as compared to previous years, as well European Society of Human Reproduction and Embryology (ESHRE) PGT
as main trends over the years? Consortium compared with previous years?
Summary answer: The 21st report shows a progressive increase in reported Summary answer: Data collection XXI, year 2018, represents valuable data
treatment cycle numbers in Europe, a slight decrease in the number of transfers on PGT activity in (mainly) Europe and reports on the main trends observed
(IVF + ICSI) with more than one embryo, stable multiple delivery rates which are the continuous increase of trophectoderm biopsy and further expan-
(DRs), unchanged outcomes for IUI cycles and higher pregnancy rates (PRs) and sion of comprehensive testing technology in PGT-SR and PGT-A.
DRs after FER compared to fresh IVF and ICSI cycles. What is known already: The ESHRE PGT Consortium was set up in 1997
What is known already: Since 1997, ART aggregated data generated by and from that time has been collecting data on PGT and PGT-A. The PGT
national registries and professional organisations have been collected, analysed database comprises the world’s largest collection of PGT / PGT-A data providing
by the European IVF-monitoring Consortium (EIM) and reported in 20 manu- a valuable resource for data mining and for following trends in PGT practice. So
scripts published in Human Reproduction and Human Reproduction Open. far, up to the year 2015, data collections were carried out in a retrospective data
Study design, size, duration: Yearly collection of European medically assisted way, from 2016 onwards a prospective data collection was in place.
reproduction and FP data by EIM for ESHRE; data on treatments performed Study design, size, duration: As the nature of PGT/ PGT-A treatments has
between January 1 and December 31 2017 in 30 European countries were pro- changed significantly over the last years and IVF cycle management and genetic
vided by either National Registries or registries based on professional analysis techniques are getting more complex, ESHRE uses an online data col-
organisations. lection system in which data are collected prospectively from oocyte retrieval
Participants/materials, setting, methods:1229 clinics offering ART ser- to analysis, embryo transfer and pregnancy / live birth. Data are collected cycle
vices in 30 countries reported a total of 840 138 treatment cycles, involving 147 by cycle on a voluntary basis.
934 with IVF, 351 588 with ICSI, 238 665 with frozen embryo replacement (FER), Participants/materials, settings, method: For the 2018 data, individual
33 721 with preimplantation genetic testing (PGT), 62 811 with egg donation centres (39) from 21 countries directly entered the data into the PGT database
(ED), 369 with IVM of oocytes and 5050 with frozen oocyte replacement through software developed by ESHRE. Data were analysed at ESHRE head-
(FOR). Data on IUI using husband/partner’s semen (IUI-H) and donor semen quarters and include all aspects of PGT/PGT-A cycles.
(IUI-D) were reported from 1170 institutions, including 141 647 treatments with Main results and the role of chance: The Consortium has analysed the
IUI-H in 24 countries and 48 918 treatments with IUI-D in 20 countries. A total PGT analyses (n=2831) performed in 2018. The indications for PGT included
of 18 540 FP interventions from 11 countries including oocytes, semen, as well inherited chromosomal abnormalities (n=553 analyses), monogenic disorders
as ovarian and testicular tissue in pre-and postpubertal patients were reported. (n=1229 analyses), aneuploidy testing for infertility (n=868 analyses), HLA typing
Main results and the role of chance: 1229 IVF clinics (94.5% of registered (n=12 analyses) and PGT for mitochondrial disorders (n=3 analyses). In addition,
clinics in participating countries) and 1170 IUI providers reported their data. In 701 clinical pregnancies and 411 deliveries have been analysed in detail. The
the 30 reporting countries, after IVF the clinical pregnancy rates (PRs) per aspi- methods used for biopsy were polar body (1.1%), cleavage stage biopsy (31.4%)
ration and per transfer were 29.5% and 39.4%, respectively (versus 28.5% and and blastocyst biopsy (67.5%; comparable with data from 2017). The method-
34.6% in 2016). Corresponding rates after ICSI were 27.2% en 39.8% (versus ology used for diagnosis is also evolving, with data set XXI showing around 10%
26.2% and 33.2% in 2016). After FER with own embryos the PR per thawing is of FISH, 37% of PCR and 53% of WGA. Within WGA 79% of the analyses were
still on the rise, from 30.9% in 2016 to 31.9% in 2017. After ED the PR per fresh done using NGS, 10% using array-CGH and in 8.5% cases SNP arrays were used.
embryo transfer was 49.1% (49.4% in 2016) and per FOR 43.1% (43.6% in 2016). The overall pregnancy rate is about 21.5%. The baby data show that it is difficult
In IVF and ICSI together, the trend towards the transfer of fewer embryos for most centres to have a detailed follow-up.
continues with the transfer of 1, 2, 3 and ≥4 embryos in 45.4%, 49.8%, 4.5% Limitations, reasons for caution: The findings apply to the 38 participating
and 0.2% of all treatments, respectively (versus 41.5%, 51.9%, 6.2% and 0.4% centres and may not represent worldwide trends in PGT. Data were collected
in 2016). This resulted in a proportion of singleton, twin and triplet DRs of prospectively, but details of the follow-up on PGT pregnancies and babies born
85.4%, 14.3% and 0.3%, respectively (versus 84.8%, 14.9% and 0.3% in 2016). were limited.
Treatments with FER resulted in twin and triplet DRs of 11% and 0.2%, respec- Wider implications of the findings: The ESHRE PGD Consortium continues
tively (versus 11.9% and 0.2% in 2016). its activities as an important forum for PGT practitioners to share data and
After IUI, the DRs remained similar at 8.7% after IUI-H (8.9% in 2016) and at exchange experiences. The information extracted from the data collections helps
12.2% after IUI-D (12.4% in 2016). Twin and triplet DRs after IUI-H were 8.5% to monitor quality issues in PGT and survey the introduction and effectiveness
and 0.3%, respectively (in 2016: 8.8% and 0.3%) and 7.0% and 0.2% after IUI-D of new PGT technologies and methods

O-051 Endometrial evaluation for all?

C.Coutifaris1,
INVITED SESSION 1
University of Pennsylvania Medical Centre, Division of Reproductive
SESSION 10: UPDATED TERMINOLOGY FOR EARLY PREG- Endocrinology and Infertility, U.S.A
NANCY ASSESSMENT
06 July 2020 Parallel 4 11:45 - 12:45 O-052 Is the male gamete relevant in ART?
P.N. Schlegel1
1
Cornell Institute for Reproductive Med., Department of Urology- Starr 900, New
O-046 Can you be sure that the pregnancy is Intrauterine? York- NY, U.S.A.
D.Jurkovic1
1
Abstract text
University College Hospital, Department of OB/GYN, United Kingdom
The critical role of female factors, especially female age, on the outcomes of
O-047 The ectopic pregnancies you cannot see with a laparoscope assisted reproductive technologies is well recognized. Fewer data have been
E. Kirk1 generated regarding the role of sperm or male factors on IVF results. Initially,
1 sperm factors were recognized to adversely affect the fertilization step of in
Royal Free Hospital London, Early pregnancy and emergency Gynaecology Unit,
vitro fertilization, with variable interpretation of the reproductive competency
United Kingdom
of the embryo derived using sperm from men with abnormal semen parameters.
With the advent of ICSI for management of male factor, the risk of fertilization
failure due to sperm abnormalities has been largely overcome. Several male-re-
INVITED SESSION lated factors, including paternal age, have been identified that affect the outcome
SESSION 11: PATIENT PRIORITIES of IVF. One pathway for male factors to adversely affect IVF results includes
sperm DNA integrity. Abnormal sperm DNA fragmentation is rarely seen in
06 July 2020 Parallel 5 11:45 - 12:55
naturally fertile men, but can be present even in the presence of normal semen
parameters. Sperm DNA damage can be evaluated by a variety of assays, and
appears to develop during sperm transport (i.e., after leaving the testis) for the
majority of men with abnormal sperm DNA fragmentation index (DFI.) It may
O-048 Genetic ancestry testing and the end of donor anonymity be associated with ejaculatory dysfunction (spinal cord injury), partial obstruction
D.Kennett1 (post-vasectomy), medications that effect ejaculatory efficiency (SSRI’s), pro-
1
University College London, Department of Genetics- Evolution and Environment, longed abstinence, advanced male age, or without identifiable cause. Abnormal
London, United Kingdom sperm DFI is associated with a marked increase in the risk of spontaneous Ab,
as well as decreased chance of pregnancy with IVF/ICSI. The evaluation and
Abstract text management of abnormal ejaculated sperm DFI will be reviewed as well as the
Over 30 million people around the world have now taken a direct-to-consumer existent data on the effect of DNA fragmentation on assisted reproductive
genetic test with companies such as AncestryDNA and 23andMe. These tests technology results.
can provide information about an individual’s genetic ancestry and health but
they also allow the user to opt in to receive matches with genetic relatives. As
a result, donor-conceived individuals can now take a DNA test to search for
their DNA relatives and identify their genetic parent. Identities can be inferred INVITED SESSION
through networks of genetic cousins even if the donor is not in the database. SESSION 13: CHALLENGING SCENARIOS IN IVF
The chances of success are increasing all the time and sometimes people are PATIENTS
matched at the outset with a half-sibling or genetic parent. Donor origins can 06 July 2020 Parallel 3 14:00 - 15:00
also be discovered unexpectedly through genetic testing if the child has not been
told that they were donor conceived. The massive growth of the consumer
DNA databases in the last few years has important implications for everyone
working in the fertility industry and for parents who are considering using egg O-053 The real place of IVF in PCOS
or sperm donation to start or complete their family. DNA testing has effectively G.Lainas1
1
ended anonymity for all donors regardless of the legislation in place at the time. Eugonia, IVF Unit, Athens, Greece
How do we deal with the challenges of this new reality? Whose responsibility
is it to provide support and counselling for those affected? How should the O-054 Low responders: From natural cycles to dual stimulation?
fertility sector adapt? F. Ubaldi1, A. Vaiarelli2, D. Cimadomo2, L. Rienzi2
Trial registration number: 1
Reproductive Medicine GENERA, Centre for Reproductive Medicine, Roma, Italy
Study funding: 2
Genera, Centre for reproductive medicine, Rome, Italy
Funding source:
Abstract text
O-049 Infertility - the political treatment “Low responders: From natural cycles to dual stimulation?”
A.Fincham1 Filippo Maria Ubaldi, MD, PhD; clinical director of GENERA, centers for reproductive
1
Fertility Europe VZW, Evere, Belgium medicine, Italy
The lack of a consensus on the definition of “poor responders” (POR) in the past
made it difficult to compare the proposed managements among each other. A
systematic review highlighted 41 different definitions of POR out of 47 random-
INVITED SESSION
ized control trials (RCTs). In this scenario, the Bologna criteria were the first
SESSION 12: ASRM EXCHANGE SESSION - concrete attempt to define POR grounded on an evidence-based approach.
CONTROVERSIES IN ART Noticeably, the predicted cumulative live birth rate per IVF cycle in patients ful-
06 July 2020 Parallel 2 14:00 - 15:00 filling these criteria is not higher than 7-8% according to the existing evidence in
the literature.

Recently, a panel of clinicians gathered together in the so-called POSEIDON Abstract text
group and suggested a novel more detailed stratification of women with a low The use of fertility preservation in children receiving gonadotoxic therapies is
response to controlled ovarian stimulation (COS). This group introduced a new often ethically challenging. Presently most countries have some type of guidelines
concept in IVF: the main aim of COS is to obtain at least one embryo with a high suggesting all patients receiving a cancer or other life-threatening diagnosis,
chance to result in a live birth, namely euploid blastocyst(s). Several data exist receive information on how that may impact their future fertility and be offered
in the literature supporting that the chance of a blastocyst to be euploid is referrals to specialist to discuss preservation. Not all types of preservation are
independent of COS, its regimen, the dose of gonadotrophins used, and the available in certain countries and in some cases, finances become a rate limiting
number of oocytes/embryos in each cohort. Therefore, the careful choice and factor as well as potential delay of treatment. Parents may feel uncertain about
tailoring of the strategy that better suits each patients is critical. their role in making future decisions about their children’s future fertility and
The obvious and mostly-used clinical strategy for women with low AFC health care clinicians may also be concerned that engaging in discussions and
and/or AMH was to increase the daily dose of gonadotrophins but unfortu- consultations may delay the lifesaving treatment needed. There are also psy-
nately without success. In fact, gonadotrophins can only support each follicular cho-social issues involved in the decision process both at the time of the diagnosis
wave in its growth, but they cannot produce follicles de novo. Natural cycle as well as in the future. There has been great controversy over the use of
has been also adopted across the years. However, it is mainly effective in posthumous assisted reproduction, particularly the use of gametes stored by a
patients with an expected number of oocytes retrievable lower than 2-3. The minor and later used by a parent to create a grandchild. There is also the unique
adoption of COS might be helpful, as also suggested by the recent ESHRE circumstance of the transgender/non-binary child and their family who may be
guidelines. considering puberty- blockers or gender affirming hormones. This presentation
Another strategy has been outlined across the years to treat time-sensitive will explore the ethical and psycho-social issues and boundaries of the use of
(oncologic and advanced maternal age) and POR patients, which is known as fertility preservation in children (minors) and explore the current related liter-
DuoStim (follicular and luteal phase stimulation in the same ovarian cycle). This ature. The objectives of this presentation are to:
strategy is supported from the evidence that 2-3 waves of follicular growth might
arise during the ovarian cycle in women, 1-2 of which in the luteal phase. Although 1. Identify current standard and experimental fertility preservation options
the ESHRE labeled this strategy still as “for research only”, several evidences have for minors
been produced along the last 5 years from our group, as well as from many 2. Discuss ethical challenges faced by parents and healthcare clinicians
other groups: 3. Explore current cases on decision-making and use of fertility preserva-
tion in minors
- we set-up the DuoStim protocol in a pilot study, 4. Compare and contract the ethics of FP on very young children (aged
- we confirmed the absence of differences in terms of fertilization, blastu- 0-7) compared to middle aged children (8-14) and older children
lation and euploidy rates between the cohorts of oocytes collected from (15-18)
the luteal and the paired-follicular phase, 5. Examine the issue of fertility preservation in transgender/non-binary
- we reported the reproducibility of the DuoStim protocol among all the children
GENERA centers
- we reported similar clinical, obstetrical and neonatal outcomes among
euploid blastocysts obtained after follicular and luteal phase stimulation
through a non-selection study design SELECTED ORAL COMMUNICATIONS
- we suggested DuoStim as a strategy to prevent drop-out in patients
SESSION 15: IN THE NAME OF THE FATHER
fulfilling the Bologna criteria, while increasing their chance of having a
live birth per intention to treat from 8 to 15% 06 July 2020 Parallel 5 14:00 - 15:00
- we conducted a systematic review that highlighted the reproducibility of
our data also in other clinics adopting slightly different protocols
- from a basic research perspective, we collaborate with Universities to
keep building evidence in favor of DuoStim safety O-057 Septum resection versus expectant management in
women with a septate uterus: a randomised controlled trial
(NTR 1676)
Clearly RCTs and cost-effective analyses are still needed but, when indicated
J. Rikken1, C. Kowalik1, M.H. Emanuel2, M. Bongers3, T. Spinder4,
and depending on the setting of each IVF clinic, DuoStim might be considered
F.W. Jansen5, A. Mulders6, R. Padmehr7, J. Clark8, H. Van Vliet9,
a valuable strategy to prevent patients’ drop-out while increasing their chance
M. Stephenson10, F. Van Veen11, B.W. Mol12, M. Van Wely11,
to identify at least one euploid blastocyst in the shortest possible time.
M. Goddijn11
1
Academisch Medisch Centrum, Center for Reproductive Medicine, Amsterdam,
The Netherlands ;
INVITED SESSION 2
University Medical Center Utrecht, Obstetrics & Gynaecology, Utrecht, The
SESSION 14: THE WAY FORWARD FOR FERTILITY Netherlands ;
3
PRESERVATION Maxima Medical Center, Obstetrics & Gynaecology, Veldhoven, The Netherlands ;
4
MC Leeuwarden, Obstetrics & Gynaecology, Leeuwarden, The Netherlands ;
06 July 2020 Parallel 4 14:00 - 15:00 5
LUMC, Obstetrics & Gynaecology, Leiden, The Netherlands ;
6
Erasmus MC, Obstetrics & Gynaecology, Rotterdamthe, The Netherlands ;
7
Avicenna Research Institute, Obstetrics & Gynaecology, Teheran, Iran ;
8
Birmingham women’s and children’s hospital, Obstetrics & Gynaecology,
O-055 Cancer during pregnancy: Outcomes for mother and child Birmingham, United Kingdom ;
F. Amant1 9
Catharina hospital, Obstetrics & Gynaecology, Eindhoven, The Netherlands ;
10
1
Netherlands Cancer Institute, The Netherlands University of Illinois. Chicago, Obstetrics & Gynaecology, Chicago, The
Netherlands ;
11
AMC, Obstetrics & Gynaecology, Amsterdam, The Netherlands ;
O-056 Fertility preservation for children: Specificities and 12
Monash Medical Center, Obstetrics & Gynaecology, Clayton, Australia
counselling for patients and parents
G. Quinn1 Study question: Does hysteroscopic septum resection improve reproductive
1
New York Univeristy, OB-GYN, New York, U.S.A. outcomes in women with a septate uterus?

Summary answer: In our study, hysteroscopic septum resection did not Study design, size, duration: Between 2014-2019, 332 egg recipients were
improve reproductive outcomes in women with a septate uterus. assigned prospectively in a ratio 1:2 to undergo hysteroscopy
What is known already: The septate uterus is the most common uterine (n=114, Hysteroscopy group) or not (n=218,Non-Hysteroscopy group).
anomaly with an estimated prevalence of 0.2-2.3% in women of reproductive These women, who performed pre-operative hysteroscopic evaluation of the
age. Women with a septate uterus are at increased risk for subfertility, pregnancy uterine cavity, also underwent fundus endometrial incision irrespectively of the
loss and preterm birth. Hysteroscopic septum resection has been applied to presence of arcuate uterus (U2a) or not (U0) with endoscopic scissor. The
improve reproductive outcome in these women, but the evidence on the effec- relation of the FEI to pregnancy outcome was investigated.
tiveness of this procedure is scarce and of low quality. We performed a ran- Participants/materials, setting, methods: The age of the women included
domised controlled trial to establish whether septum resection improves in the study ranged from 35-50 years old. The mean years of infertility was 4,6.
reproductive outcome in women with a septate uterus. Women received treatment with donor oocytes and the mean blastulation rate
Study design, size, duration: We did an international open-label randomised was 59%. They underwent embryo transfer with two blastocysts except from
controlled trial in 10 centres in the Netherlands, United Kingdom, United States 10 cases which preferred single blastocyst transfer to avoid twins.
of America and Iran. The trial is registered with the Netherlands Trial Register Main results and the role of chance: Among 114 women of the Hysteroscopy
as NTR 1676. group(Storz Bettocchi 5mm) prior to embryo transfer,33 were diagnosed with
Participants/materials, setting, methods: Women with a septate uterus U2a (partial septate, arcuate uterus), 10 with U2b (complete septate uterus)
and a history of subfertility, pregnancy loss or preterm birth were randomly and 6 women with UIa (T shape uterus). Both women with uterine abnormality
allocated to septum resection or expectant management. The primary outcome (n=49) plus the ones with normal cavity (n=65) underwent Fundus Endometrial
was live birth within 12 months after randomization. Secondary outcomes were Incision with Wolf endoscopic scissor. The rest 218 recipients did not undergo
clinical pregnancy, ongoing pregnancy, pregnancy loss, preterm birth and com- hysteroscopy prior to ET. Initial positive human Chorionic Gonadotropin (hCG)
plications following hysteroscopic septum resection. We analysed the data on test was 73.7% in the hysteroscopy group and 57,34% in the non-hysteroscopy
intention to treat basis. We estimated differences as relative risks (RR) with 95% group. Live Birth rate was statistically significantly (p=0,015) higher in the hys-
confidence intervals (CI). teroscopy group in a rate of 56,14% (n=64/114) compared to the live birth rate
Main results and the role of chance: Between October 2010 and October of 42,2% (n=92/218) in the non-hysteroscopy group.
2018 we randomly assigned 80 women (40 to septum resection and 40 to Limitations, reasons for caution: The sample size of the participants should
expectant management). Live birth occurred in 12 women who underwent be expanded in order to obtain more solid evidence of the impact of FEI in
septum resection (32.4%) and in 14 women who had expectant management pregnancy outcomes. Moreover, the time interval between the hysteroscopy
(37.8%) ((RR 0.89 (95% CI 0.56-1.41). Clinical pregnancy occurred in 22 and the ET should be taken into further consideration.
women who underwent septum resection (59.5%) and in 19 women who had Wider implications of the findings: The current findings indicate that
expectant management (51.4%) (RR 1.18 (95% CI 0.75-1.85); pregnancy loss in this particular subgroup of patients- like oocyte recipients- hysteroscopy
occurred in 10 women who underwent septum resection (43.5% of women plus FEI might increase delivery rates. Apart from the obvious benefit of
who had a conception) and in 5 women who had expectant management recognizing obscured anomalies, requiring surgical correction, it appears that
(26.3% of women who had a conception) (RR 1.56 (95% CI 0.70-3.47); ongoing even fundus incision might improve uterine receptivity and thus pregnancy
pregnancy occurred in 13 women who underwent septum resection (35.1%) outcome.
and in 14 women who had expectant management (37.8%) (RR 0.92 (95% CI Trial registration number: none
0.58-1.48); preterm birth occurred in 5 women who underwent septum resec-
tion (38.5% of women who had an ongoing pregnancy) and in 4 women who
had expectant management (28.6% of women who had an ongoing pregnancy) O-059 Improvement of symptoms after hysteroscopic
(RR 1.33 (95% CI 0.48-3.70). In women who underwent septum resection, isthmoplasty in women with abnormal uterine bleeding and
one perforation of the uterus occurred during surgery. The procedure was expected pregnancy: a prospective study
stopped immediately and the septum was removed 4 weeks later without D. Tran1, T. Nguyen2, D. Do3, H. Nguyen4, P. Thuong2
complications. 1
Ha Noi Obstetric and Gynecology hospital, Department of high risk pregnancy,
Limitations, reasons for caution: Our major limitation is the limited sam-
Ha Noi, Vietnam ;
ple size. 2
Ha Noi Obstetric and Gynecology hospital, Delivery department, Ha Noi,
Wider implications of the findings: Our results suggest that septum resec- Vietnam ;
tion does not result in better pregnancy outcomes. This procedure is not without 3
Ha Noi Obstetric and Gynecology hospital, Gynecology department, Ha Noi, Vietnam ;
risks and should not be offered to women without proper counselling. We 4
Ha Noi Obstetric and Gynecology hospital, Assisted Reproductive Center, Ha Noi,
believe an expectant management is the preferred primary treatment in women Vietnam
with a septate uterus.
Trial registration number: NTR 1676 Study question: To determine the effectiveness and safety of hysteroscopic
repair of isthmocele to treat postmenstrual spotting and the rate of natural
O-058 Hysteroscopic Fundus Endometrial Incision (FEI) in pregnancy after surgery
oocyte recipients before embryo transfer (ET) during In Vitro Summary answer: Our data supports that hysteroscopic isthmoplasty is safe
Fertilization and effective in treating isthmocele-related abnormal uterine bleeding
R. Najdecki1, E. Papanikolaou1, T. Chartomatsidou1, F. Pakaki1, What is known already: A cesarean scar defect, also called as an isthmocele,
S. Stamataki1, P. Tatsi1, E. Timotheou1, F. Chouliara1, E. Bampas1, is one of the consequences of cesarean section that may cause secondary infer-
S. Bouchlariotou1, G. Michos2, A. Athanasiadis2 tility and postmenstrual spotting.
1
Assisting Nature, IVF Unit, Thessaloniki, Greece ; Study design, size, duration: 23 patients with abnormal uterine bleeding
2
Aristotle University, 3rd Department Ob Gyn, Thessaloniki, Greece and expected pregnancy who had an isthmocele with a residual myometrium of
≥2.5mm, measured during transvaginal untrasound were performed hystero-
Study question: Does Hysteroscopic Fundus Endometrial Incision (FEI) in scopic repair of isthmocele
oocyte recipients before embryo transfer increase pregnancy and live birth rates? Participants/materials, setting, methods: 23 patients with abnormal uter-
Summary answer: Oocyte recipients who underwent FEI before ET showed ine bleeding and expected pregnancy who had an isthmocele with a residual
statistically higher live births rates. myometrium of ≥2.5mm,were performed hysteroscopic repair of isthmocele
What is known already: Controversial results are published regarding the using a step-wise approach: identification of relevant anatomy; resection of the
necessity of hysteroscopy plus uterine scratching or not before ET. For oocyte cephalad edge of fibrosis; resection of the caudad edge of fibrosis; and ablation
recipients, who might have previously undergone several failed IVF procedures, of the isthmocele base. Complications of surgery were observed and the sym-
the importance of correct evaluation of uterine cavity is of paramount impor- toms of postmentstrual spotting and prevalence of natural pregnancy were
tance since there are limited psychological resources to sustain potential failure. monitored for 6 months after surgery.

Main results and the role of chance: No complications of surgery as Trial registration number: 19-024
bleeding, uterine perforation, infection was noticed. 11 (47.8%) patients had
resolution of their symptoms. In the remaining cases, 7 (30.4%) patients had an
improvement of symtoms with decreasing duration of postmentstrual spotting
from 8.1 ±1.7 days to 4.3 ± 0.8 days (P < 0.05), whereas 5 (21.8%) patients did
INVITED SESSION
not obtain any relief. 7 (30.4%) patients had natural pregnancy after surgery
6 months. SESSION 16: BREAKING NEWS IN CURRENT PRACTICE
Limitations, reasons for caution: The sample size is not enough large 06 July 2020 Parallel 6 14:00 - 15:00
Wider implications of the findings: Hysteroscopic isthmoplasty should be
used to treat isthmocele-related abnormal uterine bleeding
O-060 Conception and Reproductive Outcomes in Asherman O-061 Fresh or frozen? - First results of e-freeze trial from UK
Syndrome after hysteroscopic adhesiolysis
A. Maheshwari1
S. Baradwan1, D. Alharbi2, M. Bashir3, A. Saleh3, D. Al-Jaroudi4 1
1
University of Aberdeen, United Kingdom
HealthPlus Fertility and Women’s Health Center- Jeddah- Saudi Arabia.,
Department of Obstetrics and Gynecology, jeddah, Saudi Arabia ;
2
King Fahad Medical City- Riyadh- Saudi Arabia, Department of Obstetrics and O-062 Recommendations for good practice for the use of time-
Gynecology., Riyadh, Saudi Arabia ; lapse technology
3
King Fahad Medical City- Riyadh- Saudi Arabia, Research Services Administration- D. Montjean1, S. Apter2, T. Ebner3, T. Freour4, Y. Guns5,
Department of Biostatistics- Research Center., Riyadh, Saudi Arabia ;
4
B. Kovacic6, N. Le Clef7, M. Marques8, M. Meseguer9,
King Fahad Medical City- Riyadh- Saudi Arabia, Reproductive Endocrine and I. Sfontouris10, R. Sturmey11, G. Coticchio12
Infertility Medicine Department., Riyadh, Saudi Arabia 1
Hopital Saint-Joseph, Service de médecine et biologie de la reproduction,
Marseille, France
Study question: Dose the reproductive methods could influence pregnancy 2
Livio, Livio, Stockholm, Sweden
outcome in women with Asherman’s syndrome ? 3
Kepler Universitätsklinikum, Department of Gynecology- Obstetrics- and
Summary answer: The spontaneous cumulative conception rates following Gynecological Endocrinology, Linz, Austria
hysteroscopic adhesiolysis were higher than those for the IVF-ICSI, whereas the 4
CHU de Nantes, Medecine de la Reproduction, Nantes, France
live birth and miscarriage rates were similar. 5
UZ Brussel, Center for Reproductive Medicine, Brussels, Belgium
What is known already: Hysteroscopic adhesiolysis anatomically restores 6
Univerzitetni klinicni center Maribor-,
the uterine cavity in cases of Asherman’s syndrome. however, the extent of Department of Reproductive Medicine and Gynecologic Endocrinology-, Maribor,
endometrial fibrosis could lead to implantation failure and miscarriages. The main Slovenia
purpose of treating infertile women with Asherman’s syndrome is improve both 7
European Society of Human Reproduction and Embryology, Guidelines,
the conception rate and the live birth rate. Few studies have evaluated the Grimbergen, Belgium
reproductive outcome after hysteroscopic adhesiolysis. 8
CEMEARE, cemeare, Lisbon, Portugal
Study design, size, duration: A retrospective cohort study that included 41 9
Instituto Valenciano de Infertilidad, IVF Laboratory, Valencia, Spain
women diagnosed with Asherman syndorme who attended Women’s Specialized 10
Hospital, King Fahad Medical City, from December 2010 to December 2016 11
Hull York Medical School- University of Hull,
and presented with a history of infertility or recurrent pregnancy loss. Patients Centre for Atherothrombosis and Metabolic Disease-, Hull, United Kingdom
were followed up for 2 years to monitor pregnancy. Details of reproductive 12
9.baby Family and Fertility Center, 9.baby Family and Fertility Center, Bologna,
methods whether with spontaneous conception or assisted reproductive tech- Italy
nology were recorded. The main outcome measure was identification of repro-
ductive methods and pregnancy outcome. Abstract text
Participants/materials, setting, methods: all women diagnosed with Traditional embryo morphology assessment is performed at static time points
Asherman syndorme who attended Women’s Specialized Hospital, King Fahad and implies interruption of embryo culture conditions. In a TLT incubator, images
Medical City, presented with a history of infertility or recurrent pregnancy loss. of embryo development are recorded at regular time intervals, which facilitates
Patients were followed up for 2 years to monitor pregnancy. Details of repro- embryo monitoring. Indeed, it allows embryologists to assess embryo develop-
ductive methods whether with spontaneous conception or assisted reproductive ment thoroughly in a dynamic fashion without removing them from the incubator
technology were recorded. The main outcome measure was identification of and thus maintaining constant culture conditions. Although TLT has been widely
reproductive methods and positive pregnancy tests that might have ended in implemented since its release in 2010, there was no recommendation on how
miscarriage or ectopic, live birth, or no pregnancy. to introduce this technology in an IVF laboratory and no review of other aspects
Main results and the role of chance: The overall conception rate was of its use. To address this need, a working group was constituted. It included
(53.6%, n=22) after hysteroscopic adhesiolysis. The live birth rate was (34.2%, 11 members of different nationalities with internationally recognized experience
n=14) the miscarriage rate was (14.6%, n=6) and the ectopic pregnancy rate in clinical embryology and basic science embryology. The stakeholders of this
was (4.9%, n=2). The conception rate was significantly (P=0.037) higher in project have reviewed the literature and collected published surveys and man-
spontaneous conception compared with IVF- ICSI, 80% (12 out of 15) com- ufacturer information up to January 2019. Once a consensus was found on the
pared to 44.4% (8 out of 18), respectively. The live birth and miscarriage rate content of the recommendation manuscript, a draft was released on ESHRE
were reported by 53.3% (8 out of 15), 20% (3 out of 15) of the spontaneous website for review by ESHRE members. The paper was published in HRopen
conception group compared to 27.8% (5 out of 18), 16.7% (3 out of 18) of (https://academic.oup.com/hropen/article/2020/2/hoaa008/5809428) and
the IVF-ICSI group, respectively. This was statistically not significant. the recommendations are available on ESHRE website (https://www.eshre.eu/
Limitations, reasons for caution: Among the limitations of this study is the Guidelines-and-Legal/Guidelines/TLT). The working group listed 11 recom-
small size of the sample, as it was limited to one Hospital, which could affect the mendations on what to do before introducing TLT in an IVF laboratory. These
generalizability. Another limitation is the study’s retrospective nature, second statements include an assessment of the pros and cons of acquiring a TLT
look hysteroscopy was not performed, and the menstrual pattern after the system, selection of relevant morphokinetic parameters, selection of an appro-
procedure was not documented. priate TLT system with technical and customer support, development of an
Wider implications of the findings: The spontaneous cumulative concep- internal checklist and education of staff. This paper also addresses more general
tion rates following hysteroscopic adhesiolysis were higher than those for the aspects of TLT introduction in IVF laboratory such as the potential benefit of
IVF-ICSI, whereas the live birth and miscarriage rates were similar. TLT especially regarding embryo quality assessment and the identification of

parameters with biological/clinical outcomes. This document discusses in what organise the cortical cytoskeleton by linking filamentous actin to the apical mem-
extent TLT helps in embryo selection/deselection for transfer and how it brane of cells during EMT, was stimulated by the activation of PRL-JAK2 signal-
allowed the development of algorithms thanks to the analysis of data generated ling. The expression of EMT-related genes, such as transforming growth factor
during the past decade. A description of the current state of TLT is provided β1, snail1, and twist1, was also promoted by PRL treatment. PRL-treated
and the question whether to share TLT data with patients is tackled. Besides, embryos exhibited higher mRNA expression of ITGA3, ITGB1, ITGAV, and
the paper presents the non-clinical/biological interests and benefits of having ITGB3 than non-treated embryos did. Focal adhesion kinase and paxillin are
TLT in IVF laboratory, which emcompass training/teaching, quality control and recruited by integrins and are involved in cytoskeletal and adhesion assembly
the management of staff time and work-flow. Overall these recommendations and organisation. There were more nascent adherent cells expressing focal
are mostly based on clinical and technical expertise. The paper provides technical adhesion kinase and paxillin in PRL-treated embryos than in non-treated
advice, but leaves any decision on whether or not to use TLT to the individual embryos.
centres. Limitations, reasons for caution: The results may vary between in vivo and
in vitro conditions. Further clinical studies are thus required to explore the clinical
efficacy of PRL supplementation in the culture. This can be done by assessing
the pregnancy outcomes after a single blastocyst transfer.
SELECTED ORAL COMMUNICATIONS Wider implications of the findings: To our knowledge, this is the first report
SESSION 17: CELLULAR CHARACTERISTICS OF EMBRYO demonstrating PRLR expression in human embryos after compaction and
DEVELOPMENT showed that culture medium supplementation with PRL improves blastocyst
adhesion by promoting EMT and integrin-based focal adhesions. Therefore, PRL
06 July 2020 Parallel 1 15:15 - 16:30
treatment during embryo culture would be advantageous for improving preg-
nancy outcomes following blastocyst transfer.
Trial registration number: not applicable
O-063 Prolactin receptor expression and its role in cytoskeletal O-064 Histone variant H3.3 chaperone complex, Hira, is essential
reorganisation in focal adhesion in human blastocysts for male pronucleus formation in mouse and human
K. Ezoe1, T. Miki1, A. Yabuuchi1, T. Kobayashi2, K. Kato2 R. Smith1, A. Saunderson2, S. Pickering3, J. Tait4, J. Thong3,
1
Kato Ladies Clinic, R&D Division, Tokyo, Japan ; 2Kato Ladies Clinic, Gynecology, R. Anderson4, C.J. Lin4
Tokyo, Japan 1
University of Edinburgh, MRC Centre for Reproduction Health, Edinburgh, United
Kingdom ;
Study question: What is the gene expression pattern of prolactin receptor 2
Royal Infirmary of Edinburgh, Simpson Centre for Reproductive Health,
(PRLR) in human pre-implantation embryos and what are its functions during Edinburgh, United Kingdom ;
embryonic development and adhesion process? 3
Royal Infirmary of Edinburgh, Edinburgh Fertility and Reproductive Endocrine
Summary answer: Human embryos express PRLR at the morula and blasto- Centre, Edinburgh, United Kingdom ;
cyst stages and PRLR signalling stimulates blastocyst adhesion by promoting 4
University of Edinburgh, MRC Centre for Reproductive Health, Edinburgh, United
integrin-based focal adhesions. Kingdom
What is known already: Predecidualisation is one of the initial processes in
endometrial stromal cell differentiation that occurs during embryo implantation Study question: Is loss-of-function of H3.3 chaperone Hira complex account-
and placentation. During this process, the decidual cells express prolactin (PRL), able for the abnormal single pronucleus (1PN) phenotype?
a decidual marker gene, prior to implantation. PRL is thus thought to influence Summary answer: H3.3 Chaperone Hira complex is essential for male pro-
preimplantation development and the subsequent blastocyst implantation in the nucleus formation in both mice and humans.
uterus. It has been reported that PRL and PRLR are not expressed in human What is known already: The Hira complex comprises Hira, Cabin1, and
embryos at the eight-cell stage. However, their expression patterns at stages fur- Ubn1. In mouse, it is responsible for H3.3 incorporation into the male genome,
ther along the first cell lineage and implantation are poorly understood. chromatin reconstruction, and male pronucleus formation after fertilisation.
Study design, size, duration: A total of 318 discarded human vitrified four- Hira-mutant zygotes show a 1PN phenotype due to failure of formation of the
cell stage embryos donated for research by consenting couples were used in this male pronucleus. An abnormal 1PN phenotype is commonly found following
study. The study was approved by the Institutional Review Board. The embryos assisted reproductive technologies (ART) in IVF clinics: however, the etiology is
were randomly allocated into two groups to be cultured in medium with (n = unknown and no investigation of whether the other subunits in the complex are
125) or without PRL (n = 193). The rates of blastocyst development and adhe- required for zygote formation in mouse or human has been carried out.
sion, outgrowth area, cytoskeletal reorganisation, and nascent adhesion forma- Study design, size, duration: Mouse model: We have generated new loss-
tion were compared between the groups. of-function mouse models of Hira, Cabin1, and Ubn1 respectively.
Participants/materials, setting, methods: Human vitrified-warmed Human samples: An HFEA Research Licence allowed us to collect abnormal
embryos were cultured to the blastocyst stage in SAGE 1-Step medium one-pronucleus zygotes from the Edinburgh Fertility and Reproductive Endocrine
with or without 100 mg/ml of PRL. The PRL and PRLR expressions in Centre, during 2018-2019. We obtained consent from 144 couples and collected
embryos at the four-cell, eight-cell, morula, and blastocyst stages were 21 1PN zygotes from 16 patients.
assessed by quantitative RT-PCR and immunofluorescence staining. The Participants/materials, setting, methods: We used mouse models to
blastocysts were plated on fibronectin-coated dishes and cultured for 96 conditionally knockout Hira and Cabin1 in oocytes using Zp3-Cre, and mor-
h to evaluate outgrowth competence. The expression of epithelial-to-mes- pholino microinjection to study the knockdown of Ubn1. We studied the con-
enchymal transition (EMT)- and focal adhesion-related genes in blastocysts sequences of the loss-of-function on the formation of the 1PN phenotype.
were analysed. Embryos were generated using a combination of ART (microinjection, IVM/
Main results and the role of chance: PRLR mRNA expression increased IVF/IVC). Histology (H&E, IHC), cytological (confocal imaging) and molecular
significantly after embryo compaction and blastulation. The increased expression analyses (qRT-PCR, IF, Proximity Ligation Assay) were then applied.
levels of the PRLR protein at the morula and blastocyst stages were also demon- Human zygotes had IF and confocal imaging applied to them.
strated by immunofluorescence staining. The supplementation of the embryo Main results and the role of chance: Results of mouse studies: We demon-
culture medium with PRL did not improve the rate of embryonic development strated that all the subunits of the Hira complex are maternal factors present in
to the blastocyst stage and their morphological grade. On the contrary, blasto- oocytes, and they are incorporated into male chromatin after fertilisation and
cyst outgrowth was significantly increased in embryos cultured with PRL. The interact with each other. It is important to note that loss-of-function Hira com-
phosphorylation of JAK2, downstream of the prolactin receptor family, was plex oocytes formed abnormal 1PN after fertilisation.
markedly higher in the PRL-treated embryos than in those cultured without PRL. Results of Human studies: In the human abnormal 1PN zygotes we collected,
Furthermore, the mRNA expression of ezrin-radixin-moesin proteins, which we observed that Hira chaperone molecules failed to incorporate into male

chromatin. This finding is based on the detection of IF signal in male chromatin successful conception. In women with more children, other factors compensate
[100% of Hira (n=4); 100% of Ubn1 (n=4), and 50% of Cabin1 (n=4)]. Since for the declined oocyte quality typical to their age.
the human data agreed with the mouse data, we proposed that the mechanism Limitations, reasons for caution: We have measured telomere length
of action is likely conserved. in leukocytes, and it remains to be shown whether it reflects oocyte telomere
Limitations, reasons for caution: We obtained consent from only 144 length. In addition, the association of telomere length with oocyte quality
couples, and collected 21 1PN zygotes. Ethical and HFEA regulations prevented and infertility should be studied more directly in a subfertile population
us from collecting normal 2PN human zygotes. However, we were able to collect of women.
morphological abnormal but cytological normal samples as controls to demon- Wider implications of the findings: Our findings biologically support previous
strate that Hira incorporates into male chromatin in the human zygote. demographic reports and explain the known link between extended fertility and
Wider implications of the findings: The Hira complex is critical during longevity. We suggest that longer telomeres, associated with extended fertility,
fertilisation in both mice and humans. The feasibility of rescuing 1PN zygotes reflect improved general health and reproductive fitness. We propose that telomere
using overexpression or nuclear transfer approaches as an alternative to the length should be further explored as a possible novel biomarker of oocyte quality.
3 parent-IVF strategy could potentially be developed into a novel person- Trial registration number: Shaare Zedek Medical Center ethical committee
alised medicine for patients who have previously produced1PN zygotes approval number: 0145-16-SZMC
after ART.
Trial registration number: Not applicable. HFEA Research Licence
Reference R0204 O-066 Cytoskeleton, ultrastructure and viability of human
biopsied embryos on day 3 vs day 5 following vitrification
O-065 The telomere link between extended fertility and longevity A. Chatzimeletiou1, N. Petrogiannis2, A. Sioga3, A. Makedos1,
J. Michaeli1, K. Rotshenker Olshinka1, N. Srebnik1, R. Smoom2, M. Filippa2, B. Tarlatzis1, E. Kolibianakis1, A. Handyside4,
N. Serruya2, S. Yanai2, O. Michaeli2, T. Eldar-Geva1, Y. Tzfati2 G. Grimbizis1
1
1
Shaare Zedek Medical Center, IVF Unit- Obstetrics and Gynecology, Jerusalem, Aristotle University Papageorgiou Hospital, IVF Unit, Thessaloniki, Greece ;
2
Israel ; Naval Hospital of Athens, IVF Unit, Athens, Greece ;
3
2
The Hebrew University of Jerusalem, Department of Genetics- The Silberman Aristotle University, Embryology Laboratory, Thessaloniki, Greece ;
4
Institute of Life Sciences, Jerusalem, Israel Illumina, Illumina, Cambridge, United Kingdom
Study question: Is extended fertility associated with longer telomeres? Study question: Are there any differences in viability, cytoskeletal abnormalities
Summary answer: Extended fertility is associated with longer leukocyte telo- and mitochondrial and other organelles’ structure/ number/ function amongst
mere length suggesting a novel biomarker of oocyte quality. embryos biopsied on day3 vs day5 following vitrification?
What is known already: Telomeres are nucleoprotein complexes that protect Summary answer: No statistically significant differences are observed in via-
the ends of eukaryotic chromosomes. Telomeres gradually shorten with age bility, spindle/chromosome configurations and the ultrastructure of human
and eventually cause aging-related pathologies. Longer telomeres are associated embryos biopsied on day 3 vs day 5 following vitrification.
with longevity. Telomeres have a cardinal role in meiosis and the development What is known already: Most studies in human biopsied vitrified embryos
of germ cells. Many demographic studies correlated late reproduction with gen- concentrate on assessing the success of the procedure on survival rates and clinical
eral health and longevity. We hypothesized that telomere length provides a link outcomes following transfer. Indeed several clinical trials have confirmed promising
between extended female fertility and longevity. Furthermore, while anti-mul- results. Limited studies have however examined the effects of biopsy and vitrifi-
lerian hormone (AMH) levels indicate the number of remaining follicles, we cation on spindle structure/chromosome alignment in human embryos, and only
hypothesized that telomere length might indicate the quality of the oocytes and in animal models cell viability using fluorescent markers and the ultrastructure of
the ability to conceive and deliver a genetically-healthy offspring. the cellular organelles in vitrified biopsied embryos have been investigated in detail.
Study design, size, duration: A prospective experimental study evaluating: This is the first study to examine in human embryos biopsied at 2 different stages
(1) leukocyte mean telomere length and AMH levels in 30 women age 43-48y cell viability, cytoskeleton and ultrastructure before and after vitrification.
who naturally conceived and delivered healthy babies, and 30 age-matched con- Study design, size, duration: 240 Day3 biopsied embryos that developed to
trols, previously fertile, who do not use contraceptives but failed to conceive blastocysts but were rejected for transfer following PGT-M or PGT-A were
after the age of 41y; (2) leukocyte mean telomere length in 20 women age 30-35y divided into 2 groups: A) 120 blastocysts treated for Viability, Cytoskeletal and
who delivered their first child compared to 20 age-matched women who deliv- TEM analysis(fresh n=20,n=20,n=20 and following vitrification/warming
ered their 6th to 10th child. Study conducted 2018-2019. n=20,n=20,n=20), B) 120 embryos were rebiopsied at the blastocyst stage and
Participants/materials, setting, methods: Blood samples obtained (1) treated for Viability, Cytoskeletal and TEM analysis(fresh n=20,n=20,n=20 and
study group: at delivery and 5-6 months later, and from the control group, and following vitrification/warming n=20,n=20,n=20). Also, 60 vitrified blastocysts
(2) at delivery of the first or ≥6th child. Erythrocytes were lysed and high molec- biopsied only on Day5 were treated for Viability, Cytoskeletal and TEM analysis.
ular weight genomic DNA was extracted from leukocyte by a standard protein- Participants/materials, setting, methods: Viability was assessed by CFSE/
ase K/phenol method. Average telomere length measured by Southern blot, PI staining. Cytoskeletal analysis by confocal laser scanning microscopy was per-
analyzed by TeloTool software. AMH levels were measured (in the study group, formed following immunostaining with α-tubulin, γ-tubulin and acetylated-tubulin
5-6 months postpartum) by an automated assay (Elecsys Cobas, Roche). antibodies in combination with DAPI or /PI. For ultrastructure assessment by
Main results and the role of chance: In study 1, mean age, 45y, and demo- transmission electron microscopy(TEM), blastocysts were fixed in gluteralde-
graphic characteristics were similar between the study and control groups. hyde, incubated in osmium, aqueous uranyl acetate, dehydrated through ethanol
Average telomere length in the study group was 9700 bp, significantly longer series and immersed in Epon. The Ultrathin lead citrate stained sections were
than that of the control group, 9270 bp (p<0.05). When plotting telomere length examined in a JEOL-TEM-2000-FXII microscope. The study was conducted in
according to parity, in women with less than 9 children, the difference in telomere an academic hospital.
length between cases and controls increased to over 1000 bp (p=0.009). While Main results and the role of chance: Viability staining with CFSE/PI before
in the subpopulation of women with ≥9 children, the difference was insignificant. vitrification revealed that all blastocysts biopsied on day5 had damaged cells at the
This led us to examine the immediate effects of pregnancy and delivery, and the position of cutting (range 4-10cells), while 95% of the day3 biopsied embryos that
long-term effect of parity on telomere length (study 2). Both short and long developed to the blastocyst stage had 0 cells damaged at the herniating
term effects were found insignificant. AMH levels were similar in the study and site. Following vitrification and subsequent warming, additional PI stained cells
control groups (averaged 0.80 and 0.82 ng/ml, respectively). Our results indicate (range 5-20) were evident in other parts of the day5 biopsied blastocysts and in
clear correlation between extended fertility and longer telomeres, particularly similar incidence to vitrified blastocysts that were biopsied on day3. Cytoskeletal
in the subpopulation of women with up to 8 children, which is unlikely to be analysis by confocal laser scanning microscopy revealed that the majority of spindles
found by chance (p=0.009). We propose that in this population of women, the examined in both fresh and vitrified biopsied embryos were normal. However in
quality of the oocytes, reflected by longer telomeres, is a major factor enabling the vitrified groups a higher incidence of spindle abnormalities was observed which

was not significantly different between Day3 biopsied embryos (33/98 DC-1 rates and the time from insemination to reaching the 5-cell, 8-cell, blastu-
(33.7%) abnormally shaped spindles and 5/98 (5.1%) multipolar and monopolar lation, and expanding blastocyst stages between sERC+ and sERC- oocytes. No
spindles and day5 biopsied embryos (42/128 (32.8%) abnormally shaped spindles correlation was observed between sERC+ status and blastocyst utilisation rate
and 7/128 (5.5%) multipolar and monopolar spindles (p>0.05). Transmission elec- [sERC+ oocyes: 46.2%, 96/209, sERC- oocytes: 48.7%, 2,111/4,331, adjusted
tron microscopy (TEM) analysis revealed similar ultrastracture between the day 3 odds ratio: 1.01, 95% confidence interval (CI): 0.75-1.35]. Study 2: The LBRs
and day 5 biopsied embryos before vitrification. However, compared to fresh following SET in sERC+ and sERC- oocytes were 21.7% (10/46) and 18.6%
blastocysts vitrified blastocysts had more characteristic lipofuscin droplets (repre- (546/2942), respectively. There was no significant correlation between the
sentative of apoptosis) and a higher number of vacuoles and distension of sERC+ status and LBR (adjusted odds ratio: 1.35, 95%CI: 0.79-2.42).
mitochondria. Furthermore, all of the babies derived from sERC+ oocytes were healthy.
Limitations, reasons for caution: The blastocysts used in this study were Limitations, reasons for caution: The retrospective design and small sample
all diagnosed with either chromosomal abnormalities or single gene defects size are the major limitations. The rate of sERC+ oocytes was very low during
following PGT-A or PGT-M. the study period. Therefore, these were the only cases we could identify. Further
Wider implications of the findings: This is the first study to compare via- studies are needed to explore the mechanism by which sERCs influence PN
bility, ultrastructure and spindle/chromosome configurations in day3 and day5 behaviour and morphokinetics.
biopsied embryos following vitrification. The similarities observed in ultrastucture Wider implications of the findings: Our results demonstrated that sERC
reflect similar patterns of metabolism, while lipofuscins indicate ‘rescue’ pro- did not adversely affect embryonic and pregnancy outcomes. Therefore, it
cesses for the embryos, which in their attempt to fully recover following vitrifi- was suggested that sERC+ oocytes could be used for IVF treatment. We also
cation, eliminate damaged/abnormal cells. found that sERCs correlated with PN behaviour and morphokinetics in ear-
Trial registration number: not applicable ly-cleavage stage. Further detailed investigations are required to confirm these
findings.
O-067 Smooth endoplasmic reticulum clusters in oocytes Trial registration number: not applocable
influence pronuclear behaviour and morphokinetics at early-
cleavage stage but have no negative impact on embryonic and
pregnancy outcomes
S. Ueno1, T. Miki2, U. Kazuo1, T. Okimura1, A. Yabuuchi2, SELECTED ORAL COMMUNICATIONS
T. Kaobayashi3, K. Kato3 SESSION 18: CELLULAR AND MOLECULAR MARKERS OF
1
Kato Ladies Clinic, IVF Laboratrory, Tokyo, Japan ; OVARIAN AGEING
2
Kato Ladies Clinic, R&D division, Tokyo, Japan ; 06 July 2020 Parallel 2 15:15 - 16:35
3
Kato Ladies Clinic, Gynaecology, Tokyo, Japan
Study question: Do smooth endoplasmic reticulum clusters (sERCs) influence

pronuclear behaviour at fertilisation, morphokinetics, blastocyst development,
and live birth following single embryo transfer (SET)? O-068 Which accumulation is better in poor ovarian responders
Summary answer: Smooth ERCs influence pronuclear behaviour at fertilisa- stratified by POSEIDON, egg or blastocyst?
tion and morphokinetics at early-cleavage stage, but have no adverse impact on T.H. Chuang1, J.Y. Hsieh1, C.T. Kuo1, M.J. Lee1, H.L. Wang1,
embryonic and pregnancy outcomes. H.H. Lai1
What is known already: sERC-positive (sERC+) oocytes and/or cycles in 1
Stork Ladies Clinic, Stork Fertility Center, Hsinchu, Taiwan R.O.C.
which sERC+ oocytes were retrieved are reported to be associated with poor
clinical outcomes following IVF-ET. However, the effects of sERCs on embryonic Study question: Which accumulation is better in poor ovarian responders
and pregnancy outcomes remain unclear. The endoplasmic reticulum (ER) is a stratified by POSEIDON, egg or blastocyst?
major internal storage site of calcium ions, and the ions stored in the ER should Summary answer: Both egg accumulation or blastocyst accumulation achieve
be mobilized properly during fertilisation for further embryonic development. comparable clinical outcomes in poor ovarian responders.
Therefore, it is hypothesized that sERCs may be associated with aberrant pro- What is known already: The patients have poor responses to ovarian stim-
nuclear (PN) behaviour at fertilisation and morphokinetics during embryo devel- ulation (POR) generally resulted in low quantity of retrieved oocytes, fewer
opment; however, the number of studies supporting this hypothesis is limited. available embryos, and thus reduced pregnancy rate. Attributing to the improve-
Study design, size, duration: A retrospective study of 6,010 cycles (mean ment of the vitrification technique in human eggs and embryos, accumulation of
age: 38.3±3.5 years old), including 294 cycles with sERC+ cycles was conducted eggs/embryos becomes an effective and safe way to increase the number of
between 2014 and 2018. Study 1: PN size and behaviour at fertilisation, direct available embryos and the opportunity of a successful pregnancy.
cleavage rate in 1st cytokinesis (DC-1), morphokinetics during embryo develop- Study design, size, duration: In this retrospective study, 955 oocyte pick-up
ment, and blastocyst utilisation rates were compared between sERC+ and, sERC- (OPU) cycles were involved from January 2017 to October 2019, consisting of
oocytes obtained from sERC- cycle. Study 2: Live birth rates (LBRs) following 409 cycles from 110 patients with egg accumulation (EAc) and 546 cycles from
SET were compared between sERC+ and, sERC- oocytes from sERC- cycle. 172 patients with blastocyst accumulation (BAc). The definition of POR was
Participants/materials, setting, methods: Oocytes were inseminated by ICSI based on the POSEIDON criteria (Humaidan et al., 2016). These patients
and were cultured in either a dry incubator (ASTEC, Japan) or a time-lapse incubator decided to undergo one of the accumulative strategies by themselves after con-
(Vitrolife, Sweden). Morhpokinetics were analysed using EmbryoViwer (Vitrolife). sulting their doctors about the pros and cons.
Wilcoxon rank sum test and chi-square test were used to compare the PN size, DC-1 Participants/materials, setting, methods: The inclusion criteria of patients
rate, and morphokinetics between sERC+ and sERC- oocytes. Chi-square test and were age ≥35 years, AMH <1.2 ng/mL, and retrieving more than one oocyte in each
multivariate logistic regression analysis were used to compare blastocyst utilisation OPU cycle. The metaphase II (MII) oocytes were cryopreserved in the EAc group,
and LBRs at >22 weeks of pregnancy between sERC+ and sERC- oocytes. and the blastocysts grading above BC according to Gardner system were cryopre-
Main results and the role of chance: Study 1: The time (hours) required served in the BAc group via vitrification (Cryotech®, Japan). The clinical outcomes
from insemination to PN appearance (male: 4.27±1.28 vs. 4.49±1.47, female: were calculated in the patients underwent frozen-thawed blastocyst transfer.
5.10±1.43 vs. 5.30±1.55), PN alignment (7.90±2.51 vs. 8.65±2.95), PN fading Main results and the role of chance: The mean retrieved MII oocytes per
(22.3±3.7 vs. 23.4±4.2), 2-cell (25.1±4.2 vs. 26.1±4.3), 3-cell (35.1±5.4 vs. case (EAc: 8.2±4.2, BAc: 7.4±6.2, p<0.05) and the mean obtained blastocyst per
35.6±6.2), and 4-cell (37.4±6.9 vs. 38.0±6.5) stages was significantly faster in case (EAc: 3.3±2.6, BAc: 2.9±2.6, p<0.05) were significantly higher in the EAc
the sERC+ oocytes than in the sERC- oocytes (P < 0.05). The difference in PN group. In contrast, significantly higher fertilization rate (EAc 71.7%, BAc: 76.0%,
size (between male and female) immediately before PN fading was significantly p=0.03) was observed in the BAc group. No difference was found in good blas-
larger in the sERC+ oocytes than in the sERC- oocytes (105.5 ± 106.1μm2 vs, tocyst formation rate (EAc: 50.3%, BAc: 51.2%, p=0.73) between the two groups.
72.8 ± 67.2μm2, P < 0.05). However, there were no significant differences in Of reproductive outcomes, the biochemical pregnancy rate (BPR), clinical

pregnancy rate (CPR), and implantation rate (IR) between the EAc and the BAc Limitations, reasons for caution: Oxidative phosphorylation and ROS
groups were comparable: [BPR] 52.9% vs. 58.1% (p=0.60); [CPR] 37.1% vs. 27.9% damage were demonstrated in all stages of follicle development, but sample size,
(p=0.29); [IR] 22.0% vs. 26.0% (p=0.53). Of PGT-A transfer cycles, there was in combination with the microscopy techniques used, limited our statistical anal-
no significant difference between the two groups. ([IR] 40.0% vs. 37.7%, p=1.0) yses based on maternal age groups.
Limitations, reasons for caution: The retrospective nature is the main Wider implications of the findings: We demonstrated that oocytes and
limitation of this study. granulosa cells at all stages of folliculogenesis have active mitochondria and
Wider implications of the findings: Though the laboratory outcomes showed generate ROS, contributing to protein and lipid damage, which can accumulate
differences between the patients with embryo and egg accumulation, the clinical during ageing. Future studies on the contribution of oxidative phosphorylation
outcomes of cryotransfer displayed comparable in the two populations. to the decline in fertility of women of advanced maternal age seem warranted.
Trial registration number: not applicable Trial registration number: not applicable
O-069 Human ovarian ageing can be caused by Reactive Oxygen

O-070 Serum dehydroepiandrosterone sulphate (DHEA-S)
Species (ROS)-induced damage in primordial follicles
concentration is a predictive factor for clinical pregnancy
M. Smits1, R. Wüst2, F. Dijk3, S. Dorrestein1, M. Kleijn1, in women with poor ovarian response undergoing in vitro
M. Goddijn1, R. Houtkooper4, S. Mastenbroek1, G. Hamer1 fertilization (IVF)
1
Amsterdam UMC- University of Amsterdam, Center for Reproductive Medicine, A. Fuentes1, K. Sequeira1, A. Muñoz1, A. Tapia-Pizarro1
Amsterdam, The Netherlands ; 1
2 IDIMI - Hospital San Borja Facultad de Medicina- Universidad de Chile,
VU University, Department of Human Movement Sciences, Amsterdam, The
Reproduccion Humana, Santiago- RM, Chile
Netherlands ;
3
Amsterdam UMC- University of Amsterdam, Pathology, Amsterdam, The
Netherlands ; Study question: Is it possible to predict clinical pregnancy rate (CPR) in women
4
Amsterdam UMC- University of Amsterdam, Laboratory Genetic Metabolic with poor ovarian response (POR) undergoing IVF?
Diseases, Amsterdam, The Netherlands Summary answer: Serum Dehydroepiandosterone sulfate (DHEA-S) concen-
tration showed to predict clinical pregnancy in women with POR undergoing IVF
Study question: Can ovarian ageing be caused by life-long oxidative phos- What is known already: Serum DHEA-S levels have been shown to decrease
phorylation, and therefore reactive oxygen species exposure, in oocytes and with age. In parallel, the probability of pregnancy also decreases. The purpose
granulosa cells? of this study was to identify a minimum serum DHEA-S level that correlates with
Summary answer: Oxidative phosphorylation, and reactive oxygen species acceptable CPR in women with POR undergoing IVF
(ROS)-induced damage, are visible during all stages of folliculogenesis, suggesting Study design, size, duration: Prospective cohort study involving 70 women
life-long ROS-exposure could indeed contribute to ovarian ageing. followed at the tertiary referral center, IDIMI, Universidad de Chile, from 2017-2018
What is known already: Oocyte quality declines with increasing female age. Participants/materials, setting, methods: A cohort of women with POR
The reason for this is unknown. Oogenesis mainly occurs before birth after which (n = 50) and a control group (n = 20) undergoing IVF were studied. Serum
oocytes have to remain dormant in primordial follicles until they are ovulated, levels of DHEA-S, AMH, FSH, estradiol as well as Antral follicle count (AFC) on
a process that may take decades. Damage caused by ROS released from mito- day 3 of ovarian stimulation were assessed. DHEA-S concentrations in follicular
chondria during oxidative phosphorylation might be involved in the decline in fluid at the time of oocyte pick-up were also measured. Clinical pregnancy out-
oocyte quality. Although based on little experimental evidence, the current comes were correlated with the assesed parameters. Correlations and ROC
thinking is that, to prevent generation of ROS, primordial follicles only use gly- analyses were perfomed to predict clinical pregnancy
colysis for energy supply. We have now assessed mitochondrial activity and Main results and the role of chance: There was no difference between the
ROS-induced damage in oocytes and granulosa cells throughout all stages of two groups in day 3 FSH and estradiol. Mean age and duration of ovarian stim-
folliculogenesis. ulation were significantly higher in women with POR than controls (35.7±0.55
Study design, size, duration: Using the Dutch Pathology Registry (PALGA), vs. 32.84±1,19 and 11.28±0.27 vs. 10.31±0.36 respectively) while day 3 AFC,
we collected ovarian biopsies stored in the Netherlands between 1991 and 2018. Serum DHEA-S, Follicle-to-Oocyte Index (FOI) and serum AMH concentrations
Biopsies were excluded in case of ovarian malignancies, polycystic ovarian syn- were significantly lower in the POR group than control (6±0.36 vs. 12.47±1.73,
drome or preventive resection due to BRCA mutation carrier. In total, 39 ovarian 160.57±12.52 vs. 218. ±25.30, 0.59±0.66 vs. 1.42±0.22, and 0.76±0.14 vs.
biopsies of individual patients, ranging in age from 18 to 45 years were received, 2.50±0.72; respectively). Serum DHEA-S concentration strongly correlated
of which 34 contained at least one follicle. with those in follicular fluid (Pearson index = 87.5%). The ROC analysis for
Participants/materials, setting, methods: Ovarian biopsies were used for DHEA-S vs. pregnancy showed an area under the curve of 81.1% with a sensi-
high resolution microscopic analysis of mitochondrial structure, activity and ROS- tivity of 87.5% and a specificity of 80.3 % with a cutoff value of 221.4 µg/dL
induced damage to proteins, lipids and DNA. Antibodies used were: TOMM20, Limitations, reasons for caution: This prospective pilot study significantly
anti-phosphorylated pyruvate dehydrogenase (PDHA), anti-3-Nitrotyrosin, implicates endogenously circulating levels of DHEA-S in the clinical pregnancy
anti-4-Hydroxynonenal and anti-8-Hydroxy-2’-deoxyguanosine. Deconvoluted rates. Although our study has a power of 81%, more systematic studies/RCTs
Z-stack images of follicles at different developmental stages were taken on a with larger sample size are required to confirm circulating DHEA-S levels as an
Leica SP8 fluorescence microscope, and analyzed by Leica Application Suite X. efficient and robust predictor of clinical pregnancy rates
IBM SPSS Statistics 25.0 was used for statistical analysis. Wider implications of the findings: This study underlines the significance
Main results and the role of chance: For the first time, we showed phos- of optimum levels of endogenous DHEA-S for successful implantation. Our
phorylated pyruvate dehydrogenase activity in oocytes and granulosa cells in results suggest that the so called adrenopause of any origin seems to be associ-
all stages of follicular development: from primordial follicles to tertiary follicles. ated with poor results in IVF. This information can be crucial during the pre-IVF
This indicates that pyruvate is actively converted into acetyl-CoA, and trans- counseling of these women
ported into the citric acid cycle, rather than used for glycolysis. We observed Trial registration number: not applicable
ROS-induced cellular damage in oocytes of primordial follicles, in women of
all reproductive ages. No age-related differences in immunofluorescence inten-
O-071 Longevity pathways are involved in human ovarian ageing
sity ratios between mitochondria (TOMM20 antibody intensity) and mitochon-
drial activity (phosphorylated PDHA antibody intensity) were observed. But G. Janssens1, M. Smits2, M. Goddijn2, G. Hamer2, R. Houtkooper1,
ROS-induced damage in lipids and proteins seemed to show an increasing trend S. Mastenbroek2
1
with age, although not statistically significant. The accumulation of ROS damage Amsterdam UMC- University of Amsterdam, Laboratory Genetic Metabolic
could impair oocyte quality, leading to decreased chances of becoming preg- Diseases, Amsterdam, The Netherlands ;
2
nant, and increased chances of pregnancy loss as seen in women of advanced Amsterdam UMC- University of Amsterdam, Center for Reproductive Medicine,
maternal age. Amsterdam, The Netherlands

Study question: Are gene expression pathways known to be involved in gen- Study question: Do poor responders present with different levels of cfDNA
eral aging, particularly longevity pathways, also involved in human ovarian ageing? concentration and integrity, as well different levels of inflammatory and growth
Summary answer: Growth, metabolism, and cell cycle progression related markers in follicular fluid?
pathways that are known to be involved in general ageing appear to play a role Summary answer: Poor responders present with similar cfDNA levels and
in ovarian ageing. integrity, compared to normal responders in natural cycles, albeit with lower
What is known already: Oocyte quality declines with advancing maternal age, follicular fluid levels of G-CSF.
a process generally referred to as ovarian ageing. Genetic pathways that modulate What is known already: Embryo developmental potential is associated with
the rate of general, somatic cell, ageing have been researched intensively. Pro- oocyte quality and maturation status. Follicular fluid includes a wide pallet of
longevity genes slow down the process of ageing and increase life span, while hormones, growth factors and members of the Transforming Growth Factor-
anti-longevity genes accelerate ageing and therefore decrease life span. Ovarian beta superfamily, interleukins, cytokines, reactive oxygen species, anti-apoptotic
ageing does not follow the same time line as general ageing. It is not known whether factors, peptides, amino-acids, sugars, and prostanoids, in conjunction with cir-
generally recognized longevity genes also play a role during ovarian ageing. culating cell-free DNA. These markers observed in follicular fluid affect the
Identification of ovarian ageing pathways can lead to new hypotheses and possible ovarian folliculogenesis process, and subsequent embryo development and
treatment options for subfertility caused by ovarian ageing. pregnancy outcome. A better understanding of the pathophysiological processes
Study design, size, duration: We analyzed a dataset of individual gene expres- involved in poor ovarian response (POR) is of added value.
sion profiles of 38 Germinal Vesicle (GV) oocytes of 38 women aged between Study design, size, duration: A total of 88 patients undergoing IVF, were
25 and 43 years old. Correlations between female age (calendar age and biological enrolled in this prospective single-center observational study, from September
age, i.e. dosage of follicle stimulating hormones, antral follicle count (AFC)) and 2018 to July 2019. Forty-four of these patients were identified as poor respond-
longevity pathways were investigated using a database of known longevity ers according to Bologna Criteria, whereas the remaining 44 were classified as
pathways. normal responders. All patients underwent natural cycles with fresh embryo
Participants/materials, setting, methods: Transcripts of 38 GV oocytes transfers. Clinical pregnancy was similarly recorded.
were used for individual gene expression analysis. R version 3.5.1 was used to Participants/materials, setting, methods: Follicular fluid was collected as
process and analyze data. GeneAge database build 19 was used to obtain mouse part of the oocyte retrieval process. Levels of cfDNA were quantified via
ageing related genes. Human to mouse orthologs were obtained using the R Q-PCR employing the ALU115 and ALU247 primers, associated with necrotic
package biomaRt. Correlations and significance between gene expression data and apoptotic and necrotic events respectively. The Q247/Q115 ratio rep-
and ages was tested for using Pearson’s product moment correlation coefficient resents cfDNA integrity. Follicular fluid levels of inflammatory markers-
using ranked expression data. Distributions were compared using a two-sample Interleukin-15 (IL-15) and Corticotropin-Releasing Hormone (CRH)- and of
Student’s t-tests. growth factors – Insulin-like Growth Factor 1 (IGF-1), Vascular Endothelial
Main results and the role of chance: 46 anti-longevity genes showed a Growth Factor (VEGF) and Granulocyte Colony-Stimulating Factor (G-CSF)
positive correlation with female calendar age and FSH dosage administered were evaluated via ELISA.
during ICSI treatment and a negative correlation with AFC. 84 pro-longevity Main results and the role of chance: In the normal responders group a
genes were negatively correlated with calendar age and FSH dosage, while pos- positive correlation between CRH and IL-15 was established (p<0.001), as well
itively correlating with AFC. as between G-CSF and progesterone (p=0.03). A negative correlation between
In general, pro- and anti-longevity genes changed in opposing directions with E2 and cfDNA integrity was also observed (p=0.005). In the poor responders
advancing maternal age in oocytes in a manner that, in somatic cells, represents group a positive correlation was established between VEGF and ALU 115
ageing and a limited life span, and in oocytes could reflect the potential or the (p=0.006) as well as ALU 247 (p=0.01). Negative correlation was established
quality of the oocyte. between VEGF and FSH (p=0.02). IGF-1 was negatively correlated with AMH
Notably, the anti-longevity genes include many ‘growth’ related genes involved (p=0.04) and CRH (p=0.008). A positive correlation CRH and IL-15 was also
in the MTOR pathway, such as EIF5A2, EIF3H, EIF4E, and MTOR. The pro-lon- observed (p=0.005). Between the two groups no statistically significant differ-
gevity genes include many cell cycle progression related genes involved in DNA ence was observed regarding the levels of IL-15, CRH, IGF-1 or VEGF. The levels
damage repair (e.g. XRCC6, ERCC2, and MSH2) or cell cycle checkpoint regula- of G-CSF were statistically significant higher in the normal responders’ group
tion (e.g. ATM, BRCA1, TP53, TP63, TP73 and BUB1B). when compared to the poor responders’ group (p=0.003). No statistically sig-
Limitations, reasons for caution: Using mature oocytes instead of GV nificant difference was observed between the groups regarding cfDNA concen-
oocytes may potentially provide different results. No correction for multiple tration or cfDNA integrity. Cycles that resulted in clinical pregnancy presented
testing was carried out because a small set of longevity related genes were with a lower cfDNA integrity compared to the cycles not leading to clinical
selected a priori for the analysis. pregnancy both in the normal and in the poor responders’ group (Normal:
Wider implications of the findings: Growth, metabolism, and cell cycle 0.07±0.04 vs 0.25±0.17, p<0.001; Poor: 0.10±0.06vs0.26±0.12 p<0.001
progression related pathways that are known to be involved in general ageing respectively). Cycles with successful blastocyst formation presented with higher
appear to play a role in ovarian ageing. We suggest that interventions known to cfDNA levels in the normal responders’ group (0.38±0.26vs0.82±0.66 p=0.03).
modulate these processes could benefit women suffering from ovarian ageing. Limitations, reasons for caution: Limitations of our study refer to the
Trial registration number: not applicable limited size of the studied population, as well as lack of data referring to the
all-inclusive patients’ history, namely body max index, exercise activity or smok-
ing, which may be identified as possible sources affecting cfDNA levels.
O-072 Investigating apoptotic, inflammatory and growth Wider implications of the findings: Poor and normal responders present
markers in the follicular fluid of poor responders undergoing with similar levels of inflammatory, apoptotic or necrotic markers. Lower levels
natural cycle IVF treatment of G-CSF associated to poor responders indicate G-CSF’s role in folliculogene-
N. Nitsos1, K. Sfakianoudis2, P. Tsioulou3, A. Glava3, E. Maziotis3, sis/follicular maturation. Levels of cfDNA and integrity, correlated with clinical
P. Giannelou3, S. Grigoriadis3, A. Rapani3, A. Nezos3, K. Pantos2, outcome, highlighting their role as possible biomarkers. A conclusive verdict
M. Koutsilieris3, N. Vlahos4, G. Mastorakos5, M. Simopoulou3 requires larger studies.
1
Genesis Athens Clinic, Microbiology Laboratory, Athens, Greece ; Trial registration number: Not applicable
2
Genesis Athens Clinic, Assisted Conception Unit, Athens, Greece ;
3 O-073 Transcriptome analysis of human granulosa cells after
National and Kapodistrian University of Athens, Physiology, Athens, Greece ;
4 conventional controlled ovarian stimulation versus mild ovarian
Aretaieion Hospital- Medical School- National and Kapodistrian University of
stimulation in poor ovarian responders
Athens, Second Department of Obstetrics and Gynecology, Athens, Greece ;
5
Aretaieion Hospital- Medical School- National and Kapodistrian University of X.P. Liu1, C.C. Zhou1, J.J. Li1, T.B. Wu1, H.S. Mai1, X.Y. Liang1,
Athens, Unit of Endocrinology- Diabetes Melitus and Metabolism- 2nd Department R. Huang1
1
of Obstetrics and Gynecology- Aretaieion Hospital- Medical School- National and Reproductive Medicine Center, 6th Affiliated Hospital-Sun Yat-sen University,
Kapodistrian Uni, Guangzhou, China

Study question: What is the difference on the transcriptome of human gran- microbiome by analysing bacterial 16S ribosomal RNA with a focus
ulosa cells after mild ovarian stimulation and conventional controlled ovarian on Lactobacillus population. Lactobacillus-dominated microbiota (LDM, defined as
stimulation in poor responders? >90% Lactobacillus species) in the endometrium was reported to be associated
Summary answer: Different stimulation protocols for poor responders with favorable reproductive outcome, while non-LDM (<90% Lactobacillus species)
induced alterations in the granulosa cells that may affect immune processes, was found to decrease implantation, clinical pregnancy, and ongoing pregnancy.
cytokine interactions and the pathway of oocyte development. Study design, size, duration: The prospective cohort study consisted of
What is known already: No consensus is available regarding the optimal 134 women with RIF (defined as at least three previous failed in vitro fertilization
decision between mild stimulation and conventional controlled ovarian stimula- (IVF)-embryo transfer (ET) attempts) from July in 2018 to October in 2019 at
tion (COS) protocols for poor responders. Currently, studies comparing the our infertility center. EMMA/ALICE testing were suggested to all patients who
efficacy of COS versus mild stimulation have generally focused on clinical preg- had failed ET three times or more. Ninety-eight patients underwent EMMA/
nancy outcomes. Until now, none of studies have discussed the impact of such ALICE (Study group) before additional transfer, 36 patients with history of RIF
two different stimulation regimens on the cellular physiology of follicular cells in continued ET without both examinations (Control group).
poor responders. There is no referable evidence that based on follicular physi- Participants/materials, setting, methods: During the patients’ luteal phase,
ology to support which stimulation protocol is optimal for them. endometrial biopsies were performed for EMMA/ALICE testing, and treatment
Study design, size, duration: This is an experimental study which contained was given based on the results. The primary outcome measure was the cumu-
48 poor responders according to Bologna criteria. lative clinical pregnancy rate after two additional ET. Clinical pregnancy was
Participants/materials, setting, methods: Forty-eight patients who met defined by visualization of a gestational sac. Statistical analysis was performed
the Bologna criteria of poor ovarian response were allocated into two groups using unpaired t-test and chi-square contingency.
as mild stimulation or COS. Twenty-seven patients allocated to the mild stimu- Main results and the role of chance: Patients characteristics such as age,
lation group, while 21 patients in the COS group. Granulosa cells are collected body mass index, duration of infertility, and anti-mullerian hormone (AMH) were
on the oocyte pick-up day, and RNA was extracted and sequenced using Illumina comparable between the groups. The mean number of unsuccessful implanta-
Hiseq technology. Some of the differently expression genes were validated by tions were comparable between the groups (Study group: 5.33±3.03 vs. Control
real-time quantitative PCR. group: 5.22±2.60, P=0.841). The rate of the patients with the history of mis-
Main results and the role of chance: In summary, a total of 192 genes were carriage was significantly higher in Study group than that of Control group (Study
up-regulated, and 233 genes were down-regulated, in the mild stimulation group group: 48.0% vs. Control group: 19.4%, p<0.010). According to the results of
compared with the conventional controlled stimulation group. Gene Ontology EMMA, 44 patients (44.9%) with normal microbiota (LDM) didn’t receive pro-
analysis and Kyoto Encyclopedia of Genes and Genomes analysis revealed that biotic treatment. The other 54 patients (55.1%) received probiotic therapies. In
the differently expression genes between the two groups were mainly involved addition, among non-LDM patients, abnormal microbiota was detected in 14
in cytokine-cytokine receptor interactions, TGF-beta signaling pathway, the bio- patients (14.3%) in EMMA, and ALICE detected significant amounts of patho-
logical function of regulation activity and immune response, which associated genic bacteria in other 6 patients. Those 20 patients had antibiotics correspond-
with the development of the follicle and the oocyte. ing to the detected bacteria as well as probiotic treatment. As results, the
Limitations, reasons for caution: The sample size was relatively small in cumulative clinical pregnancy rate in Study group was significantly higher than in
our study. There is a need for larger number of patients. Control group (Study group: 54.1% vs. Control group: 27.8%, p<0.010).
Wider implications of the findings: The two different ovarian stimulation However, there was no significant difference in the ongoing pregnancy rate (Study
protocols for poor responders cause dissimilar biological functions in granulosa group: 44.9% vs. Control group: 27.9%, p=0.073) and the miscarriage rate (Study
cells, which have influence on the development of the oocytes and subsequent group: 15% {8/53 cases} vs. Control group: 0% {0/10 cases}, p=0.332).
IVF outcome. Most importantly, our findings indicate that the mild ovarian stim- Limitations, reasons for caution: The main limitation of our study is the
ulation may be more benefit to the competence of the oocytes. lack of randomization, although it is prospective. As the sample size is small,
Trial registration number: not applicable further prospective studies are needed to confirm the efficacy of
EMMA/ALICE.
Wider implications of the findings: Dysbiotic microbiota could be found
in patients more than half using the NGS technique. Personalized treatment
SELECTED ORAL COMMUNICATIONS recommendations based on the EMMA/ALICE results can improve IVF outcome
SESSION 19: RIF AND ENDOMETRIAL FACTORS: DOES IT of RIF and repeated pregnancy loss. Moreover, broad-spectrum antibiotics treat-
MATTER? ments can be avoided, reducing the physical and economic burdens on patients.
06 July 2020 Parallel 3 15:15 - 16:30
O-075 Composition of the endometrial microbiome is associated
to reproductive outcomes in IVF patients
O-074 Efficacy of endometrial microbiome metagenomic I. Moreno1, I. Garcia-grau2, D. Bau3, D. Perez-Vilarrolla3,
analysis and analysis of infectious chronic endometritis on in vitro M. Gonzalez-Monfort2, D. Valbuena4, F. Vilella5, C. Simon6
fertilization outcome in women with recurrent implantation 1
Igenomix, Research, Boston, U.S.A. ;
failure 2
Igenomix, Research Department, Valencia, Spain ;
N. Iwami1, M. Kawamata1, N. Ozawa1, T. Yamamoto1, 3
Igenomix, Bioinformatics Department, Valencia, Spain ;
E. Watanabe1, M. Mizuuchi1, O. Moriwaka1, H. Kamiya1 4
Igenomix, Clinical Department, Valencia, Spain ;
1 5
Kamiya Ladies Clinic, Center of reproduction, Sapporo, Japan Igenomix Foundation / INCLIVA, Research Department, Valencia, Spain ;
6
Valencia University- Harvard University, Department of Obstetrics and
Study question: Following results of endometrial microbiome metagenomic Gynecology, Valencia- Boston, Spain
analysis (EMMA) and analysis of infectious chronic endometritis (ALICE), is there
an effect post treatment for the patients with recurrent implantation failure (RIF)? Study question: Is there an association between the composition of the endo-
Summary answer: The clinical pregnancy rate of the patients who underwent metrial microbiota and the reproductive outcomes in infertile patients undergoing
EMMA/ALICE testing was significantly higher than that of the patients who did in vitro fertilization (IVF)?
not undergo testing. Summary answer: The composition of the endometrial microbiota investi-
What is known already: Chronic endometritis (CE) is persistent endometrial gated the cycle prior to embryo transfer differs between patients with successful
inflammation mainly caused by bacterial infections. CE is found in about 30% of versus unsuccessful clinical outcome.
infertile women, and 60% of patients with RIF, so pathogenic flora identification What is known already: The investigation of bacterial communities in the
is the first step of treatment. Using next generation sequencing (NGS) technology, female reproductive tract using molecular methods has revealed the existence
EMMA/ALICE testing can determine the composition of the endometrial of a continuum microbiota that extends from the vagina to the upper genital

tract. Despite similarities between vaginal and uterine microbiota, the endome- Groene Hart Hospital Gouda, dr. C.A.H. Janssen, ; Meander Hospital, Mr. E.A.
trial and vaginal microbiota are different in 20% of patients. The analysis of Brinkhuis, ; Noordwest Ziekenhuisgroep, locatie Gemini, Dr. E.R. Groenewoud ;
bacterial composition in endometrial microbiota during the window of implan- St. Antonius Hospital, dr. J. van Disseldorp, ; Wilhelmina Hospital Assen,
tation has shown an association of absence of pathogens and high abundance Mr. M.W. Glas, ; Medisch Centrum Kinderwens, Dr. A.M. van Heusden, ;
of Lactobacilli with IVF success, while the presence of pathogenic bacteria Albert Schweitzer Hospital, Ms. M.L. Bandell, ; Leiden University Medical Centre,
-non-Lactobacilli dominated microbiota-, is associated with poor reproductive Ms. N.F. Klijn, ; Haga Hospital, Dr. Q. Pieterse, ; Haaglanden Medical Centre,
outcomes in assisted reproductive treatments (ART). Ms. S. van der Meer,
Study design, size, duration: This is a prospective observational multicenter
study including a total 452 patients. The study was performed in 13 reproductive Study question: Does mid-luteal endometrial scratching prior to the 2nd stim-
centres located in Europe, North America, South America and Asia. The duration ulation cycle increase live birth rates (LBR) in women with one failed IVF/
of the study was 30 months and the recruitment period extended for 1.5 years ICSI cycle?
starting from August 4, 2017. Summary answer: During follow-up, non-significant 4.0-4.7% increases in
Participants/materials, setting, methods: Patients undergoing IVF LBRs were observed for women that underwent scratching. These differences
receiving personalized embryo transfer with frozen blastocysts in a hormonal should be further explored as to their clinical significance.
replacement therapy in whom endometrial fluid was collected at the time of What is known already: Previous trials on endometrial scratching have been
the ERA test the cycle prior to embryo transfer. Taxonomic data were gen- reported to have methodological limitations and high risk of bias making it unclear
erated by sequencing of the 16S rRNA gene using the Ion 16S metagenomics whether this procedure improves pregnancy rates. Even the recent large PIP trial
kit (Thermofisher) and analysed using compositional data to transform suffers from the fact that a very heterogeneous study population was included
scale-invariant values and avoid bias derived of technical different between and the timing of scratching was not standardized. Moreover, previous trials only
batches. reported the results of a single transfer following randomization. An RCT in a
Main results and the role of chance: The interim analysis in 177 patients with more homogeneous population which standardizes timing and method of
different clinical outcomes including no pregnancy, (n=82), ongoing pregnancy + scratching, and with a follow-up of 12 months may provide the best information
live birth (n=69), biochemical pregnancy (n=10), or clinical miscarriage (n=16) on the effectiveness of scratching.
revealed differential compositional data of the endometrial microbiota collected Study design, size, duration: A multicenter, non-blinded, randomized con-
in the cycle prior to the embryo transfer. Based on the analysis of bacterial genera trolled trial was conducted between January 2016 and July 2018 in the
with an abundance >2-fold that the average, 29 specific taxa were identified to Netherlands. Women were allocated 1:1 to endometrial scratching or no inter-
discriminate between the microbiota profile of women who did not become preg- vention, using a web-based system that ensured allocation concealment. Using
nant and those whose transfers resulted in live births. When the compositional an expected difference in LBR after the IVF/ICSI cycle following randomization
data were interrogated for taxa with >2-fold abundance and filtering from those of 9% (39% vs. 30%, respectively), the sample size was set at 900 participants
genera most differentially represented between these two categories, Citrobacter, (80% power and two-sided alpha of 0.05).
Haemophilus, Kocuria, Gemella, Enterococcus, Gardnerella, Atopobium and Aeromonas Participants/materials, setting, methods: Participants with a failed first
were differentially identified discriminating between the successful or unsuccessful IVF/ICSI cycle (≥1 embryo transfer) were eligible. Endometrial scratching was
outcome groups. Patients that did not become pregnant presented higher abun- performed using an endometrial biopsy catheter in the mid-luteal phase prior
dance of Enterococcus and Haemophilus; patients with biochemical pregnancies to ovarian stimulation. The primary outcome was LBR from the fresh embryo
presented increased abundance of Enterococcus, Gardnerella, Haemophilus and transfer post-randomization; secondary outcomes included cumulative preg-
Kocuria; while patients suffering clinical miscarriages presented higher abundance nancy outcomes up to ongoing pregnancy (OP) (achieved within 12 months after
of Enterococcus. The confounding factors analysed to ruled out any bias of the randomization) leading to LB. This abstract reports the complete case results
reported results were body mass index, age of patient, endometrial receptivity for the primary and secondary 12-month follow-up outcomes.
analysis (ERA) results and preimplantation genetic diagnosis of embryos. Main results and the role of chance: In total, 936 women were included
Limitations, reasons for caution: These are interim results from 177 out (468 scratch/468 control). Baseline and infertility characteristics were compa-
of 452 patients recruited. Because the endpoint of the study was live birth, final rable between the groups. The participation rate was 89% (936/1054 eligibles).
results will only be complete when all the babies will be born. A conservative approach was used, in which participants who were lost-to-
Wider implications of the findings: The differential endometrial microbiota follow-up were assumed not to have conceived.
composition associated to different clinical outcomes in patients undergoing IVF The LBR after the post-randomization treatment (i.e. 2nd cycle) was 22.6% in
might have profound implications for understanding the microbiology of intra- the scratch and 18.6% in the control group (RR 1.21 [95%CI 0.94-1.56]) with a
uterine environment and its implication in unknown causes of infertility and corresponding absolute risk difference (ARD) of 4.0% [95%CI -1.2% to +9.2%].
implantation failure. For the 12-month follow-up, the cumulative LBR was 42.1% vs. 37.4% (RR 1.13
Trial registration number: 03330444 [95%CI 0.96-1.32]) with a corresponding ARD of 4.7% [95%CI -1.7% to +11.1%].
Important strengths include the fairly homogeneous study population, standard-
O-076 Twelve-month follow-up results of a randomized controlled ized scratching method, 12-month follow-up period, large sample size and high
trial studying endometrial scratching in women with one failed participation rate. The 12-month follow-up period is unique and offers the possi-
IVF/ICSI cycle (the SCRaTCH trial) bility to study longer-term effects of endometrial scratching. The high participation
N. Van Hoogenhuijze1, H.L. Torrance1, SCRaTCH study group(2), rate is probably due to the fact that endometrial scratching is not offered as part
M.J.C. Eijkemans1, F.J.M. Broekmans1 of clinical care in the Netherlands. This decreases the risk for selection bias and
1
University Medical Center Utrecht, Gynaecology, Utrecht, The Netherlands ; improves the inference for the effect of endometrial scratching for daily practice.
2
Radboud University Medical Centre, dr. K. Fleischer, ; Academic Medical Centre Limitations, reasons for caution: This analysis was based on complete data
Amsterdam, dr. F. Mol, ; Isala Fertility Clinic, dr. G. Teklenburg, ; Jeroen Bosch from 97-99% of the participants, which means the final results (available March
Hospital, dr. J.P. de Bruin, ; Maastricht University Medical Centre, dr. J.E. den 2020) may differ slightly. Despite this high percentage of complete data, complete
Hartog, ; Vrije Universiteit Medical Centre Amsterdam, Prof. dr. C.B. Lambalk, ; case analysis reduces precision and may introduce bias. Therefore, in the final
Maxima Medical Centre Veldhoven, dr. J.W.M. Maas, ; University Medical Centre dataset residual missing data will be imputed.
Groningen, dr. A. Cantineau, ; Erasmus Medical Centre Rotterdam, Prof. dr. J.S.E. Wider implications of the findings: Twelve-month follow-up results showed
Laven, ; Sint Franciscus Gasthuis, dr. M. van Hooff, ; Onze Lieve Vrouwe Gasthuis that a non-significant LBR improvement of 4.0% after the post-randomization
Amsterdam, dr. E. Moll, ; Amphia Hospital, Dr. J. Visser, ; Fertility clinics Twente, treatment in the scratch group is maintained, resulting in a non-significant 4.7%
dr. M. Verberg, ; Diakonessenhuis Utrecht, dr. P.A. Manger, ; Catharina Hospital improvement after 12 months. Further analyses in this dataset and an international
Eindhoven, dr. M.M.E. van Rumste, ; Deventer Hospital, Ms. L.F. van der Voet, ; individual participant data-analysis (IPD) will focus on understanding these findings.
Maasstad Hospital, dr. R.H.F. van Oppenraaij, ; Sint Elisabeth-Twee Steden Trial registration number: This trial was prospectively registered in the
Hospital, dr. J. Smeenk, ; Gelre Hospital Apeldoorn, Ms. M.A.F. Traas, ; Ter Gooi Dutch Trial Register (Nederlands Trial Register) under number ‘NL5193’ (old
Hospital, dr. C.H. de Koning, ; Reinier de Graaf Gasthuis, Dr. J.C. Boxmeer, ; number: ‘NTR 5342’)

O-077 Ovarian stimulation alters the cervical microbiota O-078 Uterine peristalsis during implantation period; experience
S. Mackens1, S. Vieira-Silva2, S. Santos-Ribeiro3, of 4,800 patients with 3 or more failure of embryo transfers
J. Centelles-Lodeiro2, A. Racca1, G. Falony2, R. Tito2, H. Tournaye1, H. Matsubayashi1, K. Kitaya1, K. Yamaguchi1, Y. Ohara2,
J. Raes2, C. Blockeel1 M. Doshida2, T. Takeuchi2, T. Ishikawa2
1 1
UZ Brussel, Centre for Reproductive Medicine, Brussel, Belgium ; Reproduction Clinic Osaka, Obstetrics and Gynecology, Osaka, Japan ;
2 2
Vlaams Instituut voor Biotechnologie VIB - KU Leuven, Center for Microbiology, Reproduction Clinic Tokyo, Obstetrics and Gynecology, Tokyo, Japan
Leuven, Belgium ;
3
IVI RMA Lisbon, Reproductive Medicine, Lisbon, Portugal Study question: Is uterine peristalsis frequently observed in patients with
recurrent implantation failure?
Study question: Does ovarian stimulation (OS) have an impact on cervical Summary answer: Uterine peristalsis was frequently (47.2%) observed in
microbiota composition and diversity? patients with recurrent implantation failure mostly in the whole uterine cav-
Summary answer: OS significantly influences the cervical microbiota com- ity with “lower→upper→lower” direction.
position and increases its diversity. What is known already: Uterine peristalsis caused by uterine contraction is
What is known already: Disturbances of the female genital microbiota are thought to be one of the risk factor for implantation failure, because the uterus
associated with female sexual health complications such as an increased risk for is quiescent at the time of implantation period.Previous studies suggested more
sexually transmitted infections and with obstetrical complications such as than 2 or 3 waves/min may be a threshold for implantation failure. Although
preterm delivery. Recently, it has been suggested that women undergoing IVF/ those reports focused on frequency and direction of the uterine contraction,
ICSI are particularly prone to genital dysbiosis and that an abnormal microbiota there were no reports regarding intensity and location of the uterine contraction.
composition and an increased species diversity might affect post-treatment Therefore, we investigated intensity and location as well as frequency and direc-
pregnancy rates. However, before introducing reproductive tract microbiota tion of the uterine contraction in the largest number of patients with recurrent
analyses in clinical practice to predict IVF/ICSI outcome, confounders need to failure of embryo transfers.
be investigated more thorough. For example, the impact of OS remains to be Study design, size, duration: Transvaginal ultrasonography scans of uter-
elucidated. ine peristalsis were performed at the mid luteal phase in 4,800 patients with 3
Study design, size, duration: This analysis was part of a prospective obser- or more failure of embryo transfers in two clinics between 2013 and 2019. The
vational cohort study investigating the potential effect of the female reproductive transvaginal probe (6 to 10 MHz) was introduced into the vagina as gently as
tract microbiota on IVF/ICSI outcomes and included 106 women each providing possible to avoid stimulating the uterine cervix. After scanning mid-sagittal plane
two samples between 2016 and 2019. Samples from the cervical microbiota of the uterus, the probe was fixed as steady as possible while 3 min, video was
were collected by swabbing at two different timepoints: prior to OS (baseline) recorded simultaneously.
and at the moment of oocyte retrieval (post-OS). Participants/materials, setting, methods: The video images were ana-
Participants/materials, setting, methods: Caucasian women, aged lyzed at 10 time the normal speed using Quick Time Player by a single observer.
<40y, planned for a first or second IVF/ICSI cycle performing OS in an antagonist Frequency, intensity, location and direction of the uterine contractile activity
protocol followed by fresh single blastocyst transfer were included. Cervical were recorded and evaluated. Intensity was divided into 3 categories; move-
microbiota samples were retrieved by swabbing, flash-frozen in liquid nitrogen ment with the whole endometrium (strong), with the middle and the surface
and stored at -80°C. Microbiota profiles were obtained by amplicon sequencing of the endometrium (medium), and just the surface of the endometrium
(16SrRNA V4) using the gold-standard DADA2 pipeline. Correlations to micro- (weak). Direction was complicated with many patterns (e.g.,
biota profiles were performed by constrained principal coordinates analysis at lower→upper→lower).
genus level (cPCoA with Aitchinson distance). Main results and the role of chance: Of 4,800 patients (average age, 37.5),
Main results and the role of chance: OS led to a significant shift in cervical 2,534 (52.8%) did not show any uterine peristalsis, 2,266 (47.2%) had uterine
microbiota (n=106x2, paired cPCoA, R2=0.006, p=0.016). Also, microbial peristalsis. In the peristalsis group, frequency was 57.1% for 1 to 3 (times/3 min),
diversity significantly increased during OS (n=106x2, paired t-test, t-ratio=4.80, 28.6% for 4 to 6, 10.7% for 7 to 9, and 3.7% for 10 or more. Intensity was almost
p=5.4E-6). We evaluated whether the menstrual cycle phase at the moment equal among 3 categories (strong 30.4%, medium 38.8%, weak 30.8%). Most uter-
of baseline sampling confounded this association between OS and the shift in ine peristalsis was observed in the whole uterine cavity (82.8%), whereas those
diversity and observed that, in our dataset, this was not correlated (Kruskal- in the upper, middle and lower part of the uterus were 8.7%, 1.2% and 7.3%,
Wallis, p=0.72). The cervical microbiota profiles were typed into 4 different respectively. In terms of direction, about half (52.1%) of uterine peristalsis was
community-types (CTs), two of them having the characteristic dysbiotic observed as “lower→upper→lower”, followed by “upper→lower→upper”
high-species diversity. Focusing on 72 patients having achieved live birth (n=39) (15.8%), “lower→upper” (13.8%), “upper→lower” (13.3%), and unfo-
versus not having reached a +hCG (n=33) after a fresh embryo transfer, CTs cused (4.9%). Pregnancy outcome of patients (N=29) who had strong uterine
were not significantly associated to outcome at both timepoints (Fisher’s test, peristalsis with 10 or more was retrospectively evaluated after taking piperidolate
baseline p=0.63, post-OS p=0.27). However, CTs shifted significantly from hydrochloride (150 mg/day). Patients with live birth or ongoing pregnancy with
baseline to post-OS (n=144, Fisher’s test, p=0.03). In cervical microbial diver- 28 weeks or more were 13 (44.8%), those with biochemical pregnancy or mis-
sity, also no significant difference was observed between the two outcomes carriage were 7 (24.1%), and those without pregnancy were 9 (31.0%).
(live birth vs not pregnant), at baseline (n=72, Kruskal-Wallis, ES=0.16, p=0.18), Limitations, reasons for caution: Since this is a retrospective observational
nor after OS (n=72, Kruskal-Wallis, ES=0.13, p=0.28). However, an increase study, a prospective randomized study is necessary to determine the cutoff value
in diversity was confirmed for both outcome groups, with this increase being that should be treated for uterine peristalsis in patients with recurrent implan-
slightly higher for the negative outcome (n=33x2, paired t-test, t-Ratio=3.50, tation failure.
p=0.001) than for the live birth outcome (n=39x2, paired t-test, t-Ratio=3.11, Wider implications of the findings: These data suggest that uterine peri-
p=0.004). stalsis was frequently (47.2%) observed in patients with recurrent implantation
Limitations, reasons for caution: Human-associated microbial communities failure, mostly in the whole uterine cavity with direction as “lower→
have a high inter-person variability and even with a reasonably scaled study of upper→lower”. However, we have to determine the cutoff value that should
106 women, statistical significance is difficult to reach. Caution is warranted as be treated. Further studies will be required.
non-significant associations do not prove non-correlations. Trial registration number: N/A
Wider implications of the findings: Interest has increased in predicting
IVF/ICSI outcome using reproductive tract microbiota analysis. The impact
of potential confounders however needs to be assessed first. We found OS
to be a significant driver of microbial compositional and diversity variation, SELECTED ORAL COMMUNICATIONS
labelling it as a potential confounder that deserves more attention in future SESSION 20: REPRODUCTIVE (EPI)GENETICS 1
research. 06 July 2020 Parallel 4 15:15 - 16:30

O-079 Chromosome errors in human eggs shape natural separation of sister chromatids (PSSC) and reverse segregation (RS) events,
fertility respectively. These results were validated directly in human oocytes of two
J. Gruhn1, A. Zielinska2, V. Shukla1, R. Blanshard3,4, A. Capalbo5,6, independent patient cohorts, as well as a third cohort of pre-implantation
D. Nikiforov7,8, A.C.H. Chan1, L. Newnham4, I. Vogel1, R. McCoy9, embryos.
G. Hartshorne10, K. Lukaszuk11,12,13, C.Y. Andersen7, M. Schuh2, Limitations, reasons for caution: None of our samples represent a com-
E. Hoffmann1,4 pletely ‘normal’ setting, due to either in-vitro maturation or hormone stimulation.
1 However, the U-curve was reproducible in three independent datasets, including
Univeristy of Copenhagen, Center for Chromosome Stability, København N,
clinically relevant mature MII oocytes from gonadotrophin-stimulated women.
Denmark ;
2 Wider implications of the findings: Our findings suggest that age-dependent
Max Planck Institute for Biophysical Chemistry, Department of Meiosis,
chromosomal errors originating directly in oocytes may shape the curve of
Göttingen, Germany ;
3 natural fertility in humans.
Illumina Inc., Clinical Genomics Group, Fulbourn, United Kingdom ;
4 Trial registration number: not applicable
University of Sussex, Genome Damage and Stability Centre- School of Life
Sciences, Sussex, United Kingdom ;
5 O-080 preimplantation genetic testing for aneuploidy: evaluation
University of Rome, Dipartimento di Scienze Anatomiche- Istologiche- Medico-
of age, indication and embryological parameters for detection of
Legali e dell’Apparato Locomotore- Sezione Istologia ed Embriologia Medica,
different types of aneuploidy
Rome, Italy ;
6
IGENOMIX, n/a, Marostica, Italy ; S. Ecemis1, K. Boynukalin2, M. Gultomruk3, E. Turgut4, M. Bahceci4
1
7
Univeristy of Copenhagen, Laboratory of Reproductive Biology- The Juliane Marie Bahceci IVF Ankara Centre, Gynecology- Obstetrics and Infertility, Ankara,
Centre for Women- Children and Reproduction- Copenhagen University Hospital Turkey ;
2
and Faculty of Health and Medical Sciences, København N, Denmark ; Bahceci IVF Fulya Centre, Gynecology- Obstetrics and Infertility, Istanbul,
8
Università degli studi di Teramo, Unit of Basic and Applied Biosciences, Teramo, Turkey ;
3
Italy ; Bahceci IVF Fulya Centre, Reproductive Genetics, Istanbul, Turkey ;
4
9
Johns Hopkins University, Department of Biology, Baltimore, U.S.A. ; Bahceci IVF Fulya Centre, Gynecology- Obstetrics and Infertility, Istanbul,
10
University of Warwick, Warwick Medical School- University of Warwick Centre Turkey
for Reproductive Medicine- University Hospital Coventry, Warwick, United
Kingdom ; Study question: Do any patient and embryo characteristics allow us to predict
11
INVICTA, Fertility and Reproductive Center, Gdansk, Poland ; aneuploidy or the type of aneuploidy (single, double, segmental or complex)?
12
Medical University of Warsaw, Department of Gynecological Endocrinology, Summary answer: Female age was the sole significant factor affecting the
Warsaw, Poland ; ploidy status of the embryo. Clinical and embryological features did not reach
13
Medical University of Gdansk, Department of Obstetrics and Gynecological to statistical significance.
Nursing- Faculty of Health Sciences, Gdansk, Poland What is known already: The correlation of morphology and morphokinetic
parameters of embryos with embryo ploidy status have been evaluated in many
Study question: How do the rate and causes of aneuploidy within human studies. Still the evidence is scarce and correlation statement is moderate. The
oocytes change throughout the reproductive lifespan? embryos those were generated from the same cohort of a patient might be
Summary answer: Distinct types of chromosomal errors drive aneuploidy affected by treatment related external factors in a same manner. The type of
in human oocytes in an age-dependent manner, thereby regulating reproductive aneuploidy such as single (trisomy, monosomy, segmental), double (monosomy/
capacity in young girls and older women. monosomy, monosomy/trisomy, trisomy/trisomy, segmental/segmental, seg-
What is known already: Aneuploidy affects an exceptionally high number of mental/aneuploidy) or complex has not been studied in detail.
human conceptions, resulting in congenital disorders or pregnancy loss. Most Study design, size, duration: This is a single center retrospective observa-
focus has been on women of advanced maternal age by studying oocytes tional study performed between January 2016 and January 2019. The study
donated during fertility treatment. What is less appreciated is that aneuploidy includes the data analysis of 1793 blastocysts with conclusive comprehensive
rates in clinically-recognized pregnancies are also elevated in teenagers that are chromosome screening (CCS) by Next Generation Sequencing (NGS) of tro-
by default excluded from IVF based studies. We currently do not understand phectoderm (TE) biopsies obtained from 591 patients. Female age under 35
whether the elevation in teenagers is due to errors in oocytes, sperm, or embry- years included in the study.
onic mitoses. This study provides insight into the mechanisms behind the inci- Participants/materials, setting, methods: PGT-A was offered to infertile
dence of aneuploidy across the entire human reproductive lifespan. couples with recurrent implantation failure(RIF) and repetitive miscarriages (RM),
Study design, size, duration: This is a multicenter study based on >3000 previous history of aneuploidy, severe male factor, and for patient anxiety. TE
oocytes collected from 268 girls and women from two independent cohorts. biopsy was performed on day 5 or day 6 to hatching blastocysts and Gardner
Small antral follicles (cohort 1) were collected directly from ovarian tissue of classification was used as morphological grading. Correlations between patient,
unstimulated girls and women undergoing ovarian cortex cryopreservation prior treatment and embryological parameters and euploidy status were tested in
to chemotherapy (9.1-38.8 years). Mature and immature oocytes (cohort 2) Generalized Linear Mixed Model using Binomial Distribution as Probability
were collected from gonadotrophin-stimulated women being treated in in-vitro distribution.
fertilization (IVF) clinics (20-43 years). Main results and the role of chance: The median female age of this patient
Participants/materials, setting, methods: Oocytes were obtained from population was 32 (30-33). Of the 1793 biopsied blastocysts, 800 were aneu-
women covering a majority of the reproductive lifespan (9-43 years) by in-vitro ploidy, 963 were euploid. Regarding the day of biopsy, the euploidy rate were
maturation of small antral follicles (cohort 1) or immature oocytes (cohort 2) significantly different between day 5 and day 6 biopsied embryos
to the mature meiosis II (MII) stage. The oocyte and polar body were isolated (830/1304(63.7%) vs 281/489(57.5%), p=0.016 respectively). Aneuploid blas-
and underwent whole genome amplification, followed by both Next Generation tocysts showed poor quality ICM [15.4%(105/682) vs 9.1%(101/1111);
Sequencing (NGS) and SNP array analysis. Chromosome segregation errors p<0.001] and TE [75%(512/682) vs 56.3%(626/1111); p<0.001]. The frequency
were identified by NGS based chromosome content. of different abnormal chromosomal patterns as single, complex aneuploid and
Main results and the role of chance: The rate of chromosomal abnor- chaotic patterns were not related to the day of biopsy (p = 0.166) as well as to
malities in oocytes follows a U-shaped curve, suggesting that aneuploidies at the ICM (p=0.12) and TE (p=0.89) score. We observed an increasing probability
both young and advanced ages are female in origin. Unexpectedly, specific for aneuploidy with female age of 3% per year (p<0.015) even though the patient
segregation errors showed different age-dependencies, therefore providing a population was <35 years of age. No statistically significant relation was found
quantitative explanation for the U shape. Increased aneuploidy in young girls between aneuploidy and the other clinical and laboratory findings.
and women (<20) was preferentially associated with whole-chromosome non- Limitations, reasons for caution: The study is limited by its retrospective
disjunction (MI NDJ) events. Whereas women of advancing maternal age (≥33) nature. Higher sample size or a prospective design within the morphokinetic
showed centromeric or more extensive cohesion loss through premature parameters could be used in future studies to corroborate the current findings.

Wider implications of the findings: This study reports that conventional O-082 The application of more stringent parameters for mosaic
morphologic parameters are insufficient for predicting the euploid embryo and classification in blastocysts-stage primplantation genetic testing
the type of aneuploidy. Also the patient characteristics other than female age for aneuploidies reduces false positive mosaic rates without
are not predictive. infertility diagnosis or PGT-A indications does not characterize comprising true detection
the ploidy status. Future research to identify non-invasive biomarkers should be L. Girardi1, C. Patassini1, M. Fabiani1, S. Caroselli1,
performed. M. Serdarogullari2, O. Coban2, N. Findikli3, K. Boynukalin4,
Trial registration number: none M. Bahceci4, R. Chopra5, R. Navarro6, M. Poli1, C. Simón7,8,9,10,
C. Rubio LLuesa11, A. Capalbo1,7
1
O-081 New evidence on mosaic developmental potential: Igenomix, Reproductive Genetics, Marostica, Italy ;
2
multicentric study of 822 mosaic embryos diagnosed by British Cyprus IVF Hospital, Embryology Laboratory, Nicosia, Cyprus ;
3
preimplantation genetic testing with trophectoderm biopsy Bahceci Fulya IVF Centre, IVF Laboratory, Istanbul, Turkey ;
4
Bahceci Fulya IVF Centre, Infertility Clinic, Istanbul, Turkey ;
F. Spinella1, E. Greco2, A. Victor3, M.G. Minasi2, F. Barnes3, 5
Igenomix FZ LLC, Dubai Health Care City, Dubai, United Arab Emirates ;
C. Zouves3, J. Grifo4, E.H. Cheng5, S. Munnè6, A. Biricik1, 6
Igenomix, Igenomix R&D, Valencia, Spain ;
M. Surdo1, M. Baldi1, A. Ruberti2, F. Fiorentino1, M. Viotti3 7
Igenomix Foundation, Reproductive genetics, Valencia, Spain ;
1
Genoma Group srl, Molecular Genetics Laboratories, Rome, Italy ; 8
Baylor College of Medicine, Department of Obstetrics and Gynecology,
2
European Hospital, Centre For Reproductive Medicine, Rome, Italy ; Houston-TX, U.S.A. ;
3
Zouves Fertility Center, Fertility Center, Foster City, U.S.A. ; 9
Harvard University, Harvard School of Medicine- Department of Obstetrics and
4
New York University, Langone Fertility Center, New York, U.S.A. ; Gynecology, Boston, U.S.A. ;
5
Lee Women’s Hospital, Lee Women’s Hospital, Taichung, Taiwan R.O.C. ; 10
Valencia University- and INCLIVA, Department of Obstetrics and Gynecology,
6
Cooper Genomics, Cooper Genomics, Livingston, U.S.A. Valencia, Spain ;
11
Igenomix, PGS Research, Valencia, Spain
Study question: Is the clinical outcome of mosaic embryos influenced by
chromosomal constitution? Study question: How do mosaic diagnostic thresholds setting affect the accu-
Summary answer: Reproductive potential of mosaic embryos is affected by the racy of Next Generation Sequencing (NGS)-based preimplantation genetic
complexity of and the number of aneuploidy cells present in trophectoderm (TE) biopsy. testing for aneuploidies (PGT-A)?
What is known already: Chromosomal mosaic embryos are characterized Summary answer: When single trophectoderm biopsy is tested, wide mosaicism
by the presence of chromosomally different cell lines within the same embryo. thresholds (i.e.,20-80%) increase false positive calls compared to more stringent
While the transfer of these embryos is now offered as an option for women who ones (i.e.,30-70%) without improving true detection rate.
undergo in vitro fertilization (IVF), several concerns remain. For instance, the What is known already: Highly sensitive NGS-based technologies for PGT-A
limited data on pregnancy outcome and the possibility that intra-biopsy mosaicism allows precise identification of intermediate chromosome copy number alter-
in the TE is a poor predictor of the ploidy status of the ICM. Therefore, some ations potentially associated to chromosomal mosaicism in trophectoderm
argue that mosaicism should be not reported until a clear classification of such biopsies. Nevertheless, differences in technical validation procedures and in
embryos in relation with their reproductive potential has been defined. detection thresholds employed for diagnostic calls could lead to incorrect clas-
Study design, size, duration: We collected the clinical outcomes of 822 sification of normal and abnormal embryos into the mosaic category. Overcalling
mosaic embryos transferred in women underwent IVF between May 2016-May mosaicism in trophectoderm biopsies lowers PGT-A accuracy, ultimately affect-
2019. All embryos were cultured to blastocyst stage; trophectoderm (TE) biopsy ing patients treatment outcome from both a clinical and a psychological stand-
was performed on Day-5 of development or Day6/7 for slow growing embryos. point. In this study, we evaluated diagnostic predictivity of different mosaicism
The clinical outcome obtained after transfer of mosaic embryos with different classification criteria by employing blinded analysis of chromosome copy number
chromosomal constitution was compared with each other and with that obtained values (CNV) in multifocal blastocyst biopsies.
from a control group of 3781 euploid blastocysts. Study design, size, duration: The accuracy of different mosaicism diagnostic
Participants/materials, setting, methods: Preimplantation genetic testing cut-offs was assessed comparing chromosomal CNV in intra-blastocysts multi-
(PGT) was performed using high resolution next generation sequencing (NGS) focal biopsies. Enrolled embryos were donated for research between January
methodology. TE biopsies were classified as mosaic if they had 20%-80% abnormal and December 2019. The Institutional Review Board at the Near East University
cells. For statistical analysis mosaic embryos were divided in groups based on mosaic approved the study (project: YDUl20l9l70-849). Embryos showing euploid/
levels and chromosomal constitution detected in TE: single mosaic aneuploidy aneuploid mosaicism in their clinical trophectoderm (TE) biopsy (n=36) and
(monosomy/trisomy; SM), double mosaic chromosomes (monosomy/trisomy euploid embryos (n=23) were disaggregated into 5 portions: the inner cell mass
or combination, DM), complex mosaic aneuploidy (>2 different aneuploidies; CM) (ICM) and 4 TE biopsies. Overall, 295 specimens were analysed.
and mosaic segmental aneuploidy (single and double deletion/insertion >5Mb, MS). Participants/materials, setting, methods: Fifty-nine donated blastocysts
Main results and the role of chance: The embryos were plotted in 10% incre- were warmed, allowed to re-expand and disaggregated in TE biopsies and ICM.
ments, representing a progressive increase in the proportion of aneuploid cells in the PGT-A analysis was performed using Ion ReproSeq PGS kit and Ion S5 sequencer
TE, and linear regression showed a statistically significant decline in rates of implan- (ThermoFisher). Sequencing data were blindly analysed with Ion-Reporter soft-
tation and ongoing pregnancy/birth (regression function with respective slopes -0.42 ware. Intra-blastocyst comparison of raw NGS data was performed employing
and -0.55, P=0.0381 and 0.0099). Regarding chromosomal constitution, MS had the different thresholds commonly used for mosaicism detection. CNV for each
best outcomes, followed by the group with one affected chromosome, followed by chromosome were reported as mosaic, according to 30-70% and 20-80% criteria.
the group with two affected chromosomes, followed by the complex group (implan- Categorical variables were compared using Fisher’s exact test.
tation P<0.0001, ongoing pregnancy/birth P<0.0001). MS showed a significantly Main results and the role of chance: To minimize the impact of technical
poorer clinical outcomes compared to the euploid control group (implantation 51.3% over biological variation, intermediate CNV were classified as confirmed mosaic
vs 61.1%, P=0.0004; ongoing pregnancy/birth 42.6% vs 52.7%, P=0.0003). according to the following criteria: 1) detection of the same mosaic alteration
Limitations, reasons for caution: Additional clinical data must be obtained in at least 3 biopsies or in 1 additional biopsy over the 50% threshold, or 2)
to evaluate the contribution of each different chromosome before this approach detection of a reciprocal mosaic pattern involving the same chromosome. If the
can be evaluated as an additional tool to choose mosaic embryos for transfer. same alteration was uniformly detected (>50%) in all biopsies, the embryo was
Wider implications of the findings: The study provides the largest dataset classified as uniform aneuploid. When the high mosaicism threshold was con-
of transferred mosaic embryo outcomes reported to date. This compiled analysis sidered (50-70%), the aneuploidy finding, mosaic or uniform, was confirmed in
conclusively shows that embryos with different pattern of chromosomal mosa- 82.5% of patterns (14/17; 95%CI=54.6-96.2). In particular, 35.3% of cases
icism have a distinct set of clinical outcomes. This findings should be considered (6/17; 95%CI=3.0-16.8) were uniform aneuploid. For the low mosaicism cate-
for genetic counseling. gory, 30-50%, putative mosaicism was confirmed in only 5.3% of the cases (3/57;
Trial registration number: None 95%CI=1.10-14.62). When 20-50% threshold was applied, a significantly higher

number of false mosaic alterations were observed, and the confirmation rate One possible mechanism to explain the increased mosaicism in RobTC may
dropped to 1.8% (n=3/168;95%CI=0.37-5.13; P<0.001). In particular, the inclu- be the involvement of the centromere in balanced translocation carrier
sion of very-low mosaicism (20-30%) results only added false positive results embryos. This involvement may lead to errors in mitotic divisions during
but no true mosaic case (66.1%;n=111/168;95%CI=58.38-73.19;P<0.001). In embryo development. Embryos of RobTC have been shown to exhibit a
the euploid embryos group, none of the results obtained in the 39 cases of mitotic interchromosomal effect that enhances genetic instability during early
20-30% and in the 30-50% range were confirmed. development (Alfarawati et al., 2012). Also, 30% of RobTC were diagnosed
Limitations, reasons for caution: The study involved only blastocysts initially as having a severe male factor.
diagnosed as euploid or mosaic. Uniform aneuploid embryos were not evaluated Limitations, reasons for caution: The limitation of our study was being a
at this stage. This approach involved the analysis of mosaicism thresholds at the retrospective analysis. A prospective follow-up study with a larger sample size
embryo level and future studies will need to evaluate these criteria in relation is needed to verify the outcome of this abstract.
to clinical predictive values following embryo transfers. Wider implications of the findings: As the possibility of finding an
Wider implications of the findings: Based on an embryo re-biopsy model, euploid or balanced carrier embryo is relatively lower for translocation
single TE biopsy results showing low mosaicism, particularly the very-low range carriers, the transfer of a mosaic embryo may be a realistic option. Higher
(20%-30%) shouldn’t be considered as mosaicism diagnoses. The application of incidence of mosaicism in Robertsonian/ male translocation carriers may pro-
the lower stringency threshold for mosaic classification (i.e.,20%) leads to mis- vide an insight to understanding the mechanism of mosaicism in human
classification of embryos, increasing false positive calls and lowering accuracy of embryos.
PGT-A analysis. Trial registration number: -
O-083 Incidence of mosaic embryos in translocation carriers:

Are translocation carriers more predisposed to generate mosaic
embryos? SESSION 21: IMPACT OF NEW TECHNOLOGIES ON HUMAN
M. Cetinkaya1, M.A. Tufekci1, C. Cinar Yapan1, S. Kahraman1 REPRODUCTION
1
Istanbul Memorial Hospital, Assisted Reproductive Technologies and Reproductive 06 July 2020 Parallel 5 15:15 - 16:30
Genetics Center, Istanbul, Turkey
Study question: Is the mosaicism ratio influenced by the sex of the translo-
cation carrier and the type of translocation in preimplantation genetic testing O-084 A construct of mesenchymal stem cell-derived exosomes/
for structural rearrangements (PGT-SR)? collagen scaffold promotes endometrium regeneration and
Summary answer: The mosaicism ratio is significantly higher in Robertsonian fertility restoration through macrophage immunomodulation
translocation carriers (RobTC) (7x) and in male translocation carriers (3x). L. Xin1
What is known already: Embryonic mosaicism is defined as the presence 1
Zhejiang University, School of Medicine, Hangzhou, China
of karyotypically distinct cell lines within an embryo and can be detected with
Next Generation Sequencing (NGS) at a 20-80% rate. High incidence of mosa- Study question: Can a construct of mesenchymal stem cell-derived exo-
icism has been reported in preimplantation embryos, with the blastocyst somes/collagen scaffold provide a new treatment for Asherman’s syndrome?
mosaicism being between 4-24% (Harton et al., 2017). Although mosaic Summary answer: A construct of mesenchymal stem cell-derived exosomes/
embryos have a chance to implant (Munne et al., 2017; Spinella et al., 2018) collagen scaffold promotes endometrium regeneration and fertility restoration
their pregnancy rate is lower, with an increased miscarriage rate. However, through macrophage immunomodulation
female and male translocation carriers, whether Robertsonian or reciprocal, What is known already: Endometrial traumas caused by recurrent curet-
have not been yet analyzed regarding the incidence of generating mosaic tage, caesarean section and myomectomy always result in intrauterine
embryos. adhesions (IUAs) and infertility. Umbilical cord-derived mesenchymal stem
Study design, size, duration: This retrospective study was based on 143 cell (MSC)-based therapies have shown some promise achievements in the
PGT-SR cycles initiated between January 2017 and September 2019. A total treatment of IUAs for their easy collection, high proliferation and low
number of 573 blastocysts were tested for PGT-SR by ReproSeq on Ion Torrent immunogenicity. However, the potential tumorigenicity, low infusion and
S5 (Thermo Fisher Scientific) following trophectoderm biopsy. low retention of MSCs are still controversial and the clinical application of
Participants/materials, setting, methods: The number of blastocysts MSCs is limited. In contrast, MSC-derived exosomes exhibit a similar func-
tested from female and male translocation carriers were 284 and 289, respec- tion to their source cells and are expected to overcome these
tively. The number of blastocysts tested from reciprocal translocation carriers limitations.
was 471 whereas 102 were from RobTC. The mean female age in all groups was Study design, size, duration: A total of approximately 200 SD rats were
similar and statistically non-significant (32.8 and 31.2 for female and male and involved and received different treatments. Uteri were examined at 1,3,7, 30
33.3 and 31.7 for Robertsonian and reciprocal translocation carriers). Chi-square and 60 days after surgery. We evaluated the effects of CS/Exos on the regen-
test was used for categorical group comparisons. eration of damaged endometrium and the restoration of fertility, as well as
Main results and the role of chance: The overall mosaicism rate for exploring the healing mechanism from the aspect of macrophage immunomod-
PGT-A was 11% (8239 trophectoderm biopsies). The euploid/ balanced trans- ulation by functional miRNAs in vivo and in vitro.
location rate of Robertsonian and reciprocal translocation carriers was 32.4% Participants/materials, setting, methods: we designed a construct of exo-
and 20.4%, respectively (p=0.008). The frequency of mosaic embryos gener- somes and a collagen scaffold (CS/Exos) for endometrium regeneration, and
ated by RobTC was seven times higher than carriers of reciprocal transloca- investigated the regeneration mechanism through macrophage immunomodulation.
tions (21.6% and 3.2%; p<0.0001). PCR,ELISA, IF, IHC, HE staining, Masson’s staining and RNA-seq were involved.
Also, male translocation carriers in general were three times more prone to Main results and the role of chance: The CS/Exos transplantation potently
have mosaic embryos than female translocation carriers (9.3% and 3.5%; induced (i) endometrium regeneration, (ii) collagen remodeling, (iii) increased
p=0.046). The euploid/ balanced translocation rate was identical for male and the expression of the estrogen receptor a/progesterone receptor, and (iv)
female translocation carriers (22.5%). restored fertility. Mechanistically, CS/Exos facilitated CD163+ M2 macrophage
When the subgroups were analyzed, female RobTC were found to generate polarization, reduced inflammation, and increased anti-inflammatory responses in
nearly seven times more mosaic embryos than female reciprocal translocation vivo and in vitro. By RNA-seq, miRNAs enriched in exosomes were the main
carriers (15.4% and 2.3%; p<0.001). Moreover, male RobTC engendered nearly mediator for exosomes-induced macrophage polarization. Overall, we demon-
six times more mosaic embryos than male reciprocal translocation carriers strated that CS/Exos treatment facilitated endometrium regeneration and fer-
(23.7% and 4.2%; p<0.0001). tility restoration by immunomodulatory functions of miRNAs.

Limitations, reasons for caution: The results of this research are inade- O-086 Embryo development of reconstituted zygotes using
quate to apply directly in human because human uterine and rat uterus differ haploid androgenotes obtained from ooplast-mediated male
in structure and function. Further large animal experiments, such as monkeys, genome cloning
are needed. A. Petrini1, P. Xie1, A. Trout1, R. Setton1, Z. Rosenwaks1,
Wider implications of the findings: Our research highlights the therapeutic G. Palermo1
prospects of CS/Exos for the management of IUAs. 1
Weill Cornell Medicine, The Ronald O. Perelman and Claudia Cohen Center for
Trial registration number: not applicable Reproductive Medicine, New York, U.S.A.
O-085 Differentiation of stromal cells into theca cells: Study question: Is it feasible to clone the male genome by utilizing haploid
understanding theca cell formation in human ovaries androgenic blastomeres as male gametes to reconstitute diploid zygotes sup-
P. Asiabi1, M.M. Dolmans1, J. Ambroise2, A. Camboni1, porting full preimplantation development?
C. Amorim3, Andrade1 Summary answer: Haploid androgenotes are capable of complementing female
1 pronuclei, allowing reconstitution of diploid zygotes that completed a normal
Pôle de Recherche en Gynécologie- Institut de Recherche Expérimentale et
preimplantation development comparable to normally fertilized conceptuses.
Clinique, Université Catholique de Louvain- 1200, Brussels, Belgium ;
2 What is known already: In cases of severely impaired spermatogenesis in
Centre de Technologies Moléculaires Appliquées- Institut de Recherche Expérimentale
which only a limited number of gametes is available, or in cases of gametal
et Clinique-, Université Catholique de Louvain- 1200, Brussels, Belgium
3 heterozygosity for heritable diseases, male gamete cloning has been sought and
Université Catholique de Louvain (UCL) -Institut de Recherche Expérimentale et
attempted. Male genome cloning can be achieved through an ooplast-mediated
Clinique Laboratoire de Gynécologie - 1200, Brussels, Belgium
approach that generates androgenic embryos, which cleave to the 2-, 4-, or 8-cell
Study question: Is there a subpopulation of precursor theca cells (PTCs) in stage while maintaining the identical genotype as the injected spermatozoa and
human ovarian cortex or all can ovarian stromal cells differentiate into theca can function as a male gamete. In addition, the propagation of the male genome
cells (TCs)? allows pre-fertilization genetic testing.
Summary answer: Around 43% of ovarian stromal cells (SCs) can differentiate Study design, size, duration: Haploid male embryos were generated and
into TCs, which indicates the presence of PTCs in human ovaries. allowed to cleave to duplicate the male genome. Resulting androgenotes were
What is known already: There are very few studies on the origin of TCs in isolated immediately after first cleavage and treated with DNA polymerase
mammalian ovaries. Progenitor TCs have been described in neonatal mice ova- inhibitor to pause cell cycle before the S phase. Monopronucleated recipient
ries, which can differentiate into TCs under the influence of factors from oocytes oocytes were fused with androgenotes to reconstitute diploid embryos.
and granulosa cells. On the other hand, studies in large animal models have Constructs were maintained in culture up to 96h in a time-lapse system to assess
reported that SCs isolated from the cortical ovarian layer can also differentiate all steps of preimplantation development. Piezo-actuated ICSI was performed
into TCs with the right stimuli. on untreated oocytes as control.
Study design, size, duration: After obtaining informed consent, ovarian Participants/materials, setting, methods: Metaphase II oocytes from
biopsies were taken from eight menopausal women (53-74 years of age) under- B6D2F1 mice were treated with cytochalasin B and enucleated by excising the
going laparoscopic surgery for gynecologic disease not related to the ovaries. spindle herniation under OosightÔ visualization. Resulting ooplasts were injected
SCs were isolated and then in vitro cultured either in basic medium (G1) or with spermatozoa from the same strain and allowed to reach the 2-cell stage.
enriched with growth factors, follicle-stimulating hormone and luteinizing hor- Resulting androgenotes were exposed to 1.5 mM aphidicolin until reconstitution
mone (G2) for 8 days. and fused with monopronucleated recipient oocytes from another cohort using
Participants/materials, setting, methods: To confirm whether SCs Sendai virus. Cleavage parameters of resulting zygotes were obtained by time-
were able to differentiate into TCs, relative mRNA levels for LHCGR, StAR, lapse comparing to ICSI conceptuses as control.
CYP11A1, CYP17A1, HSD3B1 and 2 were assessed. Immunohistochemistry Main results and the role of chance: A total of 59 oocytes were all success-
(IHC) was also performed for their protein detection and a specific marker fully enucleated. The resulting 59 ooplasts were injected with sperm heads; 44
for theca interna cells (CD13). Finally, we analyzed the ultrastructure of the survived (75%), and all developed a single male pronucleus 4-6h post-ICSI. After
cells before (D0) and after (D8) in vitro culture, and DHEA and progesterone culturing up to 16h, 42 (95%) constructs cleaved to the 2-cell stage, yielding 84
levels in the medium using transmission electron microscopy and ELISA haploid androgenotes. Parthenogenic activation by calcium ionophore was suc-
respectively. cessful in 93% of the recipient oocytes, confirmed by the extrusion of the second
Main results and the role of chance: Quantitative PCR results showed polar body and the appearance of a single female pronucleus. A total of 42 haploid
a significant (p<0.05) increase in mRNA levels of HSD3B2 in G1 and G2 and androgenotes were subzonally inserted with Sendai virus and successfully fused
CYP17A1 in G1 after 8 days of in vitro culture. IHC results confirmed expres- with corresponding activated oocytes at a rate of 92.9%. Untreated oocytes were
sion of each enzyme involved in the steroidogenic pathway at the protein inseminated by piezo-actuated ICSI serving as control, yielding 30 zygotes. The
stage. However, only G2 exhibited a significant proportion (43%) of CD13- cleavage of reconstructed embryos into the 2-cell stage (89.7%), 4-cell stage
positive cells compared to cells soon after isolation (0%) and in vitro-cultured (87.2%), morula compaction (84.6%), and blastocyst formation (76.9%) was com-
in basic medium (0%). Ultrastructural analyses showed a distinct difference parable to control ICSI conceptuses (86.7%, 83.3%, 80.0%, and 80.0%, respec-
between the two groups versus D0. Linear trends displayed a significant rise tively), with no noticeable morphokinetic differences.
(q<0.001) in dehydroepiandrosterone (DHEA) and progesterone concen- Limitations, reasons for caution: While this approach is successful in a
trations in medium in G2 culture with time. Similarly, G1 exhibited a signif- mouse model, its application to human reproduction requires serious consider-
icant upturn (q<0.001) in DHEA, but not progesterone, which remained the ation. Major concerns include the retention of imprinting of the male genome
same throughout the culture period. Statistical comparisons of progesterone during the development of the pseudo-blastomere. Other biological hindrances
and DHEA levels on day 8 for G1 versus G2 were q<0.05 and q=0.33 include the role of the human sperm centrosome and heteroplasmy.
respectively. Wider implications of the findings: This technique offers the possibility of
Limitations, reasons for caution: While our results indicate the presence replicating a male gamete and may improve our understanding of reprogramming
of PTCs in the human ovary, as after in vitro culture, 43% of cells express TC immature germ cells as gametes. Male genome cloning may benefit patients with
genes and proteins and are able to synthesize steroids. It is now necessary to heritable diseases who need pre-fertilization gamete screening or the use of genomic
identify specific markers for PTCs to confirm our findings. editing to provide healthy or genome-corrected functional pseudo-gametes.
Wider implications of the findings: Results from current experiments are Trial registration number: not applicable
a promising step towards understanding TC ontogenesis in the human ovary.
Moreover, in vitro-generated human TCs can be used for studies on drug screen- O-087 Use of intraovarian platelet rich plasma does not increase
ing, as well as to understand TC-associated pathologies, such as androgen-se- the ovarian reserve markers,ovarian response or IVF outcome in
creting tumors and polycystic ovary syndrome. Bologna poor responders.
Trial registration number: not applicable S. Singh1, S. Singh1, N. Ashraf1, R. Basheer1, M. Ashraf1

1 1
Craft Hospital And Research Center, Reproductive Medicine, Thrissur, India La Fe University Hospital- Joint first authors, Women´s Health Area, Valencia,
Spain ;
Study question: Does administration of autologous intraovarian platelet rich 2
La Fe University Hospital- Joint first authors, IVI Foundation-Health Research
plasma (PRP) increase the ovarian reserve markers,ovarian response or IVF Institute La Fe, Valencia, Spain ;
outcome in Bologna poor ovarian responders (POR)? 3
La Fe University Hospital, Women´s Health Area, Valencia, Spain ;
Summary answer: There is no benefit of autologous intraovarian PRP in 4
La Fe University Hospital, IVI Foundation-Health Research Institute La Fe,
increasing AMH,AFC,ovarian response to ovarian stimulation or IVF outcome Valencia, Spain ;
in Bologna POR women. 5
La Fe University Hospital, Hematology Department, Valencia, Spain ;
What is known already: Poor ovarian responders are the most difficult 6
La Fe University Hospital, Radiology Department, Valencia, Spain ;
subgroup of patient to treat in ART.The challanges in treatment includes high 7
IVI- RMA- Rome, IVI-RMA Rome, Rome, Italy
risk of cycle cancellation,low number of oocytes/embryos and very low preg-
nancy rates.Many interventions have been tried in these women including andro- Study question: Could reproductive outcomes and menopausal symptoms
gen supplementation,LH supplementation,growth hormone etc,but none has of POI women be improved by autologous stem cell ovarian transplant (ASCOT)
been clearly shown to be beneficial in improving the take home baby rate.PRP or Granulocyte-colony stimulating factor (G-CSF) stem cell-mobilization?
has been utilised as a regenrative therapy by many specialities of medicine since Summary answer: Both ASCOT and G-CSF mobilization promoted follicle
it is rich in growth factors and cytokines.It’s use in reproductive medicine has growth and raised AFC in 60% POI women; obtaining after COS 4 MII-oocytes,
been tried in endometrial regeneration,but data in poor responders for improving 2 embryos, and 1 pregnancy.
treatment outcome is lacking. What is known already: Premature ovarian insufficiency (POI) is characterized
Study design, size, duration: A prospective observational cohort pilot study by oligo-amenorrhea due to follicular depletion in young women, where the only
was performed over a period of 18 months on 30 POR reproductive aged practical option is egg/embryo donation, as their spontaneous pregnancy rate is
women diagnosed based on Bologna criteria at craft hospital,Kerala,India.All the very low(<4%). We recently described that bone marrow derived stem cell
women previously had atleast 1 failed IVF at our center and had details of pre- (BMDSC) infusion promotes follicular growth by increasing ovarian vascularization,
viously performed ovarian reserve testing available within last 3 months.All the stromal cell proliferation, and reducing cell death in POI mice models and xeno-
couples had detailed counselling about the experimental nature of the procedure grafted human ovarian tissue. Based on this, ASCOT technique has been already
and written consent was obtained.Internal ethical committee approval was taken. tested in poor responders improving ovarian function biomarkers (AMH and
Participants/materials, setting, methods: PRP prepared from autologus AFC) in 81.3% of women and allowing a total of 6 pregnancies and 3 healthy babies
blood following standard protocols in mid-late follicular phase was injected under Study design, size, duration: Randomized prospective pilot study started
ultrasound guidance into the ovarian cortex under conscious sedation, using a in 2018 at La Fe University Hospital, Valencia. The study involves 20 POI women,
35 cm 17 G needle at 2 sites in each ovary. After a period of 2 months,antral according to the following inclusion criteria: ≤38 years, ≥4months oligo/amen-
follicle count (AFC) and anti-mullerian hormone (AMH) were reassesed and 2nd orrhea, serum FSH>25IU/l.
IVF was performed by antagonist protocol.The results of post-PRP values of Patients were randomized into two study arms: 1) Mobilization of bone
AFC,AMH and IVF outcome were compared to the pre-PRP value. marrow stem cells to peripheral blood by a 5-day treatment with G-CSF (10ug/
Main results and the role of chance: A total of 30 POR women (mean age kg/day subcutaneous injection), where cells remain circulating; 2) ASCOT:
+/- SD: 38.7 +/- 4.6 ) were included in the study.The ovarian reserve markers after G-CSF mobilization, stem cells were collected and transplanted into
and IVF outcome prior to administration of intraovarian PRP ( pre-PRP group) one ovary.
was compared to the the parameters obtained after intraovarian PRP ( post-PRP Participants/materials, setting, methods: To date, 10 patients have been
group) The pre-PRP and post-PRP group has similar AFC (4.2 +/- 2.6 vs included, 4 of them were randomized to the G-CSF arm and 6 to the ASCOT.
3.8+/- 1.8; p>0.05) and AMH (0.46 +/- 0.26 vs 0.49 +/- 0.22; p>0.05). When After intervention, patients were monitored during 6-month for endocrine func-
compared to the IVF cycle performed pre-PRP administration,the post-PRP tion, serum levels of FSH, AMH and estradiol, antral follicle count (AFC) was
group showed no difference in the number of growing follicles of >/= 14 mm assessed by ultrasound scan. Patients’ basal levels were established before
(3.7 +/- 2.2 vs 4.0 +/- 2.4) , number of MII oocytes (3.2 +/- 1.2 vs 3.5 +/-1.7), treatment.
fertilization rate (2.8 +/- 1.2 vs 2.6 +/- 1.7) and cleavage stage embryos (1.8. When follicular growth was detected (AFC increased in at least 1 follicle)
+/- 1.1 vs 2.0 +/- 1.4) respectively; p>0.05 for all). In 5 women (16.6%),cycle controlled ovarian stimulation was initiated following standard protocols.
had to be cancelled due to lack of follicular growth after 5 days of gonadotrophin Main results and the role of chance: Preliminary results in 10 patients,
administration. 3 women (10%) did not have any embryos to be transferred showed follicular development in both arms when compared to basal levels;
and hence the cycle was cancelled.Embryo transfer was performed in total of these follicular growth waves were detected 90-140 days after treatments.
22 women.Clinical pregnancy rate of 13.3 % (4/30) was achieved with a live In the G-CSF group, AFC increased in 50% of recruited patients (2/4) while
birth rate of 10% (3/10). in the ASCOT this improvement was detected in 66.6% of women (4/6).
Limitations, reasons for caution: This study has limitation of being obser- Statistically significant decrease of FSH levels was not recorded, but two
vational cohort study with the women serving as their own historical controls. women in the G-CSF group(50%) and 1 in the ASCOT(16%) showed a two-fold
The study also has a small smaple size.The data presented here were analysed decrease, concluding the follow-up period(82.7±8.1 to 24.1±17.4 and 34.9 to
2 months after the administration of PRP.Whether waiting for a longer duration 18.1; respectively).
would have changed the outcome is not known. After G-CSF mobilization, 2/4 women, initiated COS, with a total of 4
Wider implications of the findings: Autologus intraovarian PRP adminis- punctured follicles, 2 MII obtained and 1 embryo vitrified. Embryo transfer
tration has not been shown to improve AMH,AFC,ovarian response or IVF was performed but pregnancy was not achieved; this patient currently has
outcome in Bologna poor responder women.It’s use should be restriced strictly regular menses after the follow-up period. In the ASCOT group, 4 of 6 patients
to research settings to prevent women being exposed to this unnecessary inter- initiated COS cycles. Oocyte pick-up was performed in 70% of initiated COS;
vention till further evidence to the contrary through well designed randomised where 3 MII and 1 GV were retrieved. One 3-day embryo was vitrified and
controlled trials. transferred, having an ongoing pregnancy(20 weeks). These women were
Trial registration number: Not applicable unable to undergo COS/oocyte pick-up before treatments due to absence of
antral follicles.
Within menopausal symptoms, hot flashes and vaginal dryness improved in
O-088 Bone marrow derived stem cells restore ovarian function 50% of women, while 40% recovered menses (spotting).
and fertility in premature ovarian insufficiency women. Interim Limitations, reasons for caution: These are descriptive preliminary results
report of a randomized trial: mobilization versus ovarian obtained in the first 10 recruited women that should be confirmed at the end
injection of the trial. Our study lacks from a real control group, without treatment, though
N. Pellicer De Castellvi1, S. Herraiz2, M. Romeu3, S. Martinez3, poor prognosis POI patients should not be left untreated, as their only repro-
A. Buigues4, I. Gomez-Seguí5, J. Martinez6, A. Pellicer7 ductive option is egg donation.

Wider implications of the findings: ASCOT in POI women can be a poten- Limitations, reasons for caution: We were unable to adjust for some poten-
tial therapy for women with no option of having offspring with their own oocytes. tially important predictors, e.g. body mass index (BMI), smoking and alcohol
This study has shown a future use of stem cells in POI women, even without intake in women, as well as measures of ovarian reserve such as antral follicle
direct ovarian infusion, which suggest potential development of less invasive count. These were not available in the linked HFEA dataset.
procedures in the future. Wider implications of the findings: This is the first model to estimate the
Trial registration number: NCT03535480 overall chance of a livebirth over three further cycles for a couple who have
already undergone a complete IVF cycle. The model can be used as a counselling
tool to help couples make an informed decision regarding further treatment.
SESSION 22: UPDATES ON ART OUTCOMES, BARRIERS AND O-090 Assisted reproductive technology in Africa: a five-year
PREDICTIONS: AN INTERNATIONAL OVERVIEW analysis of data from the African Network and Registry
L. Potgieter1, P. Archary1, I. Smit1, O. Ashiru2, E. Gwet-Bell3,
06 July 2020 Parallel 6 15:15 - 16:35
S. Dyer1
1
Groote Schuur Hospital, Obstetrics and Gynaecology, Cape Town, South Africa ;
2
Medical ART Centre, Not applicable, Lagos, Nigeria ;
O-089 Predicting cumulative livebirth from the second complete 3
Clinique Médicale Odyssée, Not applicable, Douala, Cameroon
cycle of IVF: a population-based study of 49,314 couples
M. Ratna1, S. Bhattacharya2, D. McLernon1 Study question: What were utilization, outcomes and practices of assisted
1 reproductive technology (ART) in Africa between 2013 and 2017?
University of Aberdeen, Institute of Applied Health Sciences, Aberdeen, United
Summary answer: Utilization remained low; fresh autologous cycles predom-
Kingdom ;
2 inated, with favorable pregnancy rates but accompanied by high rate of multiples
University of Aberdeen, School of Medicine- Medical Sciences and Nutrition,
and low reporting of delivery outcomes.
Aberdeen, United Kingdom
What is known already: Global access to infertility treatment requires global
Study question: Can we develop an IVF prediction model to estimate the chance understanding of regional realities, challenges and differences. Many African
of cumulative livebirth in couples embarking on a second complete cycle of IVF? countries, the majority of which are low and lower-middle income countries,
Summary answer: Yes, our prediction model can estimate individualised carry a triple burden of infertility, namely high infertility disease prevalence,
chances of cumulative livebirth over three additional complete cycles. negative socio-cultural consequences, and poor access to effective treatment.
What is known already: Existing models can estimate the individualised Tubal and severe male factor are the leading causes of infertility in Africa, often
cumulative chance of livebirth following one or more complete IVF cycles in requiring ART. Availability of ART has been low and treatment outcomes and
couples embarking on their first IVF treatment. There are currently no prediction practices were not monitored regionally prior to 2013. The African Network
models in IVF which can estimate the individualised cumulative chance of livebirth and Registry for ART (ANARA) commenced data monitoring in 2013 with
over multiple complete cycles in women who decide to continue IVF after com- increasing participation.
pleting their first cycle. Existing cumulative prediction models also fail to account Study design, size, duration: Retrospective registry data pertaining to the
for couples who decide to stop treatment after a complete cycle. Models that years 2013 - 2017 were collected cross-sectionally over a period of three years.
do not adjust for discontinuation may give predictions that are too optimistic. All ART centres known to exist on the African continent were invited to partici-
Study design, size, duration: For model development, a population-based pate through mailing campaigns, and in conjunction with local workshops and
cohort was used of 49314 women treated in IVF clinics across the UK from 1999 conference presentations. ANARA online software or standardized data forms
to 2008 using their own oocytes and their partners’ sperm. Model external adjusted from the International Committee for Monitoring ART were used for
validation was performed on data collected from 36170 women who underwent data collection. Both retrospective summary and cycle-based data were collected
treatment in UK IVF clinics from 2010 to 2017. and analysed.
Participants/materials, setting, methods: All UK IVF treatments were Participants/materials, setting, methods: Centre participation and data
obtained from the Human Fertilisation and Embryology Authority (HFEA) data- submission were voluntary. Data sets were received and if necessary, transferred
base. Using a discrete time logistic regression model, we predicted the cumulative to standard data forms. All data were checked for mathematical errors and
probability of livebirth from the second to the fourth complete cycle. We corrected as far as possible but not further validated. Data were pooled by year,
adjusted for couple characteristics at the second cycle and outcome of first cycle. country and then at regional (Africa-wide) level; and analyzed using descriptive
Inverse probability weighting was used to account for treatment discontinuation. statistics. Most results and trends were reported at regional level only, with the
Discrimination was assessed using c-statistic and calibration was assessed using exception of number of procedures and ART utilization.
calibration-in-the-large and calibration slope. Main results and the role of chance: Registry participation increased from
Main results and the role of chance: Of 49314 women,12408 (25.2%) had 40 ART centres in 13 countries in 2013 to 47 centres in 17 countries in 2017.
a livebirth after their second complete cycle. Cumulatively, 17394 (35.3%) had Data are based on 153 461 procedures. Fresh autologous IVF/ICSI predominated
a livebirth over three further complete cycles. Women who had a livebirth in in over 70% of procedures, with autologous frozen embryo transfer and fresh
their first complete cycle had over twice the odds of a livebirth compared to and frozen oocyte donation accounting for 17% and 7% respectively. ICSI was
women who did not get pregnant in their first complete cycle (Odds Ratio: 2.18 performed in over 90% of autologous cycles. Fresh embryo transfer was
(95% Confidence Interval: 2.15, 2.21)). favoured overall.
As an example, consider a 32-year-old woman with two years of unexplained The transfer of two and three embryos predominated for all years and all
infertility who had a livebirth after her first complete cycle. If she starts her procedures. This practice remained constant with no observable increase in
second complete cycle one year later, she has a 48% predicted chance of a single embryo transfers. There was however a trend towards fewer transfers of
second livebirth resulting from this second complete cycle. Over three complete four and more embryos. The majority of autologous cycles were conducted in
cycles her cumulative predicted chance of livebirth is 75%. For a woman with women younger than 35.
similar characteristics who suffered a pregnancy loss in her first complete cycle During the period of observation, the average pregnancy rate per aspiration
the cumulative predicted chance of livebirth is 72% while for a woman who did was 33% for fresh IVF/ICSI, while the pregnancy rate per thaw in frozen autol-
not get pregnant it is 66%. ogous cycles fluctuated between 25% and 39%. These rates were accompanied
The c-statistic for the development and validation cohorts were 0.647 (0.641, by a mean multiple delivery rate of 27% and 23% respectively. Overall, only 61%
0.652) and 0.626 (0.612, 0.632) respectively. Both calibration-in-the-large of pregnancies were followed up to delivery.
(Intercept: 0.0351 (0.0149, 0.0553)) and calibration slope (0.804 (0.769, 0.840)) The evaluation of cycle-based data demonstrated a profound difference
showed under-prediction in validation cohort. However, after recalibration the between elective and non-elective single embryo transfer in terms of pregnancy
fit was much improved. rates (47% vs 22%).

Limitations, reasons for caution: Registry participation varied, limiting data O-092 Is the number of oocytes retrieved associated with time
representation in some countries and for the continent overall. There were large to conception leading to live birth? A population-based analysis of
variations regarding completeness of data. Means to validate data have not yet 221,073 cycles
been established. Y.J. Law1, N. Zhang1, R. Paul1, O. Fitzgerald1, K. Harris1,
Wider implications of the findings: ART monitoring has been successfully G. Chambers1, C. Venetis1
initiated in Africa. Efforts are now directed at expanding registry participation, 1
University of New South Wales, Centre for Big Data Research in Health, Sydney,
increasing capacity for the collection of prospective cycle-based data with use Australia
of appropriate software, and improving pregnancy follow up. Registry data
are essential for improvement strategies regarding access to and quality Study question: Does retrieving a higher number of oocytes after ovarian
of care. stimulation prolong the time to conception leading to live birth in women of
Trial registration number: Not applicable different ages?
Summary answer: Retrieval of higher numbers of oocytes does not prolong,
O-091 Pregnancy outcomes and lifetime fecundity – but rather, shortens the time to conception leading to live birth in women of all ages.
a nationwide, registry-based cohort study What is known already: Higher numbers of oocytes retrieved have been
A.M. Kolte1, D. Westergaard2, Ø. Lidegaard3, S. Brunak2, shown to be independently associated with more euploid embryos, subsequently
H.S. Nielsen4 leading to higher cumulative live birth rates. However, the euploidy rate (euploid/
1
Copenhagen University Hospital Rigshospitalet, Recurrent Pregnancy Loss total embryos available) may vary depending on the number of oocytes retrieved
Unit- Fertility Clinic 4071, Copenhagen, Denmark ; and female age. With more oocytes and eventually embryos available for transfer,
2
University of Copenhagen, Novo Nordisk Foundation Center for Protein Research, particularly in older women, there may be a lower chance of selecting a euploid
Copenhagen, Denmark ; embryo and therefore, an increased time to conception leading to live birth
3
Copenhagen University Hospital Rigshospitalet, Dept. Gynaecology 4231, (TCLB). Whether the number of oocytes retrieved is associated with TCLB has
Copenhagen, Denmark ; never been investigated.
4
Copenhagen University Hospital Hvidovre, Dept. Gynaecology and Obstetrics, Study design, size, duration: This is a large retrospective population-based
Hvidovre, Denmark cohort study using data from the Australian and New Zealand Assisted
Reproduction Database. Overall, 116,579 women undergoing 221,037 autologous
Study question: How do different pregnancy outcomes and maternal age at aspiration cycles between January 2009 to December 2015 were included in the
first pregnancy influence the predicted total number of live born children (lifetime analysis. All fresh and frozen embryo transfers resulting from the associated aspi-
fecundity)? ration cycle were included in the analysis until one live birth occurred or all embryos
Summary answer: A first ectopic pregnancy or pregnancy loss significantly were used. Cycles with no oocytes retrieved were excluded from this analysis.
reduced lifetime fecundity. Recurrent pregnancy loss (≥3 consecutive losses) Participants/materials, setting, methods: Time to conception leading to
also reduced lifetime fecundity. live birth was assessed in two ways: the number of days (from oocyte pick-up
What is known already: Several studies have investigated the impact of to the date of confirmation of pregnancy) and the number of intended embryo
pregnancy loss, ectopic pregnancies, molar pregnancies and still birth on the next transfers needed to achieve a live birth. Competing risks regression analysis and
pregnancy. To our knowledge, no large-scale studies have described the effect cumulative incidence functions were used to evaluate the association between
of prior pregnancy outcomes on lifetime fecundity. oocyte yield and the TLCB. All analyses were also stratified by female age while
Study design, size, duration: Nationwide register-based cohort study com- controlling for important confounders.
prising 458,475 women with at least one pregnancy in the Danish Medical Birth Main results and the role of chance: Utilising time in days as the time variable,
Registry or the Danish National Patient Registry from 1977 until 2017. the number of oocytes retrieved remained a significant positive predictor of TCLB
Participants/materials, setting, methods: We investigated lifetime after adjusting for female age, parity, type of embryo transferred, cycle count,
fecundity in all women who lived in Denmark between their 20th and 45th insemination method and type of infertility. Compared to the reference group of
birthday who had been pregnant at least once. Analyses were based on age 10-14 oocytes retrieved, the subdistribution hazard ratio (SHR) of achieving a
at first pregnancy, outcome of the first pregnancy (live birth, ectopic preg- conception leading to a live birth increased with the number of oocytes retrieved
nancy, pregnancy loss, still birth or molar pregnancy) and number of preg- across all age groups. Indicatively, for women aged 35-39, the SHRs (95% CIs) were
nancy losses, both total number and maximum number of consecutive 1-3 oocytes: 0.44 (0.42-0.47), 4-9 oocytes: 0.71 (0.69-0.73), 10-14 oocytes: 1.00,
pregnancy losses. Lifetime fecundity was estimated using a Generalized Linear 15-19 oocytes: 1.16 (1.12-1.20), 20-24 oocytes: 1.35 (1.29-1.41), ≥25 oocytes:
Mixed Model. 1.41 (1.33-1.50). When the number of intended embryo transfers was used as the
Main results and the role of chance: Any adverse outcome in the first time variable, SHRs (95% CIs) were 1-3 oocytes: 0.46 (0.43-0.48), 4-9 oocytes:
pregnancy significantly reduced lifetime fecundity compared to live birth. A 0.72 (0.71-0.74), 10-14 oocytes: 1.00, 15-19 oocytes: 1.14 (1.11-1.18), 20-24
first ectopic pregnancy had the largest effect (1.16 children on average, 1.11- oocytes: 1.32 (1.26-1.37), ≥25 oocytes: 1.37 (1.30-1.44). The cumulative incidence
1.22, 95% CI) compared to a woman with a first live (1.95 children on average, curves for all female age subgroups supported that with higher number of oocytes
1.86-2.03, 95% CI), both with the first pregnancy at age 30. Recurrent preg- there is a higher cumulative incidence of conception leading to live birth at all time
nancy loss (≥3 consecutive losses) also decreased lifetime fecundity signifi- points (either using days or intended embryo transfers as the time variable). A
cantly (1.57 children on average, 1.5-1.65, 95% CI) compared to women with sensitivity analysis including only term live births also yielded similar results.
no pregnancy losses (1.92 children on average, 1.83-1.97, 95% CI). Limitations, reasons for caution: This study is based on retrospective
Conversely, the total number of pregnancy losses among women <35 year observational data, thus not all confounders or clinical decisions may be
at first pregnancy had no impact. Sex of the first-born child did not impact accounted for. Furthermore, causality should not be inferred using these data.
lifetime fecundity. Wider implications of the findings: This study demonstrates that retrieval
Limitations, reasons for caution: It is a limitation that pregnancy of higher numbers of oocytes not only does not prolong the TCLB, but instead,
losses handled outside of hospitals are not registered in the Danish registries. seems to lead to higher cumulative incidence of conception leading to live birth
Other factors other than pregnancy history likely also influence lifetime in all female age groups. This information is important in informing future clinical
fecundity. decisions and furthering patient education.
Wider implications of the findings: In this unique dataset, we show that Trial registration number: Not applicable
adverse pregnancy outcomes have a significant impact on lifetime fecundity. This
highlights the importance of taking prior pregnancy outcomes into account in O-093 Variability and ovarian dysfunction of the menstrual
family planning, and the necessity of more research into the pathophysiology of cycle – a prospective analysis of the German Natural Family
ectopic pregnancies and recurrent pregnancy loss. Planning database including 43,999 menstrual cycles
Trial registration number: N/A P. Herrmann1, T. Freundl-Schuett2,3, L.M. Wallwiener3,
P. Frank-Herrmann1,3, T. Strowitzki1,3, A. Freis1,3

1
Universitäts-Frauenklinik, Department of Gynecologic Endocrinology and Fertility O-094 Motivational factors and barriers for infertile patients and
Disorders, Heidelberg, Germany ; their partners to seek consultation and treatment
2
Heinrich Heine University Duesseldorf, Department of Obstetrics and A. Domar1, R. Vassena2, M. Dixon3, M. Costa4, E. Vegni5,
Gynaecology, Duesseldorf, Germany ; J Guiglotto6, M. Markert7, J. Boivin8
3
German Society of Gynecological Endocrinology and Fertility Medicine DGGEF 1
Boston IVF, Domar Center for Mind/Body Health, Waltham, U.S.A. ;
e.V., Section Natural Fertility, Heidelberg, Germany 2
Clinica Eugin, Obstetrics and Gynaecology, Barcelona, Spain ;
3
University of Toronto, ANOVA Fertility and Reproductive Health, Toronto,
Study question: To evaluate if individual variability of cycle length and ovula-
Canada ;
tion, as well as the presence of luteal phase deficiency are more frequent than 4
International Evangelical Hospital, Reproductive Medicine, Genova, Italy ;
currently assumed 5
Università Statale di Milano, Dept of Health Sciences, Milan, Italy ;
Summary answer: Intraindividual cycle length as well as fertility parameters 6
W2O Group, Marketing Research & Strategic Insights, Philadelphia, U.S.A. ;
differ distinctly subject to age, emphasizing the importance of individual assess- 7
Ferring Pharmaceuticals, Health Economics & Outcomes Research, Copenhagen,
ment during the ongoing cycle.
Denmark ;
What is known already: As the chances to conceive spontaneously vary 8
Cardiff University, School of Psychology, Cardiff, United Kingdom
dramatically during the menstrual cycle, determination of peak fertility remains
crucial for couples trying to conceive. Recent reviews and guidelines have
Study question: What are the key drivers and barriers for infertile patients
shown that there is still a lack of knowledge concerning the spontaneous
and their partners to see an infertility specialist and subsequently undergo
menstrual cycle. Moreover, the American Society for Reproductive Medicine
treatment?
scrutinizes the clinical relevance of luteal phase deficiency in the etiology of
Summary answer: Patients waited three years seeking infertility diagnosis.
infertility. The symptothermal method combines observation of the periovu-
There were significant differences in perceived healthcare provider services
latory temperature rise and cervical mucus changes and determines ovulation,
among respondents seeking treatment versus not seeking treatment.
as well as the onset and the end of the fertile phase according to the dou-
What is known already: Individuals and couples frequently wait years prior
ble-check principle.
to seeking out medical advice and treatment for infertility, decreasing their
Study design, size, duration: 43,999 menstrual cycle charts of 1923 women
chances of treatment success. The causes of delays to diagnosis, treatment and
aiming for conception or contraception were collected prospectively between
pregnancy are not well established amongst patients or their partners.
1985-2019. Ovulation and the fertile window were determined applying the
Study design, size, duration: An online, international, 30-minute quantitative
Sensiplan method, using estrogen- as well as progesterone-parameters and an
survey collected data from 15th March–17th May 2019. The study included 1,944
evidenced symptothermal algorithm based on the extended Holt-rules. Ovulation respondents across nine countries: United States, Canada, United Kingdom,
was determined as the day before temperature rise. Luteal phase deficiency France, Germany, Italy, Spain, Australia and China. The survey was developed
(LPD) was defined as hyperthermic phase < 10 days and anovulation as the in English and translated to local language. All translations were validated by
presence of a monophasic temperature curve. national linguists.
Participants/materials, setting, methods: Patterns of menstrual cycles Participants/materials, setting, methods: The survey assessed average
(MC; n=12,612 cycles of 1051 women) were analyzed after excluding the use time to treatment, patient and partner perspectives on the treatment journey,
of oral contraception within the last three months. We included women aged and drivers for, and barriers to, infertility treatment. Participants were either
18 - 44 years who contributed data consisting of 12 cycle charts. Women after (a) infertile patients (N=1037) or (b) partners to infertile patients (N=907; but
breast feeding, birth or miscarriage were included as soon as the first ovulatory not necessarily partners of the patient sample), who were at different stages
cycle with sufficient luteal phase was detected. Statistical analysis was per- of the treatment journey. Average age was 35.8 (SD=9.66) years, 56%
formed using single factor variance analysis, chi-squared-tests and logistic (N=1095) were female, 67% (N=1119) were married and 91% (N=1773) were
regression. heterosexual.
Main results and the role of chance: Mean cycle length (CL) was Main results and the role of chance: Across countries, average time to
29.7±7.6 days (d). 62.5% of women had cycle length variation (CLV) of >7d diagnosis was 3.2 years (SD=2.4), followed by an average of 2.0 years (SD=2.1)
within 12 cycles. Respective to age, women aged 35-39 years (y) presented attempting natural conception before consultation, and an average of 1.6 years
with the lowest amount of CLV>7d (52.2%), whereas women aged 18-24y (SD=1.4) of treatment before successful couples achieved pregnancy.
showed CLV in 67.0%. Ovulation occurred most frequently on d15 (12.8%). The most frequently reported driver for considering treatment in respondents
54.8% of women had a difference of >7d between the earliest and the latest with a consultation (N=1025) was an equal desire within couples to have a child
day of ovulation, with a significant decrease subject to age (18-24y: 62.2%; (41%), followed by a willingness to do anything to become parents (38%). Among
25-29y: 56.5%; 30-34y 58.2%; 35-39y: 40.5%; 40-44y: 30.2%, p<0.001). partners (N=356), 28% cited transparency of information from health care
Average luteal phase length was 12.1±2.3d. 11.2% of MC presented with professionals about treatment expectations as an important driver. Of respon-
luteal phase deficiency (LPD). 17.6% of women had ≥3 cycles with LPD, dents not seeking consultation following diagnosis (N=352), the most frequent
whereas 46.9% did not show any LPD. Presence of LPD decreased according reason was perceived cost (38%).
to age: 14.4% (18-24y) vs. 8.3% (35-39y) (OR=0.983, 95%CI: 0.972-0.994, Post consultation, only 32% of respondents not seeking treatment (N=207)
p=0.002). Incidence of LPD rose constantly if ovulation occurred later reported that their healthcare professional offered services, such as supportive
(OR=1.084, 95%CI: 1.074-1.095, p<0.001). Cycles with an ovulation later services, compared to 61% of respondents seeking treatment (p<0.001).
than d22 showed LPD in 25.8%. Anovulation occurred in 2.6% of MC, irre- Similarly, 37% of respondents not seeking treatment agreed that their practi-
spective of age. In cycles consisting of <25d or >35d, relative amount of tioner had offered mental health services, compared to 62% among those receiv-
anovulatory cycles increased up to 8.6% and 6.9%, respectively. Ovulatory ing treatment (p<0.001). The most frequently reported barriers for those not
cycles with sufficient luteal phase occurred in 61.0% (CL<25d), 88.9% seeking treatment were cost (42%, N=87) and determination to conceive nat-
(CL=25-35d) and 82.0% (CL>35d). urally (31%, N=64). Of the 95 couples who discontinued treatment, 35% (N=33)
Limitations, reasons for caution: Data regarding possible comorbidities, discontinued due to financial impact.
such as hyperprolactinemia, were not collected. However, to reduce the impact Limitations, reasons for caution: This study was an anonymous, quantitative
of possible confounders, only women who contributed at least 12 cycles were online questionnaire, there was no response validation by healthcare profession-
included. als, with potential to be a non-representative sample. Additionally, respondents
Wider implications of the findings: Individual assessment of the current were unable to ask clarifying questions. Recruitment of individual participants
fertile window is crucial for couples trying to conceive. Our data demonstrate meant it was not possible to run sub-analyses of patient-partner pairs.
that current innovations (such as the majority of mobile apps) claiming to support Wider implications of the findings: Respondents delayed consultation and
conception using a rudimentary calendar method based on previous cycles are treatment for years, which can negatively impact chances of pregnancy.
not suitable to reliably indicate the fertile days. Motivational coherence in the wish for a child was a key driver for treatment.
Trial registration number: not applicable Costs were a main barrier. There were large differences in perceived supportive

service offerings among respondents seeking treatment versus not seeking but the wise phrasings also remains relevant considering today´s challenges in
treatment. human reproduction, with low total fertility rates (TFR) around 1.5 in Europe.
Trial registration number: Not applicable The question today is how we can assist more women and men to fulfil their
reproductive life-plans in terms of number and spacing of children.
The lecture will propose a 3-step strategy. A) Women and men below 20 years
evidently should have their main focus on contraception, but public campaigns and
INVITED SESSION school education should create an initial awareness their fertility and avoidance of
SESSION 23: RECENT ADVANCES IN ENDOMETRIOSIS sexually transmitted diseases B) In the age groups 20 to 30 there should still be a
focus on public campaigns, but another effort could be to “integrate pro-fertility
06 July 2020 Parallel 2 17:00 - 18:00
thinking” into family planning. To phrase Seifer et al. (2015) by “putting family back
into family planning”. In family planning individual assessment and counselling is
part of the consultation, and it could include simple tools for self-assessment of
reproductive risks, like the FertiSTAT colour-coded sheet C) In the age of 30
O-095 Management of pelvic pain in women actively trying to society could offer nulliparous women and men pro-fertility-focussed individualized
conceive assessment, education and counselling. Thirty is the age where many nulliparous
S. As-Sanie1 women really gets motivated for individual pro-fertility guidance.
1 The lecture will focus on point C, the approach offered in the Fertility
Assessment and Counselling Clinic (FAC) that has initially been outlined by
O-096 Organoids as 3D models for endometrium and Hvidman et al. in 2015. In short, it offers a free 30 minutes consultations with
endometriosis specialist in Reproductive Medicine for singles and couples. Women have assess-
ment of risk factors for impaired fertility graded in green, yellow and red scores,
H. Vankelecom1
1
including measurements of antral follicle counts and pelvic pathology by ultra-
Cluster of Stem Cell and Developmental Biology, Development and Regeneration, sound, and a blood sample for AMH. The consultations always include review
Leuven, Belgium of educational graphs on age and female fertility. Men are reviewed for known
risk factors for male subfertility, and a sperm analysis is made “on-site”. Couples
Abstract text
with known infertility are NOT seen at the FAC clinic, which use the facilities of
The endometrium is the first contact site of the embryo and crucial for human
the Fertility Clinic, but operates separately. Individuals do not need any referral
reproduction. Mechanisms underlying the tissue’s monthly remodeling during
and the activity is funded by the public regional health system. During the last 8
the menstrual cycle and its embryo receptivity remain far from understood, as
years the clinic has had a total of around 4.000 consultations.
well as how these processes go awry during endometrium pathology. This limited
The mean female age is around 33 years. Lower in couples and higher in single
understanding is primarily due to a lack of research models reliably recapitulating
women. The main motive for seeking the consultation was that 70% of the
endometrium biology and disease in nature and heterogeneity.
females wanted an estimate on how long they could postpone childbearing.
Organoid technology provides an innovative tool to grow mini-tissues in cul-
Around 40% are single women and 30% use contraception. Follow-up studies
ture. We established 3D organoid models from both healthy and diseased endo-
have shown that both women and men remembered the consultation several
metrium which reproduce key features of the original epithelium. The organoids
years and that the score sheets to some extend can predict natural conceptions.
show long-term expansion capacity while remaining genomically, transcriptom-
Many women saw the consultation as a catalyst for change, in general towards
ically and functionally stable. The endometrium-derived organoids phenocopy
advancing their attempts to conceive. The men felt empowered after the fertility
physiological responses to reproductive hormones and mimic the menstrual
counseling because they were equipped with concrete information that could
cycle in a dish. The organoids derived from endometriosis (as well as endometrial
inform their parenthood plans and decisions.
cancer) recapitulate characteristics of the patients' diseased tissue and faithfully
We argue that 30 years is the time where nulliparous women a) consider
capture the clinical heterogeneity of the disease. Finally, the endometrial disease
themselves susceptible to the condition of not being able to achieve their desired
organoids reproduce the original lesion when transplanted in immunodefi-
family size and b) still have 5 years ahead without major decline in age-related
cient mice.
subfertility and c) believes that specific action to become pregnant can help them.
Taken together, we created new organoid models that provide powerful and
innovative tools to decipher the mechanisms underlying endometrium biology
and pathology, and at the same time serve as screening platform to test (new)
O-098 Getting fertility education on the national curriculum
drugs, even in a patient-personalized manner.
Trial registration number: - A. Balen1
1
Study funding: - The Leeds Centre for Reproductive Medicine,
Funding source: - Professor of Reproductive Medicine and Surgery, Leeds, United Kingdom
Abstract text
Getting fertility education on the national curriculum
INVITED SESSION Professor Adam Balen
SESSION 24: PROMOTING FERTILITY AWARENESS IN YOUR Professor of Reproductive Medicine, Leeds Teaching Hospitals, UK
OWN BACKYARD In 2016, when I was chair of the British Fertility Society I founded the Fertility
06 July 2020 Parallel 3 17:00 - 18:00 Education Initiative( FEI) with aim of improving people’s knowledge of fertility
and reproductive health in the UK. We brought a number of partner organisa-
tions to the project, including the RCOG, Faculty of Sexual and Reproductive
Healthcare, Sex Education Forum, Sexpression, Teenage Pregnancy Knowledge
O-097 Educate, assess and counsel – who, how and when? Exchange, Fertility Network UK, Fertility Fest and Public Health England. Key
A.N. Andersen1 members of the national committee are the deputy chairs Professor Joyce
1
Copenhagen University Hospital, The Fertility Clinic- section 4071, Copenhagen, Harper (Institute for Women’s Health, University College London) and Professor
Denmark Jacky Boivin (School of Psychology, Cardiff University).
We first held a “Fertility Health Summit” which created a lot of publicity in the
Abstract text national media. Our vision is to ensure that people have a greater understanding
The United Nations definition of reproductive health from 1995 stated that and awareness about fertility and reproductive health, so they can make an informed
people should have “the capability to reproduce and to decide if, when and how choice about their own fertility journey, or that of others they may have an impact on.
often to do so”. This was written at a time where focus was on contraception, In brief, our three key aims are:

1. Understanding human fertility: receptors, pathways activation, second messengers as well as morphological
a) Human Reproduction characteristics of the spermatozoa. Many of these features have been disclosed
b) Male and female reproductive health, including the reproductive life in the last years, but more research efforts are needed in order to fully under-
cycle; fertility and infertility; signs, symptoms and preventable causes stand the complexity of the fertilization process. Based on this knowledge, in
of fertility issue; and planning for a healthy pregnancy the last three decades, tests have been developed to evaluate the sperm ability
2. Understanding modern families: to accomplish many of the functions required for a correct fertilization, including
a) Societal and cultural variations in family building the integrity of DNA. Some of these tests have potential clinical value both
b) Routes to parenthood; for heterosexual, LGBTQ+ and single peo- for natural and assisted reproduction, as they predict sperm fertilizing ability
ple with and without fertility issues; assisted conception techniques and ability to support development of embryos characterized by better quality
for family building; other routes to parenthood (such as adoption, and implantation with good sensitivity and specificity. However, some of these
fostering, step-families); and living a life without children texts are complex, poorly standardized, expensive and require equipment that
3. Understanding current reproductive technologies: To help educate peo- may not be present in laboratories for routine semen analysis. Hopefully, pro-
ple about what reproductive technologies can and cannot do and how gresses in technology will lead to development of easy to perform and inex-
they might impact on how human beings are made in the future. pensive tests.
We are developing age-appropriate educational material, some of which is

already available on the BFS website and a series of animations, The first Your O-100 Routine and new diagnostic approaches for male infertility:
Fertility Matters https://youtu.be/ETwDCKBaYd4 and the second Fertility Are new genetic markers near primetime?
Technologies Shaping Modern Families https://youtu.be/dOi08g3CLOc cov- D.J. Lamb1
ering modern ways of forming families and routes to parenthood for hetero- 1
Weill Cornell Medicine, Urology, New York, USA
sexual, LGBTQ+ and single people with and without fertility issues.
In 2019, the draft statutory guidance on sex education (now known as
Relationship and Sex Education, RSE) and health education was presented to
the UK government for debate. This followed a highly publicised public con- SELECTED ORAL COMMUNICATIONS
sultation that we contributed to on behalf of the Fertility Education Initiative. SESSION 26: THE DAY AFTER. FERTILITY PRESERVATION AND
We were delighted that the new guidance includes the need to educate young EMBRYO TRANSFER IN PATIENTS WITH CANCER DIAGNOSIS.
people about ‘the facts about reproductive health, including fertility and the
potential impact of lifestyle on fertility for men and women’. The bulk of the 06 July 2020 Parallel 5 17:00 - 18:00
guidance naturally deals with general health and wellbeing, the foundation of
healthy relationships and all aspects of physical, emotional, mental, sexual and
reproductive health and wellbeing. There is also reference to understanding
the various forms of sexuality and sexual relationships and ‘that others’ fam- O-101 Fertility preservation in men affected by cancer: a
ilies, either in school or in the wider world, sometimes look different from retrospective analysis of their experiences around fertility
their family, but that they should respect those differences and know that counseling and evaluation of needs regarding additional support
other children’s families are also characterised by love and care’. There is tools.
emphasis on age-appropriate information and when specific topics should be V. Ehrbar1, G. Sartorius1, C. Urech1, C. Rochlitz2, S. Tschudin1
discussed. 1
University Hospital Basel, Gyn. Socialmedicine and Psychosomatics, Basel,
The inclusion of information on fertility in the guide is a huge step forward
Switzerland ;
for fertility education, which until now has been largely overlooked, poorly taught 2
University Hospital Basel, Medical Oncology, Basel, Switzerland
and not even properly covered in most biology syllabi, let alone PHSE (personal,
social, health and economic education) or RSE lessons. Study question: What are the needs of male cancer patients regarding an
online information tool based on their experience with fertility preservation?
Summary answer: Male cancer patients consider counselling by professionals
regarding fertility preservation as very helpful and would favor complementing
INVITED SESSION
support tools.
SESSION 25: THE FUTURE OF ANDROLOGY What is known already: Impairment of fertility is a common sequela of
06 July 2020 Parallel 4 17:00 - 18:00 the nowadays available and often highly efficient options to treat cancer. Thus,
discussing fertility preservation (FP) with all young cancer patients before start
of treatment is crucial. The few existing studies in male cancer patients show
that appropriate information about FP increases counseling satisfaction and
O-099 Advanced testing, the advantage of additional knowledge prevents regrets about decisions regarding FP. According to research in female
E. Baldi1, M. Muratori2, L. Tamburrino3, S. Marchiani4 cancer patients, online decision aids are a helpful and efficient complement to
1
University of Florence- Italy, Experimental and Clinical Medicine, Florence, Italy ; fertility counseling. Support tools for men, however, are scarce and predom-
2
University of Florence, Experimental and Clincal Biomedical Sciences, Florence, inantly in English. Little is known about their needs regarding this kind of
Italy ; support.
3
University of Florence, Experimental and Clinical Medicine, Florence, Italy ; Study design, size, duration: The study consisted of a quantitative and a
4
University of Florence, Experimental and Clinical Medicine, Florence, Italy qualitative part, i.e a retrospective questionnaire-based cross-sectional survey
followed by focus groups. Recruitment was ongoing for 30 months in three
Abstract text fertility centers in Switzerland. Participants had a history of cancer within the
In order to fertilize and support embryo development, spermatozoa require last five years and were invited regardless of their decision regarding FP.
several functional attributes, most of which cannot be revealed by a routine Participants/materials, setting, methods: Participants (N=72) completed
semen analysis. Indeed, semen analysis reveals only two functional sperm prop- the online questionnaire specifically developed for the given context. The survey
erties needed for fertilization, progressive motility and normal morphology. In consisted of questions about the experiences and needs of male cancer patients
vivo, spermatozoa must penetrate cervical mucus, undergo the complex process regarding counseling and regarding information provision concerning FP.
of capacitation, develop a special type of motility known as hyperactivation Subsequently three focus groups (N=12) were conducted, where the topics
necessary to penetrate oocyte vestments, undergo acrosome reaction, fuse covered by the survey were discussed more profoundly.
with the oolemma and, finally, deliver an intact DNA to the oocyte in order to Main results and the role of chance: Mean age of the participants of the
achieve syngamy. Each of these functions requires the endowment of molecules, online survey was 32.94 (SD 8.38), most of them are living in a relationship

(70.8%) and 30.6% have at least one child. Predominant type of cancer was Out of 133 adolescence, four (3%) gave consent to their parents to use their
testicular cancer (55.6%), followed by lymphomas (16.7%) and leukemia (13.9%). sperm in case of death, while 97% (129 cases) ordered to discard their sperm
All but two participants decided to undergo FP by semen cryopreservation. The in such a case.
moment, when they were informed about FP was for 68.1% of the participants Limitations, reasons for caution: Despite the size of the group and the
before start of cancer treatment and for 29.2% during treatment, respectively; social diversity of its members, the findings might still reflect a local reality.
whereas 2.8% did not recall having been informed at any time. The professional Wider implications of the findings: These results indicate that despite the
who provided the information was predominantly an oncologist (41.9%) or value of parenthood in the Israeli society, most single men cryopreserving sperm
urologist (30.6%). Participants rated the significance of the counseling as very in face of morbidity/mortality danger do so for their own future live parenthood
high with a mean of 4.2 (maximum of 5) and experienced the professionals as are not interested in posthumous reproduction
supportive (4.37 out of 5). Analysis of the focus groups showed that support Trial registration number: Not applicable.
tools such as the existing decision aids for women would be welcome. The
majority of the participants (70.8%) stated that they would use such a tool, which O-103 Considerations regarding (non-)use of frozen oocytes or
should ideally comprise structured information provision, patient stories, and embryos for embryo transfer after fertility preservation in young
deeper insights in topics such as sexuality, fertility as well as contact information breast cancer survivors.
to peers and professionals. E. Butalid1, I. Vriens2, R. Van Golde3, J. Derhaag4, B. Van Bree3,
Limitations, reasons for caution: Participants answered the questionnaire C. De Die - Smulders5, V. Tjan- Heijnen2, L. Van Osch5
retrospectively; therefore, a recall bias needs to be considered. The final sample 1
MUMC+, Obstetrics and Gynaecology, Utrecht, The Netherlands ;
of participants consisted uniquely of patients who got a referral to fertility coun- 2
MUMC+, Oncology, Maastricht, The Netherlands ;
selling. Consequently, these results are not generalizable for all male cancer patients. 3
MUMC+, Obstetrics and Gynaecology, Maastricht, The Netherlands ;
Wider implications of the findings: These data complement the preexisting 4
MUMC+, Embryology, Maastricht, The Netherlands ; 5MUMC+, Genetics,
studies focusing on female cancer patient’s needs regarding FP and support by Maastricht, The Netherlands
a male perspective. They highlight that male patients would profit from a support
tool, as well. In general, more research is required in the specific domain of Study question: What are female breast cancer survivors’ considerations
psychological impact on male patients. regarding (non)use of frozen oocytes or embryos for embryo transfer after
Trial registration number: NCT03876366 fertility preservation prior to breast cancer treatment?
Summary answer: Most women report a strong intrinsic motivation to pursue
O-102 Single men’s attitudes towards posthumous reproduction natural conception. Time pressure to become pregnant was the most mentioned
does not necessarily match accepted social trends. consideration to perform embryo transfer.
A. Stein1, E. Altman1, M. Rotlevi1, A. Deutsch1, Y. Shufaro2 What is known already: Fertility preservation is used in young women under-
1
Beilinson Medical Center, Sperm bamk and Male infertility, Petah Tikva, Israel ; going chemotherapy for early-stage breast cancer. Several groups have reported
2
Beilinson Medical Center, Infertility and IVF Unit, Petah Tikva, Israel on the return rate of women for their cryopreserved oocytes or embryos after
fertility preservation, varying from 0% to 33%. Insight into the women’s under-
Study question: What’s the attitude of single men cryopreserving sperm in lying reproductive considerations and their preferred mode of conception in
the face of serious morbidity/mortality risks to the use of their sperm for this specific situation, are, however largely lacking.
posthumous reproduction? Study design, size, duration: This qualitative study investigated women’s
Summary answer: Most of the single men who cryopreserved their sperm deliberation of reproductive options after fertility preservation and breast cancer
in face of a mortality/morbidity risk object to posthumous reproduction using treatment. Interviews were planned until saturation of themes had been
their sperm. achieved. Sixteen women and three male partners took part in semi-structured
What is known already: Requests for posthumous reproduction are growing, interviews between August 2017 and August 2018.
raising an ethical debate and prolonged litigation, especially when written instruc- Participants/materials, setting, methods: Interviews were conducted with
tions were not left by the patients. The issue of the progenitors’ intention to women who had oocytes or embryos cryopreserved prior to breast cancer
procreate (or not) after death is the key to ethically based decision making. In treatment, at the Maastricht University Medical Center between October 2008
the Israeli society procreation is a very strong value, so in the absence of written and March 2015. To achieve heterogeneity, women who had had embryo trans-
instructions the patients’ relatives often claim for posthumous reproduction fer, who had tried to conceive without embryo transfer and who had not tried
based on the presumed will of the deceased in accordance to the mainstream to become pregnant after breast cancer diagnosis were included. Grounded
social trend. theory approach was used for analysis.
Study design, size, duration: Prior to sperm cryopreservation, single men Main results and the role of chance: Mean age at breast cancer diagnosis
were asked to sign a structured form declaring their consent or refusal for usage was 29.5 years (range 23-38). Mean time after diagnosis was 4.6 years (range
of their cryopreserved sperm in case of future mortality. Addressing this issue 2-8). There was a strong intrinsic motivation to pursue natural conception, which
and signing the form after counselling is a mandatory requirement for sperm was strengthened by psychological, practical and physical considerations.
cryopreservation. Adolescent patients signed the form with their parent / guard- Reported psychological considerations included that trying spontaneously felt
ian. The patients were from different religious and cultural origins. more relaxed and would save ‘the back-up option’ (i.e. the frozen oocytes or
Participants/materials, setting, methods: Five-hundred-thirty-four single embryos). A practical consideration was that hospital visits or medicines were
man; 401 adults and 133 adolescents, referred for sperm cryopreservation prior not needed. Time pressure to become pregnant quickly was a major issue among
to medical/surgical treatment in a tertiary hospital. All the adolescents and women who considered embryo transfer, which was considered as a faster
296/401 (74%) adults were diagnosed with malignant disorders. The remaining method. Time pressure was experienced due to interruption of adjuvant endo-
105 adults were facing fertility endangering procedures. The choices in the form crine therapy or the feeling that conception was already delayed because of
were to destroy the sperm or to allow posthumous use by a female partner or breast cancer treatment. The wish to use pre-implantation diagnosis (PGD)
other people of their choice. treatment for hereditary breast cancer disease was another consideration to
Main results and the role of chance: Five hundred thirty four single men opt for embryo transfer. Furthermore, women considered the physician’s advice
signed the form providing instructions in case of mortality. One hundred thirty as a strong influence to choose for either mode of conception. Women did not
three were 13-18 years old (age 15.9 ± 1.16 years) and the rest (401) were have any regret regarding the fertility preservation procedure and regarded the
adults (age 27.4 ± 8.06 years). presence of oocytes or embryos as a back-up plan as psychologically reassuring.
Out of the adult group 5 patients (1.2%) authorized their sibling to use their Limitations, reasons for caution: During the study participants were in
sperm, 23 (5.7%) willed their sperm to their informal female partners, and 16 different phases of family planning. This possibly is causing recall bias in the
(4.0%) permitted their parents to use their sperm posthumously. The significant reporting of original considerations. Some already completed their family for
rest 357 (89.0 %) ordered to destroy their cryopreserved sperm in case of their some years. Others were still using adjuvant endocrine therapy and may not
expiry. oversee future considerations.

Wider implications of the findings: The lack of regret and the psychological and the value clarification sheet for recognizing their personal values was of great
reassurance of a back-up plan could be supportive for women who opt for value. All patients would have liked to use the DA if this was available. To improve
fertility preservation. Since time pressure and PGD were the most reported the DA, the following was suggested: clarify navigation through DA, make tables
considerations for embryo transfer, well-thought-out counseling about the mode and graphs more readable, personalize DA more by letting choose which FP
of conception should be done by a reproductive specialist. options to read about, and emphasize that the DA does not replace oncofertility
Trial registration number: n/a consultations.
Limitations, reasons for caution: Bias could have occurred because
O-104 Development and testing of an online fertility preservation most interviewed patients had a strong wish to conceive prior to deci-
decision aid for female cancer patients sion-making. Patients who have doubts about their wish to conceive and
M. Van den Berg1, C. Beerendonk1, A. Bos2, M. Boshuizen3, refrain from a treatment may make their decisions based on different infor-
D. Determann3, R. Van Eekeren4, C. Lok5, E. Schaake6, mation and values.
E. Witteveen7, M. Wondergem8, D. Braat1, R. Hermens9 Wider implications of the findings: After revising the DA according to the
1
Radboud University Medical Centre, Obstetrics and Gynaecology, Nijmegen, The improvement suggestions, the tool is ready for implementation into routine
Netherlands ; 2University medical center Utrecht, Obstetrics and Gynaecology, clinical practice. Future studies should evaluate if the DA reduces decisional
Utrecht, The Netherlands ; conflict and decision regret regarding fertility preservation decision-making.
3
Patientplus, Medical writer, Utrecht, The Netherlands ; Trial registration number: not applicable
4
Rijnstate hospital, Surgical oncology, Arnhem, The Netherlands ;
5
Antoni van Leeuwenhoek, Gynaecology, Amsterdam, The Netherlands ;
6
Antoni van Leeuwenhoek, Radiotherapy, Amsterdam, The Netherlands ;
7 INVITED SESSION
University medical center Utrecht, Medical oncology, Utrecht, The Netherlands ;
8
Amsterdam UMC- locatie VU, Hematology, Amsterdam, The Netherlands ; SESSION 27: FRONTIERS IN DEVELOPMENTAL BIOLOGY
9
Radboud University Medical Centre, IQ Healthcare, Nijmegen, The Netherlands 06 July 2020 Parallel 6 17:00 - 18:00
Study question: What are female cancer patients’ experiences with an online
developed fertility preservation decision aid (DA) tailored to cancer type and
associated cancer treatments?
Summary answer: Female cancer patients considered the DA very helpful in O-105 The mRNA translational program and the control of
decision-making, particularly the personalized information and the value clarifi- nuclear and cytoplasmic events
cation sheet for recognizing personal values in decision-making. M. Conti M.D.1, F. Franciosi2, N. Costermans3
What is known already: Decision-making regarding future fertility is very 1
UCSF, Department of OB/GYN- CRS, San Francisco- CA, U.S.A.
difficult and complex for female cancer patients. The decision has to be made 2
Reproductive and Developmental Biology Laboratory,
in a very short time frame in a period with great emotional distress. As a con- Department of Health- Animal Science and Food Safety- University of Milan, Milan,
sequence, patients experience decisional conflict regarding this decision. To Italy
support female cancer patients and to decrease their decisional conflict, fertility 3
Center for Reproductive Sciences,
preservation (FP) DAs are being developed. However, in order to make a well-in- Department of Obstetrics Gynecology and Reproductive Sciences, San Francisco,
formed decision, patients need personalized information tailored to their cancer U.S.A.
type and treatment. DAs that provide information based on cancer type and
treatment are not available yet. Abstract text
Study design, size, duration: The DA was systematically developed by a The mRNA translational program and the control of nuclear and cytoplasmic
multidisciplinary steering group in 2019 using recommendations published by events 20/02/2020: Changed by ESHRE Chairs
Coulter and the International Patient Decision Aid Standards. Patients’ and pro- After a prolonged phase of growth and accumulation of cytoplasmic and
fessionals’ needs in decision-making were identified by in-depth interviews with nuclear components, a fully-grown mammalian oocyte undergoes a series of
female cancer survivors and oncofertility professionals. Patients’ experiences more rapid changes in preparation for fertilization and acquisition of totipotency.
with the DA were evaluated with semi-structured interviews with female cancer All these changes are thought to be essential to produce a “good quality egg”
survivors by using the think aloud method and by questioning the acceptability, that supports embryo development after fertilization. Although the events asso-
usability, comprehensibility, and readability of the DA. ciated with nuclear maturation are relatively well described and linked to gen-
Participants/materials, setting, methods: The DA steering group (N=21) eration of an egg of the correct ploidy, much less is known about the molecular
consisted of representatives of healthcare professionals working in female onco- changes in the cytoplasm of an oocyte. To investigate these processes, we have
fertility care throughout the Netherlands, patient association representatives focused on the program of maternal mRNA translation taking place during
(adolescent and young adult, breast, gynaecological and haematological cancer), meiotic maturation and at the oocyte-to-zygote transition. Since transcription
researchers with expertise in shared decision-making and medical writers. After is silent at these stages, gene expression is entirely dependent on RNA transla-
an iterative process of reviewing and revising with the steering group, the DA tion. Taking advantage of a novel RNAseq strategy to quantify mRNA translation,
was evaluated with female cancer survivors recruited from a tertiary hospital we have generated a blueprint with genome-wide resolution of the changes in
and patients’ associations. translation during both mouse oocyte growth and meiotic maturation. Analysis
Main results and the role of chance: Patients and professionals expressed of these data show that a switch in the pattern of maternal mRNA translation
a need for information tailored to cancer type with associated cancer treatments takes place at the time of oocyte reentry into the meiotic cell cycle. Translation
and infertility risks in the DA. Furthermore, the following information was con- of mRNAs coding for proteins required during oocyte growth, including those
sidered important: infertility risks associated with cancer treatment; burden/ involved in ribosome and mitochondrial biogenesis, ceases. Conversely, trans-
risks and pregnancy chances of fertility preservation treatment; consequences lation of mRNAs for cell cycle components, for chromatin remodeling, and for
of the decision for future fertility; and patients’ personal values in decision-mak- the transcription machinery that will be used in the embryo becomes acti-
ing. These preferences complemented with the national fertility preservation vated. Preliminary data show that similar regulation occurs in human oocytes
guideline formed the basis for the development of the DA. although with different timing. Translational inhibition of specific mRNAs includ-
Several face-to-face and online meetings were held with the steering group ing those coding for the histone variant H3.3 causes delayed developmental
to discuss and review the structure and content. After reaching consensus, 17 defects in the zygote and in the early embryo. In the same vein, defective trans-
female cancer survivors and patients’ representatives were interviewed while lation of cell critical cycle components yields aneuploid oocytes and compro-
using the DA. mised pregnancy. We show that oocyte manipulations known to decrease
Patients were satisfied with the content and lay-out of the DA and considered oocyte quality, including denudation or disruption of endocrine or paracrine
it very helpful in decision-making. In particular, the cancer-specific information signaling in the somatic compartment, also affect the program of maternal mRNA

translation. Remarkably, preliminary data strongly suggest that translation of O-106 Extended in vitro culture of human embryos beyond the
maternal mRNAs is also disrupted in oocytes during maternal aging. implantation stages
Taken together, these data demonstrate that execution of the maternal mRNA M.Popovic1
translation program during oocyte maturation is essential to produce a good 1
Eugin Clinic, Barcelona, department of research and development, Spain
quality egg and suggest that translational defects are a cause of the compromised
developmental competence observed during maternal aging. Supported by
NIH P50 HD055764 and R01 GM116926.

human
reproduction

O-109 ESHRE as partner in current and future EU projects
K. Lundin1
INVITED SESSION 1
Sahlgrenska University hospital, Reproductive Medicine, Göteborg, Sweden
SESSION 28: REVISITING EARLY EMBRYO DEVELOPMENT
O-110 The future of EU legislation on tissues and cells
07 July 2020 Parallel 1 08:30 - 09:30 D. Fehily1
1
European Commission, DG SANTE, Brussels, Belgium
O-107 Dynamics of the epigenetic landscape in blastocyst

development and stem cell models
P. Rugg-Gunn1 INVITED SESSION
1
Babraham Institute, Epigenetics, Cambridge, United Kingdom SESSION 30: NURSE OR MIDWIFE LED E-HEALTH CARE
INTERVENTIONS
Abstract text 07 July 2020 Parallel 3 08:30 - 09:30
Epigenetic processes are essential for regulating the spatiotemporal patterns of gene
expression in development. Upon fertilisation, terminally differentiated gametes
undergo extensive and genome-wide epigenetic reprogramming as they become
totipotent zygotes. These events establish distinctive chromatin states and are O-111 Development and evaluation of a preconception health
thought to enable the efficient activation of zygotic transcription. As the embryo online learning module for nurses in primary care
develops, differences in chromatin patterns are established between embryonic and K. Hammarberg1
extraembryonic tissues that reinforce lineage specification decisions. Excitingly, the 1
Monash University, School of Public Health and Preventive Medicine, Melbourne,
recent development of genome-wide, epigenetic profiling methods that can examine Australia
low cell numbers, including single cells, have opened up the possibility of directly
investigating the links between chromatin states and lineage specification and also Abstract text
the molecular basis for proposed mechanisms such as lineage priming. It has long been known that a woman’s health and health behaviours during
Another exciting development in the field, and one that complements our pregnancy are crucial for pregnancy health and the health of the newborn. There
expanding knowledge of human embryos, is the progress that has been made in is now growing evidence that poor maternal and paternal health before concep-
using human pluripotent stem cells (hPSCs) as in vitro models to understand the tion can result in difficulties conceiving, pregnancy complications, impaired fetal
early stages of human embryo development. Recent work has captured hPSCs in growth, poor birth outcomes, and short and longer-term health problems for
multiple states that are broadly termed naïve and primed. Both cell states can children [1].
self-renew, but are functionally and molecularly distinct. Naïve hPSCs largely reca- Nurses and midwives who provide primary health care are ideally placed to
pitulate the transcriptome and epigenome of pre-implantation embryos, and primed opportunistically discuss pregnancy intention and promote preconception health
hPSCs are similar to early post-implantation embryos. This is an important distinc- with people of reproductive age. However, we have established in two separate
tion because these two developmental stages differ enormously in gene regulation studies conducted in collaboration with nurses and midwives, that although
and in epigenetic hallmarks such as X-inactivation status and DNA methylation. nurses and midwives believe it is part of their role to promote fertility and
The research in my laboratory is focused on understanding the mechanisms of preconception health in their practice, they rarely do so because they lack the
epigenetic and gene regulatory changes as hPSCs transition between the two states, knowledge, skills, and resources to initiate conversations about optimising health
with the aim of applying that information to more precisely control cell fate deci- before conception [2, 3]. Almost all nurses and midwives in these studies agreed
sions and to better understand human development. Some of our work has also that more information and education about preconception health would increase
uncovered major differences in developmental strategies that exist between humans their confidence to discuss it with their patients.
and other species in the control of epigenetic events, such as X-chromosome ‘Your Fertility’ is an Australian government funded fertility and health pro-
regulation. I will discuss our recent progress on these topics in my seminar. motion program. In collaboration with the Australian Primary Health Care
Nurses Association (APNA), and using co-design where nurses and midwives
O-108 Cell cycle in the preimplantation embryo: mechanisms,
contributed to the development work, members of ‘Your Fertility’ developed
errors, consequences and cell fate
an evidence-based online learning module to assist nurses and midwives
J. Vermeesch1 working in primary health care settings promote preconception health in their
1
Leuven University hospital, department of Human Genetics, Leuven, Belgium practice. We are currently conducting an evaluation where participants com-
plete a brief online survey before and after completing the learning module.
This will allow us to assess if the online learning module improves knowledge
about and attitudes towards promoting preconception health in clinical prac-
INVITED SESSION tice. The second survey will also include questions about participants’ per-
SESSION 29: BUILDING BRIDGES TOWARDS ceptions of the acceptability, salience, and comprehensibility of the
HARMONISATION learning module.
07 July 2020 Parallel 2 08:30 - 09:30 In my presentation I will describe the development and evaluation of this
preconception health online learning module for nurses in primary care.
BK-OUP-ESHRE2020_ABSTRACTS-200150-Tuesday.indd 47 22/06/20 8:43 PM

1
1. Stephenson J, Heslehurst N, Hall J, Schoenaker DAJM, Hutchinson J, Ghent University Hospital, Department for Reproductive Medicine, Gent, Belgium ;
2
Cade JE, Poston L, Barrett G, Crozier SR, Barker M et al. Before the Apricity, Apricity, Paris, France
beginning: nutrition and lifestyle in the preconception period and its
importance for future health. The Lancet 2018;391: 1830-1841 Study question: To compare the ploidy prediction capability of traditional
2. Hammarberg, K., et al., Knowledge, attitudes and practices relating to statistics versus machine learning when assessing human embryo
fertility among nurses working in primary health care. Australian Journal morphokinetics.
of Advanced Nursing, 2016. 34(1): p. 6-13. Summary answer: Compared to traditional statistics, using deep learning
3. Hammarberg, K. and L. Taylor, Survey of Maternal, Child and Family algorithms increases prediction of euploidy in human embryos from 61% to 72%.
Health Nurses’ attitudes and practice relating to preconception health What is known already: Use of traditional statistics to predict PGT outcome
promotion. Australian Journal of Primary Health, 2019. 25(1): using embryo morphokinetics has been attempted by various groups, with con-
p. 43-48. flicting results as to whether it is possible to find significant morphokinetic markers
to predict ploidy status of embryos. Conflicting results may be due to differences
in approach. The main limitation is that regression analysis cannot exploit intercon-
O-112 The midwife-led development of e-based pre-conception
nections between predictors, whilst artificial intelligence approaches can. To our
care programme
knowledge, this is the first attempt to compare the efficacy of prediction of tradi-
I. Delbaere1 tional statistics with machine learning approaches in IVF. Moreover, there are limited
1
VIVES University of Applied Sciences, Health care, Kortrijk, Belgium studies using machine learning predictors to assess morphokinetic parameters.
Study design, size, duration: Traditional statistics and machine learning meth-
Abstract text ods were used to assess the same retrospective dataset, (Jan2016-Dec2019)
As in most European countries, a large proportion of Flemish women start to consisting of 445 biopsied embryos (129cycles) analysed for morphokinetic
late with the intake of folic acid preconceptionally. A study of Hoppenbrouwers development using time-lapse imaging (EmbryoScopeTM). Trophectoderm biopsy
et al. addressing this issue, was the motivation for the Flemish Minister of was performed on day5 (D5) or 6 (D6). PGT for structural rearrangements or
Welfare, Public Health and Family to launch a website on preconception care aneuploidy screening was performed by Next Generation Sequencing(NGS).
‘gezondzwangerworden.be’ (getting pregnant healthy) in 2015. The embryos were classified as either euploid [n=222 (178 D5, 44 D6)] or as
For the preparation of this website, preconceptional and prenatal guidelines displaying a chromosomal anomaly [n=223 (168 D5, 55 D6)].
were assessed by means of AGREE II. Topics for the website were selected by an Participants/materials, setting, methods: Input included morphokinetics
internal committee of 5 experts and an external committee of 16 experts. Content (17 absolute and 7 time-interval timepoints), patient (such as age) and cycle
was developed, based on guidelines and content validation was carried out by (number of injected oocytes, 2pn fertilized, embryos biopsied) characteristics.
40 experts. Multiple logistic regression (MLR) analysis was performed on log-transformed
When the website was launched, there was press communication and pro- morphokinetics parameters (SPSS) and on the entire dataset. For Machine
fessional societies of midwives, obstetricians and family medicine were informed. learning; a random forest model (RFM) was used (80% of the dataset as training
Unfortunately, there were no resources for large communication strategies. set and 20% as testing set). ROC analysis was performed to analyse the perfor-
Nevertheless, the website is visited by a growing number of couples and health mance of the two approaches.
care providers (currently 800 visitors daily). However, the intended effect on Main results and the role of chance: Using MLR (data displayed as geo-
folic acid intake remains unclear. metric mean), only the following inputs were significantly different in embryos dis-
Recently, the website was transformed formally. An included lifestyle test playing a chromosomal numerical and/or structural anomaly versus embryos with
enhances interaction with visitors and enables tailored advise. Although this an euploid profile respectively: t9+[72.61h versus 70.96h;p=0.007], tSC [82.60h
website with preconception advise includes information about the effect of age versus 83.17h;p=0.05], tM [92.26h versus 92.47h;p=0.047], the time interval for
on fertility, this is not the most optimal platform to increase fertility awareness, the duration of compaction (tcomp) [8.32h versus 7.56h;p=0.043] and patient age
in that couples visiting the website mostly already decided to start a family. In (33.8 ± 4.6y vs 32.5 ± 4.0 years;p=0.031) achieving an AUC of 61% (CI: 56%-67%).
order to inform people about the impact of age on fertility, we developed an Using machine-learning, it was found that each absolute time point needed
interactive tool ‘klaarvoorkinderen.be’ (ready for children). The aim of this tool to be normalized by its previous morphokinetic parameter to reduce redundancy.
is to encourage individuals and couples to reflect about parenthood on the one The ranking of importance in predicting ploidy was tsC, t9+, t7, age, tM, t3, t6
hand and to inform them on the other hand. The tool was developed in coop- and tPB2 (importance greater than 0.05). The following parameters approached
eration with professionals of different disciplines and was finetuned after focus significance (tB, tPNf, tSB, importance greater than 0.04). The remaining param-
group discussions with the target audience. Within these focus groups in different eters, on their own, were not significantly predictive of ploidy. The top 7 param-
age groups, men and women were involved, as well as people with a desire to eters explained only 37% of the ploidy prediction, emphasizing the importance
start a family and people who did not want children. of using multiple datapoints for prediction of ploidy. This may explain discrep-
In Europe, a growing number of e-based preconception tools with evi- ancies between previous papers in attempting to find a relationship between
dence-based information is now available to inform the public about optimal morphokinetics and ploidy.
preparation for a healthy pregnancy. These websites can be useful for Machine learning achieved an AUC of 72% (CI: 68%-77%) in the testing data
healthcare providers as well, as a tool of guidance in preconception set which was significantly higher than achieved with traditional statistics.
consultations. Limitations, reasons for caution: Dataset derived from a single clinic:
generalization of results for other clinics was not assessed. Future work will
include deep learning methodology on larger datasets with more diverse data-
points, using a data sharing hub, increasing data size, complexity and diversity to
optimize the capabilities of machine learning in ploidy prediction.
SESSION 31: PREDICTIVE ALGORITHMS IN CLINICAL Wider implications of the findings: Machine-learning raised the accuracy
EMBRYOLOGY from 61% (weak) to 72% (acceptable) compared to MLR. Given the vast amount
07 July 2020 Parallel 1 10:00 - 11:30 of variables impacting ART outcomes, traditional statistical methods should be
replaced with machine learning and artificial intelligence approaches, becoming
the new established standard for modelling prediction in ART.
Trial registration number: BC-07095
O-113 Compared to traditional statistics, using machine learning
algorithms increases prediction of euploidy in human embryos
O-114 A universal algorithm is available in last generation time-
from weak (61%) to acceptable (72%)
lapse incubators: embryo score provided by the KIDScoreD5
S. De Gheselle1, J. Chambost2, K. Declerck1, C. Jacques2, is strongly correlated with chromosomal status and clinical
I. De Croo1, C. Hickman2, K. Tilleman1 outcomes.

L. Bori1, F. Meseguer1, M.A. Valera Cerdá1, L. Alegre2, A. Tejera2, I. Miyatsuka1, K. Shimizu1, T. Trisitichoke1, A. My, Le1, N. Enatsu2,
J. Remohí3, M. Meseguer2 M. Inubushi2
1 1
IVIRMA, Research laboratory, Valencia, Spain ; NextGeM Inc., Data science, Tokyo, Japan ;
2 2
IVIRMA, IVF laboratory, Valencia, Spain ; Hanabusa Women’s Clinic, Reproductive Medicine, Kobe-Hyogo, Japan
3
IVIRMA, Co-President, Valencia, Spain
Study question: Can an ensemble artificial intelligence (AI) system which uses
Study question: Is the inbuilt software in EmbryoScope and EmbryoScopePlus both images and non-image features enhance the accuracy of embryos viability
systems useful to identify embryos with normal chromosomal status and high prediction?
potential to achieve a live birth? Summary answer: Compared to conventional embryologist morphology
Summary answer: The embryo score provided by KIDScoreD5 algorithm is assessments and image-based AI algorithms, predicting embryo viability using
highly different depending on chromosomal status and the likelihood of achieving ensemble AI models yielded 30% and 4% improvement in accuracy respectively.
a pregnancy and a live birth. What is known already: Recent studies have demonstrated the ability of AI
What is known already: Time-lapse technology has allowed embryologists to and computer vision to outperform traditional morphological assessments at
develop selection algorithms with morphological and morphokinetic parameters. predicting the likelihood of clinical pregnancy using images of human embryos.
Numerous models have been described, but no one has yet been sufficiently Existing algorithms, however, focus exclusively on using embryo images while
consolidated for universal use. The EmbryoScope and EmbryoScopePlus systems neglect other non-image inputs such as patients inspection history and embryos
include a selection method, KIDScoreD5, which classifies the embryos in categories hormonal profiles. An ensemble AI model which allows for mixed inputs can
based on cleavage time points and blastocyst appearance. The version 2 (v2) simultaneously evaluate both image and non-image data, thus enhancing the
considers PN, t2, t3, t4, t5, tB and trophectoderm quality. Later, the Inner Cell accuracy of identifying embryo viability by positive fetal heartbeat.
Mass quality was added in version 3 (v3). To our knowledge, this is the first time Study design, size, duration: A retrospective analysis using 19,342 static
that these newest versions are validated with such a large sample size. Day-5 blastocyst images with related inspection histories from 9,961 infertile
Study design, size, duration: This retrospective analysis including 22,461 patients undergoing IVF or ICSI treatment at Hanabusa Women’s Clinic between
embryos from 2017 to 2019 was performed in IVI Valencia. Embryos were cultured January 2011 and August 2019 was conducted. Hormonal profiles are available
in the time-lapse systems EmbryoScope and EmbryoScopePlus and routinely eval- for 1,358 embryos. Since the ensemble AI model requires both patients inspec-
uated by senior embryologists according to the ASEBIR criteria. The EmbryoViewer tion histories and embryos hormonal profiles as inputs, development of such
software automatically detected morphological and morphokinetic parameters. models was restricted to this subset of 1,358 embryos.
If some error occurred, it was manually modified. Then, embryos were graded Participants/materials, setting, methods: All 19,342 blastocyst images are
using the KIDScoreD5 algorithm in different scores from low to high quality (1-9.9). from single embryo transfer with known pregnancy outcomes. Images were con-
Participants/materials, setting, methods: The KIDScoreD5 algorithm was verted into grayscale and rescaled into a resolution of 800x600 pixels. Two types
tested with 7,857 embryos for v2 and 14,604 for v3. The embryo score was of algorithms were evaluated: an image-only model and an ensemble model which
compared with the morphological grade assigned by embryologists, from A to combines deep learning algorithms for image inputs and machine-learning algo-
D and excluded embryos. The correlation between the score of 3,311 embryos rithms for non-image inputs. Due to limited sample size (1,358), predictive accu-
that underwent preimplantation genetic testing with their normal or abnormal racy for the ensemble model was evaluated by the 10-fold cross-validation method.
chromosomal status was also studied. Finally, the association between the Main results and the role of chance: Out of 19,342 Day-5 blastocysts
embryo score and clinical outcomes was analyzed in 3,296 Known Implantation (7,717 with fetal heartbeat), 1,358 embryos containing all the required image
Data (KID) embryos. and non-image inputs are reserved for testing purpose. The remaining 17,984
Main results and the role of chance: The comparison between the embryo images were split into training (~90%) and validation (~10%) sets for the devel-
score provided by the KIDScoreD5 and the category assigned by embryologists opment of an image-only AI model. The ensemble AI model, on the other hand,
showed a direct association*. The means in V3 were 8.2 ± 1.2 for A; 5.7 ± 1.4 were trained and tested using only 1,358 embryos in the testing dataset.
for B; 3.6 ± 1.2 for C, 2.2 ± 1.0 for D and 1.8 ± 0.7 for excluded embryos. Accuracy is used as the main measure to evaluate the performance of AI
Regarding the chromosomal status, embryos with normal content had signifi- algorithms and defined as the percentage of both viable and non-viable embryos
cantly* higher score than abnormal ones. The score means and standard devia- correctly identified by the AI models.
tions for the newest version were 4.6 ± 1.8 for abnormal embryos and 5.3 ± All blastocyst images were graded by experienced embryologists according
1.9 for normal ones. Embryos with higher marks achieved significantly* more to the Gardner scoring system. The prediction accuracy of the embryologist
implantation rate and live birth rate in both versions. Following results belong morphology assessment is calculated as the percentage of grade 3-6 embryos
to V3 and are presented per quartiles of similar sample size. The implantation which resulted in positive pregnancy with fetal heartbeat and is set as the bench-
rates were 41.0% for score < 5.3, 54.2% for score 5.4-6.4, 59.3% for score mark rate at 49.8%.
6.5-7.4 and 67.9% for score >7.5. The live birth rates were 20.2% for score The accuracy rate achieved by the image-only AI model was 62.7%. The
< 5.3, 25.1% for score 5.4-6.4, 40.3% for score 6.5-7.4 and 48.6% for score >7.5. accuracy rate achieved by the ensemble AI model was 65.2%, representing an
In addition, V3 was capable of distinguishing between implanted and improvement in prediction accuracy of 4% when evaluating against image-only
non-implanted good quality blastocysts (A+B)*. The score means were 6.8 ± 1.5 AI model and 30% when evaluating against the visual inspection method per-
for implanted good quality embryos and 6.4 ± 1.6 for non-implanted ones. formed by embryologists.
*p<.05 Limitations, reasons for caution: Samples were collected from one clinic
Limitations, reasons for caution: This project is limited by its retrospective which can limit our results’ reproducibility. The accuracy of the ensemble AI
and single-center nature. Multicenter validation would be necessary to corrob- model is limited by the small number of embryos that we used. Non-image
orate the universal use of the KIDScoreD5 algorithm included in last generation features in the current model do not include information related to male factor
time-lapse incubators. data and the embryo culture process.
Wider implications of the findings: This study showed the capability of Wider implications of the findings: Even with limited training dataset, our
the KIDScoreD5 in distinguishing between potential embryos with similar mor- results indicate that we can improve the accuracy of predicting good-quality
phological characteristics. Therefore, embryo score could help embryologists embryos by incorporating relevant non-image features. Future models may allow
to make decisions. Recently, time-lapse technology has taken a step forward for embryo quality prediction at even earlier embryonic stages prior to syngamy.
towards automated annotations. The combination of universal selection models Trial registration number: not applicable
and automatism could improve the embryo selection.
Trial registration number: not applicable O-116 Camera-agnostic self-annotating Artificial Intelligence (AI)
system for blastocyst evaluation
O-115 Artificial Intelligence (AI) system combining both images M. VerMilyea1,2, J.M.M. Hall3,4, S. Diakiw4, A. Johnston3,4,
and non-image inputs can improve the accuracy of human embryo T. Nguyen4, M.A. Dakka4, A. Lim5, W. Quangkananurug6,
viability prediction D. Perugini4, A.P. Murphy4, M. Perugini4

1
Ovation Fertility, Laboratory, Austin, U.S.A. ; S. Wang1, L. Chen1, C. Zhou2, D. Zhang2, H. Sun1
2
Texas Fertility Center, IVF Laboratory, Austin, U.S.A. ; 1
The Affiliated Drumtower Hospital, Reproductive Medicine Center, Nanjing-
3
Australia/Australian Research Council Centre of Excellence for Nanoscale Jiangsu, China ;
BioPhotonics, The University of Adelaide, Adelaide, Australia ; 2
GrowthEngine Information Technology Co.- Ltd., artificial intelligence laboratory,
4
Presagen, Life Whisperer, Adelaide, Australia ; Beijing, China
5
Alpha Fertilty Centre, IVF Laboratory, Petaling Jaya, Malaysia ;
6
Safe Fertility Center, IVF Laboratory, Bangkok, Thailand Study question: In deep learning based human blastocyst image classification,
where are the essential features from?
Study question: Can computer vision image annotation techniques be used Summary answer: In deep learning based human blastocyst image classifica-
alongside machine learning to provide reliable blastocyst evaluation that is robust tion, the essential features are from trophectoderm(TE) , inner cell mass(ICM)
to different camera or microscope types? and zona pellucida(ZP).
Summary answer: AI that combines automated embryo annotation, trained What is known already: Deep learning is a kind of algorithms that use artificial
on optical microscope images alone, generalises to yield high accuracy and con- neural networks as a framework to learn features from data. Deep convolutional
sistency for time-lapse derived images. neural networks have rapidly become dominant in medical image automatic
What is known already: Recent studies have shown that AI and computer analysis. There have been related automatic grading studies in the field of repro-
vision can improve embryo selection and accurately predict clinical pregnancy from duction, but features used in deep leaning based medical image classification
images of human embryos at a fixed time point (e.g. Day 5). These results are basis have not been explained.
expanded to consider techniques that are robust to camera and microscope type, Study design, size, duration: This study retrospectively analyzed the image
and objective focal length, including snapshots taken from cameras used in time- of 1025 blastocysts from March 2017 to August 2018 at the center for
lapse incubators. Computer vision detection and segmentation techniques are able Reproductive Medicine of Affiliated Drum Tower Hospital of Nanjing University
to improve the distribution of AI ranking scores, showing consistent accuracy when Medical School in China.By mapping different grading to good quality blasto-
using EmbryoScope or (preliminary) GERI time-lapse incubator data, with only cysts(graded 4 or higher,ICM or TE is not C)and poor quality blastocysts(graded
2.2% sample deviation of accuracy across six different focal lengths. less than 4,or ICM or TE is C).We randomly divided the images into train-
Study design, size, duration: The original Life Whisperer model (VerMilyea ing(80%), validation(10%) and test(10%) set.
et al, 2020), was retrained using extensive image augmentation, 2,530 non-time- Participants/materials, setting, methods: 10957 images of blastocysts
lapse incubator microscope images of Day 5 blastocyst embryos, and related were obtained by excluding images of incomplete or fuzzy embryos.The embryo
clinical pregnancy outcomes, from four US laboratories, two Australian labora- classification model based on the convolutional neural network VGG-16 was on
tories and one New Zealand laboratory. The AI includes embryo-detection and the training set based on transfer learning. Receiver operating characteristic
segmentation to maximize generalizability across different imaging modalities. (ROC) Curve and Area Under Curve (AUC) are used as performance measure
The AI was applied to double-blind datasets of optical microscope, EmbryoScope of the classification model. Grad-cam algorithm is used to visualize the classifi-
(Malaysia/Thailand) and GERI images from the US. cation features of the model, and it is displayed in the form of thermal diagram.
Participants/materials, setting, methods: 3,470 separate optical micro- Main results and the role of chance: An independent test set of 1,104
scope, 221 EmbryoScope, and 38 GERI images from patients undergoing fertility images of blastocysts acquired at 116±1h after fertilization was used to evaluate
treatment at 12 IVF laboratories in five countries were used to train, validate the quality assessment model of blastocysts, which achieve an accuracy rate of
and test the AI accuracy, distribution, robustness to camera/microscope type, 91.74%. The AUC of the image classification model of blastocyst can reach
and objective focal length. Only images of Day 5 blastocysts for which pregnancy 0.970. Using Grad-cam algorithm to visualize all blastocyst stage embryo images
(heartbeat at first scan) outcome was known, were used. This study was deter- on the test set, it was found that key features relied on by the classification model
mined exempt from IRB review by Sterling IRB, USA (#6467). were on TE, ICM, and ZP.
Main results and the role of chance: This is the first study to show that AI Limitations, reasons for caution: In order to further verify the features
trained on standard Day 5 microscope images can generalize to time-lapse incubator relied on in image classification of the model, multi-center blastocyst images
images, demonstrating the robust and camera agnostic nature of this approach. are needed.
The AI accuracy for prediction of clinical pregnancy (fetal heartbeat) was Wider implications of the findings: Automatic evaluation of blastocyst
65.4% when averaged over a blind test set of Day 5 blastocyst images from 10 images helps to reduce the variability of embryologist assessments. Using visu-
IVF clinics in four countries. The sensitivity of the AI was 86.2%, while the alization techniques to explain classification features can help embryologists
specificity varied depending on the curation of the dataset. New blinded datasets understand the principles and functions of deep learning, and to promote the
of Day 5 single images of blastocyst stage embryos from the EmbryoScope and development of deep learning technology in clinical application.
GERI time-lapse systems were then assessed with the AI. The AI generalized Trial registration number: H2019011
well to the time-lapse derived images and consistent overall accuracy (59.1%)
was achieved, with a sensitivity of 77.9%. Multiple focal lengths were also con- O-118 The migration speed of nucleolar precursor bodies in
sidered and showed only a 2.2% deviation of accuracy. Together these results pronuclei is a predictor of human embryo development
suggest that this method of pre-processing and automated annotation, as well T. Inoue1,2, S. Taguchi2, M. Uemura3, K. Miyazaki2,
as AI trained on a globally diverse dataset, creates a generalizable AI that is robust Y. Yamashita2
to camera type and focal setting. 1
Hyogo College of Medicine, Department of Emergency- Disaster and Critical
Limitations, reasons for caution: The GERI data set is small and therefore Care Medicine, Nishinomiya- Hyogo, Japan ;
analysis of the distribution of scores from this set should be expanded. 2
Umeda Fertility Clinic, Department of Gynecology, Osaka, Japan ;
Additional consideration of camera effects and focal lengths from both 3
Kobe Gakuin University, Faculty of Rehabilitation, Kobe, Japan
EmbryoScope and GERI devices across a wider range of clinics should also be
considered. Study question: Does the migration speed of nucleolar precursor bodies
Wider implications of the findings: Constructing AI that is robust to image (NPBs) in male and female pronuclei (mPN and fPN) affect human embryo
variation represents an advancement in the computer vision field. Applying these development?
techniques to blastocyst-stage embryos demonstrates that AI can be robust and Summary answer: In the zygote, having potential to develop into a blastocyst/
generalizable to different clinical environments. This suggests that AI in the clinical baby, the migration speed of NPBs was faster, which is a novel predictor of
embryology setting is practical and scalable, regardless of hardware. embryo development.
Trial registration number: Not Applicable What is known already: NPBs are not considered as simple nucleolar com-
ponents transmitted from an oocyte to an embryo, and they could participate
O-117 Automated evaluation system based on artificial in genome remodeling during embryo development. In the zygote, pericentro-
intelligence and visualization technology can effectively improve meric, and centromeric heterochromatin surrounds most NPBs in a ring-like
the accuracy of blastocyst evaluation shape. NPBs are essential only shortly after fertilization, suggesting that they

1
might actively participate in centromeric chromatin establishment. Several studies University of New South Wales, Centre for Big Data Research in Health & School
have shown that NPBs are dynamic, and the characteristic NPB pattern may of Women’s and Children’s Health, Sydney, Australia ;
2
change within a short time during the syngamy process on time-lapse imaging. Flinders Fertility, Department of Reproductive Medicine, Adelaide, Australia ;
3
However, the relationship between NPB migration speed and embryo develop- University Hospital Geelong, Barwon Health, Geelong, Australia ;
4
ment is unclear. Monash University, Department of Obstetrics and Gynaecology, Clayton,
Study design, size, duration: After ICSI, migration of 262 NPBs from Australia ;
5
47 zygotes (12 patients) were analyzed, and embryonic development was Merck Serono S.p.A, Medical Affairs Fertility EMEA, Rome, Italy ;
6
prospectively observed until blastocyst (blastocyst: n=25, arrest: n=22). Merck KGaA, Global Clinical Development Fertility, Darmstadt, Germany ;
7
The relationship between NPB migration and live birth was retrospectively EMD Serono- Inc, R&D Global Biostatistics- Epidemiology & Medical Writing,
analyzed from take-home-baby (n=10) and negative-clinical-pregnancy Billerica, U.S.A. ;
8
patients (n=15) in frozen-thawed single ICSI-derived blastocyst transfer Merck KGaA, Global Medical Affairs Fertility, Darmstadt, Germany
cycles. The zygotes were cultured in a time-lapse incubator (Geri+; CO2,
6%; O2, 5% at 37°C and 80±20% humidity), and images were recorded every Study question: Which GnRH antagonist protocol maximises ongoing preg-
5 minutes. nancy rate (OPR) when considering the type of GnRH antagonist, the protocol
Participants/materials, setting, methods: The mPN and fPN were of initiation, and use of pre-treatment or not?
identified by appearance location in a zygote (fPN appearance: just below Summary answer: In pairwise and network meta-analysis of combined studies
polar bodies). The central coordinates of mPN, fPN, and 2–5 NPBs/PN with GnRH antagonists (ganirelix and/or cetrorelix), OPR was maximised with-
were measured by Kinovea (motion capture software). Their central coor- out pre-treatment and after a fixed protocol.
dinates were confirmed/revised every image and were decided. The migra- What is known already: Several GnRH antagonist protocols are currently
tion distance of NPBs between two sequential images was calculated as the used for ovarian stimulation in the context of assisted reproductive technologies
standard of central coordinates of PN. Thereafter, the migration speed of (ART). These are based on the use of the most commonly available GnRH
NPBs was calculated. antagonists (cetrorelix and ganirelix), in either a fixed or flexible modality and
Main results and the role of chance: The migration speed of NPBs was with or without pre-treatment with oral contraceptive pills (OCPs). However,
significantly faster in the blastocyst developed group than in the arrested group the effectiveness and safety of the type of GnRH antagonist (cetrorelix or gan-
(mPN: 4.61±1.26 vs. 3.37±0.87 μm/h, P<0.001, fPN: 4.06±1.15 vs. 3.34±0.97 irelix), type of protocol (fixed day 5/6 or flexible protocol) and the type of
μm/h, P=0.024). The migration speed of NPBs in mPN was correlated with pre-treatment (no pre-treatment [NP] or pre-treatment with OCPs, estrogens,
that of NPBs in fPN (r=0.65, P<0.001). The timing of blastocyst formation was progestins) have not been properly evaluated.
correlated with the migration speed of NPBs in mPN (r=−0.546, P<0.002) and Study design, size, duration: A systematic literature search was performed
had a correlation tendency with that of NPBs in fPN (r=−0.367, P=0.05). In the on 8th December 2018 in MEDLINE, EMBASE, CENTRAL, SCOPUS and Web
arrested group, 68.2% embryos arrested until day 3. In univariate logistic analysis, of Science, according to PRISMA guidelines. Qualitative and quantitative synthe-
the blastulation was related to the migration speed of NPBs in mPN and fPN, ses of all direct (pairwise) comparative data were performed using the ran-
tPNf, tPNf-tPNa, t2, t2-tPNf, t3-t2, t3-tPNf, t5-t4, and t5-tPNf. In multivariate dom-effects model. A frequentist network meta-analysis was also performed to
logistic analysis, the factor associated with blastocyst development was the leverage the power of data from randomized studies that compared various
migration speed of NPBs in mPN (OR: 5.14, 95%CI: 1.20–21.90, P=0.027). The GnRH antagonist protocols with the long agonist protocol, while checking
migration speed of NPBs in mPN in take-home-baby patients was significantly whether the underlying methodological assumptions hold.
faster than that in negative-clinical-pregnancy patients (4.64±0.67 vs. 3.75±0.79 Participants/materials, setting, methods: Randomized controlled trials
μm/h, P=0.008). In contrast, the speed in fPN was not significantly different performed on women undergoing controlled ovarian stimulation (COS) for ART
between both groups (4.30±1.24 vs. 3.80±0.92 μm/h, P=0.254). The migration treatment that compared different GnRH antagonist protocols (direct data) or
speed of NPBs in mPN was correlated with that of NPBs in fPN (r=0.603, a GnRH antagonist protocol with a long GnRH agonist protocol (indirect data)
P=0.001). were included. Studies with a day 2/3/4 fixed antagonist protocol were excluded.
Limitations, reasons for caution: We could not analyze the migration of The effect size of choice for dichotomous outcomes was the relative risk (RR)
NPBs in the z-axis direction. When NPBs were large in number or drastically and uncertainty around these estimates was expressed using 95% confidence
moved, NPB tracking could not be performed. Although our results did not intervals (CI).
completely explain the relationship between NPB migration and embryo devel- Main results and the role of chance: The searches resulted in 7,042 studies.
opment, the findings should help in elucidating the relationship. Seventy-two studies were included in the systematic review and sixty-nine in the
Wider implications of the findings: The migration speed of NPBs is a novel network meta-analysis. Studies were of low-to-moderate quality. There was no
predictor of blastocyst development. NPB migration speed may add clinical value difference in clinical pregnancy rates when comparing cetrorelix with ganirelix
for embryo selection, which may be associated with live birth, and consequently, (RR: 0.99, 95% CI: 0.74 to 1.33; I2=0%; 2 RCTs; 258 participants; low-quality
the time to live birth could be shorter. The predictor could be an attractive evidence). OPR was not reported for this comparison. Lower OPR was observed
marker for non-invasive embryo selection. after a flexible protocol compared with the fixed day 5/6 protocol (RR: 0.74,
Trial registration number: not applicable 95% CI: 0.58 to 0.96, I2=0%; 5 RCTs; 703 participants; moderate quality evi-
dence) and after OCP pre-treatment compared with NP (RR: 0.79, 95% CI: 0.64
to 0.97; I2=0%; 4 RCTs, n=1,244 participants; moderate quality evidence).
Network meta-analysis of 41 RCTs (n=8,009 women) showed that the use
SELECTED ORAL COMMUNICATIONS of the fixed-OCP protocol resulted in a significantly lower OPR compared with
SESSION 32: WHICH ARE THE OPTIMAL OVARIAN STIMU- fixed-NP protocol (RR: 0.84, 95% CI: 0.71 to 0.99; moderate-quality evidence).
LATION PROTOCOL? A flexible-NP protocol may result in lower OPR compared with the fixed-NP
07 July 2020 Parallel 2 10:00 - 11:30 protocol; however, this was not statistically significant (RR: 0.85, 95% CI: 0.72
to 1.00; moderate-quality evidence). By comparing all available protocols,
the surface under the cumulative ranking suggested that the fixed-NP protocol
is most likely to result in the highest OPR.
Limitations, reasons for caution: Limited data for some subgroups (pre-treat-
O-119 What is the optimal gonadotropin releasing hormone ment with estrogens or progestins and type of ovarian response) and for live birth;
(GnRH) antagonist protocol during ovarian stimulation for assisted heterogeneity in the criteria used for flexible protocols; limited data directly com-
reproductive technologies (ART)? A pairwise and network meta- paring cetrorelix with ganirelix and, therefore, limited evidence to consider them
analysis as equivalent in combined studies comparing GnRH antagonist protocols.
C. Venetis1, A. Storr2, S.J. Chua3, R. Wang4, B. Mol4, E. Gravotta5, Wider implications of the findings: In the general population, the fixed day
S. Longobardi6, X. Yin7, T. D’Hooghe8 5/6 antagonist protocol without any pre-treatment should be considered the

optimal antagonist protocol. More trials are required to compare the effective- ovarian-stimulation period was 9-10 days whereas in Idiopathic-group-(4) was
ness of ganirelix and cetrorelix separately, and identify the optimal protocol in reached with time slots ≤8 days (27.94±27.16; P-value:0.006). Finally, miscarriage
women with high or low response. rate was statistically significant lower (14.00±34.24; P-value:0.049) in Idiopathic-
Trial registration number: Merck KGaA, Darmstadt, Germany group (4) with shorter stimulation periods (≤8 days).
Limitations, reasons for caution: Reduced number of ICSI-cycles in some
O-120 Influence of the duration of GnRH-antagonist ovarian- of studied groups. Baseline levels of FSH and Estradiol (E2) hormones should
stimulation protocol on IVF outcomes in patients with be included.
anovulation, endometriosis, premature ovarian failure (POF) and Wider implications of the findings: To perform a prospective and random-
idiopathic infertility. ize study including baseline levels of FSH and E2 in order to standardize the
J. Gómez1, J.V. Martínez-Sanchis2, I. Iniesta Mirón2, optimal time slot of GnRH-antagonist-stimulation for patients with different
P.J. Fernández-Colom3, J.M. Rubio Rubio4 types of infertility.
1 Trial registration number: k
Hospital Universitari i Politècnic La Fe, Laboratory of Human Assisted
Reproduction, Valencia, Spain ;
2 O-121 Comparison of different progestin regimens for pituitary
Hospital Universitari i Politècnic La Fe, Laboratory of Reproduction, Valencia,
suppression during ovarian stimulation for assisted reproductive
Spain ;
3 technology, a systematic review
Hospital Universitari i Politécnic La Fe, Laboratory of Reproduction, Valencia,
Spain ; B. Ata1, E. Turkgeldi2, P. Alexandru2, S. Guler Cekic2, S. Yildiz2
4 1
Hospital Universitari i Politecnic La Fe, Laboratory of Reproduction, Valencia, Koc University, Obstetrics & Gynecology, Istanbul, Turkey ;
2
Spain Koc University Hospital, Obstetrics and Gynecology, Istanbul, Turkey
Study question: To evaluate the effect of the length of ovarian-stimulation Study question: Do progestins differ in preventing premature ovulation during
protocol on oocyte maturation, fertilization, clinical pregnancy, miscarriage and ovarian stimulation for assisted reproductive technology?
live births in four different infertile pathologies. Summary answer: All progestins seem to effectively prevent premature ovu-
Summary answer: Duration of GnRH-antagonist-ovarian-stimulation proto- lation in ART cycles. Low-quality evidence suggests that progestins can effectively
col had an effect on oocyte-maturation rate in all groups and also pregnancies, prevent premature ovulation in ART cycles.
live-births and miscarriages in endometriosis and idiopathic infertility patients. What is known already: Progesterone secreted by corpus luteum in normal
What is known already: In last years, Gonadotropin-Releasing Hormone menstrual cycle supresses luteinizing hormone (LH) secretion from the pituitary
(GnRH)-antagonist protocols have become in first choice for ovarian-stimulation during the luteal phase. Studies showed that progestins can also supress LH in
in infertile women due to its shorter stimulation duration and lower associated freeze all assisted reproduction technology (ART) cycles. It is important to inhibit
risk to suffer an ovarian hyperstimulation syndrome (OHSS). It has been demon- pituitary LH secretion before oocyte pick up and generally gonadotropin (GnRH)
strated that follicular growth, oocyte maturation and endometrial development antagonists are used to inhibit the LH surge. Progestins are as effective as GnRH
are affected by the length of gonadotropin stimulation. Very short or long expo- antagonists in supressing pituitary LH secretion.
sure to gonadotropins could have a detrimental effect on oocyte maturity stage Study design, size, duration: This is a a systematic review of studies com-
and endometrial receptivity, leading to poor embryo development and lower paring the effectiveness of progestins in preventing premature ovulation during
pregnancy rates. There is a lack of studies which evaluate these correlations ovarian stimulation for ART. We searched several electronic databases, trial
taking into account the type of female infertility. registers, and websites from the date of inception until June 1, 2019 for studies,
Study design, size, duration: Retrospective study which included a total of which compared; i) two different progestins or ii) two different doses of the
3589 ICSI-cycles from January 2015 to December 2019. All patients were stim- same progestin for pituitary suppression in ART. Only studies published in English
ulated by similar GnRH-antagonist stimulation protocol for different period of as a full text article were included.
time depending on patient’s characteristics. Patients were distributed in four Participants/materials, setting, methods: Five randomized trials and
groups according to two selection criteria: female infertility pathology cohort studies involving a total of 2404 women., which compared; i) two different
(1.Anovulation (n=492); 2.Endometriosis (n=449); 3.POF (n=722); 4.Idiopathic progestins or ii) two different doses of the same progestin were included. The
(n=1926) and three different time slots of ovarian-stimulation (≤ 8 days; 9-10 primary outcome was live birth rate (LBR) per woman. Secondary outcomes
days and ≥ 11 days) into each group. were live birth or ongoing pregnancy (LB/OP) per woman and per embryo
Participants/materials, setting, methods: Mean age of all patients were transfer (ET), ongoing pregnancy, clinical pregnancy, positive pregnancy test,
34.47 which ranged from 19 to 40 years old. GnRH-antagonist protocol numbers of oocytes and metaphase-two oocytes, duration of stimulation and
(Orgalutrant or Cetrotide) was combined to recombinant Follicle-Stimulating gonadotropin consumption.
Hormone (FSH: ranged 150-300 IU) and human Menopausal Gonadotrophin, Main results and the role of chance: Medroxyprogesterone acetate (MPA),
(hMG: 6500 IU). Oocyte retrieval was scheduled 36 hours (h) following human dydrogesterone (DYG) and micronized progesterone (MIP) were compared in
Chorionic Gonadotropin (hCG) and inseminated by ICSI at 39-40 h. Embryo combinations of two in these studies. None of the studies compared all three
transfers were performed on day 2 or 3 taking into account good-scored progestins together. Three studies compared two different progestins: MPA versus
embryos based on morphological parameters. DYG, DYG versus MIP and MPA versus MIP; two studies compared two different
Main results and the role of chance: We found a significant increase in dosages of the same progestin 4 versus 10 mg of MPA and 100 mg versus 200 mg
oocyte maturation rate in all studied groups with longer periods of ovarian of MIP. The primary outcome was not reported in most studies however there
stimulation (1.Anovulation (P-value: 0.016): ≤ 8 days: 72.22a±19.51; 9-10 days: were no differences between progestins for secondary outcomes. All progestins
75.22ab±12.90; ≥ 11 days: 78,86b±13,79; Endometriosis (P-value: 0.011): ≤ 8 seem to effectively prevent premature ovulation in ART cycles. Lower doses of
days: 71.81a±24.11; 9-10 days: 78.05b±14.04; ≥ 11 days: 80,00b±14.17; 3.POF the same progestins were also similarly effective with higher doses.
(P-value: 0.049): ≤ 8 days: 75.18a±28.46; 9-10 days: 75.18a±15.33; ≥ 11 days: Limitations, reasons for caution: The presence of a limited number of trials/
79.64b±11.82; 4.Idiophaty (P-value<0.001): ≤ 8 days: 72.91a±10.53; 9-10 days: studies, most of which are not randomized nor accounts for every woman starting
76.37b±6.50; ≥ 11 days: 78.94c±7.48), regardless type of infertility. Fertilization stimulation are drawbacks, preventing definitive conclusions on the subject.
rate was similar in all group and periods of stimulation. Clinical pregnancy rate Wider implications of the findings: Progestins in general can become a
showed significant differences in Endometriosis-group (2) reaching the highest reasonable alternative to GnRH analogues in ART cycles when a fresh embryo
value (37.12±4.52; P-value:0.013) when patients were stimulated for 9-10 days transfer is not intended.
compared to ≤8 days: 17.86±5.50 and ≥ 11 days: 22.63±38.38. Similar trend Trial registration number: not applicable
was observed in pregnancy rate in POF-group-(3) whereas Anovulation-(1) and
Idiopathic-(4)-groups showed a slight increase with shorter stimulation periods O-122 Superiority of cumulative live birth rates after GnRH
(≤8 days) but there were not significant differences. Live birth rate was significant antagonist cycles relates to ovarian response: a cycle-specific
higher (28.41±35.02; P-value:0.011) in Endometriosis-group-(2) when analysis of data from a Japanese national registry

S.C. Jwa1, M. Takamura1, A. Kuwahara2, T. Kajihara1, O. Ishihara1 2

Monash University, Department of Obstetrics and Gynaecology, Melbourne,
1
Saitama Medical University, Department of Obstetrics and Gynecology, Saitama, Australia ;
3
Japan ; ORT iOS IPD-MA study group, , ; Bastu, Ercan, Acibadem University Faculty of
2
Graduate School of Biomedical Sciences- Tokushima University, Department of Medicine ; Bergh, Christina, Gothenburg University ; D’Hooghe, Thomas, Merck
Obstetrics and Gynecology, Tokushima, Japan Healthcare KGaA ; Friis Petersen, Jesper, North Zealand Hospital ; Jayaprakasan,
Kannamannadiar, Royal Derby Hospital ; Khalaf, Yacoub, London Bridge Hospital ;
Study question: Are cumulative live birth rates (CLBRs) in different ovarian Klinkert, Ellen, University Medical Center Groningen, The Netherlands ; La Marca,
stimulation protocols for assisted reproductive technology (ART) similar in Antonio, University of Modena and Reggio Emilia ; Vuong, Lan Ngoc, University of
groups with different ovarian responses? Medicine and Pharmacy at Ho Chi Minh City ; Lapensée, Louise, Clinique ovo ;
Summary answer: Mild ovarian stimulation (clomiphene citrate [CC] and Lensen, Sarah, Melbourne ; Magnusson, Åsa, Gothenburg University ; Allegra,
letrozole) achieves similar or superior CLBRs to GnRH antagonist protocols Adolfo, ANDROS Day Surgery Clinic ; Nyboe Andersen, Anders, Rigshospitalet ;
with suboptimal egg retrieval cycles (< 10 oocytes). Oudshoorn, Simone, University Medical Center Utrecht ; Popovic-Todorovic, Biljana,
What is known already: CLBR is regarded as the most relevant outcome of Universitair Ziekenhuis Brussel ; Youssef, Mohamed, Cairo University
IVF and has a strong relationship with the number of oocytes retrieved. To
maximize CLBRs per oocyte retrieval, GnRH antagonist protocols are adopted Study question: Can a gonadotropin dose-selection model be developed and
for normal to high responders. However, there have been few reports on selec- validated that will help guide ovarian stimulation by maximizing safety without
tion of ovarian stimulation protocols for suboptimal or poor responders in terms negatively impacting pregnancy/live birth rates?
of CLBR. Summary answer: Development and validation of a gonadotropin dosing-se-
Study design, size, duration: This is a large retrospective cycle-specific anal- lection model results in a moderately to fairly accurate prediction of treatment
ysis of data from the Japanese ART registry, which includes all ART treatment effectiveness and safety.
cycles in Japan. A total of 20,4675 fresh autologous cycles performed in 2014 What is known already: Many studies addressed the effect of individualized
and 2015 and subsequent 179,313 frozen embryo transfer cycles up to December gonadotropin dosing in IVF/ICSI using ovarian reserve tests (ORTs) including
2016 were included in the analysis. anti-Müllerian hormone (AMH), antral follicle count (AFC) and basal follicle-stim-
Participants/materials, setting, methods: The cycles analyzed were as ulating hormone (bFSH). A Cochrane review reported no evidence that indi-
follows: GnRH agonist (n=66,962), GnRH antagonist (n=56,027) and mild ovar- vidualized dosing is effective at increasing ongoing pregnancy/live birth rates
ian stimulation using CC alone (n=30,098), CC + gonadotropin (Gn) (n=41,991), (OPR/LB), but may reduce the incidence of ovarian hyperstimulation syndrome
letrozole alone (n=4,363), and letrozole + Gn (n=5,234). CLBR per oocyte (OHSS). Individual patient data meta-analysis (IPD-MA) can be used to develop/
retrieval was defined as at least one liveborn baby from one fresh and subsequent validate a universal prediction model to help choose the optimal dose that will
frozen cycles. Odds ratios (ORs) for cumulative live birth were calculated using maximize safety without a negative impact on OPR/LB and reduce costs by
generalized linear model. preventing OHSS and use of ineffective high gonadotropin doses.
Main results and the role of chance: The overall CLBR was 27.6%; the Study design, size, duration: For this IPD-MA, electronical databases were
highest being with GnRH antagonist protocols (31.2%), followed by GnRH ago- systematically searched for dosing randomized controlled trials (RCTs) in women
nist protocols (30.1%), and mild ovarian stimulation protocols (22.6%–24.4%). undergoing IVF/ICSI. Women were categorized as predicted low, normal or
Compared with GnRH antagonist protocol, all other ovarian stimulation proto- high responders based on ORTs. RCTs comparing different ovarian stimulation
cols achieved significantly lower overall adjusted ORs for cumulative live birth doses (standard, adjusted or individualized) were included if the outcomes of
per oocyte retrieved. Stratification by number of oocytes retrieved revealed interest were reported: OPR/LB and/or (interventions to prevent) OHSS.
that, among high responders (>15 oocytes), GnRH agonist protocols still had Eligibility assessment was conducted by two independent reviewers.
significantly less chance of achieving cumulative live birth than GnRH antagonist Corresponding authors of eligible studies were invited to join this
protocols (adjusted OR, 0.72, 95% CI, 0.68–0.77). Among normal responders collaboration.
(10–15 oocytes), adjusted ORs of CC + Gn and GnRH agonist protocols did Participants/materials, setting, methods: After transfer of the IPD, data
not differ significantly from those of GnRH antagonist protocols (adjusted ORs were cleaned and inconsistencies were resolved with the corresponding authors.
of CC + Gn, 1.04, 95% CI, 0.95–1.14, adjusted ORs of GnRH agonist protocols, Missing data were imputed 100 times with multiple imputation by chained equa-
0.96, 95% CI, 0.91–1.01). However, adjusted ORs of mild ovarian stimulation tions for systematically and sporadically missing multilevel data. Models were
using CC alone (adjusted OR, 1.43, 95% CI, 1.32–1.54), CC + Gn (adjusted OR, developed with logistic regression. Predictor variables were selected after pool-
1.07, 95% CI, 1.02–1.12), letrozole (adjusted OR, 1.35, 95% CI, 1.14–1.61) and ing and backwards selection (using p < 0.3) and by using background literature.
letrozole + Gn (adjusted OR, 1.43, 95% CI, 1.27–1.61) showed higher ORs than Internal-external cross validation was used to assess prediction model
those of GnRH antagonist protocols in suboptimal (<10 oocytes) and poor (<5 performance.
oocytes) responders. Main results and the role of chance: A total of 14 RCTs reporting on 3,455
Limitations, reasons for caution: Because the registry provides anonymous women were included: 762 had an OPR/LB and 171 had a safety event (severe
cycle-specific data, we could not distinguish women who received multiple fresh or moderate OHSS or OHSS preventive interventions). Candidate predictor
cycles during the study period. Further, indications for selecting specific ovarian variables were age, body mass index (BMI), AFC, AMH, bFSH, parity, cycle
stimulation protocols vary across clinics, potentially causing residual confounding. length, duration of infertility, smoking, GnRH-agonist or GnRH-antagonist use
The registry lacks important confounders such as parity and indicators of ovarian and IVF/ICSI. For age and BMI, restricted cubic splines were used. Selected
reserve. predictor variables for the chance of OPR/LB (the effectiveness prediction
Wider implications of the findings: Our results suggest that for patients model) were age, BMI, AFC and IVF/ICSI. Age, AMH and bFSH were selected
expected suboptimal or poor ovarian response, mild ovarian stimulation using as predictors for safety (the safety prediction model). Starting gonadotropin
CC or letrozole may be an alternative option for selecting ovarian stimulation dose (range 75-600 IU) was forced in both models. Internal-external cross val-
in ART to achieve similar or superior CLBRs to GnRH antagonist protocols. idation resulted in 14 area under the receiver operating characteristic curve
Trial registration number: This study was supported by Health and Labour (AUC) values and confidence intervals (CIs) for the effectiveness prediction
Sciences Research Grants. model and 10 AUCs and CIs for the safety prediction model. These AUCs were
summarized in a meta-analysis forest plot. The resulting overall AUC for the
O-123 Development and validation of a gonadotropin dose- effectiveness model was 0.60 (95% CI of 0.55-0.65), which can be classified as
selection model for individualized ovarian stimulation in IVF/ICSI: moderately accurate. The resulting overall AUC for the safety model was 0.71
an individual participant data meta-analysis (IPD-MA). (95% CI of 0.64-0.79), which can be classified as fairly accurate. Heterogeneity
N. Schouten1, R. Wang2, M. Eijkemans1, B.W. Mol2, H. Torrance1, was low in both analyses: I2 was 0%.
T. Van Tilborg1, F. Broekmans1, ORT iOS IPD-MA study group.(3) Limitations, reasons for caution: Two participating groups are in the pro-
1
University Medical Center Utrecht, Department of Obstetrics and Gynaecology, cess of data transfer/cleaning, therefore these preliminary results may not reflect
Utrecht, The Netherlands ; the final results. In the following months, predictions from these models have to

be combined to develop the clinically adaptable gonadotropin dosing tool. We patient-friendly, oral administration approach may provide conveniences for egg
expect to present this dosing tool at ESHRE. donors thus, facilitating egg donation programs.
Wider implications of the findings: This tool may aid in choosing the Trial registration number: Not applicable
optimal gonadotropin stimulation dose per individual by minimizing safety risks
and maintaining effectiveness in terms of OPR/LB rates. This could potentially
reduce treatment costs, by preventing prescription of high gonadotropin doses
in low responders and preventing OHSS in high responders. SELECTED ORAL COMMUNICATIONS
Trial registration number: CRD42019115489 SESSION 33: PREDICTORS. TECHNOLOGY AND PROCESSES
IMPROVING OUTCOMES IN ANDROLOGY
07 July 2020 Parallel 3 10:00 - 11:30
O-124 Natural micronized progesterone versus a GnRH
antagonist in egg-donation cycles. An extended experience.
J.C. Castillo1, J. Guerrero1, R. Delgado2, B. Moliner1, L. Lydia1,
J. Ten1, A. Fuentes1, A. Bernabeu1, J. Llacer1, R. Bernabeu1 O-125 Live Births With a Novel ROSI Technique Using Elongating
1 Spermatids for Non Obstructive Azoospermia Patients: A
Instituto Bernabeu, Assisted Reproduction Unit, Alicante, Spain ;
2 European Cohort Study
ACCUNA, Assisted Reproduction Unit, Alicante, Spain
R. Pabuccu1, S. Sertyel2, E. Pabuccu1, K. Aydos3, A.H. Haliloglu4,
Study question: Is oral natural micronized progesterone equivalent compared D. Demirkıran2, G. Keles5, A. Tanaka6
to a GnRH antagonist with respect to the oocyte yield retrieved in egg-dona- 1
Ufuk University School of Medicine, Obstetrics and Gynecology, Ankara, Turkey ;
tion cycles? 2
Centrum Clinic Assisted Reproduction Center, IVF and Andrology, Ankara, Turkey ;
Summary answer: Oral natural micronized progesterone is comparable to a 3
Ankara University School of Medicine, Urology, Ankara, Turkey ;
GnRH antagonist in terms of total number of oocytes and mature oocytes (MII) 4
Ufuk University School of Medicine, Urology, Ankara, Turkey ;
retrieved in egg-donation cycles. 5
Centrum Clinic Assisted Reproduction Center, IVF Section, Ankara, Turkey ;
What is known already: Studies in own eggs IVF cycles shows that exogenous 6
Saint Mother Hospital., IVF Unit, Fukuoka, Japan
oral natural micronized progesterone is capable of blocking the LH surge without
compromising oocyte competence. The strategy may be also suitable for egg Study question: Can elongating spermatids be used for novel round spermatid
donors (cost effective, patient friendly, oral administration). However, scarce injection (ROSI) technique in non-obstructive azoospermia (NOA) patients when
and conflicting evidence is available with regards to its performance in the context no sperms are retrieved during a micro testicular sperm extraction (mTESE)
of ovarian stimulation in egg donors. The aim of this study is to evaluate the procedure?
efficacy of oral progesterone compared to a GnRH antagonist protocol in terms Summary answer: A novel ROSI technique using elongating (Sb2) spermatids
of oocyte yield at egg retrieval in a large sample of egg-donation cycles. and electrical oocyte activation reveals promising reproductive outcomes for
Study design, size, duration: Retrospective analysis of egg-donation cycles NOA cases with a failed mTESE
(n=1090) performed between January 2019 to December 2019 to assess the What is known already: NOA is one of the leading causes of male infertility
performance of oral natural micronized progesterone (Progeffik® 200 mg/d) and nearly one half of all mTESE procedures fail to reveal mature spermatozoa
versus cetrorelix (Cetrotide® 0.25 mg/day) in ovarian stimulation cycles trig- for ICSI. For these cases, previously different stages of spermatids have been
gered with triptoreline acetate (Decapeptyl® 0.2 mg). The clinical outcomes proposed for ICSI, however, with unsatisfactory reproductive outcomes.
per embryo transfer in matched recipients (n=598) receiving a fresh embryo Recently, healthy live births were reported following a novel ROSI technique
transfer were also assessed. without major adverse events (Tanaka et al., 2018). In brief, selection of sper-
Participants/materials, setting, methods: All participants received urinary matids after mTESE, followed by disruption of the cell membrane, separation
FSH (Fostipur® 150-225 IU/d s.c.). The oral progesterone group (n=583) con- of the nucleus and intra cytoplasmic injection of the nucleus into electrically-
comitantly received 200 mg/d micronized progesterone per os. The GnRH activated oocytes are the basic steps.
antagonist group (n=507) received cetrorelix 0 25 mg/d beginning with a leading Study design, size, duration: A cohort study was performed in a private
follicle of 14 mm. Triptorelin 0.2 mg induced the final follicular maturation. Egg IVF center from February 2018-December 2019. After urology consultation, all
collection was performed 36 hours after triggering. The main outcome was the NOA cases underwent mTESE procedure on the day of oocyte pick-up (OPU).
total number of oocytes and the number of MII retrieved at egg retrieval. Testicular tissue was examined to collect spermatozoa forms when available.
Main results and the role of chance: Overall, baseline and cycle charac- Otherwise, elongating (Sb2) spermatids were investigated to be used for the
teristics were similar between progesterone and antagonist groups with regards novel ROSI technique. After standard incubation, cleavage stage embryos
to age: 25.2 (SD 4.4) vs 25.6 (SD 4.3) years old, AFC: 18.1 (SD 6.4) vs 18.5 (SD (day 3) or blastocysts were transferred in either a fresh or frozen-thaw cycle.
7.4), BMI: 22.4 (SD 2.6) vs 22.8 (SD 2.7) kg/m2; total dose of gonadotropins: Participants/materials, setting, methods: Signed informed consent was
2088 (SD 817) vs 2086 (SD 633) IU and duration of stimulation: 9.8 (SD 1.4) obtained from all patients. After mTESE, testicular tissue was evaluated under
vs 9.9 (SD 1.3) days; respectively. The total number and MII of collected eggs an inverted microscope with contrast system to classify spermatids if no sper-
were no different: 15.8 (SD 7.5) vs 15.2 (SD 7.6), p=0.9 and 13.4 (SD 6.9) vs matozoa were seen. After selecting Sb2 forms, elongated spermatid injection
12.9 (SD 7.1), p=0.7 in the progesterone versus GnRH antagonist group, respec- was applied, using the novel technology previously described for ROSI. After
tively. In matched recipients, mean number of MII assigned were 9.5 (SD 1.3) enzymatic digestion of the testicular tissue, spermatids were isolated, cell mem-
vs 9.9 (SD 1.7) coming from the progesterone vs antagonist group, respectively branes were disrupted and finally the nucleus with the remaining cytoplasmic
(p=0.009). Fertilization rate was 78% in progesterone versus 75% in the antag- part was injected into electrically-activated oocytes.
onist group (p=0.001). Biochemical pregnancy rate was 64 versus 65% (p=0.7); Main results and the role of chance: A total of 472 couples underwent
clinical pregnancy rate 55.9 versus 56% (p=0.9) and ongoing pregnancy rate 47.7 904 cycles using elongating (Sb2) spermatids for the ROSI technique. Cases with
versus 48%, (p=0.9). the retrieval of mature spermatozoa and elongated spermatids were excluded
Limitations, reasons for caution: Retrospective design. Without a formal from the present analysis. The mean age of women and men were 33.5±5 and
sample size, findings after fertilization should be interpreted with caution. 37±5, respectively. The mean number of oocytes was 6.6±4. The fertilization
Additionally, changes in the laboratory were introduced in second half 2019, and cleavage rates were 49.9% (3024/6052) and 42.5% (2577/6052) respec-
when the vast majority of cases received progesterone for LH peak prevention. tively. The blastulation rate was 38.9 % (1005/2577). The number of cycles
Prospective, preimplantation genetic testing for aneuploidies studies on the topic reaching a transfer was 78%. The number of positive hCG tests per ET was 14.2
are warranted. % (102/714) and the number of ongoing pregnancies beyond 20 weeks was 9.6
Wider implications of the findings: These data, from a retrospective study % (69/714). A total of 8 live births have been recorded until now from this
design, suggest that exogenous progesterone is capable to block the LH surge series, and pregnancies are still undergoing. The total live birth rate from the
without compromising oocyte yield or competence. This cost/effective, series will be reported during the conference.

Limitations, reasons for caution: The selection of early spermatogenic Wider implications of the findings: Enzymatic digestion in addition to
cells demands expertise as these cells could be indistinguishable from other mechanical mincing increases sperm retrieval substantially in NOA patients fol-
round shape cells. Moreover, these cells do not properly trigger oocyte activa- lowing either a diagnostic or a therapeutic TESE.
tion, thus electrical stimulation is needed for fertilization. Although early sper- Trial registration number: Not Applicable
matogenic cells are haploid, genetic as well as epigenetic alterations might be
possible.
Wider implications of the findings: The novel ROSI technique enables a O-127 Ooplasm-Mediated Sperm Nuclear Decondensation for
proportion of males, for whom sperm donation or adoption until now was the Heritable Genome Editing of the Mammalian Male Gamete
only option, to father their own offspring. The present results support that M.S. Wang1, P. Xie1, A. Trout1, Z. Rosenwaks1, G. Palermo1
elongating spermatids as an advanced form of round spermatids can be used 1
for ICSI, resulting in healthy offspring. Reproductive Medicine, New York, U.S.A.
Study question: Can we identify a method to access the DNA of spermatozoa
O-126 The added value of enzymatic digestion to mechanical and allow gene editing by CRISPR-Cas9?
mincing in testicular sperm retrieval in non-obstructive Summary answer: An ooplasm-mediated approach is effective in allowing
azoospermia sperm decondensation for CRISPR access and successful genome editing in
V. Vloeberghs1, A. Racca1, B. Popovic-Todorovic1, I. Mateizel2, the mouse.
G. Verheyen2, H. Tournaye1 What is known already: Previous studies on heritable genome editing (HGE)
1
UZ Brussel, Obstetrics-Gynaecology, Jette- Brussels, Belgium ; have been carried out at the zygote phase; however, they faced the difficulties
2
UZ Brussel, Centre for Reproductive Medicine, Jette- Brussels, Belgium of mosaicism and embryo research ethics. These issues may be resolved by
first editing the DNA of the gametes. The challenge with spermatozoa, how-
Study question: To evaluate the added benefit of enzymatic digestionto ever, resides largely in their distinct structure—they contain tightly packed
mechanical mincing in testicular sperm retrieval for patients with non-obstructive chromatin, carry no DNA repair mechanisms, and are highly susceptible to
azoospermia (NOA). damage. Our previous attempts to directly edit spermatozoa DNA required
Summary answer: Enzymatic digestion increases the sperm retrieval from considerable resources but yielded limited results. We hypothesize that an
testicular biopsies of NOA patients by 32.3%. alternative cellular environment may be necessary to allow proper male genome
What is known already: Although there is increasing evidence that the surgical editing.
technique to recover testicular sperm in NOA makes little difference as long as Study design, size, duration: Over a period of 3 months, 88 oocytes
multiple biopsies are taken, the debate on which is the best technique continues. were divided into two groups. One group was used for ooplasm-mediated
Few studies focus on the role of the lab and even fewer evaluated the benefit sperm decondensation (OMSD) to produce haploid embryos containing only
of enzymatic tissue digestion in order to improve sperm retrieval rates. Research DNA from the male. The control group consisted of embryos produced
in this domain has been flawed by heterogeneity of either the NOA diagnosis through standard ICSI, following the established HGE approach. Both groups
or the selected patient study population. were treated with a CRISPR solution designed to knock out Tyr to create an
Study design, size, duration: A retrospective single-centre cohort study albino phenotype. Gene editing success was then compared between
from 2004 till 2017 included all patients who underwent their first diagnostic or the groups.
therapeutic testicular sperm extraction (TESE) by open biopsy. They all had Participants/materials, setting, methods: Oocytes and sperm were col-
normal standard genetic workup and subsequent histological confirmation of lected from B6D2F1 mice. OMSD oocytes were enucleated and allowed 2 hours
NOA. Patients who had a history of previous TESE or histological diagnosis of to rest. All oocytes then underwent ICSI with the addition of a CRISPR-Cas9
hypospermatogenesis were excluded. Primary outcome was sperm retrieval for solution containing pre-complexed Cas9 protein and Tyr knockout gRNA.
either cryopreservation or intracytoplasmic sperm injection (ICSI). Embryos were incubated for 24 hours, and then cryopreserved. Later, DNA
Participants/materials, setting, methods: Up to 6 biopsies were obtained was extracted and amplified for T7E1 analysis of the target site.
from each testis. In cases with no or insufficient number/quality spermatozoa Main results and the role of chance: Of the 88 oocytes, 31 were enu-
observed during 30 min after mechanical mincing, enzymatic treatment of resid- cleated for the OMSD condition, while 57 constituted the control. After under-
ual tissue pieces was performed with collagenase type IV. The results were going ICSI with CRISPR solution, 74% (23/31) of the OMSD oocytes survived
divided according to the biopsy type (diagnostic or therapeutic). In order to compared to 75% (43/57) of the control. Of those, 43% (10/23) of the
evaluate which factors predicted sperm retrieval, multivariable regression analysis experimental group developed to the 2-cell stage, while 70% (30/43) of the
was performed adjusting for the following confounding factors: age, FSH level, control progressed (p<0.05). After extraction, a 423-bp region around the
testicular volume and histology. CRISPR target site was amplified. A total of 35 embryos produced a sufficient
Main results and the role of chance: In total, 425 patients were included. concentration of DNA, while 3 OMSD and 2 control embryos failed to amplify.
Overall, in 69/425 (16.2%) men, spermatozoa for cryopreservation or ICSI Gene modification at the target site was confirmed in 29% (2/7) of OMSD
were found after mechanical mincing. In 115/356 (32.3%) patients who had embryos and 36% (10/28) of the control group. We achieved successful
no sperm following mincing, spermatozoa were found after enzymatic digestion. genome editing of spermatozoa DNA through the use of ooplasm-mediated
In order to reduce bias in sperm retrieval rate when TESE is scheduled on the sperm decondensation at a comparable rate to zygotic heritable genome edit-
day of ICSI (therapeutic) or prior to planning ICSI (diagnostic), these two ing. It is important to note that these results are likely to be underestimated,
patient populations were analyzed:316 (74.4%) in the diagnostic and 109 since any embryos with completely uniform modifications may not be detected
(25.6%) in therapeutic testicular biopsy group. There was no difference in mean by T7E1.
age (34.9 vs. 35.9 years), mean volume of testicles (10.9 vs. 10.2 ml), histo- Limitations, reasons for caution: Currently, sample size is limited. Refining
logical diagnosis (Sertoli Cell only 56.1 vs. 56%; maturation arrest 21.8 vs. of experimental techniques is needed to improve survival and development
21.2%; sclerosis and/or atrophy 19.9 vs. 18.4%) between the diagnostic vs. rates, and DNA sequencing of blastomeres will provide a more accurate eval-
therapeutic group. FSH was significantly higher in the diagnostic group com- uation of editing efficiency. Additionally, off-target effects, mosaicism, epigenetic
pared to the therapeutic group (22.3 vs. 18.5 IU/l, p=0.008). Sperm retrieval modifications, and birth rate have yet to be evaluated.
was significantly higher in the therapeutic vs. diagnostic group both after mincing Wider implications of the findings: Once editing efficiency is optimized,
(24.8% vs. 13.9%, p=0.007) and enzymatic digestion (43.1% vs. 21.5%, individual pseudo-blastomeres of OMSD embryos can be used in place of sper-
p< 0.001%). Multivariable logistic regression analysis showed that enzymatic matozoa to fertilize oocytes. Subsequently, the pre- and post-implantation devel-
digestion remained a significant predictor of sperm retrieval, when adjusting opment of offspring that contain the intended genetic modifications can be
for potential confounders. studied. Our model, if reproduced in human, may prevent the inheritance of
Limitations, reasons for caution: The main limitation of the study is its paternal mutations.
inherent retrospective design. Trial registration number: not applicable

O-128 A Novel Microfluidics Method for Reliable and Efficient O-129 Effect of microfluidic sperm separation versus standard
Sperm Sex Selection sperm washing processes on fertilization rates, blastocyst
R. Elias1, S. Cheung1, A. Parrella1, P. Xie1, D. Keating1, development and euploidy rates among all infertility patients
Z. Rosenwaks1, G. Palermo1 G. Adaniya, E.L.D.- H.C.L.D.- Ph1, C. Leisinger2, M. Freeman3,
1
Weill Cornell Medicine, The Ronald O. Perelman and Claudia Cohen Center for E. Behnke4, M. Aguirre5, T. VerMilyea6, M. Schiewe7
1
Reproductive Medicine, New York, U.S.A. Ovation Fertility, IVF Laboratory, Carmel- IN, U.S.A. ;
2
Ovation Fertility, IVF Laboratory, Baton Rouge- LA, U.S.A. ;
Study question: Can selection of Y-specific spermatozoa be achieved through 3
Ovation Fertility, IVF Laboratory, Nashville- TN, U.S.A. ;
ATP content modulation in a microfluidics system? 4
Ovation Fertility, IVF Laboratory, Cincinnati- OH, U.S.A. ;
Summary answer: Ligand activation on the sperm flagellum decreased motility 5
Ovation Fertility, IVF Laboratory, Las Vegas- NV, U.S.A. ;
of the female spermatozoa and ensured expedited Y-bearing spermatozoa selec- 6
Ovation Fertility, IVF Laboratory, Austin- TX, U.S.A. ;
tion in a microfluidics chamber. 7
Ovation Fertility, IVF Laboratory, Newport Beach- CA, U.S.A.
What is known already: Several techniques have been proposed to success-
fully select for X- or Y-bearing spermatozoa, such as centrifugation methods, Study question: Our goal was to determine if the ZyMōt microfluidic sperm
layering techniques, electrophoresis, and flow cytometry. At our clinic, we have separation device effectively improved ICSI sperm selection and subsequent
been able to carry out sex selection with an 80% success rate with the use of a embryo development applied to a general IVF patient population.
proprietary density gradient technique that requires over 3 hours of processing. Summary answer: Utilization of a ZyMōt device to process all IVF semen
However, it has recently been proposed that modulation of the Toll-like receptor samples (untested for Sperm DNA Fragmentation, SDF) revealed no overall
7/8 (TLR7/8) on the X-bearing sperm flagellum impairs motility and may there- benefit to embryo outcomes.
fore be effective in sex selection techniques. What is known already: The natural sperm filtering actions of cervical/
Study design, size, duration: In the last 5 years, we processed ejaculates uterine crypts has been mimicked by a new microfluidic sperm separation device
from 98 consenting couples undergoing treatment at our center with a custom (ZyMōt). Its microporous filter and channels help separate the most motile sperm
multilayer density gradient. The proportion of X- and Y-bearing spermatozoa with normal morphology without centrifugation. In turn, not only does the
before and after selection was assessed by fluorescent in situ hybridization (FISH). ZyMōt device claim to reduce procedure-induced reactive oxygen species (ROS)
Cycle pregnancy outcomes and offspring sex were recorded. To expedite our associated with causing SDF, it selectively isolates healthier, progressively motile
selection process while maintaining sex enrichment, we tested a new method sperm with lower SDF. Pilot studies suggest that the higher chromatin integrity
utilizing TLR7/8 ligand activation together with microfluidics. (i.e., lower SDF) attained for IVF use improves fertilization, embryo morphology
Participants/materials, setting, methods: A total of 98 couples were and blastocyst euploidy rates. We aimed to test the ZyMōt’s proposed devel-
treated at our center (IRB 1306014043) in 120 cycles. A proprietary multilayer opmental benefits.
density gradient method was used to select for sex-specific spermatozoa. We Study design, size, duration: A prospective, randomized, multicenter, sibling
confirmed successful selection by FISH analysis on at least 1,000 cells/speci- oocyte study was conducted with 86 consenting patients possessing ≥10 oocytes.
men. Pregnancy outcomes and offspring sex were assessed and compared. In Non-DFI tested ejaculates underwent a split processing treatment: 1) Control
a new selection technique, we incubated spermatozoa for 45 minutes in HTF washing procedures (density gradient separation or swim-up) or 2) Treatment
medium containing 0.3µM of the TLR7/8 ligand, prior to microfluidics - ZyMōt swim out. Each sample was then used to inseminate an equal number
processing. of sibling oocytes. Resulting blastocyst were biopsied and NGS tested. Euploid
Main results and the role of chance: Of the couples (maternal age, (non-mosaic) embryo selection for ET was randomized. Chi-squared analysis
36.8±4yrs; paternal age, 39.6±5yrs) included, 53.1% wanted a female child, was performed to assess differences(p<0.05).
while 46.9% desired a male child. The initial sperm concentration was Participants/materials, setting, methods: Upon informed consent, partner
65.4±26x106/ml, with 47.7±5% motility, normal morphology, and average sperm was processed in parallel by 1) either density gradient centrifugation
sperm aneuploidy of 3.3±4%. After multilayer density gradient selection, sperm (DGC) at OF-IN(n=26), OF-OH(n=19) and OF-LA(n=18) or a swim-up wash
concentration decreased to 24.3±14x106/ml, while motility rose to 94.5±3% procedure (OF-TN,n=23); or 2) a 0.85µ ZyMōt device. After removal of cumu-
(P<0.0001). lus/coronal cells by hyaluronidase, half of the mature oocytes were randomly
Of the 52 couples who wanted a female child, FISH assessment confirmed a allocated to ICSI with Control sperm and the other half with ZyMōt sperm.
spermatozoa enrichment of 80%, and over 80% of couples obtained a female Fertilization, embryo culture and biopsy were performed using the clinic’s stan-
embryo as assessed by PGT-A. The clinical pregnancy rate for these couples was dard protocols.
25.7% (18/70), with a delivery rate of 61.1% (11/18) of the desired sex; the Main results and the role of chance:
remaining pregnancies are ongoing.
For 46 couples who desired male offspring, FISH analysis confirmed a suc- Table. ICSI cycle embryo development outcomes
cessful enrichment for Y-bearing spermatozoa, and over 80% of couples following standard sperm washing procedures
obtained a male embryo. The clinical pregnancy rate for these couples was (Control) or the use of a microfluidic sperm
28.0% (14/50), and the delivery rate was 57.1% (8/14) of the desired sex; the
separation device (ZyMōt)
remaining pregnancies are ongoing. To test our new method, in a separate
non-clinical investigation, four additional sperm specimens were processed by
Treatment: ZyMōt Control
the proposed ligand activation technique, yielding >80% enrichment, confirmed
by FISH, that was comparable to the more time-consuming multilayer density
Number of Oocytes 787 777
gradient technique.
Limitations, reasons for caution: Although the TLR7/8 ligand mechanism Fertilization rate (% 2PN) 604 (76.7%) 592 (76.2%)
is not absolutely clear, its effect on the flagellum is reversible and therefore does Blastocyst rate (%>2BB) 296 (49.0%) 282 (47.6%)
not affect acrosomal function or viability. Nevertheless, the capacity of sperma-
Euploidy rate 165/283 (58.3%) 151/265 (57.0%)
tozoa selected by this technique to support embryonic development needs
further investigation. Mosaicism rate 25/283 (8.8%) 31/265 (11.7%)
Wider implications of the findings: We were able to enhance the pro-
portion of female and male embryos with our current multilayer density gra-
dient technique requiring >3 hours. However, the proposed method There were no statistical differences observed between the ZyMōt or Control
incorporating ligand activation and microfluidics would lower this processing sperm processing methods. Although some normal variation between labs was
time to <1 hour. We are refining this method in order to achieve >80% observed, there were also no differences between the control DGC and swim-up
selection. methods. Clinical pregnancy outcomes are still being evaluated, but preliminary
Trial registration number: not applicable results reveal no advantage to the ZyMōt treatment. To date, intrauterine

pregnancies are trending upward(p>0.1) for the Control groups (21 of 30, 70%) O-131 Exome-wide screen for somatic mutations in
in contrast to 18 of 37 (48.6%) in the ZyMōt group. endometriosis lesions
Limitations, reasons for caution: Male factor patients with known elevated K. Ward1, R. Chettier2, H.M. Albertsen2
SDF were not selected for this multi-center trial. Therefore, we can not speak 1
Juneau Biosciences- LLC, Administration, Salt Lake City, U.S.A. ;
to the potential improvements ZyMōt may elicit. However, it is possible that 2
Juneau Biosciences, Research and Development, Salt Lake City, U.S.A.
selection of normal, progressive sperm for ICSI, plus the repair capacity of
oocytes, is sufficient to promote normal development. Study question: Which genes undergo somatic mutation during the develop-
Wider implications of the findings: We do believe the ZyMōt device may ment and progression of endometriosis lesions; do these genes differ from those
have a useful application in infertile men with elevated SDF, yet it is equally that predispose women to develop endometriosis?
possible that an ICSI sperm selection alone allows for the immobilization and Summary answer: Endometriosis lesions develop somatic mutations in several
injection of highly progressive, morphologically normal sperm as seen in this genes. The mutations identified in lesions differ from germline variants that pre-
study of typical infertility patients. dispose women to develop endometriosis.
Trial registration number: OF191218B What is known already: Endometriosis is a clinically heterogenous condition
affecting 10% of women. It is well established that germ-line mutations may
O-130 Testicular cell communication mediated by extracellular predispose some women to develop endometriosis, however penetrance,
vesicles expressivity, and pleiotropy of the “initiating” genes implicated to date are not
E. Fok1, K.H.K. Choy1, S.Y. Chan1, W. Lam1 understood. Clinical heterogeneity is even observed within a single patient as
1
The Chinese University of Hong Kong, School of Biomedical Sciences- Faculty of an affected woman may have some lesions that are progressive, invasive, and
Medicine, Hong Kong, Hong Kong possibly metastatic or malignant while other lesions are not. Recent studies have
shown that somatic mutations accumulate during the clonal evolution of indi-
Study question: Is extracellular vesicle (EV) presence in the mouse testis? If vidual endometriosis lesions.
yes, do these testicular EVs mediate cell communication in mouse testis? Study design, size, duration: This study is a retrospective cohort association
Summary answer: EVs were present in mouse testis. Testicular EVs were study. 2,932 DNA samples from unrelated women with surgically-con-
uptake by both germ cells and somatic cells and they carry novel small RNA cargos. firmed endometriosis (GERMLINE) and 274 tissue blocks containing endome-
What is known already: Spermatogenesis requires a precisely orchestrated triosis lesions (LESION) underwent whole exome sequencing. Fisher’s Exact
cellular communication networks. EV is known to be ubiquitously released by Test was used to determine excess burden (alpha <0.001).
eukaryotic cells. EV carries various cargo, including genomic DNA, mRNA, Participants/materials, setting, methods: DNA was extracted using standard
miRNA and proteins, that can be uptake by target cells and regulate their cellular methods. Exome sequencing was performed using an Ion Proton Instrument,
processes, indicating that EV is an important and effective way of cell-cell com- AmpliSeq Exome Capture, and the Torrent Variant Caller. Missense and truncat-
munication. EVs secreted by the prostate and the epididymis have been charac- ing variants having a minor allele frequency <0.005 in the gnomAD database were
terized and shown to play important roles in sperm maturation and functions. considered for analysis. We calculated somatic burden for each gene by first count-
However, the presence of EVs in the testis and their involvement in spermato- ing protein-altering variants identified in only one patient per group. We also calcu-
genesis remain elusive. lated cumulative genetic burden for each gene by adding all coding variants identified.
Study design, size, duration: We collected testis samples from mouse and Main results and the role of chance: A mean of 63 “singleton” variants per
isolated testicular EVs using a newly established tissue dissociation method. patient were detected in the GERMLINE samples and 65 singleton variants per
Participants/materials, setting, methods: The physical properties of tes- patient in the LESION group. 11 genes showed excess somatic mutations in the
ticular EVs were assessed by dynamic light scattering and electron microscopy. LESION samples when compared to the GERMLINE results (MIEF1, MSX1,
Testicular EVs were validated by Western blot of established EV markers and FAM149A, VEZT, FAM160B1, PCDHA6, RIBC1, IRAK4, HYAL2, AQP4, and
Golgi marker. The small RNA cargo was profiled by RNA-sequencing and verified DSE). Singleton variants in these genes were observed at a 6.3 to 22 fold excess
by real-time PCR. The uptake of testicular EVs was examined by the fluores- in the LESION samples when compared with the expectations from the
cence-labelling of EVs. GERMLINE samples. None of these genes are among the top 1000 FLAGs
Main results and the role of chance: Testicular EVs were isolated from the ranking (frequently mutated genes). VEZT has been previously implicated as an
mouse testis using the newly established tissue dissociation method. Electronic endometriosis associated gene through GWAS studies.
microscopy results showed the hallmark cup-shape morphology of EVs with Limitations, reasons for caution: The sample size is modest and replication
sizes ranged from 50 – 200 nm. The presence of hallmark EV markers CD63, of these results in additional cohorts is warranted. It is very unlikely formalin
CD81 and CD9 and the negligible amount of Golgi marker suggest clean isola- fixation or paraffin embedding caused the observed mutations, but untreated
tion of testicular EVs with minimal membranous organelle contamination. Small LESION samples are currently being tested in our laboratory.
RNA sequencing (RNA-seq) revealed RNA species commonly observed in EVs, Wider implications of the findings: Discovery of gene mutations underlying
including rRNA, tRFs, snoRNA, miRNAs and piRNAs. Importantly, we have endometriosis progression may lead to new pathophysiology insights, improved
identified three novel miRNAs that were not reported in the testis transcrip- diagnostics, and novel treatment approaches.
tome. To study the type of testicular cells that uptake testicular EVs, we injected Trial registration number: Not applicable
PKH-labelled testicular EVs into the interstitial space or seminiferous tubules.
O-132 The landscape of endometrial transcriptome in ovulatory
The results showed that EVs were uptake by Sertoli cells and all stages of
obese PCOS women
germ cells.
Limitations, reasons for caution: The physiological function of testicular V. Salamun1,2, E. Vrtačnik Bokal1, B. Peterlin3, A. Maver3,
EVs awaits further investigation. N. Kočevar3, O. Maloku4, T. Papler Burnik1
Wider implications of the findings: The testicular EVs may provide novel 1
University Medical Centre Ljubljana, Department of Human Reproduction-
insight into our understanding of sperm production and inheritance. Division of Obstetrics and Gynecology- University Medical Centre Ljubljana-
Trial registration number: N/A Ljubljana- Slovenia, Ljubljana, Slovenia ;
2
Medical faculty of Ljubljana, University ljubljana, ljubljana, Slovenia ;
3
University Medical Centre Ljubljana, Department of Genetics- University Medical
Centre Ljubljana- Ljubljana- Slovenia, Ljubljana, Slovenia ;
SELECTED ORAL COMMUNICATIONS 4
University Medical Centre Ljubljana, Department of Genetics University Medical
SESSION 34: ENDOMETRIOSIS - PATHOGENESIS AND Centre Ljubljana- Ljubljana- Slovenia, Ljubljana, Slovenia
DIAGNOSIS
Study question: To determine the coexisting influence of obesity and PCOS
07 July 2020 Parallel 4 10:00 - 11:30
on the endometrial transcriptome in ovulatory obese PCOS patients during the
window of implantation in spontaneous cycles.

Summary answer: Enrichment analyses revealed significant disruptions of Summary answer: 3D-TVUS has a high detection rate for the studied pathol-
inflammation, insulin signaling, fatty acid metabolism and lipotoxicity pathways ogies, is less invasive and more cost-effective diagnostic tool.
in ovulatory obese PCOS endometrium compared to controls. What is known already: Both HYS and 3D-TVUS are currently used for
What is known already: Obese and PCOS women have a reduced rate of diagnosis of uterine malformations and submucosal fibroids. Their accuracy and
spontaneous conception even when their cycles are ovulatory indicating other the indications are operator-dependent and largely variable, between 25% and
factors like the uterine environment. The mechanism of this intricate interplay 99% depending on the circumstances and the instruments used.
in which obesity modifies already different PCOS endometrium is not fully clar- Study design, size, duration: Prospective observational study. From April
ified. There are only a few analyses of the endometrial transcriptome in which 2010 to December 2018, 672 patients were enrolled as they met the inclusion
the impact of obesity and/or PCOS on the endometrium was elucidated. The criteria for the study.
most relevant was the study of Belver et al. in which obese women showed a Participants/materials, setting, methods: Patients found to have submu-
different endometrial gene expression compared to controls with pronounced cosal uterine fibroids or uterine congenital anomalies on a routine pelvic trans-
changes of transcriptome in a combination of obesity and PCOS. vaginal ultrasound were prospectively studied. All patients underwent 3D-TVUS,
Study design, size, duration: Prospective transcriptomic analysis of endo- HYS and pelvic MRI, each one performed by an expert operator. Women with
metrium using the RNA-seq method was done in university-affiliated infertility endometrial polyps or suspected endometrial or uterine cancer were
hospital between February 2017 and February 2018. Clinical information and excluded. Median age was 32 years (range 23-66 years).Specificity, sensitivity,
endometrial samples were collected from 14 infertile patients that were divided accuracy and k-index of 3D-TVUS and HYS were recorded and compared to MRI.
in 2 groups: ovulatory obese PCOS ( ovul-O-PCOS) and control group. The Main results and the role of chance: Submucosal fibroids were confirmed
inclusion criteria for the ovul-O-PCOS group were obesity, PCOS, and infertility by MRI in 441 out of 672 (65.6%) women; uterine malformations were reported
and for the control group were normal body mass and tubal or unexplained in 231 (34.4%) women. Both 3D-TVUS and HYS showed great sensitivity in
infertility. uterine malformations and submucosal fibroids detection, however 3D-TVUS
Participants/materials, setting, methods: 14 patients were invoved in the showed the highest specificity in detection of uterine malformations, while HYS
study according to the study criteria. Ultrasound detection of follicular ovulation/ showed the highest specificity in detecting submucosal fibroids. Moreover,
anovulation with determination of progesterone levels on the day of endometrial 3D-TVUS had a slightly higher accuracy in the diagnosis of uterine malformations
biopsy. Endometrial biopsy collected between 21 and 23 of menstrual cycle. compared with HYS (96.8% vs 93.7% respectively), while k index was excellent
RNA isolation (RNeasy Kit, Qiagen, Germany), sequencing, library prepara- for both diagnostic tools. The main limitation of HYS was in differential diagnosis
tion and subsequent RNAseq data analyses were done. of fundal submucosal fibroids and uterine septum/arcuate uterus.
Main results and the role of chance: According to functional and upstream Limitations, reasons for caution: All procedure were performed and
analyses the most important biological processes in ovulatory obese PCOS reviewed by expert operators with state of the art diagnostic tools, so the results
compared to control group were related to inflammation (TNFR1 Signalling reported in the study might not reflect detection rates by average operators in
Pathway as the most important), insulin signaling and energy consumption everyday practice.
(Insulin Receptor signaling and PI3/AKT pathway,) fatty acid metabolism Wider implications of the findings: 3D-TVUS is a valid tool in primary
(Stearate Biosynthesis I Pathway and Palmitate Biosynthesis I Pathway) and detection of uterine malformations and submucosal fibroids. It could replace
lipidotoxity. diagnostic HYS and/or laparoscopy in diagnosis of uterine malformations, both
Upstream analysis perfectly captured the pathophysiological mechanisms where intervention is not required or to guide the surgeon in preparation for
found through functional analysis of ovul-O-PCOS endometrium. Most of the the surgery.
upstream regulators were involved in some of the already mentioned mecha- Trial registration number: n/a
nisms: inflammation (CASP1, EGR1, TGFb, ZAP 70), glucose metabolism and
energy homeostasis regulation (insulin, insulin receptor, GSK3, MYC, IGF1),
adipokines & lipid regulators (ADIPOQ, clofibrat as a lipid-lowering agent, O-134 Integration of clinical factors and biomarkers for non-
PPARD, PPARGC1B). Additionally, the accentuated effect of estrogen (estrogen) invasive diagnosis of endometriosis
and impaired progesterone effect (tretinoin and altretinoin- progesterone antag- A. Vanhie1, D. O2, C. Tomassetti1, C. Meuleman1, P. Mestdagh3,
onists) was also found. T. D’Hooghe2
Limitations, reasons for caution: Since the comorbidity of obesity and 1
University Hospital Leuven, Leuven University Fertility Center, Leuven, Belgium ;
PCOS was investigated, we found the duality of the pathology as the strong 2
KU Leuven, Development and Regeneration, Leuven, Belgium ;
point of the study. The second strong point was strictly defined study population. 3
UGent, Center for Medical Genetics, Gent, Belgium
The most important limitation could be the sample size.
Wider implications of the findings: Obesity adds the additional burden of Study question: Can integration of clinical data and ultrasound findings with
lipotoxity of free fatty acids in endometrium to already known factors of inflam- biomarkers result in accurate combined clinical-biochemical diagnostic models
mation and insulin resistance in the O-PCOS comorbid state. The only modifiable for endometriosis?
factor is weight loss that could represent a potential mechanism for normalisation Summary answer: In this proof-of-concept study we show that combining
of endometrium towards controls. clinical factors and biomarkers results in a diagnostic model for that outperforms
Trial registration number: KME 0120-491-2017 the individual factors or biomarkers.
What is known already: Diagnosis of endometriosis solely based on presence
O-133 Three-dimensional ultrasonography versus diagnostic of symptoms has a low specificity and sensitivity. Transvaginal ultrasound can
hysteroscopy in the differential diagnosis of intra-uterine benign diagnose endometrioma’s and deep endometriotic nodules, but does not rule
disease. A single institution experience. out peritoneal endometriosis. Cancer antigen 125 (CA-125) levels have been
A.M.L. Gambaro1, M. Monti2, M.G. Piccioni2, A. Musella2, shown to be elevated in some women with endometriosis. However, the sen-
V. Colagiovanni2, E. Marcoccia2, M.C. Schiavi2, C. Fazi2, L. Muzii2, sitivity and specificity of CA-125 for endometriosis is low and serum CA-125
L. Manganaro3 levels alone have no value as a diagnostic tool. Recently, circulating miRNAs have
1
Guy’s and St Thomas’ NHS Foundation Trust, Assisted Conception Unit, London, been described to be differentially expressed in patients with endometriosis,
United Kingdom ; suggesting that they could be used for the non-invasive diagnosis of
2
Sapienza University of Roma, Department of Obstetrical-Gynecological Sciences endometriosis.
and Urological Sciences-, Roma, Italy ; Study design, size, duration: Retrospective case-control study in a cohort
3
Sapienza University of Roma, Department of Radiological Sciences, Roma, Italy of 222 patients (151 women with endometriosis and 71 controls)
Participants/materials, setting, methods: Clinical and ultrasound data
Study question: Comparison of diagnostic accuracy of three-dimensional were extracted from the electronic patient record. Plasma was available in the
transvaginal ultrasonography (3D-TVUS) versus hysteroscopy (HYS) in the biobank of the Leuven University Fertility Center. CA-125 was measured on a
assessment of uterine malformations and submucosal fibroids. Roche Modular E170 instrument using a commercial assay (Roche Diagnostics,

Germany). RNA was extracted with the miRNeasy Plasma Kit and miRNA 0.3 pmol/mL blood). Of note, estradiol:estrone ratios, approximately one in
expression analysis was done by RT-qPCR using Qiagen miScript assays. Models blood, were around three in tissue, indicating active local synthesis.
were build using multivariate logistic regression and validated by a 10-fold cross 17-hydroxy-progestogens and androstenedione were over 4-fold higher in endo-
validation. metriotic lesions than eutopic tissue (p<0.05). The activity of HSD17B1 was
Main results and the role of chance: We developed a diagnostic model comparable between eutopic and endometriotic tissues.
which finally consisted of 2 clinical factors (dysmenorrhea and dyschezia), 4 Regarding corticosteroids, active cortisol was 4-fold higher in endometriosis
biomarkers (hsa-miR-20a-5p, hsa-miR-21-5p, hsa-miR-28-5p and CA-125) and than in the eutopic tissue (p<0.001), whereas inactive cortisone was 2.5-fold
1 interaction factor (hsa-miR-20a-5p*dyschezia). The diagnostic model had an lower in endometriosis (p<0.001). HSD11B1 (activation to cortisol) and
AUC of 79% with a high sensitivity (82%) and moderate specificity (65%) at HSD11B2 (deactivation to cortisone) mRNA levels were in line with the corti-
optimal cut-off (determined by the Youden-index). In the 10-fold cross validation costeroid levels: HSD11B1 mRNA was higher in endometriosis, and the opposite
diagnostic performance of the model dropped to an AUC of 66%. was observed for HSD11B2 compared with the eutopic endometrium (p<0.001
Limitations, reasons for caution: The 10-fold cross-validation shows that for both enzymes). The levels of compounds acting as precursors for cortico-
an important drop in diagnostic performance could be expected when applying steroid synthesis (i.e., 21-hydroxyprogesterone; 11-deoxycorticosterone) were
this model in an independent cohort. higher in endometriosis compared with the eutopic tissue (p<0.05), and a number
Wider implications of the findings: Our data indicate that combined clin- of enzymes involved in the generation of active compounds from these precur-
ical-biochemical diagnostic models perform better than the individual fac- sors were expressed in both eutopic endometrium and endometriotic tissue.
tors. This implies that the classic design of endometriosis biomarkers studies Limitations, reasons for caution: This is a retrospective study, that included
should be reconsidered towards an approach where the bar for each individual patients with all stages of disease and with manifestation of different symptoms
biomarker is lowered while keeping the overall requirement of high sensitivity in a pooled analysis. Sub analyses in relation to type of endometriosis and kind
and specificity. of symptoms of the patient were hampered by the small patient population.
Trial registration number: not applicable Wider implications of the findings: Steroid levels differed between tissue
and blood. HSD17B1 activity was measured in most cases (irrespective to patient
O-135 Local steroid metabolism in eutopic endometrium and and tissue location), hence, inhibition of HSD17B1 is a valid strategy for lowering
corresponding endometriotic lesions: intra-patient variability the local estrogen production in endometriosis. Sustained generation of anti-in-
S. Xanthoulea1, B. Delvoux1, P. Koskimies2, M.R. Häkkinen3, flammatory cortisol may relate to the chronic inflammation associated with
J. Koskivuori3, S. Auriola3, A. Vanhie4, C. Tomassetti4, A. Romano1 endometriosis.
1 Trial registration number: NOT APPLICABLE
Maastricht University Medical Center, Gynaecology, Maastricht, The Netherlands ;
2
Forendo Pharma, Ltd., Turku, Finland ;
3 O-136 Exploring the delays to diagnosis of endometriosis in the
University of Eastern Finland, School of Pharmacy, Kuopio, Finland ;
4 United Kingdom; a triphasic mixed-methods study
KU Leuven, Obstetrics and gynaecology, Leuven, Belgium
B. Karavadra1, E. Morris1
Study question: Is the local steroid metabolism (including estrogens, progesto- 1
Norfolk & Norwich University Hospital, Obstetrics & Gynaecology, Norwich,
gens, androgens and corticosteroids) different between the normal endometrium United Kingdom
of patients and the endometriotic tissue from different anatomical locations?
Summary answer: Steroid levels differ between normal and endometriotic Study question: What are the reasons for the delay to diagnosis of endome-
tissue. Irrespective of the location, endometriosis shows active synthesis of triosis from a patient and healthcare worker perspective?
estrogens and sustained corticosteroid levels. Summary answer: Delay was attributed to patient factors (symptom recog-
What is known already: Endometrial tissue produces locally steroids, relevant nition, seeking help), healthcare system issues (waiting times, referral pathways)
in endometriosis: estrogens control lesion establishment, progestogens oppose and clinician experiences (normalisation, general vs specialist gynaecologist).
estrogen action, androgens are estrogen precursors and corticosteroids suppress What is known already: The average time to diagnosis of endometriosis in
inflammation. These events are related to endometriosis symptoms such as the United Kingdom is 7.5 years and It is widely known that a delayed diagnosis
infertility and pain and offer novel therapeutic targets aimed at blocking local can have a significant impact on women quality of life and activities of daily living.
estrogen generation, such as 17beta-hydroxysteroid dehydrogenase 1 In addition, repeated medical consultations prior to diagnosis can have a signif-
(HSD17B1). icant burden financially on the healthcare system.
To elucidate to what extent steroid metabolism is implicated in endometriosis, Study design, size, duration: A triphasic study design was adopted and it
and what inter- and intra-patient variability exists, we profiled major steroids in blood involved an online questionnaire (1252 participants), semi-structured interviews
and tissue (normal endometrium and endometriosis) of patients; we also determined with 16 participants with endometriosis and separate focus groups with 15
the expression levels of major enzymes involved in local steroid metabolism. healthcare workers (General Practitioners, nurses and gynaecologists). A mixed
Study design, size, duration: This is a retrospective study using biobanked methods approach was used whereby the findings were analysed by content
frozen patient material. Eutopic endometrium, multiple endometriotic lesions analysis and grounded theory qualitative methodology. For the online question-
from each patient, and peripheral blood of 14 women (seven in the secretory naire, women throughout the UK responded and for the qualitative aspects of
and seven in the proliferative phase) with histologically confirmed endometriosis the study, purposive sampling was used.
were analysed. Endometriotic lesions originated from the uterosacral ligament/ Participants/materials, setting, methods: Phase one involved developing
Pouch of Douglas, bladder, ovarian fossa, rectum/rectosigmoid. Patients had an online questionnaire exploring women’s experiences of being diagnosed with
stage I (n=1), II (n=9), III or (n=3), or stage IV (n=1) endometriosis (ASRM endometriosis and this was distributed via the Endometriosis UK charity website.
classification). In phase two, the findings from the phase one study were used to design a
Participants/materials, setting, methods: Plasma, eutopic endometrium qualitative study exploring the finer details of delays in diagnosis and analysed
(n=14) and endometriotic lesions (n=39) were obtained and stored following by grounded theory. Phase three involved focus groups with healthcare workers
the EPHect/WERF guidelines (https://endometriosisfoundation.org/ephect/). to explore their opinions on the findings from the phase two study.
Patients with endometriosis were not under hormonal medication for six months Main results and the role of chance: In phase one, 1252 women replied
prior to the biopsy. RNA expression was determined by whole RNA-sequencing. to the online questionnaire with an average delay to diagnosis of 5.5 years. Delay
Levels of major steroids were measured by Liquid Chromatography-Mass was attributed to patient factors (symptom recognition, seeking help), healthcare
Spectrometry (LC-MS). HSD17B1 activity was measured in tissue lysates (cell- system issues (waiting times, referral pathways) and healthcare worker experi-
free extracts) by High Performance Liquid Chromatography (HPLC). ences (normalisation, general vs specialist gynaecologist). In phase two, 16
Main results and the role of chance: Estrogens (estrone, estradiol) were women took part in individual interviews. They provided significant detail into
non-statistically significantly higher in eutopic and endometriotic tissues com- delays by highlighting the triadic combination of coping mechanisms, health seek-
pared with blood (estradiol: 1.0 pmol/g eutopic; 3.2 pmol/g endometriotic; ing behaviours and the process of adjustment to symptoms. Phase three findings
0.4 pmol/mL blood – estrone: 0.3 pmol/g eutopic; 1.1 pmol/g endometriotic; from 15 healthcare workers provided insight into the preconception’s healthcare

workers had ‘about the endometriosis patient’ and the challenges of recognising Wider implications of the findings: Our study has shown an alteration of
abnormal symptoms. the oocyte microenvironment in women with endometriosis associated with
Mixed-method’s enabled a positivist and constructivist stance to be taken high follicular fluid levels of MCP-1 allowing the identification of a subgroup of
when exploring a complex problem like delay. The quantitative aspect enabled endometriosis patients with a potentially worse prognosis.
insight into the general factors attributed to delay, and the subsequent qualitative Trial registration number: Not applicable
phase enabled in-depth exploration of these factors and the complex interaction
between them.
Limitations, reasons for caution: The findings are based on participants in
the United Kingdom (UK) and the respective healthcare system; therefore, the SELECTED ORAL COMMUNICATIONS
reasons for the delay to diagnosis may vary outside the UK. SESSION 35: IMPACT OF ART ON HEALTH OUTCOMES OF
Wider implications of the findings: This study provides new in-depth insight CHILDREN
to the health-seeking behaviours, coping mechanisms and complex interaction 07 July 2020 Parallel 5 10:00 - 11:30
of factors contributing to delayed diagnosis. It is hoped that the findings from
this study will be used to encourage collaboration between professional bodies
and patient groups to design a pre-diagnostic tool to facilitate diagnosis.
Trial registration number: Not applicable O-138 Differences in the landscape of mitochondrial DNA
variants between ART and spontaneously conceived individuals
O-137 Elevated levels of monocyte chemotactic protein-1 (MCP- links to maternal infertility and low birth weight
1) in the follicular fluid reveals different populations among J. Mertens1, F. Zambelli2, F. Belva3, A. Van Montfoort4,
women with severe endometriosis. E. Couvreau de Deckersberg1, M. Bonduelle3, K. Stouffs3,
P.E. Bouet1, J.M. Chao de la Barca2, H. El Hachem3, S. Blanchard4, S. Seneca3, H. Van de Velde5, C. Blockeel5, K. Barbé6, K. Sermon1,
P. Jeannin4, P. Reynier2, P. May-Panloup5 C. Spits1
1 1
Department of Reproductive Medicine, Angers University Hospital, Angers, France ; Vrije Universiteit Brussel, Reproduction and Genetics, Brussels, Belgium ;
2 2
Department of Biochemistry and Genetics, Angers University Hospital, Angers, Clinica Eugin, Clinica Eugin, Barcelona, Spain ;
3
France ; UZ Brussel, Center for Medical Genetics, Brussels, Belgium ;
3 4
Department of Reproductive Medicine, Clemenceau Medical Center, Beirut, Maastricht University Medical Center, Department of Obstetrics & Gynaecology-
Lebanon ; GROW School for Oncology and Developmental Biology, Maastricht, The
4
Department of Immunology and Allergology, Angers University Hospital, Angers, Netherlands ;
5
France ; UZ Brussel, Center for Reproductive Medicine, Brussels, Belgium ;
5 6
Department of Reproductive Biology, Angers University Hospital, Angers, France Vrije Universiteit Brussel, Interfaculty Center Data Processing & Statistics ICDS,
Brussels, Belgium
Study question: Is there a modification of the cytokine profile of the follicular
fluid (FF) of women with endometriosis undergoing in vitro fertilization (IVF)? Study question: Can mitochondrial DNA variants explain the differences in
Summary answer: In patients with endometriosis, we have identified a sub- birth weight between ART and spontaneously conceived individuals?
type with a high FF MCP-1 level, that is associated with an alteration of the Summary answer: We have found significant differences in the mitochondrial
oocyte microenvironment. genome of children born after ART, which link to a background of maternal
What is known already: Several factors implicated in the acute and chronic infertility and birth weight.
inflammatory reaction (hormones, cytokines, chemokines and markers of oxi- What is known already: Follow-up studies of children born after ART show
dative stress) are thought to be involved in the pathophysiology of endometri- an increased risk of lower birth weight and the potential development of an
osis. Indeed, an alteration of the cytokine profile has been shown in the serum, abnormal cardio-metabolic profile later in life. Efforts to find the molecular cause
the peritoneal fluid, the endometrial tissue, and in endometriomas of women of these differences have focused on the epigenome, but have yet to yield
with endometriosis. Different biological signatures have been assessed in order conclusive results. In this study, we investigated variation in the mitochondrial
to better understand the pathophysiology behind the endometriosis-related genome, which is known to be linked to cardio-metabolic disorders in the general
infertility, using proteomics, metabolomics and the analysis of the cytokine pro- population and potentially to infertility. Our hypothesis was that mitochondrial
file. However, such studies have been scarce and have yielded disparate results. DNA variants could explain the differences in birth weight between ART and
Study design, size, duration: We performed a prospective observational spontaneously conceived individuals.
study on the follicular fluid retrieved from 87 women undergoing IVF with or Study design, size, duration: We studied the full mitochondrial DNA
without intracytoplasmic sperm injection from January 2018 to February 2019 (mtDNA) of 283 ART and 189 spontaneously conceived (SC) samples collected
at the Angers University Hospital, France. in multiple centers collaborating with large fertility clinics. The DNA was
Participants/materials, setting, methods: The patients were divided into extracted from different tissues (ART: 116 blood, 66 placental and 101 buccal
two groups: the endometriosis group and the control group. The endometriosis samples, SC: 65 blood, 51 placental and 73 buccal samples). The DNA was
group comprised 43 patients with severe endometriosis. The control group enriched for the mtDNA by long-range PCR and sequenced on an Illumina
included 44 patients undergoing IVF for other causes and without endometrio- platform.
sis. The cytokine profile of the FF was determined by multiplex fluores- Participants/materials, setting, methods: The variant calling was per-
cent-bead-based technology allowing the measurement of 59 cytokines. formed using mtDNA server and MuTect, with a threshold of >1.5% load, versus
Main results and the role of chance: Univariate analysis showed a significant the reference NC_012920.1. The results were correlated to maternal age, indi-
increase of nine cytokines in the FF in the endometriosis group compared to cation for ART, culture medium, gestational age, and birth weight corrected for
the control group, while Monocyte Chemoattractant Protein 1 (MCP-1) was gestational age and gender. Statistical analysis was done using SPSS. For the
the only variable retained in the multivariate analysis. We identified two sub- complex data of the heteroplasmic variants, an orthogonally rotated factor anal-
groups of patients in the endometriosis group: MCP-1- group (n=23) which had ysis was used to reduce the dimensionality of the studied dependent variables.
FF MCP-1 levels comparable to the control group and MCP-1+ (n=20) which Main results and the role of chance: Haplogroup U4 was overrepresented
had significantly higher FF levels. Only patients in the MCP-1+ group had a sig- in the ART group (ART: 5.3% vs. SC: 0.5%, p=0.004). ART individuals have more
nificantly altered cytokine profile in the FF, and had a significantly higher serum not previously described homoplasmic variants (ART: 31.4%, SC: 21.7%,
estradiol level (p=0.002) and a significantly lower number of oocytes recovered p=0.021). ART Individuals whose mother was infertile more frequently carried
(p=0.01) compared to the MCP-1- and the control group. homoplasmic tRNA variants than SC or individuals with infertile fathers
Limitations, reasons for caution: One potential limitation of our study is (p=0.041). ART individuals carry higher heteroplasmic loads in the coding region
the lack of assessment of serum MCP-1 levels, which could have been compared (p=0.039) and component analysis, using rank randomized tests, indicated that
to those in the follicular fluid. they carry a different mtDNA variant composition from SC individuals (p=0.015),

which is driven by variants inducing non-synonymous changes and in the rRNA Participants/materials, setting, methods: From the national health regis-
regions. These heteroplasmic variants were highest in individuals born from tries in Denmark and Finland, we identified all children diagnosed with Prader-
infertile mothers, followed by male factor infertility when compared to the SC Willi syndrome (n=143); Silver-Russell17 syndrome (n=69); Beckwith-Wiedemann
group (significant trend, p=0.017). Maternal age nor gestational age were linked syndrome (n=105) and Angelman syndrome (n=72) born between 1990/1994
to an increased mutational burden. Finally, both ART and SC individuals ≤10th and 2014, respectively. he median follow-up time was 8 years and 9 months for
birth weight percentile more frequently carried homoplasmic tRNA variants ART children and 11 years and 9 months for NC children.
(p=0.032). SC individuals with birth weight ≤10th and ≤25th percentile carried Main results and the role of chance: We identified a total of 389 children
more frequently heteroplasmic coding variants (p=0.032 and p=0.008). In the diagnosed with imprinting disorders, sixteen of these were conceived after ART.
ART group, only individuals from embryos that had been in Vitrolife G1 medium The overall odds ratio for the four imprinting disorders in ART children compared
followed the same pattern as SC, with individuals ≤25th percentile carrying more with NC children was 1.60 [95%CI 0.97-2.65]. Made up separately, eight ART
heteroplasmic coding variants (p=0.0232). children were diagnosed with Beckwith-Wiedemann syndrome, the odds ratio
Limitations, reasons for caution: Other factors not considered in this study for this specific imprinting disorder was 3.07 [95%CI 1.49-6.31]. The risk of
could play a role in the birth weight, which we are currently correcting for. This Prader-Willi syndrome, Silver-Russell syndrome and Angelman syndrome was
study is observational and no functional tests were performed. Finally, after strat- not increased in children conceived after ART.
ification for embryo culture medium, the sample sizes are small (N=49- 66). Limitations, reasons for caution: Imprinting disorders are rare events and
Wider implications of the findings: These results suggest a link between our results based on few ART children with imprinting disorders. The etiology
maternal infertility and mtDNA variant composition, which is transmitted to the is complex and only partly clarified, and the clinical diagnoses challenged by a
offspring. These variants correlate to birth weight, particularly in the SC individ- broad phenotypic spectrum.
uals, suggesting a universal mechanism. Embryo culture medium appears to affect Wider implications of the findings: In the existing studies results on the
birth weight through another mechanism, masking the effect of mtDNA variation. risk of imprinting disorders in children conceived after ART are ambiguous. This
Trial registration number: not applicable’ study adds that the risk of imprinting disorders in ART children is very small and
perhaps restricted to Beckwith-Wiedemann syndrome
O-139 Imprinting disorders in children born after assisted Trial registration number: not applicable
reproductive technology (ART): a Nordic study from the
CoNARTaS group O-140 ICSI does not increase the odds of adverse perinatal
Ø. Lidegaard1, A.K.A. Henningsen2, M. Gissler3, S. Rasmussen4, outcomes in autologous/ donor oocyte cycles without male-factor
S. Opdahl5, U.B. Wennerholm6, A.L. Spangsmose7, A. Tiitinen8, subfertility: analysis of 121,448 singleton live births
C. Bergh6, L.B. Romundstad9, H. Laivuori10, J. Forman11, P.R. Supramaniam1, L.N. Lim1, E. McVeigh1, I. Granne1, C. Becker1,
A.B. Pinborg7 M. Mittal2
1
Rigshospitalet- University of Copenhagen, Dept. of Ob/Gyn, Copenhagen, 1
Oxford University Hospitals NHS Foundation Trust, Gynaecology, Oxford, United
Denmark ; Kingdom ;
2
Rigshospitalet- University of Copenhgen, Fertility Clinic, Copenhagen, Denmark ; 2
Imperial College Healthcare, Gynaecology, London, United Kingdom
3
THL Finnish Institute for Health and Welfare-, Information Services Department,
Helsinki, Finland ; Study question: Does ICSI increase the odds of adverse perinatal outcomes
4
Rigshospitalet- University of Copenhagen, Department of Gynaecology, in autologous and donor oocyte cycles without male factor subfertility?
Copenhagen, Denmark ; Summary answer: ICSI does not increase the odds of adverse perinatal out-
5
Central Norway Regional Health Authority- Norwegian University of Science and comes in autologous and donor oocyte cycles in the absence of male factor
Technology-, Department of Public Health and General Practice, Oslo, Norway ; subfertility.
6
Institute of Clinical Sciences- Sahlgrenska Academy- University of Gothenburg-, What is known already: An increased incidence of obstetric complications
Department of Obstetrics and Gynecology, Gothenburg, Sweden ; is recorded in pregnancies conceived through assisted reproductive treatments
7
Rigshospitalet- Copenhagen University Hospital-, Fertility Clinic, Copenhagen, (ART) compared to spontaneous conception. There is a paucity of information
Denmark ; is available on the effect of method of fertilisation, particularly ICSI, on perinatal
8
University of Helsiniki- Helsinki University Hospital -, Department of Obstetrics outcomes in the absence of male factor subfertility in either autologous or donor
and Gynecology, Helsinki, Finland ; oocyte cycles.
9
Central Norway Regional Health Authority- Norwegian University of Science and Study design, size, duration: Retrospective cohort analysis of all fresh cycles
Technology-, Department of Public Health and General Practice- Spiren Fertility conducted between 2002-2016, from the UK HFEA national database. 110,805
Clinic, Oslo, Norway ; singleton live births(LB) have been recorded from all oocyte usage, of which,
10
University of Helsinki- Institute for Molecular Medicine- Institute of Life 105,618 were from autologous oocyte cycles and 5187 from donor oocyte
Science- University of Helsinki- Helsinki University Hospital- Medical and Clinical cycles. Assuming that the adverse perinatal outcome rate for IVF is 10%, a power
Genetics- Faculty of Medicine and Health Technology- University of Tampere- calculation demonstrated that 686 LBs would need to be analysed for 80% power
Tampere Univers, , ; and 5% significance to detect a 10% difference.
11
University of Copenhagen-, Department of Biostatistics, Copenhagen, Denmark Participants/materials, setting, methods: The database was analysed for
singleton live birth rate (SLBR), stratified by recipient age, donor age, number
Study question: Is the risk of imprinting disorders increased in children con- of previous IVF treatment cycles undertaken and type of cycle (fresh IVF versus
ceived after assisted reproductive technology (ART)? fresh ICSI). Cycles complicated by male factor subfertility were excluded from
Summary answer: We found an odds ratio of 3.07 [95%CI 1.49-6.31] for the final analysis as this is known to influence the method of fertilisation. Statistical
Beckwith-Wiedemann syndrome in ART children, but no increased risk of three analysis was performed using Logistic Regression and Chi-square; p<0.05 was
other imprinting disorders. considered statistically significant.
What is known already: Earlier studies, most of them small, have suggested Main results and the role of chance: The overall IVF to ICSI ratio for all
an association between ART and imprinting disorders. Results, however, far from treatment cycles, in the absence of male factor subfertility, favoured IVF (58:42)
consistent. in 2002, with a yearly rise seen in ICSI cycles (46:54 in 2016).
Study design, size, duration: Binational register-based cohort study. All IVF increased the risk of very preterm births (<32 weeks’) (adjusted odds
children conceived by ART in Denmark (n=45 393) born between 1994 and ratio [aOR] 1.25, 99.5% confidence interval [CI] 1.10-1.42, p<0.0001) and
2014 and in Finland (n=29 244) born between 1990 and 2014 were identified. preterm births (32-36 weeks’) (aOR 1.10, 99.5% CI 1.03-1.17, p<0.0001) com-
The full background populations born during the same time periods in the two pared to ICSI treatment cycles with autologous oocytes, using term delivery as
countries were included as controls. Odds ratios of imprinting disorders in the reference category. This significance was not sustained in donor oocyte
ART children compared with naturally conceived (NC) children were cycles: very preterm births (aOR 1.17, 99.5% CI 0.70-1.96, p=0.393) and
calculated. preterm births (aOR 0.90, 99.5% CI 0.69-1.17, p=0.250). The odds of a

post-dates delivery was increased in autologous oocyte cycles employing ICSI transfer of frozen-thawed ICSI embryo with a frozen-thawed IVF embryo. The
when compared to IVF (aOR 1.05, 99.5% CI 1.01-1.10, p=0.003), however, this risk of major malformations was similar for the two groups, AOR 1.04; [0.90-
was not maintained in donor oocytes (aOR 0.94, 99.5% CI 0.75-1.19, p=0.459). 1.21]; p-value=0.57. When we investigated the risk of malformations grouped
There was no significant difference demonstrated when the birthweight per by different organ systems, we did not find higher risk among ICSI children in
method of fertilisation was analysed after adjusting for potential confounding any of the above comparisons. However, we confirmed that children born after
factors in either oocyte category ICSI have a significantly higher risk of being born with a major malformation
Limitations, reasons for caution: The accuracy of the database is dependent compared with NC children, AOR 1.30; [95%CI 1.24-1.36]; p-value<0.0001.
on the information submitted to the HFEA. Until 2007, this data was manually Limitations, reasons for caution: Due to the poorer quality of registration
captured adding the risk of data entry error. Furthermore, information on mater- of minor malformations in the national health registries, together with surveil-
nal confounding factors such as body mass index and smoking status was not lance bias, we restricted our analyses to major malformations. We do not expect
available to be evaluated. the detection of major congenital malformation to be biased by method of
Wider implications of the findings: ICSI in the absence of male factor conception.
subfertility does no increase the odds of adverse perinatal outcome in autologous Wider implications of the findings: Since the first ICSI child was born in
and donor oocyte cycles. This at present suggests that the adverse outcomes 1992, there has been concern whether the risk of congenital malformations is
previously described with ICSI is related to the significant abnormalities associ- increased among ICSI children, where often poor-quality semen has been used
ated with the male component rather than the technique. for conception. Our findings based on 39 684 Nordic ICSI children, showing no
Trial registration number: N/A differences compared to IVF, are reassuring.
O-141 The risk of major congenital malformations in children
conceived after ICSI: a Nordic cohort study O-142 The DNA methylation patterns within whole blood
A.K. Henningsen1, S. Opdahl2, U.B. Wennerholm3, A. Tiitinen4, of adolescents born from IVF are not different from those
L.B. Romundstad5, C. Bergh3, M. Gissler6,7, J. Forman8, A. Pinborg1 adolescents conceived naturally within the Raine birth cohort.
1
Rigshospitalet, Fertility Clinic, Copenhagen, Denmark ; B. Penova-Veselinovic1, P. Melton2, R.C. Huang3, J. Yovich4,
2
Norwegian University of Science and Technology, Department of Public Health P. Burton5, R. Hart6
1
and Nursing, Trondheim, Norway ; University of Western Australia, Division of Obstetrics and Gynaecology, Perth,
3
Institute of Clinical Sciences- Sahlgrenska Academy- University of Gothenburg- Australia ;
2
Sahlgrenska University Hospital, Department of Obstetrics and Gynecology, University of Western Australia, Centre for Genetic Origins of Health and
Gothenburg, Sweden ; Disease- Statistical Genetics, Perth, Australia ;
4 3
University of Helsinki- Helsinki University Hospital-, Department of Obstetrics University of Western Australia, Telethon Kids Institute, Perth, Australia ;
4
and Gynecology, Helsinki, Finland ; Curtin University, PIVET Medical Centre, Perth, Australia ;
5 5
Spiren Fertility Clinic, Spiren Fertility Clinic, Trondheim, Norway ; Edith Cowan University, Concept Fertility Centre, Perth, Australia ;
6 6
THL Finnish Institute for Health and Welfare, Information Services Department, University of Western Australia & Fertility Specialists of WA, School of Women’s
Helsinki, Finland ; and Infants Health, Perth- Western Australia, Australia
7
Karolinska Institute- Division of Family Medicine, Department of Neurobiology-
Care Sciences and Society, Stockholm, Sweden ; Study question: Do the epigenome-wide DNA methylation profiles of ado-
8
University of Copenhagen, Department of Biostatistics, Copenhagen, Denmark lescents born from in-vitro fertilisation (IVF) differ from the epigenome of nat-
urally conceived counterparts from a birth cohort?
Study question: Is the risk of major congenital malformations higher in children Summary answer: There is no significant difference in the DNA methylation
conceived after ICSI compared with children conceived after IVF? profiles of adolescents born from IVF when compared to their naturally con-
Summary answer: We did not find any difference in the risk of being born ceived, age-matched counterparts.
with a major congenital malformation in children conceived after ICSI compared What is known already: Epigenetic changes are heritable modifications that
with IVF. may alter gene expression without changing the DNA sequence. Growing evi-
What is known already: It is well known that children born after ART have dence suggests that the adverse health outcomes reported in IVF-born offspring
a higher risk of being born with a major congenital malformation compared with might have underlying epigenetic mechanisms. Both, features of infertile couples,
children born after natural conception (NC). It is still not clear whether the risk as well as the IVF procedure itself have been shown to alter the epigenetic sig-
of congenital malformations is higher in children conceived after ICSI compared nature in offspring and placental tissue. As most studies investigated DNA meth-
with IVF, as studies so far present diverging findings. It is also uncertain if cryo- ylation changes in cord blood/placenta tissue, and a recent study reported that
preservation of the embryos influences the risk of malformations in the offspring. these changes are mitigated by adulthood, it is essential to further investigate
Study design, size, duration: Nordic cohort study based on national registry potential effects of IVF on the DNA methylation profiles in adolescents using
data from Denmark, Norway and Sweden. The analyses were based on single- whole blood.
tons born after ART (n=98 026) and NC (n=4 804 844) between 1992 and 2015. Study design, size, duration: The Growing Up Healthy Study (GUHS) is a
Participants/materials, setting, methods: The study included all singletons, prospective study that recruited 303 adolescents to compare various long-term
both live-born and stillborn, conceived after ICSI (n=39 684); IVF (n=58 342) health outcomes and DNA methylation profiles with age-matched counterparts
and NC (n=4 804 844). We differentiated between children conceived after from a birth cohort (Generation 2 of the Raine Study). GUHS assessments were
transfer of fresh embryos versus frozen-thawed embryos. Malformations were conducted in 2013-2017. The effect of IVF on DNA methylation levels of 238
coded in all countries using the International Classification of Diseases (ICD). adolescents mean age 16±1.67 (52.94% male) was compared to 1188 naturally
The European Concerted Action on Congenital Anomalies and Twins conceived, age-matched controls, 17.25±0.58 (50.93% male) from the
(EUROCAT) was used to differentiate between major and minor malformations. Raine Study.
Minor malformations were not included in the analyses. Participants/materials, setting, methods: Genomic DNA from whole
Main results and the role of chance: I the multiple regression analyses, we blood was used to generate the epigenome profiles of 238 GUHS adolescents
adjusted for maternal age, parity, year of birth, child’s sex, body mass index, and 14 technical replicates using the Infinium Methylation Epic Bead Chip that
smoking and country. We found no difference in the risk of being born with a measures and quantifies approximately 850,000 DNA methylation probes. These
major congenital malformation, when comparing children conceived after ICSI generated profiles were compared to the DNA methylation profiles from the
with children conceived after IVF, adjusted odds ratio (AOR) 1.06; [95% confi- Raine Study that was quantified using Illumina 450K platform. DNA methylation
dence interval (CI) 0.98-1.13]; p-value=0.09. Neither could we detect any dif- profiles for both studies were normalized using the same quantile normalization
ference in the risk of major malformations when comparing children born after procedure (BMIQ) for comparative purposes.
transfer of a fresh ICSI embryo versus a frozen-thawed ICSI embryo, AOR 1.11; Main results and the role of chance: We defined adolescence by an age
[95%CI, 0.98-1.26]; p-value=0.09. We also compared children conceived after range between 13 and 19.9 years. The average age of the generated DNA

methylation profiles in the GUHS cohort was 16.06 years. The DNA- disorders; 9.8% for endometriosis; 8.1% for male factor causes and 8.5% for
methylation profiles of the Raine Study participants have an average age of unexplained infertility. Compared to unexplained infertility cause, the risk of PTB
17.25 years. After quality control and filtering, a total of 401,022 DNA meth- was significantly higher with ovulatory disorders (adjusted odds ratio-aOR 1.31,
ylation probes overlapped between the two studies. These DNA methylation 95% CI 1.21, 1.41); tubal disorders (aOR 1.25, 95% CI 1.18, 1.34) and endo-
probes were then each investigated and compared between the IVF born GUHS metriosis (aOR 1.17, 95% CI 1.05, 1.29). There was no significant difference in
and naturally conceived Raine study adolescents. We tested for an association the risk of PTB with male factor causes compared to unexplained infertility (aOR
between these groups applying Firth’s bias reduced logistic regression against 1.25, 95% CI 1.18, 1.34).
the outcome of IVF vs naturally conceived. Potential confounders, namely the The incidence of LBW based on infertility causes was 10.5% for ovulatory
DNA methylation probe as well as technical variation due to different meth- disorders; 9.4% for tubal disorders; 9.8% for endometriosis; 8.4% for male factor
ylation platforms used between studies, were adjusted for within-study batch causes and 8.5% for unexplained infertility. Compared to unexplained infertility
effects. After adjustment none of the compared DNA methylation probes cause, the risk of LBW was significantly higher with ovulatory disorders (aOR
reached a Bonferroni correction of 1.24E-0.7 (0.05/402,022) for statistical 1.29, 95% CI 1.20, 1.40); tubal disorders (aOR 1.12, 95% CI 1.05, 1.20) and
significance. Overall, a small minority, 3850 (0.96%) of the analyzed 401,022 endometriosis (aOR 1.11, 95% CI 1.0, 1.24). The risk of LBW was significantly
DNA methylation probes showed nominal significance with a p-value <0.05, lower with male factor causes compared to unexplained infertility (aOR 0.94,
most likely to be false positives after controlling for cross-study comparisons. 95% CI 0.89, 0.99).
1,810 Differentially Methylated Regions (DMR’s) were identified between the Limitations, reasons for caution: Although the analysis was adjusted for
cohorts; however none reached statistical significance after correcting for mul- several important confounders, there was no information on medical history of
tiple testing. women during pregnancy to allow adjustment. Limitations with observational
Limitations, reasons for caution: With contradictory findings on small data would apply to this study including residual confounding.
sample size studies to date, there is a need to better understand the complex Wider implications of the findings: This is the first study to address causes
outcomes and effects of IVF manipulations on the epigenome of offspring. To of infertility affecting perinatal outcomes of PTB and LBW. The information is
infer an association with higher statistical and biological significance, data from important and needs to be understood further.
various groups should be pooled for a meta-analysis. Trial registration number: Not applicable
Wider implications of the findings: Our results support previous findings
that the epigenome profiles of IVF born adolescents are stable, and not affected
by the techniques used in IVF. The increased cardiometabolic risk factors O-144 The risk of cerebral palsy in ART children has more than
observed in adolescents born from IVF treatment may result from other non- halved over two decades – a Nordic collaborative study on 55,233
DNA methylation related epigenetic influences. liveborn children
Trial registration number: NHMRC Project grant number 1042269
ThisSpangmose
A.L. abstract has been
, L. Hee chosen
, M. Gissler2for
, A. the press
Tiitinen , S. Opdahl4,
1 1 3
L.B. Romundstad , C. Bergh , U.B. Wennerholm ,

5 6 6
O-143 Female causes of infertility are associated with higher programme

A.K. Henningsen and
1
, A.will not1 be made public until the
Pinborg
risk of preterm birth and low birth weight: analysis of 120 902 1abstract is presented.
Fertility Clinic Rigshospitalet, Copenhagen University Hospital, Copenhagen,
singleton live births following IVF Denmark ;
M.S. Kamath1, B. Antonisamy2, S.K. Sunkara3 2
Information Services Department- Finnish Institute for Health and Welfare THL,
1
Christian Medical College and Hospital, Reproductive Medicine, Vellore, India ; Department of Neurobiology- Care Sciences and Society- Karolinska Institute,
2
Christian Medical College and Hospital, Biostatistics, Vellore, India ; Helsinki and Stockholm, Finland ;
3 3
King’s College London-, Division of Women’s Health- Faculty of Life Sciences and Helsinki University Hospital and University of Helsinki, Department of Obstetrics
Medicine-, London, United Kingdom and Gynecology, Helsinki, Finland ;
4
Norwegian University of Science and Technology, Department of Public Health
Study question: Does cause of infertility affect perinatal outcomes of preterm and Nursing, Trondheim, Norway ;
birth (PTB) and low birth weight (LBW) following In vitro fertilisation (IVF) 5
Norwegian Institute of Public Health, Spiren Fertility Clinic- Center for Fertility and
treatment? Health, Trondheim, Norway ;
Summary answer: The risk of PTB and LBW was significantly higher with 6
Institute of Clinical Science- Sahlgrenska Academy, Department of Obstetrics and
female causes of infertility – ovulatory disorders, tubal disorders and endome- Gynecology, Gothenburg, Sweden
triosis compared to unexplained infertility.
What is known already: Women with infertility have an increased risk of Study question: Has the risk of cerebral palsy (CP) changed over a 20-year
adverse perinatal outcomes. Risk of adverse perinatal outcomes is also higher period in children born after assisted reproductive technology (ART)?
following assisted reproductive treatments (ART) compared to spontaneous Summary answer: Over two decades, the risk of CP in ART-children
conceptions. Infertility can result from female and or male factors or unexplained decreased with more than 50%, mainly due to single embryo transfer resulting
when the cause cannot be delineated by standard investigations. Given that in lowering twin rates.
infertility and ART are contributory to the adverse perinatal outcomes, it is a What is known already: During the last two decades we have seen a con-
matter of interest to delineate if the specific cause of infertility influences the siderable reduction in twin birth rates after ART treatment in Europe herein the
perinatal outcomes following IVF treatment. Nordic countries. In the Nordic countries, the twin births rates in ART pregnan-
Study design, size, duration: Anonymous data were obtained from the cies have declined from almost 25% two decades ago to less than 5%, compared
Human Fertilization and Embryology Authority (HFEA). The HFEA has collected with 2–3% in the background population of spontaneously conceived (SC)
data prospectively on all ART cycles performed in the UK since 1991. Data from pregnancies. Concomitantly the preterm birth (PTB) rate has diminished con-
1991 to 2016 comprising a total of 120 902 singleton live births following IVF ± siderably. PTB increases the risk of CP, which is one of the most severe com-
ICSI for sole causes of infertility were analysed for PTB and LBW. Cycles having plications in ART-children with long term consequences for the children and
more than one cause of infertility and multiple births were excluded. their families.
Participants/materials, setting, methods: Data on all women undergoing Study design, size, duration: A Nordic registry-based cohort study using
stimulated IVF ± ICSI treatment cycle were analysed to compare perinatal out- data from Denmark (birth years 1994–2010) and Finland (1990–2010) including
comes of PTB and LBW among singleton live births based on the cause of two cohorts: 55,233 ART-children and 2,327,350 SC-children. Among the ART-
infertility (ovulatory disorders, tubal disorders, endometriosis, male factor, unex- children, 37,404 were singletons, 17,057 were twins and 772 were higher order
plained). Logistic regression analysis was performed adjusting for female age multiples. We investigated the risk of CP in the following time periods: birth
category, period of treatment, previous live births, IVF or ICSI, number of year 1990–1994, 1995–1999, 2000–2004 and 2005–2010. Children were fol-
embryos transferred and fresh or frozen embryo transfer cycles. lowed up until 2014.
Main results and the role of chance: The incidence of PTB following IVF Participants/materials, setting, methods: Data originated from the
based on infertility causes was 10.8% for ovulatory disorders; 10.5% for tubal CoNARTaS cohort (Committee of Nordic ART and Safety) containing

information from national ART- and patient registries. CP was defined as G80 Study design, size, duration: Between April 27th and May 19th, 2020, 1549
ICD-10-diagnoses registered in the patient registry before the age of ten. Risk women were tested for SARS-CoV-2. Before these tests were performed, a
of CP was compared for ART vs. SC children, singletons and twins using logistic symptomatic triage had been carried out in which patients were asked by tele-
regression models. Adjustments were made for maternal age, parity, child’s sex, phone for the presence of symptoms of the virus or if they had been in contact
country and birth year as well as plurality (if applicable). with someone suspected or confirmed to have been infected with the disease.
Main results and the role of chance: Overall, 307 (0.6%) ART- and 5,911 Only patients classified as negative triage attended the clinic for further testing.
(0.3%) SC-children were diagnosed with CP. During the study period the crude Participants/materials, setting, methods: IgG and IgM antibody against
risk of CP in ART-children decreased consistently from 0.9% (1990–1994) to SARS-CoV-2 in plasma samples were tested using enzyme linked immunosorbent
0.4% (2005–2010), while the risk remained unchanged for SC-children (0.3%). assay (ELISA) kits (Epitope Diagnostics, USA). Women with asymptomatic triage
For ART-singletons the risk of CP decreased from 0.7% (1990–1994) to 0.3% and negative IgM started a treatment; on the 5th-8th day of stimulation for retrieval
(2005–2010), but remained stable for ART-twins (0.7%), SC-singletons (0.2%) or 3-5 days before a frozen embryo transfer, a nasopharyngeal RT-PCR was
and SC-twins (0.8%). Throughout the study period the adjusted risk of CP was performed in order to avoid an active infection during the treatment.
higher for ART- versus SC-children (adjusted odds ratio [aOR] 1.93 [95%CI Main results and the role of chance: Study was performed in 17 private
1.71;2.17]). The risk remained increased after further adjustment for plurality clinics belonging to IVIRMA group. Serological testing was carried out in 1549
(aOR 1.18 [95%CI 1.04;1.34]). The risk of CP was higher for ART-singletons patients before starting a treatment. Seroconversion rate for IgG was 3.8%
(aOR 1.32 [95% CI 1.10;1.57]) but similar for ART-twins compared with their (n=59) and for IgM was 0.7% (n=11). As previously discusses, only those patients
SC counterpart. Analyses stratified on birth year showed a consistent decrease in whom the recent presence of the infection had been ruled out (IgG+/IgM- or
in risk of CP over time for ART- versus SC-children (1990–1994: aOR 2.88 IgG-/IgM-) continued with the treatment. Before embryo transfer, both in fresh
[95%CI 1.81;4.32]; 2005–2010: aOR 1.34 [95%CI 1.12;1.61]). In 2005–2010 and frozen cycles, a molecular determination of the virus was carried out and
the risk of CP, after further adjustments for plurality, was no longer statistically we observed a 0.06% (n=1) rate of positive RT-PCR.
significant (aOR 0.96 [95%CI 0.79;1.15]). Additionally, the risk of CP decreased The possibilities of having a patient with negative IgG and IgM and a positive
substantially over time for ART-singletons compared with SC-singletons (1990– RT-PCR is due to the period between the pre-symptomatic or asymptomatic
1994: aOR 2.53 [95%CI 1.20;4.63]; 2005–2010: aOR 1.21 [95%CI 0.88;1.62]). shedding and the first time at which the production of IgM starts, which can be
Limitations, reasons for caution: Observational studies may have inade- up to 12 days. Even though a negative RT-PCR could be considered enough as
quate adjustment for potential confounding factors. Despite adjustment for some immunity is not well established, there is no clear evidence that past infections
major confounders, residual confounding cannot be excluded. could be re-infected, and IgG seem to be neutralizing. Serological testing of IgM
Wider implications of the findings: Limiting the number of twins born after prevents patients from starting endometrial priming or controlled ovarian stim-
ART treatment lowers the risk of CP in the ART population. Multiple embryo ulation decreasing the risk of cancelling the cycle while a positive IgG avoids
transfer is still the standard care in many countries. Our findings emphasize that repeating RT-PCR for each oocyte retrieval or embryo transfer
single embryo transfer should be encouraged worldwide. Limitations, reasons for caution: The low prevalence of infected patients
Trial registration number: ISRCTN11780826 after the triage makes these number very approximate. The utility of these tests
will worsen as the virus incidence decreases.
Wider implications of the findings: In this current scenario, serological
testing does not seem to be a cost-effective strategy to avoid asymptomatic
INVITED SESSION carriers of the virus for starting a treatment, but it still detects a low number of
asymptomatic patients. Lower occurrence of the virus may suggest the no need
SESSION 36: COVID-19-LESSONS LEARNED
for a later screening.
07 July 2020 Parallel 6 10:00 - 11:30
O-147 Seroconversion of Immunoglobulins to SARS-CoV-2 of
healthcare workers in nine European fertility units
O-145 Modelling the COVID-19 epidemic and implementation of S. Nelson M.B.B.Ch- Ph.D.1, S. Ehnert2, T. Child3, G. Trew4
population-wide interventions in Italy 1
University of Glasgow, Level 2 New Lister Building, Glasgow, United Kingdom
2
G. Giordano1 VivaNeo Laborgiagnostik A member of The Fertility Partnership, Laborgiagnostik,
1 Düsseldorf, Germany
Universita de Trento, Dipartimento di Ingegneria Industriale, Italie 3
The Fertility Partnership, Oxford Fertility, Oxford, United Kingdom
4
The Fertility Partnership, Boston Place, London, United Kingdom
O-146 Is COVID-19 symptomatic triage enough? The limited
value of serological testing Study question: To assess SARS-CoV-2 seroconversion in healthcare workers
C. González-Ravina1, V. Vergara2, M. Cruz2, N. Prados3, in nine European IVF units on recommencement of clinical activity.
A. Requena2 Summary answer: A large proportion of staff remain susceptible to SARS-
1
IVI Sevilla, Andrology and Endocrinology laboratories, Sevilla, Spain CoV-2 infection with no evidence of seroconversion. Comprehensive risk mit-
2
IVI Madrid, Reproductive Medicine, Madrid, Spain igation strategies are essential for continued staff welfare.
3
IVI Sevilla, IVF Laboratory, Sevilla, Spain What is known already: The diagnosis of COVID-19 is based on the detec-
tion of the SARS-COV2 virus using RT-PCR from nasopharyngeal samples at
Study question: Do serological and molecular tests increase the detection of the time of active infection. Most patients infected with SARS-COV-2 develop
patients with SAR-CoV-2 after a negative triage? antibodies (Ab) against SARS-COV2 proteins. Regulatory approved CE marked
Summary answer: Serological testing has limited value in detecting SARS- commercial ELISAs are now available to assess seroconversion, yet the preva-
CoV-2 once triage has been negative if molecular testing is being performed lence of SARS-CoV-2 antibodies amongst healthcare workers in IVF units is
before oocyte retrieval or embryo transfer unknown.
What is known already: Fertility community is uncertain about how to opti- Study design, size, duration: Prospective cohort study of 175 staff members
mally provide care to infertile patients, without compromising safety, once the from 8 European IVF clinics and one diagnostic laboratory (5 Germany, 3
activity is re-established. The key principle established by the scientific societies Austria), sampled over a two-week period with paired SARS-CoV-2 antigen
for resuming activity is that anyone attending a clinic should be triaged negative. testing and blood sampling for serology.
More precise methodology has been developed such as immunological tests that Participants/materials, setting, methods: Staff returning to work in the
inform us of the state of the disease and molecular tests (RT-PCR) that measures nine European clinics of The Fertility Partnership received paired oropharyngeal
viral load. These tests may help us to identify asymptomatic and pre-symptomatic antigen testing for RT-PCR and blood sampling for serology. RT-PCR for SAR-
carriers that could have not been detected only with triage and patients with CoV-2 was performed in accordance with WHO guidelines. Detection of SARS-
past infections that may need no further tests. CoV-2 antibodies was performed by the Abbott Diagnostics SARS-CoV-2 IgG

assay on an Abbott Architect i2000 according to the manufacturer’s instructions. day uses patient age, number of follicles at baseline scan and follicle count by
This qualitative assay detects IgG binding to an undisclosed epitope of the SARS- size for the current scan. The model to predict over-response uses age and
CoV-2 nucleocapsid protein. number of follicles of a given size.
Main results and the role of chance: Of the 175 staff members tested 0 In contrast, models using data from a baseline scan are not highly predictive of
(95%CI 0.0 to 2.1%) had evidence of SARS-CoV-2 as detected by RT-PCR. In the trigger administration day (the mean MSE of models using data only from
contrast 6 healthcare workers 3.4% (95%CI 1.6 to 7.3%) had antibodies against baseline scans is 3.83 +/- 0.24). Over-response can be predicted at baseline
SARS-CoV-2. There was no evidence of clustering within the clinics with four using AFC and age with moderate accuracy (mean AUROC 0.77 +/- 0.01).
of the nine facilities having only one staff member affected (prevalence estimates These results suggest that, if there is a significant need to reduce patient contacts
ranging from 5.6% to 12.5%), and one clinic having three staff members affected per cycle in order to facilitate safe access to care, priority for patients attending
(9.1% (95%CI 3.1% to 23.6%). for scans after day 7 of the treatment cycle should be considered.
Limitations, reasons for caution: Understanding viral and host interactions Limitations, reasons for caution: This is a single-centre retrospective study.
during acute and convalescent phases are critical to interpret both the timing of RCTs with healthy live birth primary outcome should guide changes to IVF care.
initial seroconversion after exposure to SARS-CoV-2, and the subsequent dura- However, for safety in extraordinary times, clinics may have to adjust protocols
tion of antibodies. At present studies regarding temporal seroconversion have immediately, with no time to await optimal evidence. These results may provide
been developed in conjunction with assay development, limiting the long-term help for those making these challenging decisions.
conclusions of seroconversion. Wider implications of the findings: These results are timely and prospective
Wider implications of the findings: A low proportion of healthcare workers evaluation of streamlined follicular tracking for COS monitoring may be war-
in nine different sites had seroconverted and had evidence of SARS-CoV-2 ranted. Previous small studies have shown that minimal monitoring protocols
antibodies. A large proportion of staff are still susceptible to infection and appro- did not adversely impact outcomes. If IVF can safely be made less onerous,
priate infection control procedures are essential to continue to mitigate risk and without compromising success, this could help reduce burden related treatment
ensure staff welfare with the resumption of clinical services. drop-out.
O-315 Streamlining follicular monitoring during controlled O-316 Maternal-fetal vertical SAR-CoV-2 (COVID-19) viral
ovarian stimulation? A data-driven approach to efficient IVF care transmission during pregnancies is possible and currently cannot
in the new era of social distancing be dismissed
I. Robertson1, F.P. Chmiel2, Y. Cheong1 G. Bahadur1,2, R. Homburg2, C. Singh3, M. Bhat4, P.A. Doreski5, J.
1
University of Southampton, Human Development and Health, Southampton, Huirne6, A. Muneer7
1
United Kingdom North Middlesex University Hospital Trust, Reproductive Medicine Unit, London,
2
University of Southampton, IT Innovation- Electronics and Computer Science, United Kingdom
2
Southampton, United Kingdom Homerton University Hospital, Homerton Fertility Centre-, - London E9 6SR,
United Kingdom
Study question: What is the optimal follicle tracking ultrasound strategy for 3
North Middlesex University, Neonatal and Peadiatric Unit-3, London N18 1QX,
controlled ovarian stimulation (COS) in order to minimise face-to-face interac- United Kingdom
tions but retain predictive power for both trigger timing and over response? 4
University Hospital Crosshouse-, Ayrshire Fertility Unit-, Kilmarnock- KA2 0BE-
Summary answer: Data from follicle tracking scans on day 8, 9 or 10 can be Scotland, United Kingdom
used to make accurate predictions of trigger timing and risk of over-response. 5
FundacionRespirar-, Pulmonologist- Head of Clinical Research-, Av Cabildo 1548
What is known already: British Fertility Society guidance for restarting ART 1st A. Buenos Aires., Argentina
following Covid-19 pandemic related shutdowns recommends aiming to reduce 6
University medical centers Amsterdam- location VUmc and AMC-, Research
the number of visits for monitoring during ovarian stimulation. If reduction is institute Reproduction and development-, De Boelelaan 1081- 1081 HV
needed, prioritising attendance for scans on the most predictive cycle days is Amsterdam, The Netherlands
prudent. Current evidence on optimal monitoring during ovarian stimulation is 7
University College London Hospital Trust-, NIHR Biomedical Research Centre-,
sparse, and protocols vary significantly, with some centres priding themselves 250 Euston Road. London NW1 2PG, United Kingdom
on monitoring intensity. Small studies of simplifying IVF therapy by minimising
monitoring have reported no adverse effects. There are opportunities to learn Study question: Is there vertical transmission (from woman to baby antenatally
from the adaptations necessary during these extraordinary times to improve or intrapartum) after SARS-CoV-2 (COVID-19) infected pregnancy?
efficiency of IVF care in the longer term. Summary answer: SARS-CoV-2 (COVID-19) infected pregnancies leaves
Study design, size, duration: A retrospective database analysis of 9294 open the possibility of vertical viral transmission from mother to baby.
ultrasound scans performed during monitoring of 2322 IVF cycles undertaken What is known already: The majority of viruses do not cross the placental
by 1875 women in a single centre. The primary objective was to identify when barrier; but when they do, can cause serious fetal illness including birth defects,
in the IVF cycle data obtained from ultrasound is most predictive of both oocyte miscarriage, abnormalities of growth and development, neurological injuries,
maturation trigger timing and an over-response to stimulation. Ethical approval fetal demise, preterm delivery and neonatal complications. Such examples are
for this study was obtained from the University of Southampton ERGO II and Zika, CMV, rubella, HIV/AIDS viral infections in pregnancy. The expression of
NHS REC (IRAS Project ID: 275218). SARS-CoV-2 receptors angiotensin-converting enzyme 2 (ACE2) plays essential
Participants/materials, setting, methods: Anonymised retrospective roles in human infection and transmission and is found in maternal-fetal interface
database analysis of IVF/ICSI cycles at a tertiary referral IVF centre in the United and human fetal heart, liver and lung. SARS-CoV-2 vertical transmission from
Kingdom. Machine learning models built combining demographic and follicular mother to baby constitutes a major defining question in this pandemic to help
tracking data to predict oocyte maturation trigger timing and over-response. clinical practices and patients.
The primary outcome was the cycle days from which scan data yields optimal Study design, size, duration: A systematic review searching terms related
model prediction performance statistics. Random Forest Regressors were imple- to SARS-CoV-2 (COVID-19), pregnancy, neonatal complications, viral and ver-
mented as our predictive models, trained using cross-validation at treatment tical transmission was suggested. The final computerized database search was
cycle level and evaluated on the out-of-fold samples. performed on 30/04/2020. No language restriction was placed on published
Main results and the role of chance: The earliest day for which our model articles. The duration was from December 2019- 15th May 2020 for extracting
has high accuracy to predict both trigger day and risk of over-response ( >18 the literature and screening the articles for potential interest. As data source
follicles >=11mm or >=18 eggs) is cycle day 8. The model using day 8 data is was international there was considerable heterogeneity in patient exposures,
strongly predictive of the day of trigger administration MSE (mean squared error) quality control and possibly variants of SARS-CoV-2 virus.
1.70 +/-0.11. At cycle day 8, our model can predict over-response with high Participants/materials, setting, methods: A systematic review was per-
precision and recall (AUROC of 0.91 +/- 0.01). The model for predicting trigger formed in PUBMED, EMBASE, CENTRAL, WEB of SCIENCE, Web of

Knowledge, the WHO, RCOG, ESHRE, ASRM, NEJM, BMJ, Lancet, Welcome We recently pioneered genome-wide analysis of DNA methylation in single
and Cochrane Central Register of Studies, UKOSS, Office of National Statistics cells, as well as methods for joint profiling of DNA methylation, gene expression
(ONS-UK) , Department of Health (UK), Google Scholar and any references and chromatin accessibility. We are deploying these capabilities to explore effects
of relevant articles. Data relevant to SARS-CoV-2 diagnostic interpretation was of factors such as maternal age and maternal diet on the epigenetic quality of the
also collected. Mostly small case series with a high degree of heterogeneity was oocyte, and to investigate whether any anomalies detected persist in the embryo.
assembled on an excel spread sheet. In addition, with detailed knowledge of the mechanisms underlying DNA meth-
Main results and the role of chance: From 58 papers, 480 COVID-19 ylation in the oocyte, we are keen understand how such epigenetic anomalies arise.
positive pregnant women were identified and 688 babies born covered in 80
publications. Most publications (47/80, 58.7%), reported small number (<5) of
babies. From 71 papers, 10 babies were COVID-19 positive. 1st RT-PCR diag-
nostic tests were done in 356 babies, 2nd RT-PCR was done in 90 cases, IgM INVITED SESSION
tests done in 28 babies, and IgG tests done in 28 babies. From 71 studies and SESSION 38: LABORATORY SESSION - TIME-LAPSE IN 2020
356 babies, 13 tested positive in 1st RT-PCR. After 2nd RT-PCR, 7 became neg- 07 July 2020 Parallel 3 11:45 - 12:45
ative, whereas 3 babies testing negative on 1st RT-PCR became positive. On 2nd
RT-PCR, 3 babies became positive (who were initially reported as negative in
1st RT-PCR), 3 continued to remain positive (positive in 1st RT-PCR) but 3 who
were positive in 1st RT-PCR were not tested again. On the 1st RT-PCR, 44 studies
reported time of testing while 27 studies did not. Earliest RT-PCR positive test O-150 Time-lapse technology: Lights and shadows of clinical
was 2 hours while in 2 studies, 10 women tested had positive amniotic fluid, and evidence
3/11 placental swabs tested positive for SARS-CoV-2 RNA but babies remained A. Ahlstrom1
negative. Changes to placental pathology reported. 1
Livio Fertilitetscentrum Gothenburg, IVF Laboratory, Sweden
In 4 unrelated report, 100% of patients initially tested negative, turning positive
after 2nd and 3rd re-tests for SARS-CoV-2 RNA. O-151 How TLT is changing our understanding and research on
Limitations, reasons for caution: Caution should be exercised in interpret- embryo development?
ing test results; sampling methods, timing, patient infectivity level, SARS-COV-2 T. Freour1
viability tests, sensitivity and specificity. Controls are generally lacking in reports. 1
CHU de Nantes, Médecine de la Reproduction, France
Small, globally heterogeneous cases reported. Voluntary case reports submission
to research databases are heterogeneous, potentially breaching patient anonym-
ity and some clinics may withhold data.
Wider implications of the findings: When factoring in the incubation period SELECTED ORAL COMMUNICATIONS
and efficacy of the diagnostic test results, it suggests the viral infection pre-existed
SESSION 39: STRATEGIES TO IMPROVE THE OUTCOMES
the tests. Thereby it is likely that vertical transmission had occurred but validation
OF OVARIAN STIMULATION 2
and controlled experiments are needed to allow a definitive assessment to coun-
sel pregnant women on SARS-COV-2 pandemic risks. 07 July 2020 Parallel 4 11:45 - 12:55
INVITED SESSION O-152 Impact of different strategies in frozen cycles in normo

responding patients undergoing IVF/ICSI cycles: a multicenter
SESSION 37: MHR SYMPOSIUM - FUNDAMENTALS ON MAK- cohort study
ING OOCYTES
C. Siristatidis1, T. Arkoulis2, N. Christoforidis3, G. Salamalekis4,
07 July 2020 Parallel 2 11:45 - 12:45 N. Koutlaki5, V. Karageorgiou6, G. Galazios5
1
University of Athens, 3rd Department of Obstetrics and Gynecology, Athens,
Greece ;
2
Mitosis IVF Clinic, Private IVF Clinic, Piraeus, Greece ;
O-148 Major molecular players in creating oocytes from germ 3
Assisted Reproduction Unit- Embryolab, Private Clinic, Thessaloniki, Greece ;
cells 4
Medical School- National and Kapodistrian University of Athens, Assisted
J. Bowles1 Reproduction Unit- Third Department of Obstetrics and Gynecology- “Attikon
1
University of Queensland, Australia Hospital”, Athens, Greece ;
5
Medical School- Democritus University of Thrace, Assisted Reproduction Unit-
O-149 Influence of genetic and maternal factors on the egg Department of Obstetrics and Gynecology, Alexandroupolis, Greece ;
epigenome 6
Medical School- National and Kapodistrian University of Athens, Medical School-
G. Kelsey1, A. Galvao1, J. Castillo-Fernandez1, H. Demond1, National and Kapodistrian University of Athens, Athens, Greece
E. Herrera1
1
Babraham Institute, Epigenetics Programme, Cambridge, United Kingdom Study question: Do different modalities used for frozen-thawed embryo trans-
fer (FET) treatment cycles in normoovulatory patients undergoing IVF/ICSI are
Abstract text linked with different pregnancy rates?
Over the life course, the mammalian genome undergoes profound setting and Summary answer: Live birth was superior in HR cycles without GnRH ana-
resetting of epigenetic information. Germ-cell specification, gametogenesis, and logs and miscarriage was higher when progesterone was administered through
early embryo development are characterised by phases of widespread erasure gel vs. tablets.
and rewriting of DNA methylation. This extensive reprogramming largely pre- What is known already: A variety of strategies and regimens for FET have
vents transmission of epigenetic information across generations; however, repro- been used and proposed. Reports have been plenty, with no existing consensus
gramming events must also ensure correct genomic imprinting, an essential regarding the best protocol to choose or to prepare the endometrium, through
epigenetic mechanism that does depend on faithful, long-term persistence of a luteal plase “creation” or replacement. Crucial parameters for the right choice
gamete-derived methylation in the next generation. This underscores the impor- consist of control and flexibility in the timing of transfer, cost, and proper endo-
tance of understanding how methylation patterns are established in the germline metrial development, along with rates of pregnancy, miscarriage and live birth.
and the extent to which they may be influenced by adverse physiological or Currently, uncertainty exists concerning the type, adjunct or dose of regimen
environmental factors, or procedures applied in assisted reproduction. to offer, while many IVF Units use variable and mixed protocols.

Study design, size, duration: This is a four-center two-arm retrospective “flushing.” Primary analysis was based on the intention to treat: oocyte yield,
cohort study conducted at both University and Private Assisted Reproductive defined as collected mature oocyte rate, was higher (n=64/83, 77.1%) in the
Units in Greece, using data from 456 cycles with vitrification from 369 women flushing group compared to the aspiration only group (n=48/81, 49.3%, risk
during the last 5 years. difference (RD): 18.2% (95% CI 3.8 - 31.6%), p-value=0.02). In the flushing
Participants/materials, setting, methods: The following modalities were group, most oocytes were retrieved in the first 3 flushes (63/83, 75.8%).
analyzed: 1. Natural cycle (NC), using hCG as ovulatory trigger, without luteal Fertilization rate was higher in the flushing group (n=53/83, 63.9% versus
support (Group 1), 2. NC, using hCG, with luteal support (Group 2) 3. Hormone n=38/81, 46.9%; RD: 16.8% (96% CI 1.5 - 31.4%), p= 0.045). Transfer rate
Replacement cycle (HRC) plus GnRHa suppression (Group 3) 4. HRC without was also higher in the flushing group (n=52/83, 62.7% versus n=38/81,
GnRHa suppression (Group 4). The primary outcome measures were live birth 46.9%; RD: 15.71 (95% CI 0.3 - 30.3%), but the difference was not significant
and miscarriage rates. (p= 0.06). The clinical pregnancy rate per transfer was not significantly dif-
Main results and the role of chance: From all four centers, data for 456 ferent between the two groups. The median duration of the intervention was
cycles through vitrification from 369 women was provided. Live birth was higher significantly shorter with “aspiration only” (0.43 min; IQR 0.3, 0.5) versus
in Groups 2 and 3 as compared to Group 4 (by 81%), and when the stable “flushing” (2.38 min; IQR 2.0, 2.7; p<0.001). There was no significant differ-
estrogen dose was administered compared to the increasing, by 8.82 times (1.25, ence in the mean VAS pain score between the follicular flushing (3.4 ±1.8)
62.03). Regarding miscarriage, 19 events were recorded; progesterone admin- and the aspiration group (3.1 ±1.89).
istration through gel was significantly associated with a higher risk, compared to Limitations, reasons for caution: Sample size was not powered to analyse
tabs [4.77 (1.80, 12.67)]. Regarding secondary outcomes, biochemical and clinical pregnancy and live birth rates.
pregnancy rates were lower in Group 3 compared to Group 4 [0.70 (0.6, 0.82) Wider implications of the findings: Our study proved that flushing of single
and 0.70 (0.61, 0.80), respectively] and when the stable estrogen dose was follicles in Natural Cycle IVF increases the oocyte yield. In contrast to polyfol-
administered compared to the increasing [0.83 (0.72, 0.96) and 0.87 (0.77, 0.97), licular and oligofollicular IVF flushing seems to be beneficial in the monofollicular
respectively]. The rest of the comparisons yielded non-significant results. In the setting if the technique used in our study (single-lumen needle, 5 flushings with
multivariable analysis, live birth appeared to be significantly affected only by the flushing volume adaptation) is applied.
endometrial preparation modality: Group 3 experienced higher rates compared Trial registration number: NCT 02641808.
to Group 4 (6.51 (2.66, 15.93)].
Limitations, reasons for caution: Limitations of the study include its ret- O-154 Do we trust in evidence based medicine? A multicentre
rospective nature that is linked with known and unknown biases and the small retrospective analysis of 2677 first IVF/ICSI cycles before and after
cohort size. the OPTIMIST trial.
Wider implications of the findings: HRC without GnRHa and probably E. Papaleo1, A. Revelli2, M. Costa3, M. Bertoli4, S. Zaffagnini5,
NC with hCG and luteal support appears to be superior to HRC with GnRHa, F. Tomei6, F. Cantatore1, M. Reschini7, A. Rebecchi1, F. Parissone5,
concerning live birth. Miscarriage was increased in women receiving progester- M. Manno6, M. Sironi4, T. Tessari4, D. Colia3, E. Somigliana7
one gel compared to tabs. Age was a significant predictor of negative biochemical 1
S. Raffaele Hospital, Centro Scienze della Natalità, Milano, Italy ;
and clinical pregnancy rates. 2
University of Turin, IVF unit, Turin, Italy ;
International Evangelic Hospital, IVF Unit, Genoa, Italy ;
4
Azienda Ospedaliera “C. Poma”, IVF Unit, Mantua, Italy ;
O-153 Follicular flushing leads to higher oocyte yield in 5
Azienda Ospedaliera Universitaria Integrata, IVF Unit, Verona, Italy ;
monofollicular natural cycle IVF – a randomized controlled trial 6
Ospedale Santa Maria degli Angeli, IVF Unit, Pordenone, Italy ;
A. Kohl Schwartz1, I. Calzaferri1, M. Roumet2, A. Limacher2, 7
Fondazione IRCCS Ca’ Granda- Ospedale Maggiore Policlinico, IVF Unit, Milan,
A. Fink1, A. Wueest1, S. Weidlinger1, V. Mitter1, B. Leeners3, Italy
M. Von Wolff1
1
Bern University Hospital, Division of Gynecological Endocrinology and Study question: To evaluate whether the practice of individualizing r-FSH
Reproductive Medicine, Bern, Switzerland ; starting dose has been substituted and outdated after the largest RCT in ART,
2
Clinical Trial Unit Bern, University of Bern, Bern, Switzerland ; the OPTIMIST trial.
3
University Hospital Zurich, Department of Reproductive Endocrinology, Zurich, Summary answer: OPTMIST trial influenced physicians to standardize r-FSH
Switzerland starting dose and to reduce the mean starting dose in predicted POR women
undergoing COS.
Study question: Does follicular flushing increase the number of mature What is known already: Individualizing r-FSH starting dose has been subject
oocytes in monofollicular natural cycle in-vitro fertilization? of many studies over the past 20 years. Although the “one size fits all” approach
Summary answer: Follicular flushing increases the number of mature oocytes, has been discouraged for decades by most authors, van Tilborg et al. (2017) on
increases the fertilisation rate and reveals a trend towards a higher transfer rate. behalf of the OPTIMIST study group demonstrated in a large prospective RCT
What is known already: In polyfollicular IVF and in oligofollicular poor respond- that dosage individualization, in general, does not appear to improve overall live
ers, flushing neither increases the oocyte yield nor the pregnancy rate. In mono- birth rates. This implies that, while reduction of dosage may create better safety
follicular IVF the effect of flushing had so far been addressed by two studies. One in predicted high responders, the use of high dosages of r-FSH, typical of pre-
prospective study with minimal stimulation IVF demonstrated an increased oocyte dicted low responders, lacks any cost-benefit advantage and may be abandoned.
yield. One retrospective study with Natural Cycle (NC)-IVF showed an increased Study design, size, duration: We performed a retrospective analysis of
oocyte yield and an increase of the transfer rate. A prospective randomized study seven italian fertility centers including all first IVF/ICSI cycles from January 1st
specifically analysing monofollicular IVF was still missing. 2017 to December 31st 2018, before and after the OPTMIST publication on
Study design, size, duration: Prospective randomized controlled trial includ- November 2017. Patients were distributed according to their ovarian reserve
ing 164 women who were randomized for either aspiration without or with markers in: predicted POR (AFC ≤ 7 and/or AMH <1.1), predicted normo-
flushing from 2016-2019. responders (AFC 8-15 and/or AMH 1.1-3.5) and expected hyper-responders
Participants/materials, setting, methods: Infertile women 18-42 years of (AFC > 16 and/or AMH > 3.5).
age with an indication for IVF treatment at a university-based infertility unit. Participants/materials, setting, methods: 2677 patients between 18 and
Women undergoing monofollicular NC-IVF in vitro fertilization were randomized 42 years old undergoing their first IVF-ICSI cycle were included. The study group
to either follicular aspiration only or follicular aspiration directly followed by five A (before OPTIMIST) included 1325 patients who underwent their first cycle
follicular flushes at a 1:1 ratio. The intervention was done without anaesthesia, from January 2017 to December 2017. The study group B (after OPTIMIST)
using a gauge 19 single lumen needle. Flushing volume was calculated (sphere included 1352 patients who underwent their first cycle from January 2018 to
formula) based on the size of the follicle. December 2018. In both study groups data regarding patients characteristics
Main results and the role of chance: A total of 164 women were (age, BMI, AMH, AFC,), stimulation protocols and stimulation outcomes were
included; 81 were allocated to “aspiration only,” and 83 to additional collected.

Main results and the role of chance: In the study group “before” ; AMH - 3.0 ± 2.14 ng /ml 2,72±0,41 ng / ml, p=0.693440). Antral follicle count
401/1325 (30, 2%) patients were expected POR, 566 (42, 7%) expected (AFC) were significantly lower in the II group 9,2±1,2; 5,8±1,14, p=0.046535).
normo-responders, 358/1325 (27, 0%) predicted hyper-responders. In the There were no differences in the mean number of oocytes retrieved in the
study group “after” patients’ distribution was 366/1352 (27.0%), 601/1352 follicular phase group, and luteal phase group (10,33 ±2,57 versus 11,62±1,64;
(44.4%), 385/1352 (28.4%) for each group (p=NS). Comparing 2017 and p=0.676949). Similarly, no significant differences were observed in the number
2018 there was an increased prescription of standard 150UI proposed by of M II occytes (6,9±1,14 versus 8,92± 0,94; p=0.179224), stimulation duration
OPTIMIST trial, from 20.8% to 24.1% (p=0.046). Interestingly, we also (12,6±0,6; 13,5±1,1, p=0.476761), initial (243,06 ± 12,96 IU versus 233,33 ±
observed a significant reduction of overall use of starting dose >300UI from 20,73 IU, p=0.694155) and total gonadotropin dose (2635,44 ±187,75 IU versus
10.3% in 2017 to 5.6% in 2018 (p <0.01). In predicted POR undergoing their 2145,83 ±277,72 IU, p=0.158947). Estradiol (E2) and progesterone (P) levels
first COS, the mean FSH starting dose reduced from 317UI to 302 UI (p< 0.03). on the day when the ovulation stimulation was started were different among the
This approach did not determine any differences in terms of oocytes retrieved two groups (211,62±40,19 pmol/L versus 693,47±151,04 pmol/L,
in the before and the after group respectively (4.21 ± 3.1 vs 4.22 ±3.4, p<0.05;1,13±0,1 nmol/L versus 22,44±9,97 nmol/L, p<0.05). Perhaps due to
p=0.258), metaphase II oocytes when ICSI was used (3.04± 2.4 vs 3.2 ± 2.7, the regression of the corpus luteum, we observe comparable levels of E2 and
p=0.43) or total number of oocytes fertilized (IVF/ICSI) (3.0 ±2.6 vs 2.8 ± P in both groups on the 6th day of ovarian stimulation and on the day of the
2.59, p=0.42). Cumulative pregnancy rate was also comparable between the trigger administration. Premature LH surge did`t observe in the absence of
two years (20.3% vs 20.9% in 2017 and in 2018 respectively, p=0.84). GnRH-ant administration in the II group.
Limitations, reasons for caution: Retrospective nature of the study. Overall Limitations, reasons for caution: The current work is a pilot study. Larger
cumulative pregnancy rate is still partly pending at the time of data collection, prospective trials are planning.
because a proportion of non-pregnant patients (mainly hyper- and normo-re- Wider implications of the findings: Our data allow us to talk about the
sponders and mainly treated in 2018) might return in the future for frozen comparable effectiveness of ovarian stimulation of the follicular and luteal phases
embryo transfer attempts. of the cycle. Modern knowledge on folliculogenesis lets us to optimize strategies
Wider implications of the findings: Our clinical results confirm that EBM of ovarian stimulation not only from the point of flexibility.
provided by the OPTIMIST trial has somehow influenced our ART daily practice, Trial registration number: None
bringing to a reduction of starting doses without limiting outcomes. On the other
hand, adherence to the OPTIMIST indications is still generally poor.
Trial registration number: not applicable O-156 Pituitary supression is not necessary for blocking LH surge
during luteal-phase stimulation
O-155 Experience in random-start ovarian stimulation in cancer M. Cruz Palomino1, E. Henzenn1, A. Requena1
patients 1
IVI Madrid, Reproductive Medicine, Madrid, Spain
Y. Martirosyan1, T. Nazarenko, Alekseevna2, A. Birukova,
Mikhailovna1, L. Dzhanashvili , Georgievna1 Study question: Can we avoid the administration of pituitary suppressors
1
National Medical Research Center for Obstetrics- Gynecology and Perinatology during a luteal phase stimulation without affecting the ovarian response?
named after V.I. Kulakov- Ministry of Health of Russia, Research and educational Summary answer: In absence of pituitary supressors during luteal-phase
center for assisted reproductive technologies named after F. Paulsen-senior, stimulation, it is posible to block a physiological LH surge without impacting
Moscow, Russia C ; normal ovarian response
2
National Medical Research Center for Obstetrics- Gynecology and Perinatology What is known already: New stimulation approaches allow for a total dis-
named after V.I. Kulakov- Ministry of Health of Russia, the Research and articulation between the time of the menstrual cycle, ovarian stimulation start
educational center for assisted reproductive technologies named after F. and embryo transfer. Double stimulation (DuoStim) was initially designed to
Paulsen-senior, Moscow, Russ optimize clinical outcomes in poor ovarian response, but it could be also useful
in fertility preservation for non-medical reasons, especially for oocyte/embryo
Study question: What is the effectiveness of ovarian function stimulation in accumulation. Pituitary suppressors block the LH surge; however, in a protocol
the luteal phase of the cycle compared with the follicular phase in patients as specific as DuoStim, it could be assumed that these suppressors are not
underwent fertility preservation? necessary in the luteal phase because the endogenous progesterone released
Summary answer: Ovarian stimulation in the luteal phase allows us to opti- during follicular phase is sufficient to block the LH surge during the luteal phase
mize strategies of ovarian stimulation in cancer patients without compromising Study design, size, duration: Prospective and observational analysis per-
the oocyte yield. formed in IVI Madrid between September and December 2019. Participants
What is known already: The choice of ovarian stimulation protocol for were randomly assigned to each of the study groups. Participants underwent
cancer patients is based on a balance between two main factors: the limited time the same stimulation protocol in the follicular phase and for luteal phase stimu-
available for fertility preservation (FP) before the start of gonadotoxic therapy, lation, they were allocated in a control group with pituitary suppressors (n=10)
and the need to obtain a sufficient number of suitable oocytes and embryos for or in a study group, where this medication was not administered (n=10).
cryopreservation. Despite data about comparable efficacy of the ovarian stim- Statistical analysis was performed by ANOVA and Chi-squared where applicable.
ulation with GnRH-ant in the follicular phase of the cycle and stimulation in the Participants/materials, setting, methods: Follicular-phase stimulation
luteal phase, many questions remain open. was the same for both study groups; daily tablet of 10 mg of acetate of
Study design, size, duration: We performed a prospective observational study medroxyprogesterone (AMP) from first day of stimulation, 225 IU/day ecom-
at the National Medical Research Center for Obstetrics, Gynecology and Perinatology binant FSH and triggering with 0.1 mg GnRH agonist. For the control group,
named after V.I. Kulakov from February 2019 to December 2019. A total of 160 luteal-stimulation is identical to the previous one; and in the study group, the
women who had FP consultation prior to neoadjuvant chemotherapy were identified. only difference is that AMP was not administered daily from the start of the
47 patients were enrolled for the study according to inclusion criteria. stimulation. LH, estradiol and progesterone were monitored during
Participants/materials, setting, methods: Depending on the phase of the luteal-phase.
menstrual cycle when ovarian stimulation was started, two groups of patients Main results and the role of chance: As expected, and in the case of a
were identified: I - ovarian stimulation started in the early follicular (n=34) and homogeneous population such as oocyte donors, no differences were observed
II - luteal phase group (n=13). between the two study groups in follicular-phase stimulation, either with respect
We measured the number of retrieved oocytes and mature oocytes, stimu- to endocrine profile or ovarian response. For control and study group respec-
lation duration, initial and total gonadotropin dose, the parameters of ste- tively, the results were as follows: basal LH (6.11±1.6 IU vs 6.6±2.4 IU, p=0.680);
roidogenesis. Differences were tested using a two-sided Student’s t-test or a LH on the triggering day (3.1±1.8 IU vs 2.3±0.7 IU, p=0.548); progesterone on
Pearson’s Chi2 test, as appropriate. the triggering day (1.4±0.3 ng/ml vs. 1.1±0.1 ng/ml, p=0.180); retrieved oocytes
Main results and the role of chance: All patients included in the study were (16.7±3.0 vs. 20.1±5.4, p=0.389);and metaphase II oocytes (14.2±3.5 vs.
comparable in age and AMH level (33.38 ± 3.73; 33.3 ± 5.47 years, p=0.967580 16.7±4.5, p=0.496).

These results are maintained for the luteal-phase stimulation, meaning that Study design, size, duration: Countries and regions annually collect ART
the endogenous profile in the stimulated folicular-phase is capable of inhibiting data, some prospectively and others retrospectively. ICMART retrospectively
LH surge, not affecting the results derived from this second stimulation. For requested these data from all known global sources for 2016 and reviewed them
control and study group respectively, the results were as follows: basal LH for missing or incorrect data. The dataset was corrected and then analyzed
(1.6±1.3 IU vs 1.7±0.6 IU, p=0.335); LH on the triggering day (0.5±0.4 IU vs utilizing standardized definitions from The ICMART/WHO Revised Glossary
1.5±0.6 IU, p=0.300); progesterone on the triggering day (0.6±0.1 ng/ml vs. on ART Terminology, 2009 which was current at the time but is now replaced
0.4±0.1 ng/ml, p=0.398); retrieved oocytes (14.5±1.6 vs. 12.8±2.1, by The International Glossary on Infertility and Fertility Care, 2017, and previ-
p=0.575);and metaphase II oocytes (11.7±1.0 vs. 10.4±2.3, p=0.609). ously developed methods. Preliminary results are presented. China results are
Finally, it should be noted that no rescue protocol with administration of summarized and compared with other global data.
GnRH antagonist was applied in the study group, because of ovulation risk Participants/materials, setting, methods: The European IVF Monitoring
Limitations, reasons for caution: These results could be considered as an Consortium (EIM), Latin American Network of Assisted Reproduction
interim analysis, as they are framed within a pilot study prior to conducting a (REDLARA), Australian/ New Zealand Registry and ANARA submitted regional
larger study so, although the current data are encouraging, we are not able to data, and other countries contributed national data, through standardized for-
draw solid evidences due to our small simple size mats to ICMART. A few individual clinics with no registry access also contributed.
Wider implications of the findings: In a certain group of patients such as Data were reviewed, corrected, validated to the extent possible, analyzed and
oocyte donors, double stimulation implies advantages such as the possibility of summarized by ICMART using descriptive statistics. China CDC and National
achieving more oocytes in less time, optimizing the economic profitability of the Health Commission published their data in Human Reproduction.
egg donation program without compromising clinical results becuase of the Main results and the role of chance: Data collection and analysis are
absence of pituitary supresor during luteal-phase. ongoing, so presented results are preliminary. The number of ART cycles con-
Trial registration number: Not applicable tinues to increase, but utilization is still highly variable among countries and
regions. Regional and country differences persist in the age of the population
treated, number of embryos transferred, rate of multiple births, use of ICSI,
cryopreservation cycles and other factors.
INVITED SESSION The role of chance is minimal. Actual global ART results are limited to report-
SESSION 40: GLOBAL ART MONITORING ing countries and clinics representing approximately 2/3 of global cycles.
However, this is a very large sample size from which imputation of total global
07 July 2020 Parallel 2 14:00 - 14:45
results is performed. Addition of data from China improves significantly global
reporting of ART data. China results are not different than overall global data.
Limitations, reasons for caution: Some countries have limited data and
many countries have limited data validation. ICMART can perform only minimal
O-157 ICMART preliminary world report 2016 verification of submitted data. Widespread adherence to consensus definitions
G.D. Adamson1, S. Dyer2, G. Chambers3, O. Ishihara4, provided in the Glossary takes time and requires translation into multiple lan-
R. Mansour5, M. Banker6, J. de Mouzon7, M. Kupka8, guages. China data has some similar and different limitations.
F. Zegers-Hochschild9 Wider implications of the findings: ICMART World Reports standardize
1 data, track trends, enable comparisons, stimulate questions and improve ART
Stanford University School of Medicine, Reproductive Endocrinology and Infertility,
quality. Better understanding of ART increases societal acceptance and support
Palo Alto, U.S.A.
2 for equitable access and ART research. Addition of China data is an important
Groote Schuur Hospital and Faculty of Health Sciences, Reproductive Medicine
milestone in global ART data reporting.
Unit, Cape Town, South Africa
3 Trial registration number:
University of New South Wales, School of Women’s and Children’s Health
Study funding:
SWCH and Centre for Big Data Research in Health CBDRH, Sydney, Australia
4 Funding source:
Saitama Medical University, Department of Obstetrics and Gynecology, Saitama,
Japan
5 O-158 SET in a global perspective. Regional similarities and
The Egyptian IVF-ET Center, Reproductive Medicine, Cairo, Egypt
6 differences
Pulse Women’s Hospital, Reproductive Medicine, Ahmedabad Gujarat, India
7
Inserm, Reproductive Medicine, Paris, France O. Ishihara1, F. Zegers-Hochschild2, J. de Mouzon3, S. Dyer4,
8
Ludwig-Maximilians-University, Reproductive Medicine, Munich, Germany R. Mansour5, M. Banker6, G. Chambers7, D. Adamson8
1
9
Universidad Diego Portales, Program of Ethics and Public Policies in Human Saitama Medical University, Department of Obstetrics & Gynaecology,
Reproduction, Santiago, Chile Iruma-gun- Saitama, Japan
2
University Diego Portales, Clinica las Condes and Program of Ethics and Public
Abstract text Policies in Human Reproduction, Santiago, Chile
Abstract title: International Committee for Monitoring Assisted Reproductive 3
Epidémiology- research and development, Paris, France
Technologies (ICMART) Preliminary World Report on ART, 2016 4
Groote Schuur Hospital and Faculty of Health Sciences- University of Cape Town,
Study question: In 2016 what was global utilization, effectiveness and Department of Obstetrics & Gynaecology, Cape Town, South Africa
safety of ART? 5
Egyptian IVF-ET Center, Egyptian IVF-ET Center, Cairo, Egypt
Summary answer: Globally, ART utilization and data collection continue to 6
Nova IVI Fertility, Nova IVI Fertility, Ahmedabad, India
increase but with wide variations in utilization, effectiveness and safety. 7
University of New South Wales, National Perinatal Epidemiology and Statistics
What is known already: ICMART began ART global data collection in 1991. Unit- Centre for Big Data Research in Health- School of Women’s and Children’s
Utilization, effectiveness and safety have continuously improved with more Health, Sydney, Australia
cycles, higher pregnancy rates and lower multiple birth rates, the latter as a result 8
Equal3 Fertility, Equal3 Fertility, Cupertino, U.S.A.
of transfer of fewer embryos. Frozen embryo transfer (FET) and donor egg
cycles continue to increase. However, wide variations in practice and outcomes Study question: How is the transition to SET being realized in a global per-
exist globally. Over 8 million ART babies have been born. ICMART has helped spective? What are key issues for policymaking regarding the number of embryos
develop registries internationally, notably with the African Network and Registry transferred in various countries in the world?
for Assisted Reproductive Technology (ANARA). A new electronic data collec- Summary answer: The global SET rate significantly rose during the last 10
tion platform is being developed; nevertheless, data collection and quality remain years, resulting in safer ART outcomes with lower multiple pregnancy rates and
challenging. China has recently published its first comprehensive report on better infant indicators. However, different policies on SET resulted in some
assisted reproductive technology service availability, efficacy and safety in main- countries and regions continuing to have very high multiple rates. It is important
land China: 2016. to consider the use of multiple strategies that have successfully been introduced

in some countries to encourage the transition to SET and have resulted in provide, both practically and biologically. Finally, we will discuss the goals and
safer ART. needs for integration of microfluidics for cell manipulation, insemination, culture,
What is known already: Decreasing the number of embryos transferred at sampling, assays, and cryopreservation. This integration will facilitate automation
ART strongly correlates with better outcomes of infants, namely, lower multiple within the clinical andrology/embryology laboratory and will have enabling influ-
pregnancy rates, lower preterm rates and fewer low birthweight infants. ences on other evolving technologies such as machine learning and artificial
Acceptance of SET has been variable in regions and countries due to multiple intelligence for data collection and decision making in the IVF lab. These are not
factors. trivial goals and will require significant design, testing, and multidisciplinary inter-
Study design, size, duration: Retrospective analysis of the accumulated actions to be successful in the next generation of andrology/embryology prac-
ICMART world ART registry data between 2002 and 2014 which had 601,243 tical laboratory patient care.
and 1,647,777 initiated cycles reported, respectively, to determine the trend of
average number of embryos transferred, the SET rates, and their outcome
parameters by countries. Individual country data are also used for selected
counties. INVITED SESSION
Participants/materials, setting, methods: ICMART world ART registry SESSION 42: STRESS AND INFERTILITY - THE CHICKEN OR
data annually reported by contributors from regional or national registries from THE EGG
53-77 countries between 2002 and 2014.
Main results and the role of chance: The global average number of fresh 07 July 2020 Parallel 4 14:00 - 15:00
embryos transferred and the SET rate were 2.47 and 12.4% in 2002 and improved
to 1.73 and 40% in 2014. In FET cycles the results were even better at 1.43 and
61.6% in 2014. In response to the decreased number of embryos transferred,
global multiple pregnancy rates in fresh non-donor IVF and ICSI cycles dropped O-161 Just relax and it will happen?: the case for stress causing
from 28.2% (triplet 2.5%) in 2002 to 16.7% (triplet 0.5%) in 2014. However, lack of pregnancy
countries can be grouped into 3 categories, those: 1) continuously maintaining J. Boivin1
highest SET rate; 2) achieving higher SET rate during the period; 3) continuing 1
Cardiff University, School of Psychology (Cardiff Fertility Studies Research Group),
to have low SET rates throughout the period. The highest countries have SET United Kingdom
rates of approximately 80% while other countries have only 10-20% with multiple
pregnancy rates of approximately 30%.
O-162 Just relax and it will happen?: The case against stress as a
Limitations, reasons for caution: The data accumulated by ICMART from
cause of infertility
different countries are of variable quality and comprehensiveness. Additionally,
it is not possible for ICMART to validate the data. A. Lawson1
1
Wider implications of the findings: These trends are important to follow. Northwestern University, Obstetrics & Gynecology and Psychiatry, Chicago,
Clinical practices regarding number of embryos transferred are affected by cul- U.S.A.
tural values, societal financial support of ART, changes in technology such as Abstract text
PGT-A, and other factors. “Just relax. It will happen.” It’s a phrase commonly heard by women struggling
to get pregnant. Although the statement is likely intended to let women know
that others are optimistic about their chances of having a baby, at its core it
blames women for being too stressed to conceive. For generations, many have
INVITED SESSION believed that psychological stress caused physiological changes that interfered
SESSION 41: ALMER EXCHANGE SESSION - IVF LABORA- with a woman’s chances of conceiving. Although anecdotal stories of women
TORY AUTOMATION relaxing and getting pregnant have been widely shared, rigorous research exam-
ining the role of stress in the etiology of infertility is limited. What little research
07 July 2020 Parallel 3 14:00 - 15:00
exists does not support a direct link between the relationship between stress
and infertility.
If, as hypothesized, psychological stress/distress can interfere with fertility
then it could be argued that activities that produce a relaxation response could
O-159 Cons improve pregnancy chances. Indeed multiple studies have examined the positive
C. Racowsky1 effects of relaxation or relaxing activities and the cessation of stress inducing
1 activities (e.g, fertility treatment) on both fertility and the reduction of emotional
Brigham & Woman’s Hospital, Department of Obstetrics & Gynecology ASB 1 + 3
distress. However, these studies are also often not appropriately designed to
O-160 Pros rigorously answer the question about whether or not a relationship between
G.D. Smith1 relaxing and pregnancy chances exists. Because controversy exists regarding the
1 relationship (or lack thereof ) between stress and infertility, inconsistency in
6428 Medical Science I University of Michigan, Departments of OB/
counseling of patients regarding the need for stress reduction is rampant. This
GYN- Physiology- and Urology, Ann Arbor- MI, U.S.A.
presentation will address the biological and psychological issues in stress and
Abstract text infertility and will provide direction to providers for how to address these issues
Andrology and embryology laboratory practices have evolved over the last 4-5 with patients.
decades, ultimately leading to increased technologies, increased personnel work
hours per cycle, and increased success in treating infertility and preserving fertility.
The manual labor component of andrology/embryology laboratory is intensive
with many technologies requiring high degree of training and subject to technol-
ogist variability, drift, and retraining. Integration of new principles of bioengi- SESSION 43: ICSI IN 2020
neering, microfluidics, and automation have been beneficial in other areas of 07 July 2020 Parallel 1 15:15 - 16:30
everyday life, science, and medicine. In this presentation we will provide exam-
ples of scientific and social automation and discuss potential benefits. The inte-
gration of microfluidics for mechanical benefit and microenvironmental/
physiological benefit will be proposed. Discussions will focus on proof-of-prin- O-163 Advancement in the future automation of ICSI: Use of
ciple applications of microfluidics and automation to andrology, embryology, deep convolutional neural networks (CNN) to identify precise
and cryobiology as they relate to laboratory tasks and the potential benefits they location to inject sperm in mature human oocytes

J. Dickinson1, A. Meyer1, N. Kelly1, P. Thirumalaraju2, Summary answer: After applying AOA the ongoing pregnancy rates of the
M. Kanakasabapathy2, D. Kartik2, C. Bormann1, H. Shafiee3 patients per initiated treatment were enhanced, as well as a significant reduction
1
Massachuesetts General Hospital, Obstetrics and Gynecology, Boston, in the cancellation rate.
U.S.A. ; What is known already: Different authors have demonstrated better results
2
Brigham and Women’s Hospital, Division of Engineering in Medicine- Department after using AOA in couples with low or non-fertilization in previous treatments:
of Medicine, Boston, U.S.A. ; (Heindryckx et al., 2005, Heindryckx et al., 2008, Montag et al., 2012). Recently,
3
Brigham and Women’s Hospital, Department of Medicine- Harvard Medical we have seen extensively described similar benefits by the use of AOA for
School, Boston, U.S.A. couples with previous standard ICSI failure (Fawzy et al., 2018). The implemen-
tation of this technique in the current clinical routine has been possible after
Study question: Can a deep-learning artificial intelligence algorithm be used reporting no detrimental impact on neither human gametes (Ebner et al., 2015)
to accurately identify the appropriate location to perform ICSI on metaphase II nor on the offspring (Vanden Meerschaut et al., 2014).
(MII) human oocytes? Study design, size, duration: Retrospective cohort study, from four con-
Summary answer: The AI trained network can be used to accurately identify secutive years. We studied 509 oocytes from 66 patients who were treated with
the correct location on the oocyte to inject sperm for ICSI. standard ICSI, and were compared to 616 oocytes from the same cohort of
What is known already: ICSI is a procedure that includes, alignment of MII patients but using AOA.
oocytes, selection and immobilization of sperm, and injection of sperm at a Participants/materials, setting, methods: 66 patients were included in
precise location that does not interfere with the mitotic spindle. The spindle is the study generating 163 cycles; standard group was composed of 509 oocytes
located adjacent to the extruded polar body (PB) and cannot be visualized using generating 75 cycles (18 fresh cycles, 4 frozen cycles, 1 mixed and 52 cancelled
brightfield microscopy. Therefore, it is standard practice to align oocytes based cycles). AOA group included 616 oocytes resulting from the same patients,
on the location of the PB and to inject sperm 90° from this visible structure. but creating 88 cycles (31 fresh cycles, 37 frozen cycles). AOA technique
The goal of this study was to train an artificial intelligence (AI) algorithm to involves oocyte injection by spermatozoa and incubation for 10 minutes with
identify the position of the PB and the corresponding location for sperm calcium ionophore. Outcome analysis included a multivariable logistic regres-
injection. sion model.
Study design, size, duration: Using a retrospective dataset of denuded MII Main results and the role of chance: Although no differences were
oocytes, a deep CNN model was trained and tested to classify between 12 observed in relation with the day of transfer (Day 3 vs Day 5) and maternal age
classes at 4-6 hours post oocyte retrieval. The twelve classifications resembled at the time of the cycle, we performed a multivariable logistic regression including
the pattern of digits on a clock, spaced 30° apart to provide an accurate location those variables as potential bias factors. The study was performed to assess the
the extruded PB. impact of AOA on ongoing pregnancy per cycle as well as per transfer. We
Participants/materials, setting, methods: We developed a deep convo- observed that the application of AOA in our patients increased the chances of
lutional neural network that was trained with 13992 annotated images of MII a viable pregnancy by more than 4 times (OR=4.57, p=0.008) per cycle but not
oocytes. We classified the location of the PB and corresponding location for per transfer (OR=0.964, Not significant). When embryos were available for
sperm injection. The validation set containing 1920 oocyte images served to transfer AOA did not increased the chances of a viable pregnancy.
ensure the program training was complete. The developed network was evalu- Limitations, reasons for caution: The retrospective analysis of this study
ated using another independent set of 3900 MII oocyte images with known PB may be a reason to take into consideration. Another limitation was not to study
and sperm injection location classifications. the PLCƺ levels of the sperm, then we presumed that this dysfunction was
Main results and the role of chance: The deep learning CNN was able to presented in the spermatozoa and, in consequence, was the responsible for the
correctly identify the location of the PB and corresponding location for sperm fertilization failure.
injection with 98.9% accuracy with a 95% confidence interval (CI) ranging Wider implications of the findings: Our findings suggest the use of AOA
between 98.5% to 99.2% (n=3900). Furthermore, a receiver operator charac- for a particular population where the fertilization was failed in previous attempts.
teristic (ROC) revealed micro and macro area under the curves (AUC) of 1, After AOA application, the fertilization rate was enhanced, increasing the chances
which confirmed that the AI can accurately identify the position of the PB and of success per treatment. The use of AOA is comforting after checking non-ex-
the corresponding location for sperm injection. istence of detrimental impact on the offspring.
Limitations, reasons for caution: Only MII oocytes were used in this trial. Trial registration number: none
Images were obtained using a single imaging platform (EmbryoScope) at a single
timepoint. All cumulous cells were removed from the oocytes prior to training.
It is unclear whether residual cumulous cells would affect the accuracy of the O-165 Toward an ICSI chip: automated microfluidic oocyte
algorithm. denudation module
Wider implications of the findings: Advanced ICSI automation requires A. Mokhtare1, P. Xie2, A. Abbaspourrad1, Z. Rosenwaks2,
precision that cannot be achieved using tracking algorithms. Automation requires G. Palermo2
complex decisions that can be achieved using deep-learning technologies. Prior 1
Cornell University, Department of Food Science and Technology, Ithaca, U.S.A. ;
studies showed ability for CNNs to identify morphologically normal sperm. This 2
study compliments this work by accurately identifying the correct position to Reproductive Medicine, New York, U.S.A.
inject sperm into oocytes.
Trial registration number: not applicable Study question: To determine whether novel microfluidic technologies can
improve the efficiency and quality of an oocyte denudation process for intracy-
toplasmic sperm injection (ICSI).
O-164 The application of AOA-artificial oocyte activation in Summary answer: The inevitable exposure of oocytes to high mechanical
patients with previous unsuccessful attempt increases the stresses, different oxygen concentrations, and temperature variations can be
ongoing pregnancy per treatment but not per transfer; a alleviated using an automated microfluidic oocyte denudation module.
multivariable study What is known already: A microfluidic device has recently been introduced
A. Galán Rivas1, L. Alegre1, M. Meseguer1, T. Viloria1, A. Pellicer2, in the clinical field and has proven successful in selecting competent spermatozoa.
A. Tejera1 Conventional cumulus removal (CR) techniques use high mechanical stress
1
Instituto Universitario IVI Valencia, IVF Lab, VALENCIA, Spain ; (pipette flushing) with enzymatic treatment or high fluidic stresses (vortexing)
2
IVIRMA Roma, Reproductive Medicine, Roma, Italy to treat cumulus-oocyte-complexes (COC) prior to ICSI. These operations not
only impose unfavorable stresses on oocytes, but also require a highly skilled
Study question: To check the effectiveness of AOA in patients with previous embryologist. Microfluidic technologies offer novel opportunities to streamline
very low or failed fertilization rates in terms of fertilization, implantation and ART to facilitate gamete preparation, reduce mechanical and cytotoxic stress,
ongoing pregnancy outcomes. and minimize inter-operator variability. Such automated and continuous systems

may replace manual CR operation while ensuring optimal environmental oocytes that IVF blastocysts start to expand 3 to 4 hours earlier than ICSI-
conditions. generated blastocysts. When standardizing for the time of pronuclear fading,
Study design, size, duration: We developed a semi-automated oocyte the differences in early cleavage disappear between IVF and ICSI, while blastu-
denudation microfluidic chip (ODMC) controlled through microcontrollers with lation and blastocyst expansion occurs earlier for IVF embryos.
a graphical user interface (GUI). The chip consists of a 2-layer microchannel, Study design, size, duration: Prospective cohort study between November
with channel width similar to the diameter of mouse COC (200 μm). The bottom 2018 and April 2019, including 568 oocytes (30 patients) with non-male factor
layer contains bas-relief structures to gently agitate the pressure-driven laminar infertility for which half of the sibling oocytes were inseminated with conventional
flow inside microchannels without imposing excessive stress on the oocytes. IVF (n=283) and the other half with ICSI (n=285). Embryos were cultured in an
Denudation rate and embryo developmental competence were compared Embryoscope time-lapse incubator and trophectoderm biopsy was performed
between oocyte cleaning by ODMC and manual pipetting. on good-quality blastocysts. The following timings were annotated: tPNf, t2-9,
Participants/materials, setting, methods: COCs were retrieved from 3 tSC, tM, tSB, tB, tEB, cc2(t3-t2), cc3(t5-t3), s2(t4-t3), s3(t8-t5) and Blast(tSB-t2).
superovulated B6D2F1 mouse oviducts and individually dissected. ODMCs were Participants/materials, setting, methods: Univariate (p<0.20) and mul-
fabricated from polydimethylsiloxane (PDMS) by planar photolithography and tivariate (p<0.05) analysis was performed in order to find morphokinetic differ-
installed with inlet and outlet reservoirs that facilitate injection and expulsion of ences between conventional IVF and ICSI. A secondary analysis was performed
hyaluronidase (80 IU/ml) throughout ODMCs. COCs were gently oscillated which corrects for the difference in time of fertilization, by standardizing all time
inside the channels on top of the bas-relief structures and denuded by laminar lapse parameters for the time of pronuclear fading. Subgroup analysis for all day
chaotic mixing. The GUI allows users to assess denuding oocytes in real-time 5+6 biopsied blastocysts was performed. Results are presented as average ±
and unload cleaned oocytes at their discretion. SD, Odds Ratio [95% CI].
Main results and the role of chance: Murine COCs were processed by Main results and the role of chance: A total of 283 and 285 cumulus oocyte
ODMC with optimized geometry and operating parameters including oscillating complexes were assigned to conventional IVF and ICSI of which 183 (64.7%)
frequency, flow rate, channel width, and number of bas-relief structures. The and 190 (66.7%) were normally fertilized, respectively. Conventional IVF gen-
oscillating frequency and flow rate were able to tumble COCs to expose the erated 120 blastocysts that were biopsied, of which 59 (49.2%) were euploid,
entire surface to bas-relief structures to evenly strip-off cumulus cells (CCs). while 116 blastocysts were biopsied after ICSI of which 56 (48.3%) were euploid.
Using a high-speed camera recording real-time denudation, we observed that Gender distribution (male/female) for blastocysts with informative outcome
angular displacement of fluid and transverse flows surrounding COCs are the was 60/49 and 50/55, respectively.
main denudation mechanisms. Computational fluid dynamics (CFD) simulations When comparing the development of all normally fertilized zygotes between
quantified microfluidic profiles in the ODMC to that of manual pipetting (MP). IVF and ICSI in the univariate model, a significant difference was found for tPNf
CFD results showed that the maximum shear stress imposed on COCs was 40% (p=0.005), t2 (p<0.001), t3 (p=0.001), t4 (p=0.003), t5 (p=0.001), t6 (p=0.096),
less in the ODMC compared to MP at the same flow rate. Denudation efficiency, t7 (p=0.177) and Blast (p=0.004) of which only t2 remained significant in the
defined as the percentage of complete CC deprivation at the outlet reservoir, multivariate model (OR: 1.282 [1.020-1.612], p=0.033); IVF: 29.3 ± 10.4 versus
was comparable between the ODMC and MP (95% vs. 91%) without oocyte ICSI: 25.9 ± 5.1. After standardizing for tPNf, only corrected tSB (p=0.009) and
loss. To ensure that our ODMC does not compromise the developmental poten- Blast (p=0.004) showed significant differences between IVF and ICSI. However,
tial of the gametes, piezo-actuated ICSI was performed on 50 oocytes processed only Blast appeared significant in the multivariate model: OR: 0.803 [0.648-
by each method. ODMC and MP groups yielded comparable post-ICSI survival 0.994], p=0.044; IVF: 70.8 ± 8.2 versus ICSI: 72.5 ± 9.7. Taking into consider-
(84.0% vs. 82.0%), fertilization (92.9% vs. 90.2%), and blastocyst formation rates ation the standardized kinetics of biopsied blastocysts, only a difference was
(90.5% vs. 87.8%), confirming that ODMC does not have adverse effects on observed for t2: OR: 1.519 [1.045-2.206], p=0.028; IVF: 3.4 ± 2.6 versus ICSI:
preimplantation development. 3.1 ± 3.3.
Limitations, reasons for caution: PDMS is known for easy prototyping, high Limitations, reasons for caution: Only couples with non-male factor infer-
optical transparency, and biocompatibility. However, adverse effects including tility and a normal response to ovarian stimulation were included, and therefore,
small molecule adsorption and gas permeability can be disadvantageous in del- the results cannot be extrapolated to other patient populations.
icate embryology procedures. Large-scale mouse embryo assay should be per- Wider implications of the findings: Following the recent debate on the
formed to assess teratogenicity of PDMS. Clinical-grade polystyrene may be over-use of ICSI, this study shows that conventional IVF results in the same
utilized due to a lower toxicity. number of blastocysts for biopsy with similar developmental kinetics, thereby
Wider implications of the findings: This device offers a glimpse into the reinforcing the use of conventional IVF in this patient population.
potential of a fully automated embryology laboratory and is being optimized for Trial registration number: NCT03708991
ICSI. Constructing different modules in addition to COC denudation is required
for maturity recognition, injection, and storage of the conceptuses to follow O-167 Intracytoplasmic sperm injection versus conventional in
embryo evaluation when applied in conjunction with artificial intelligence. vitro fertilisation in couples with non-male factor infertility: a
Trial registration number: not applicable randomised controlled trial
Q.V. DANG1, L. Vuong2, Q. Nguyen1, T. Ho3, A. Ha1, B. Truong3,
O-166 Marginal differences in preimplantation development Q. Pham4, R. Wang5, R. Norman6, B. Mol5
between conventional IVF and ICSI in patients with non-male 1
My Duc Hospital, IVF Department, Ho Chi Minh City, Vietnam ;
factor infertility: a sibling oocyte study. 2
University of Medicine and Pharmacy at Ho Chi Minh City, Department of
N. De Munck1, A. Bayram1, A. Arnanz1, A. Abdala1, I. El-Khatib1, Obstetrics and Gynecology, Ho Chi Minh City, Vietnam ;
A. El-Damen1, L. Melado2, B. Lawrenz2, H.M. Fatemi2 3
An Sinh Hospital, IVF Department, Ho Chi Minh City, Vietnam ;
4
1
IVI RMA Middle East Fertility Clinic LLC, IVF lab, Abu Dhabi, United Arab Emirates ; My Duc Hospital, HOPE Research Center, Ho Chi Minh City, Vietnam ;
5
2
IVI RMA Middle East Fertility Clinic LLC, Gynaecology, Abu Dhabi, United Arab Monash University, Department of Obstetrics and Gynecology, Melbourne,
Emirates Australia ;
6
The University of Adelaide, Robinson Research Institute and Adelaide Medical
Study question: Are there differences in the morphokinetic behavior between School, Adelaide, Australia
conventional IVF and ICSI in cycles with pre-implantation genetic testing for
aneuploidies? Study question: Does intracytoplasmic sperm injection (ICSI) result in a higher
Summary answer: Preimplantation development is marginally, yet significantly ongoing pregnancy rate compared to conventional in vitro fertilisation (IVF) in
different between embryos generated by conventional IVF and ICSI. couples with non-male factor infertility?
What is known already: Conventional IVF results in a delayed (4 hours) Summary answer: In couples with non-male factor infertility, ICSI did not
pronuclear formation and first mitotic divisions when compared to ICSI on sibling result in a higher ongoing pregnancy rate compared to conventional IVF
oocytes. This difference disappears around the time of morula formation in What is known already: There is an increasing trend worldwide in the use
donor oocyte cycles. On the other hand, it has also been shown in autologous of ICSI for couples with non-male factor infertility. Randomised controlled trials

(RCTs) have not demonstrated better outcomes of ICSI compared to conven- What is known already: Despite the recent rise in the annual number of FET
tional IVF, but their statistical power was limited and most studies used fertilisa- cycles, a large survey of IVF clinics highlighted little consensus between IVF clinics
tion as primary outcome. Moreover, non-randomised studies suggest no benefit regarding the day of blastocyst transfer in NC-FET. Only two small retrospective
and maybe even harm from ICSI compared to conventional IVF in this population. studies have compared different days of blastocyst transfer in NC-FET. The first
Study design, size, duration: This open-label, multi-center RCT was con- compared blastocyst transfer on day five versus day six after serum LH surge;
ducted at two IVF centers in Vietnam. The protocol has been published (Dang demonstrating higher live birth rate in the day five group. However, the most
et al., 2019). A sample size of 1064 couples was calculated to demonstrate a recent study demonstrated a similar ongoing pregnancy rate when blastocysts
10% difference between ICSI and conventional IVF (power 0.90, two-sided alpha were transferred on day six compared to day seven after a serum LH level of
5%, loss to follow-up 10%). Randomisation was done using a computer-gener- ≥20 IU/L.
ated randomisation list, with a variable block size of 2, 4 or 8. Study design, size, duration: We retrospectively analysed all NC-FETs
Participants/materials, setting, methods: Couples were eligible if the between January 2017 and March 2019, before and after a protocol change (May
male partner had normal sperm count and motility (WHO, 2010) and had under- 2018) from blastocyst transfer occurring on day seven after urinary LH surge
gone ≤2 previous IVF/ICSI attempts. Women undergoing in vitro maturation (LH+7) to day 6 (LH+6). The primary outcome was ongoing pregnancy
cycles, using frozen semen or having a poor fertilisation (≤25%) in previous cycle rate ≥24 weeks (OPR). We included 558 NC-FET cycles (180 LH+6; 378
were not eligible. The primary outcome was ongoing pregnancy resulting in live LH+7). As an independent control, outcomes from medicated FET were also
birth after the first embryo transfer. Intention-to-treat analyses were used. compared between time periods (651 before, 483 after).
Main results and the role of chance: Between March 2018 and August Participants/materials, setting, methods: We included all cycles involving
2019, 1064 couples were randomly assigned to ICSI (n=532) or conventional NC-FET of vitrified-thawed, unbiopsied blastocyst(s) derived from autologous
IVF (n=532). Baseline characteristics were comparable between the two groups oocytes between January 2017 and March 2019 at Oxford Fertility. Outcomes
(mean female age 32 years, total sperm counts 190 million, sperm motility 43%). were compared using Chi-squared and multivariate binary logistic regression.
The follow-up for live birth is ongoing and will be available by June 2020. Ongoing The primary outcome was ongoing pregnancy beyond 24 weeks. Over the time
pregnancy rates after the first embryo transfer were 36.3% in the ICSI group period other aspects of clinical practice remained unchanged and, as an inde-
versus 32.7% in the conventional IVF group (RR 1.11; 95% CI 0.94-1.31; p=0.25). pendent control, outcomes from medicated FER cycles were also compared
Fertilisation rate per oocyte retrieval and total fertilisation failure rate also did between time periods.
not differ siginficantly between the two groups (55.6% versus 52.7% and 5.5% Main results and the role of chance: A total of 774 natural cycle FET cycles
versus 6.4%, respectively). The mean number of embryos and number of frozen were started during the study period and after the inclusion criteria were applied
embryos were not significantly different between the two groups, 5.9 ± 4.0 a total of 558 cycles were analysed (180 LH+6; 378 LH+7). Baseline charac-
versus 5.7 ± 4.5 and 3.9 ± 2.9 versus 3.8 ± 3.1, respectively. Mean number of teristics were similar, including age at FET and oocyte recovery, number of
embryo transferred were 1.91 ± 0.29 and 1.90 ± 0.30. Implantation rates were oocytes collected in the fresh cycle, fertilisation method (IVF or ICSI), endome-
28.5% in the ICSI group versus 28.9% in the conventional IVF group. The rates trial thickness, day of blastocyst cryopreservation (day five or six), number of
of clinical pregnancy and miscarriage were 43.0% versus 39.6% (RR 1.08; 95% embryos transferred and proportion of top-quality embryos.
CI 0.94-1.25; p=0.29) and 4.7% versus 5.1% (RR 0.92; 95% CI 0.54-1.57; Ongoing pregnancy rate (OPR) beyond 24 weeks was significantly higher when
p=0.89), respectively. blastocyst transfer occurred on LH+6 compared to LH+7 (45.0% v 29.1%, p<
Limitations, reasons for caution: Double blinding was not possible due 0.0001). Adjusting for potential confounders the adjusted odds ratio was 2.13
to the nature of the interventions. This study excluded couples with a poor (95% CI: 1.44-3.14, p<0.0001). Clinical pregnancy rate (52.2% v 32.8%; aOR
fertilisation (≤25%) in previous cycle, which limits its generalizability. The 2.28, 95% CI 1.56 – 3.31, p<0.0001) and biochemical pregnancy rate (61.7% v
results are based on preliminary analysis, as follow-up of live birth is still 42.1%; aOR 2.26, 95% CI 1.56 – 3.30, p<0.0001) were also significantly
ongoing. (p<0.0001) higher in the LH+6 group. Miscarriage rate was similar between
Wider implications of the findings: In couples with non-male factor infer- groups (26.1% v 30.2%, aOR 0.75 95% CI 0.42- 1.32, p=0.317).
tility, ICSI did not show significant benefit over conventional IVF. This questions The OPR for medicated FET was stable between time periods (38.9% after
the value of routinely use of ICSI in non-male fertility in routine assisted v 43.0% before; aOR 0.90, 95%CI: 0.70-1.16, p=0.41).
reproduction. Limitations, reasons for caution: Our study is not randomised and com-
Trial registration number: NCT03428919 pares groups before and after change in practice, rather than patients undergoing
treatment contemporaneously. The fact that OPR for our independent control,
medicated FET cycles, remained stable between the same time periods is reas-
suring. However, randomised prospective studies are needed to confirm our
SELECTED ORAL COMMUNICATIONS findings.
SESSION 44: WHAT ARE THE OPTIMAL REGIMES FOR FRO- Wider implications of the findings: We observed double the odds of OPR
ZEN EMBRYO TRANSFER? (≥24 weeks) in cycles in which blastocyst transfer occurred at urinary LH+6
compared to LH+7, which is highly significant. Moreover, the high OPR highlights
07 July 2020 Parallel 2 15:15 - 16:30
NC-FET with urinary LH testing as an effective, convenient and elegant method
of FET in ovulatory women.
O-168 Natural cycle frozen-thawed embryo transfer (FET): O-169 Pregnancy outcome and cost-effectiveness comparisons of
A comparison of outcomes following blastocyst transfer on day six artificial cycle-prepared frozen embryo transfer with or without
versus day seven after urinary luteinising hormone (LH) surge. GnRH agonist pretreatment for polycystic ovary syndrome: a
M. Noble1, J. Craig1, A. Bevan1, T. Child1,2 randomized controlled trial
1
Oxford Fertility, Institute of Reproductive Sciences, Oxford, United Kingdom ; L. Luo1, Q. Wang1
2 1
University of Oxford, Department of Women’s and Reproductive Health, Oxford, 1st affiliated hospital of Sun Yat-Sen University, Center of Reproductive Medicine,
United Kingdom Guangzhou, China
Study question: In natural cycle FET (NC-FET) is there a difference in clinical Study question: Can gonadotropin releasing hormone agonist (GnRH-a) pre-
outcome between frozen-thawed blastocyst transfer on the sixth compared to treatment improve pregnancy outcome of artificial cycle-prepared frozen
the seventh day after urinary luteinising hormone (LH) surge? embryo transfer (AC-FET) for women with polycystic ovary syndrome (PCOS)?
Summary answer: We observed double the odds of ongoing pregnancy (≥24 Summary answer: For women with PCOS, GnRH-a pretreatment prior to
weeks) when frozen-thawed blastocyst transfer occurred on day six rather than AC-FET does not improve live birth rate but increases direct costs compared
day seven after urinary LH-surge to AC-FET without GnRH-a pretreatment.

What is known already: For women with normal ovulation, randomized Wider implications of the findings: If there are no differences with mor-
controlled trials (RTCs) and meta-analyses have reported similar live birth rates phological grade between day 5 and day 6 blastocysts, it should be considered
after AC-FET with or without GnRH-a pretreatment. However, two recent to transfer day 5 vitrified blastocysts first in order to shorten time to live birth.
retrospective studies reported that AC-FET with GnRH-a pretreatment Trial registration number: not applicable
improved pregnancy outcomes for women with PCOS.
Study design, size, duration: A total of 342 women with PCOS (24–40
years of age) scheduled for AC-FET from January 2017 to November 2018 were O-171 Comparison of day-5 serum progesterone levels using
enrolled in this RCT. Of these patients, 170 received AC-FET without GnRH-a different routes of administration for artificial endometrial
pretreatment (AC group) and 172 with GnRH-a pretreatment (G-AC group). preparation: Intrapatient variation.
A total of 328 FET cycles were analyzed. Primary outcome was live birth rate M. Cerrillo Martínez1, G. Nardini1, M. Cruz2, M. Mayoral1,
and secondary outcomes were clinical pregnancy and early pregnancy loss rates. M. Toribio3, E. Labarta4, J.A. García Velasco5
A cost-efficiency analysis was also performed. 1
IVI Madrid, Reproductive Medicine, Madrid, Spain ;
Participants/materials, setting, methods: This study was conducted at 2
IVI Madrid, embriology, Madrid, Spain ;
the reproductive center of a university-affiliated hospital. Frozen embryos for 3
IVI Madrid, Nurse department, Madrid, Spain ;
transfer were obtained from the first ART cycle, and no patient received more 4
IVI Valencia, Reproductive Medicine, Madrid, Spain ;
than two cycles. All patients included received blastocyst transfer. 5
IVI Madrid- URJC, Reproductive Medicine, Madrid, Spain
Main results and the role of chance: Neither live birth rate per FET cycle
nor live birth rate per embryo transferred differed significantly between AC and Study question: Is there any variability on the day-5 serum progesterone levels
G-AC groups [92/163 (55.8%) vs. 85/165 (51.5%) and 92/234 (39.3%) vs. during artificial endometrial preparation using different routes of administration
85/245 (34.7%), respectively; both P>0.05]. Clinical pregnancy and early preg- for Luteal phase ( LP)?
nancy loss rates were also comparable between groups. The median direct cost Summary answer: Day-5 serum progesterone levels seem to be maintained
was significantly lower in the AC group compared to the G-AC group (4526.6 above 9.2 ng/mL irrespective of the route of administration.
RMB vs. 7984.0 RMB, P<0.001). The median direct cost per live birth was also What is known already: Progesterone for artificial endometrial preparation
significantly lower in the AC group (8117.6 RMB vs. 15415.1.0 RMB, P<0.001). can be administered orally, vaginally, subcutaneously or intramuscularly. Although
Limitations, reasons for caution: Indirect costs during the FET cycle and the use of different P4 preparations differ among countries and physicians, vaginal
costs during the whole pregnancy period were not analyzed. Pregnancy-related route seems to be the most preferred one, but new options for luteal support
complications and neonatal outcomes were also not analyzed. are available nowadays. Previous studies suggest that a minimum threshold of
Wider implications of the findings: For women with PCOS, endometrial day-5 serum progesterone levels needs to be reached during hormonal replace-
preparation for FET was equally effective with and without GnRH-a pretreatment therapy in order to improve ongoing pregnancy rates. However, there is
ment, but the direct cost was much higher with GnRH-a pretreatment. controversy on the efficacy of distinct types of progesterone preparations and
Trial registration number: 81701403 routes of administration. More studies are needed to fully understand the phys-
iology of progesterone in ART
O-170 Safety and IVF outcomes of Day 6 vitrified blastocyst Study design, size, duration: Prospective cross-over pilot study performed
transfers : comparison with Day 5 vitrified blastocyst transfers between January and June 2019 in IVI-RMA Global clinic, Madrid, Spain. We
with propensity score matching aimed to evaluate if there is significant intrapatient variation on the day-5 serum
D.S. Park1, S.W. Lyu1, H.M. Park1, J.H. Jeong1, W.S. Lee1 progesterone levels with the use of different routes of administration during
1
Fertility Center of CHA Gangnam Medical Center, Department of Obstetrics and hormonal replacement therapy. IRB approval 1809-MAD-063-MC
Gynecology, Seoul, Korea- South Participants/materials, setting, methods: Three different protocols were
administered to 5 participants underwent 3 consequtive cycles of HRT. LP sup-
Study question: Does day 6 (D5) vitrified blastocyst transfer (VBT) have port consisted on 1º) vaginal progesterone 400 mg/12 hours ; 2º) subcutaneous
similar live birth rate (LBR) with day 5 (D6) VBT? progesterone 25 mg/12 hours ; and 3º) intramuscular progesterone 50 mg / 24
Summary answer: D6 group has significant lower LBR than D5 group in h . After estrogen priming for 10 days, P4 was given for 5 days, with daily serum
propensity score matching study. P were performed. Linear regression model accounted for progesterone levels
What is known already: In fresh embryo transfer cycles, D6 blastocyst has (response variable) and route of administration (explanatory variable).
significantly lower pregnancy rate compared with D5 blastocyst. However, it is Main results and the role of chance: The mean day-5 serum progesterone
unclear whether this result is maintained in VBT cycles, because of conflicting levels depending on the route of administration were as follows a) 14.6 ± 5.5
studies. ng/mL; b) 47.9 ± 22.3 ng/mL; and c) 60.3 ± 65.5 ng/mL. The linear regression
Study design, size, duration: This is retrospective cohort study with pro- model showed that changing vaginal progesterone to either subcutaneous or
pensity score matching. We evaluated 1157 cycles of VBT performed between intramuscular route had a statistically significant effect on serum progesterone
January 2014 and December 2015, at the Fertility Center of CHA Gangnam levels, with an average increase of 25 ng/mL (5.4-45.1 ng/mL; p=0.01) using
Medical Center. the subcutaneous route and 32 ng/mL (13-52.7 ng/mL; p=0.001) with the
Participants/materials, setting, methods: All VBT cycles were allocated intramuscular route. The mixed effects model proved that the subcutaneous
to two groups according to the day of blastulation: There were 933 VBT cycles preparation maintained the mean progesterone level at 29.5 ng/mL (SE: 7.6,
of D5 group and 224 VBT cycles of D6 group. The propensity scores were fixed effect), while the intramuscular preparation showed a mean value of 18.1
calculated using binary logistic regression analyses based on the patient and cycle ng/mL (SE: 9.75, fixed effect). Age and body mass index did not produce a
baseline variables. The matched ratio for D5 vs D6 was 3:1, respectively. All significant effect on the results of the mixed model analysis. Despite the route
patients were underwent natural endometrial preparation. of administration, all progesterone preparations achieved a mean day-5 serum
Main results and the role of chance: After matching, there were no signif- level above the threshold of 9.2 ng/mL, which is considered the optimal cutoff
icant differences of baseline characteristics between D5 and D6 groups. D6 group according to a previous prospective cohort. Of note, one patient did not reach
had significantly lower implantation rate (IR), clinical pregnancy rate (CPR) and the recommended day-5 concentration with vaginal progesterone.
LBR (IR 44.2% vs. 53.1%, p=0.023; CPR 48.4% vs. 60.4, p=0.009, LBR 33.5% vs. Limitations, reasons for caution: This is a cross-over pilot study with pre-
51.8%, p<0.001). There were no significant differences in the ectopic pregnancy liminary data and few patients. Further studies are needed to better understand
rate (1.3% vs. 2.8%, p=0.458) and multiple pregnancy rate (29.3% vs. 30.2%, circulating progesterone levels during hormonal replacement therapy and its
p=0.816). Miscarriage rate was higher in Day 6 group (29.3% vs. 10.7%, p<0.001). impact on reproductive outcomes. Nevertheless, considering the scarcity of
Neonatal outcomes of D5 and D6 groups were not significantly different. data, our results provide important insights and draw attention to intrapatient
Limitations, reasons for caution: Although we performed analysis using variability.
propensity score matching to control for potential confounders between the Wider implications of the findings: Apparently, the three types of proges-
study groups, retrospective design itself has limitations. terone preparations lead to sufficient circulating hormone levels in the day of

1
the embryo transfer. However, in the era of personalized reproductive medicine, University of Naples, Biology, Naples, Italy ;
2
endometrial preparation and luteal phase support deserve further investigation Institute of Biochemistry and Cellular Biology, National Research Council CNR,
and patient-specific behaviors must be taken into account. Naples, Italy ;
Trial registration number: NONE 3
Reproductive Medicine Unit of Check Up Polydiagnostic Center, GEA - Gynecology
Embryology Andrology, Salerno, Italy ;
O-172 Frozen-thawed embryo transfer in natural cycles leads to 4
Andrology Unit of the “S. Francesco d’Assisi” Hospital, Local Health Authority ASL
higher clinical pregnancy rates compared to artificial endometrial Salerno, Salerno, Italy
preparation cycles
N. Aslih1, Y. Atzmon1, M. Michaeli1, A. Bilgory1, E. Shalom-Paz1 Study question: Does the content and properties of DNA binding proteins
1
Hillel Yaffe Medical Center, IVF unit, Hadera, Israel in human sperm change in response to environmental pollution?
Summary answer: We provide evidences that environmental pollution alters
Study question: Which endometrial preparation protocol in Frozen Embryo human sperm protein framework, their binding to DNA and makes these pro-
Transfer (FET) Cycles offers the optimal results? teins able to promote DNA oxidative damage.
Summary answer: Frozen-thawed embryo transfer in natural cycles leads to What is known already: The nucleohistone–nucleoprotamine transition
higher clinical pregnancy rates compared to artificial endometrial prepara- is fundamental in order to ensure condensation of the sperm nucleus into
tion cycles. a compact hydrodynamic shape and a protection of the DNA delivered
What is known already: Number of FET Cycles has risen dramatically by the spermatozoa. The exposure to many environmental contaminants
through the years due to several factors including: efficient cryopreservation produce sperm DNA damage. We have already reported a significant
strategies (vitrification), elective single embryo transfer strategies which lead to increase of sperm telomere length, a higher DNA fragmentation index,
a higher number frozen embryos and transfers per Ovum Pick Up (OPU) and and a lower antioxidant activity in males recruited in high environmental
increasing use of Pre Gestational Diagnosis and Screening (PGD/PGS) tech- impact areas. The major cause of male infertility seems to be sperm DNA
niques. The optimal protocol for endometrial preparation has yet to be deter- damage but the molecular mechanisms which lead to DNA break are not yet
mined. Artificial preparation of the endometrium is a well-established protocol fully understood.
offering a more flexible time table and patient monitoring. On the other hand, Study design, size, duration: In order to investigate the impact of environ-
raising experience with natural cycles for FET suggests non-inferior results. mental pollution on molecular alterations of human sperm, we collected, sperm
Study design, size, duration: A retrospective study. 703 FET cycles per- samples from 240 healthy males living in high and low environmental impact
formed in the years 2016-2018 were analyzed. Cycles were divided into two areas of Campania (Italy). We extracted and characterized the sperm protein
groups: Group A – artificial endometrial preparations (n=390), Group B – natural content, analyzed their DNA-binding affinity and investigated on the possibility
cycle (NC FET) (n=313). that these proteins participate to oxidative DNA damage. The study was per-
Participants/materials, setting, methods: Group A – FET with artificial formed from October 2017 to December 2019, within the EcoFoodFertility
endometrial preparation by exogenous estrogen (Estrofem®, Novo Nordisk) initiative.
and Progesterone preparations (Endometrin®, Ferring or Duphastone®, Participants/materials, setting, methods: Sperm samples were obtained
Abbott or Crinone®, Merk Serono).Group B – FET based on ovulation cycles with informed consent from 240 healthy males, not presenting other factors
included: natural ovulation cycles, modified natural cycles using hCG (Ovitrelle affecting semen quality, living in areas at high (n=160) and low (n=80) environ-
Merck-Serono) for the final triggering of spontaneous and Letrozole induces mental impact. The sperm proteins were extracted and their electrophoretic
ovulation.Data collection was performed including demographics, causes pattern characterized. Sperm proteins DNA binding ability was determined by
infertility, history of fertility treatments, current fresh cycle details and FET electrophoretic mobility shift assays and fluorescence measurements. The
cycle outcomes. involvement of sperm proteins in DNA oxidative damage was evaluated by their
Main results and the role of chance: both groups were comparable in ability to convert supercoiled to relaxed plasmid DNA.
terms of patient characteristics, demographics, infertility causes, treatment Main results and the role of chance: We found, as expected, a canon-
protocols and number of embryos transferred. Although the mean ESHRE ical protamines/histones ratio in the 95.06% of people living in low-impact
score of the transferred embryos was significantly lower in group B, we area. Differently, this result was observed only in the 16.61% of samples
found a significantly higher clinical pregnancy rate in Group B (NC FET) com- from males living in high environmental impact areas. In these latter about
pared to Group A (Artificial endometrial Preparation), 43.1% Vs. 30.9%, the 62% showed the presence of only histone, and the remaining 21.78%
P value = 0.021. a not canonical protamines/histones ratio. Moreover, also the samples
Limitations, reasons for caution: A retrospective study, included different showing a canonical protamines/histone ratio, presented a different DNA
protocols of ovulation cycle for FET. binding mode with respect to those from people living in low-impact area.
Wider implications of the findings: large Randomized Controlled Trials However, our most relevant result was the finding that, in presence of some
are needed to establish the superiority of natural cycle protocol over the artificial pollutants, sperm proteins change their protective ability, participating to
endometrial preparation protocols for FET. DNA oxidative damage. This result was particularly marked for those sam-
Trial registration number: Not Applicable ples presenting a not canonical protamine/histone ratio. The presence of
excess of copper and chromium (two heavy metals involved in Fenton-like
reaction) found in the semen of people living in high impact areas, could
explain the occurrence of DNA oxidative damage. In fact, we have already
SELECTED ORAL COMMUNICATIONS reported, in other organism, that the binding of these metals to sperm
SESSION 45: SPERMATOGENESIS SUBTLE REGULATORY proteins promote DNA break. The differences observed in our work, that
EFFECTORS are not due to chance, suggest that environmental pollution has impact, at
molecular level, on semen quality.
07 July 2020 Parallel 3 15:15 - 16:30
Limitations, reasons for caution: The altered protamine/histone ratio and
the DNA oxidative damage promoted by sperm proteins does not necessarily
imply infertility in men living in high environmental impact areas. Limitations are
due to the impossibility of carrying out in vitro fertilization tests.
O-173 Environmental pollution affects the protamine-histone Wider implications of the findings: Our molecular approaches could rep-
content in human sperm and changes their properties making resent additional tools for semen quality evaluation and represent a starting point
able to promote DNA oxidative damage: the contribution of for develop therapeutic protocols to counteract with these types of sperm
molecular investigations. alterations. Moreover, our findings give new insights into mechanisms of DNA
M. PISCOPO1, G. Lettieri1, F. Febbraio2, G. D’Agostino1, E. Mele1, damage, which is implicated also in several human diseases.
C. Cardito1, T. Notari3, L. Montano4 Trial registration number: not applicable

O-174 The super-enhancer-driven long non-coding RNAs as key 1

iBiMED - Instituto de Biomedicina, Universidade de Aveiro, Aveiro, Portugal ;
cell identity genes define the essential core regulatory circuitry 2
Research Institute in Healthcare Science, University of Wolverhampton,
in non-obstructive azoospermia through activation of immune Wolverhampton, United Kingdom ;
signaling pathway. 3
i3S - Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Porto,
“Abstract withdrawn by the authors” Portugal ;
4
Kent State University, Kent State University, Kent, U.S.A. ;
5
O-175 Beta2-adrenergic receptors are mainly expressed in the University of Pavia, Department of Chemistry, Pavia, Italy
terminal piece of the human sperm flagellum
Study question: Are bioportides capable of disrupting Phosphoprotein
J.L. Girela1, M. Francou1, A. De Juan1, J. De Juan2
Phosphatase 1 (PPP1) complexes as a strategy to modulate sperm motility?
1
University of Alicante, Biotechnology, San Vicente Del Raspeig, Spain ; Summary answer: A cell penetrating peptide (CPP) was able to deliver a
2
University of Alicante, Instituto Universitario de Estudios de Genero, San Vicente peptide as a sychnologically-organized bioportide into spermatozoa with a sig-
Del Raspeig, Spain nificant impact on motility.
What is known already: Phosphoprotein phosphatase 1 catalytic subunit
Study question: How is the expresión and distribution of beta2-adrenergic gamma 2 (PPP1CC2) is a PPP1 isoform restricted to testicular germ cells and
receptors (ADRB2) in human sperm? spermatozoa and its inhibition is essential for sperm motility acquisition. PPP1
Summary answer: Using ultra high resolution microscopy techniques we specificity and activity is regulated by the interaction with PPP1-interacting pro-
indentified that the expression of ADRB2 is mainly restricted to the terminal teins. The mechanism of sperm motility acquisition is a perfect target for a new
piece of the human sperm flagellum. male contraceptive since it affects only the post-testicular sperm maturation.
What is known already: Recent studies have shown that spermatozoa have Study design, size, duration: We designed a peptide based on the region includ-
very complex machinery for cell signalling. As part of that machinery, numerous ing the PPP1 binding motif of a sperm-specific interactor (A-kinase anchor protein
neuroreceptors are expressed in the sperm membrane. Human sperm is region- 4, AKAP4) covalently coupled to inert CPP (penetratin) as a sychnologically-orga-
alised, with several physiological processes restricted to determined areas, being nized bioportide construct. A mutated homologue was also designed as a control.
therefore expected that specific molecules involved in those processes should We evaluated the intracellular accumulation of the bioportide in normozoospermic
also be restricted to that area. Although indirect methods had determined the human sperm and its impact on sperm viability and motility at different time-points.
presence of ADRB2 in human sperm, there is no direct evidence nor the descrip- Participants/materials, setting, methods: Molecular dynamics simulations
tion of the localisation of that receptor. ADRB2 is the direct target of medica- were used to design the bioportide, that was synthetized by microwave-assisted
tions used to treat health problems like Hypertension (beta blockers) or asthma solid phase. The complex disruption was assessed by blot overlay. Fluorescent
(bronchodilators). cell imaging analysis was used to study the intracellular accumulation of the
Study design, size, duration: Sperm from young healthy donors (n =7) were bioportide in human sperm. Sperm viability was evaluated with the trypan blue
included in the study. All the samples were normozoospermic according to test and sperm motility was monitored by computer-assisted sperm analysis.
WHO 2010 criteria. As a positive control, we used the neuroblastoma cell line Main results and the role of chance: In vitro studies revealed that the
SH-SY5Y known to express ADRB2. We design the study by developing new bioportide was able to disrupt the PPP1CC2/AKAP4 complex. Exposure of
techniques that allow us to use high- resolution microscopes (i.e. Laser Scanning human sperm to the bioportide led to a significant decrease in both fast (mean
Confocal Microscopy LSCM and Field Emission Scanning Electron Microscopy decrease of 83.9 ± 15.8 % compared with negative control; 79.0 ± 24.8 %
or FE-SEM) to identify the localisation and distribution of ADRB2 compared with mutated control; p<0.01) and slow progressive motility (mean
Participants/materials, setting, methods: To identify the presence of decrease of 74,1 ± 24,1 % compared with negative control; 72,0 ± 28,5% com-
messenger and protein we used RT-PCR and Western Blot techniques respec- pared with mutated control; p<0.01) and a significant increase in the percentage
tively. Immunolabelling techniques were performed by using the same anti- of immotile sperm (mean increase of 220.4 ± 29.6 % compared with negative
ADRB2 primary antibody followed by the fluorochrome-conjugated secondary control; and 148.2 ± 68.2 % compared with mutated control; p<0.01), with no
antibody for LSCM or gold conjugated for FE-SEM. Negative controls were impact on sperm viability. No significant alterations were observed in non-pro-
performed by omitting the primary antibody. In FE-SEM images we quantify the gressive motility. No time-dependent effect was observed.
number of gold nanoparticles in the sperm cells and in the background to deter- Limitations, reasons for caution: Studies in animal models are needed to
mine the specificity of the technique. validate these results and investigate pre-clinical safety and feasibility.
Main results and the role of chance: We confirm the presence of both Wider implications of the findings: This study exploited two ground-break-
messenger and ADRB2 protein in human sperm cells in all the samples, demon- ing areas of the pharmaceutical field by combining CPP technology with pro-
strating the expression of the receptor. LSCM images showed a predominant tein-protein interaction modulators to generate a bioportide capable of blocking
distribution of the ADRB2 receptor on the Sperm flagellum, with and brighter sperm motility by disrupting a sperm-specific PPP1 complex. This approach has
signal at the terminal piece. In the FE-SEM images, the mean number of gold the potential to allow the development of a “male pill” to arrest sperm motility.
particles found in the end piece of the sperm flagellum was significantly higher Trial registration number: not applicable
than the one found in the mid and principal piece (p<0.001).
Limitations, reasons for caution: The present study only shows the pres-
ence of the receptors on the sperm plasma membrane. The activity of those
receptors should be investigated by developing bioassays based on the use of O-177 In vitro co-exposure to Benzo(a)pyrene (B(a)P) and aged
agonist and/or antagonist. A more reliable genetic overexpression or knockout cerium dioxide nanoparticles (CeO2NPs) enhances DNA damage
approach was not possible in human sperm. in human and rat gametes
Wider implications of the findings: The presence of ADRB2 in the flagellum J. Perrin1, M. Cotena2, V. Tassistro2, C. Sunyach2, J. Rose3,
of human sperm cells is in concordance with the suggested function of that M. Auffan3
receptor, the regulation of sperm motility and hyperactivation. It may also suggest 1
Ap-hm, CECOS Centre clinicobiologique d’AMP CHU La Conception, Marseille
a possible side effect of betablockers or bronchodilators with the male fertility Cedex 5, France ;
potential that needs to be investigated. 2
aix marseille university, Institut Méditerranéen de Biodiversité et d’Ecologie,
Trial registration number: Not applicable Marseille, France ;
3
CNRS, Centre Européen de Recherche et d’Enseignement en Géosciences de
l’Environnement, Aix en Provence, France
O-176 A novel approach for male contraception: modulation of
phosphoprotein phosphatase 1 complexes using bioportides Study question: Do the addition of Benzo(a)pyrene (B(a)P) enhance the DNA
J. Silva1, M.J. Freitas1, J. Santiago1, S. Jones2, S. Guimarães3, damage induced by in vitro exposure to aged cerium nanoparticles (CeO2NPs)
S. Vijayaraghavan4, G. Colombo5, J. Howl2, M. Fardilha1 in human and rat gametes?

Summary answer: In vitro gametes co-exposure to aged CeO2NPs + B(a)P J. Shah1,2, M. Venturas3,4, T. Sanchez5, A. Penzias1,2,
induced significant higher DNA damage than exposure to CeO2NPs alone in D. Needleman3, D. Sakkas2
human and rat gametes. 1
Beth Israel Deaconess Medical Center, Obstetrics and Gynecology, Boston, U.S.A. ;
What is known already: Polycyclic aromatic hydrocarbons (PAHs) are ubiq- 2
Boston IVF, Reproductive Endocrinology and Infertility, Boston, U.S.A. ;
uitous pollutants affecting millions of humans through environmental, dietary 3
Harvard University, Molecular and Cellular Biology and School of Engineering and
and occupational sources. B(a)P is a PAH, known for its carcinogenic and repro- Applied Sciences, Cambridge, U.S.A. ;
toxic potential. Higher DNA damage was observed in Cumulus - Oocyte 4
Universitat Autònoma de Barcelona, Department de Biologia Cellular- Fisiologia i
Complexes of B(a)P-exposed mice than in those of vehicle controls after oral Immunologia, Cerdanyola, Spain ;
administration. CeO2NPs are widely used as diesel additive to increase fuel 5
None, None, Boston, U.S.A.
economy; during combustion, they are associated to PAH and then released in
air. Pristine CeO2NPs affect in vitro fertilization in mice and induce significant Study question: Can non-invasive metabolic imaging via fluorescence lifetime
DNA damage in human sperm. The potential hazard due to the co-exposure imaging microscopy (FLIM) detect differences between euploid and aneuploid
CeO2NPs-B(a)P is still unknown. discarded human blastocysts?
Study design, size, duration: Pristine CeO2NPs were extracted from Summary answer: Non-invasive metabolic imaging via FLIM can detect sig-
EnviroxTM diesel additive and combusted at 850°C, (average combustion tem- nificant differences between embryos deemed aneuploid, euploid and of
perature in a diesel engine), to obtain aged CeO2NPs, which were physico-chem- unknown ploidy status.
ically characterized. B(a)P was purchased from Sigma-Aldrich. Rat gametes were What is known already: FLIM of endogenous nicotinamide adenine dinucle-
sampled in epididymis and oviducts after cervical dislocation of sexually mature otide dehydrogenase (NADH) and flavin adenine dinucleotide (FAD) autofluo-
males and 26-days-old females (after ovarian stimulation). Human frozen sperm rescence reveals the metabolic state of mouse embryos. In mouse and other
from fertile donors were purchased from Germetheque biobank (France). animal models, the metabolic state of embryos correlates with embryo viability.
Participants/materials, setting, methods: Human and rat gametes were Ploidy status is one of the primary determinants of embryo viability. Metabolic
exposed in vitro to very low concentrations of CeO2NPs (1 µg.l-1), to B(a)P alone state correlates with ploidy status in a number of non-embryonic systems, includ-
(1.13µM) and to 1 µg.l-1 CeO2NPs + 1.13µM B(a)P during 1 hour in Ferticult© ing yeast and cancer cells.
Medium + 1% DMSO + 1% S9mix at 37°C, 5% CO2. DNA damage was analysed Study design, size, duration: This was a prospective observational study
by alkaline comet assay (ACA) and quantified by Olive Tail Moment (OTM) in that included 158 discarded human vitrified blastocysts of which 110 had known
oocytes and follicle cells and by % Tail DNA in sperm. ploidy status (12 euploid; 98 aneuploid) by preimplantation genetic testing and
Main results and the role of chance: In human sperm, co-exposure to the remaining 48 embryos were of unknown ploidy status. All blastocysts were
CeO2NPs + B(a)P induced significantly higher DNA damage (mean % Tail day 5 or 6 with Gardner morphology grade A or B except 4 that were grade C.
DNA±SEM = 35.35 ± 0.75) compared to CeO2NPs alone (29.57 ± 0.68) (p < Multilevel models were used for analysis with p<0.05 showing significance.
0.0001) and B(a)P alone (18.07 ± 0.79), (p < 0.0001). Participants/materials, setting, methods: Metabolic imaging via FLIM of
In rat sperm, co-exposure induced significantly higher DNA damage (mean %Tail NADH and FAD was used in a University affiliated private IVF laboratory to
DNA±SEM = 32.4 ± 0.43) compared to CeO2NPs alone (22.73 ± 0.43) (p < study discarded human blastocysts. Eight metabolic parameters were obtained
0.0001) and B(a)P alone (13.91 ± 0.44), (p < 0.0001). from each blastocyst (4 for NADH and 4 for FAD): short (t1) and long (t2)
In rat follicle cells, co-exposure induced significantly higher DNA damage fluorescence lifetime, fluorescence intensity (I), and fraction of the molecule
(mean±SEM OTM = 8.48 ±0.35) compared to CeO2NPs alone (4.31 ± 0.30) engaged with enzyme (F). The redox ratio (Intensity of NADH)/(Intensity of
(p < 0.0001) and B(a)P alone (1.83 ± 0.34), (p < 0.0001). FAD) was also calculated for each embryo.
In rat oocytes, co-exposure (7.5±0.7) induced a significant increase of DNA Main results and the role of chance: Each blastocyst was warmed, incu-
damage compared to B(a)P alone (4.28±0.7) (p = 0.0046), but not if compared bated for 2 hours and imaged to analyze its metabolic signature. The mean ages
to CeO2NPs alone (8.43±0.8) (p >0.05). in years for the unknown ploidy, euploid, and aneuploid groups were 32.0 ± 4.9,
The significant increase in genotoxicity induced by co-exposure to CeO2NPs 37.0 ± 4.6, and 36.9 ± 3.7, respectively. In comparing blastocysts from the
and B(a)P suggest the implication of the so-called “Trojan Horse” mechanism aneuploid, euploid, and unknown groups, the FLIM parameters of NADH-I
by which nanoparticles can act as a carrier of another chemical compound by (p<0.0001), NADH-t2 (p<0.02), FAD-I (p<0.0001), FAD-t1 (p<0.001), and
promoting the interaction with cells. This hypothesis is supported by the results redox ratio (p<0.0001) showed significant metabolic differences. When com-
obtained in oocytes, which show a potentialized effect of B(a)P due to the paring ploidy status to embryo day, a number of significant metabolic differences
co-exposure with CeO2NPs. were seen in the FAD parameters. FAD-F, FAD-t1 and FAD-t2 all showed sig-
Limitations, reasons for caution: These results cannot be extrapolated to in nificant differences (p<0.01) when comparing euploid day 5 versus day 6, aneu-
vivo toxicity of CeO2NPs + B(a)P after inhalation, but demonstrate that interactions ploid day 5 versus day 6, and unknown day 5 versus day 6. Furthermore, euploid
between the mixture CeO2NPs + B(a)P can significantly and synergistically enhance day 5 versus day 6 showed significant differences in redox ratio (p<0.0001) while
the DNA damage in human and rat sperm and in rat follicle cells and oocytes. unknown ploidy day 5 versus day 6 showed significant differences in redox ratio
Wider implications of the findings: These results show for the first time and FAD-I (p<0.02).
that interactions between aged CeO2NPs and B(a)P synergistically potentiated Limitations, reasons for caution: The study was performed using discarded
toxicological effects on human and rat gametes. This information should help frozen human blastocysts, which may differ metabolically from non-discarded
further our understanding of the combined toxicity of PAH and nanoparticles, human embryos. Only a small number of blastocysts deemed euploid (12) were
as PAH are the most ubiquitous organic compound pollutants in ambient air. assessed. Additional data is required to validate the results.
Trial registration number: non applicable Wider implications of the findings: FLIM has revealed significant metabolic
differences between discarded human blastocysts associated with ploidy status.
It is unclear if these associations are correlative or causative (and, if so, if ploidy
impacts metabolism or if metabolism impacts ploidy). Further studies are needed
SELECTED ORAL COMMUNICATIONS to determine if FLIM can assist in clinical embryo selection.
SESSION 46: REPRODUCTIVE (EPI)GENETICS 2 Trial registration number: not applicable
07 July 2020 Parallel 4 15:15 - 16:30
O-179 Clinical outcomes after transfer of 110 euploid and 11
mosaic blastocysts identified using non-invasive preimplantation
genetic testing for aneuploidy
O-178 Non-invasive metabolic imaging with fluorescence lifetime J. Zhang1, M. Olcha1, M. Elzaky1, M. Jaremko1, A. Chavez2
imaging microscopy (FLIM) detects differences in ploidy of human 1
New Hope Fertility Center, Reproductive Endocrinology, New York- NY, U.S.A. ;
blastocysts 2
New Hope Fertility Center, Reproductive Endocrinology, Mexico City, Mexico

Study question: What is the biochemical pregnancy rate after single embryo parental-proband trios and the offspring monogenic disorder was inherited from
transfer (SET) of euploid and mosaic embryos identified via non-invasive pre- only one spouse (both the wives). We obtained 12 and 5 embryos for the 2
implantation genetic testing for aneuploidy (PGT-A) families, respectively. Parental-proband trio haplotyping and PCR-Sanger
Summary answer: Transfer of 110 euploid and 11 mosaic embryos resulted sequencing would validate the result of patient-only haplotyping.
in a 63.6% and 36.3% positive beta-human chorionic gonadotropin level (ß-hCG), Participants/materials, setting, methods: Embryos were trophectoderm
respectively biopsied in Day5 blastocyst stage, and used for Qiagen REPLI-g Kit whole genome
What is known already: PGT-A has been widely used around the world to amplification (WGA). High molecular weight genomic DNA was extracted from
identify ploidy status of embryos. The knowledge obtained can help assist clini- the spouse patient peripheral blood, and used for MGIEasy stLFR Library Prep
cians and patients with prioritization of embryos for transfer. The transfer of Kit (Item No.1000005622) according to instructions. Regular WGS libraries from
single euploid embryos has be shown to improve per cycle pregnancy rate, reduce WGA DNA and family members bulk DNA, together with spouse patient stLFR
miscarriage rate, and prevent transfer of embryos harboring certain genetic anom- libraries were used for DNBSEQ-G400 sequencing with 15X depth each sample
alies such as Down Syndrome. The primary modality for obtaining DNA from and haplotype analysis.
blastocysts has been trophectoderm biopsy, which requires removal of embryonic Main results and the role of chance: According to the co-barcoding infor-
cells. Recently, the advent of non-invasive methods have been developed which mation in the spouse patient(both the wives) stLFR sequencing data, we phased
allow embryonic DNA to be amplified by analysis of culture media only. the pathogenic mutationCOL4A5 c.1834G>C and COL1A1 c.769G>A in the
Study design, size, duration: Retrospective data analysis of biochemical 2 families, and confirmed the pathogenic maternal haplotype linkage with the
pregnancy rates after 110 euploid and 11 mosaic embryos were transferred mutation allele by patient-only haplotyping. Combining the pathogenic haplotype
between August 1st 2019 to December 30th 2019. information and embryonic informative SNP alleles, we determined the genetic
Participants/materials, setting, methods: Ploidy information on 24 chro- status of monogenic disorders for each embryo. In COL4A5 family, 7 embryos
mosomes was derived by testing spent media collected on Day 5, 6, or 7. Media were affected with pathogenic maternal haplotype and 5 embryos were normal.
was processed on site using Non-Invasive Chromosomal Screening (NICS) In COL1A1 family, 2 embryos were affected and 3 embryos were normal. The
platform (Yikon Genomics), utilizing multiple annealing and looping-based ampli- patient-only haplotyping results were validated by parental-proband trio haplo-
fication cycles (MALBAC) for whole-genome amplification followed by next-gen- typing and were 100% concordant. PCR-Sanger sequencing of mutationCOL4A5
eration sequencing. Initial ß-hCG levels were analyzed 7-10 days after SET. ß-hCG c.1834G>C and COL1A1 c.769G>A in the 2 families, respectively, showed
levels were measured with an automated immunoassay analyzer (TOSOH). Chi 100% concordance with the patient-only haplotyping. PGT for aneuploidy (PGT-
squared analysis was used to compare biochemical pregnancy rates. A) was performed by MPS. In COL4A5 family, 2 aneuploidy embryos and 10
Main results and the role of chance: A total of 70 out of 110 (63.6%) euploidy embryos were tested. In COL1A1 family, 3 aneuploidy embryos and
euploid SETs and 4 out of 11 (36.3%) mosaic SETs resulted in a positive ß-hCG. 2 euploidy embryos were determined. The MPS PGT-A results were validated
As expected, the pregnancy rate after euploid embryo transfer was significantly by SNP-array and showed 100% concordant. Three embryos inCOL4A5 family
higher than mosaic embryo transfer (p=0.03). and one embryo in COL1A1 family were both euploidy and with normal mater-
Limitations, reasons for caution: A small study size is the major limitation. nal haplotype that were available for implantation.
Additionally, caution should be taken when generalizing results as there are many Limitations, reasons for caution: The patient-only PGT-M haplotyping by stLFR
other clinical and embryologic practice parameters that could affect pregnancy MPS is applicable in the families that only one spouse carry the pathogenic mutation.
rates in general. Otherwise, parents-only PGT-M haplotyping by stLFR MPS can be performed.
Wider implications of the findings: This data helps demonstrate that a Wider implications of the findings: The patient-only PGT-M haplotyping
non-invasive PGT-A platform can be utilized successfully in a private setting by stLFR MPS provides an effective PGT solution for patients with IVF artificial
without compromising clinical outcomes. Furthermore, the use of a non-invasive insemination by donor (AID) or patients with unavailable spouse. Both
methods to identify embryo ploidy may reduce risk to the embryo, however this patient-only and parents-only haplotyping provide complement to the current
has yet to be determined PGT methodology and can be clinically applied according to the actual
Trial registration number: not applicable situations.
O-180 Single tube long fragment read (stLFR) with massively
parallel sequencing achieved patient-only haplotyping for
preimplantation genetic testing of monogenic disorders O-181 First report about preimplantation genetic testing for
S. Chen1, X. Cheng2, D. Chen2, H. Yan2, J. Xia2, P. Liu2, X. Yin2, the Xq24 microdeletion related to X-linked mental retardation,
F. Chen2, C. Xu1 Nascimento type in a woman with recurrent pregnancy loss
1
The International Peace Maternity and Child Health Hospital- School of I. Lebedev1, E. Soloveva1, D. Zhigalina1, L. Minaycheva1,
Medicine- Shanghai Jiao Tong University, Reproductive Genetics Department, O. Kanbekova1,2, A. Nemtsova2, K. Tolmacheva1, E. Fonova1,
Shanghai, China ; N. Skryabin1, A. Kashevarova1, M. Lopatkina1, I. Stepanov2,
2
MGI- BGI-Shenzhen, R&D center, Shenzhen, China L. Nazarenko1, A. Svetlakov1
1
Research Institute of Medical Genetics, Tomsk National Research Medical Center,
Study question: Can massively parallel sequencing (MPS) offer an approach Tomsk, Russia C.I.S. ;
for preimplantation genetic testing of monogenic disorders (PGT-M) haplotyping 2
Tomsk Regional Perinatal Center named after I.D. Evtushenko, ART Department,
with only a patient? Tomsk, Russia C.I.S.
Summary answer: Single tube long fragment read (stLFR) with MPS achieved
patient-only haplotyping for PGT-M. Study question: Can the presence of inherited chromosomal microdeletions
What is known already: Several MPS-based PGT methods have been devel- associated with microdeletion syndromes be effectively detected in human pre-
oped including low-depth MPS such as OnePGT, MARSALA and haploseek as implantation embryos by current PGT approaches?
well as high-depth MPS. Parents-child trios or parents with additional family Summary answer: Combining STR-haplotyping (PGT-M) and array-CGH
members are required for PGT-M haplotyping with low-depth MPS. Moreover, (PGT-A) provides high-resolution detection of inherited chromosomal
a reference embryo can be used for phasing, that parents-only haplotyping with- microdeletions less than 300 kb in size in human preimplantation embryos.
out proband can be achieved by relative high-depth MPS. PGT-M haplotyping What is known already: Copy number variations (CNVs), including chromo-
for parents-only has been reported to be performed by stLFR previously. somal microdeletions and microduplications less than 5 Mb in size, have significant
However, PGT-M haplotyping for patient-only in some cases that the spouse is impacts on human pathology as they are strongly associated with intellectual dis-
uncertain or unavailable have not been reported. ability, developmental delay and congenital malformations. The average resolution
Study design, size, duration: We recruited 2 families that had undergone in of current PGT-A and PGT-SR technologies, including NGS and array-CGH, is
vitrofertilization (IVF) and traditional PGT-M treatment. Each participant was on approximately 5-10 Mb, ruling out the possibility of preimplantation genetic testing
an informed-choice basis and signed a written consent. Both families were with for the most severe microdeletion and microduplication syndromes. Moreover,

8
segmental aneuploidies lack any genotype-phenotype correlations at the blastocyst Department of Gynecology and Obstetrics, Horsens Regional Hospital, Horsens,
stage. However, some CNVs have low penetrance and can be inherited from Denmark ;
9
healthy parents, providing a chance to be detected in preimplantation embryos. Department of Gynecology and Obstetrics, Randers Regional Hospital, Randers,
Study design, size, duration: Here, we report about preimplantation genetic Denmark ;
10
testing of a 270-kb Xq24 microdeletion associated with X-linked syndromic Department of Gynecology and Obstetrics, Aalborg University Hospital, Aalborg,
mental retardation, Nascimento type (OMIM 300860) by combining STR- Denmark ;
11
haplotyping and array-CGH in a couple with recurrent pregnancy loss. A previ- Department of Gynecology and Obstetrics, Kolding Regional Hospital, Kolding,
ously investigated woman and her mother are carriers of the Xq24 microdeletion, Denmark
as detected by array-CGH and confirmed by quantitative real-time PCR, but did
not demonstrate the classical syndromic phenotype due to extremely skewed Study question: To investigate cell-based non-invasive prenatal testing
X-chromosome inactivation (sXCI). (cbNIPT) following preimplantation genetic testing for monogenic disorders
Participants/materials, setting, methods: A 30-year-old woman had 8 (PGT-M) as an alternative to invaisve chorionic villus sampling (CVS).
pregnancies; one was terminated by a medical abortion; six were spontaneous Summary answer: CbNIPT had a high specificity of fetal cell isolation (95 %).
abortions, including two males with a Xq24 microdeletion and one female with- Successful diagnosis in agreement with PGT-M and CVS was achieved in all
out a microdeletion but random XCI. The woman also gave birth to a healthy seven cases.
girl without microdeletion and random XCI. PGT was performed using WGA What is known already: Although the risk of misdiagnosis associated with
(REPLI-g MDA, Qiagen), nested-PCR for STR markers within deleted, 0.8 Mb preimplantation genetic testing (PGT) is low (< 1 %), the consequences are
upstream and 1.0 downstream regions and array-CGH (GenetiSure Pre-Screen severe. Hence, prenatal analysis is recommended folllowing PGT, but only
Microarray, Agilent Technologies). approximately 50 % of patients opt for the procedure. The current golden
Main results and the role of chance: To develop a system for preimplan- standard for prenatal analysis is chorionic villous sampling, which is assocated
tation STR-haplotyping PGT setup was performed as for monogenic disorders. with discomfort and a small risk of miscarriage (< 1 %). It is possible that a more
Family DNA samples were used to haplotype normal and mutant comfortable and non-invasive alternative would increase the proportion of
X-chromosomes. Seventeen polymorphic STR markers were selected and patients receiving prenatal analysis were recommended, thereby increasing the
tested in the PGT-M setup, and 12 were approved as informative. The system chance of detecting the rare case of misdiagnosis.
was validated in single cells. Nine oocytes were fertilized by ICSI, and 8 zygotes Study design, size, duration: Desciptive case-series. Seven patients achieving
were obtained in the IVF programme with standard ovarian stimulation. pregnancy following PGT-M had a blood sample taken on the day of CVS, from
Trophectoderm biopsy was performed for 4 blastocysts on Days 5-6 followed which potential fetal cells were isloated using a subset of fetal specific cell surface
by WGA by MDA, which was successful for 3 samples. STR-haplotyping com- markers within 48 hours. Fetal cells were analyzed by STR marker analysis and
bined with testing for AMEL and SRY genes revealed a hemizygous Xq24 direct mutation detection and compared to the original PGT-M analysis of the
microdeletion in male embryo, heterozygous carrier female embryo and normal transferred embryo and the CVS result. Patients were enrolled between
female embryo. Array-CGH revealed monosomy 4 in female carrier embryo November 2018 and August 2019.
sample. Standard array-CGH data processing did not find microdeletion but Participants/materials, setting, methods: The following was evaluated
detailed analysis allowed us to obtain concordant results with regard to the for the isolated potential fetal cells:- Number of potential fetal cells isolated in
Xq24 microdeletion despite small size rearrangement. The same microdeletion total, the range and median per case - Fraction of cells receiving a diagnosis
arr[hg19] Xq24(118518668_118836030) like in the patient was also detected including range and median per case - Fraction of successfully diagnosed cells of
in hemizygous male and heterozygous female carrier embryos. Normal female fetal origin including range and median per case - Fraction of fetal cells receiving
embryo was recommended for transfer. a diagnosis including range and median per case - Agreement between fetal cells,
Limitations, reasons for caution: Only inherited chromosomal microdele- original PGT-M analysis and CVS analysis.
tions can be detected by combining PGT-M and PGT-A. De novo microdeletions Main results and the role of chance:
and microduplications are still missed using PGT approaches. STR-haplotyping
requires a preliminary step for selecting heterozygous markers. Risks of hetero-
zygous for X-chromosome microdeletions female embryos transfer are unknown Median
and should be further estimated. N (%) Range
per case
Wider implications of the findings: Our data indicate the successful detection
of clinically significant chromosomal microdeletion by combining PGT-M and PGT- Fetal cells isolated 29 5
A. Implementation of PGT-A reduces the risk of aneuploid embryo transfer. The
Fraction of cells 21/29 (72 %) 1-6 2
presented approach provides new strategies for PGT in women with recurrent
receiving a diagnosis
pregnancy loss and sXCI. This study was supported by RFBR (18-015-00437a).
Trial registration number: not applicable Fraction of successfully 20/21 (95 %) 1-6 2
diagnosed cells being
O-182 Cell-based non-invasive prenatal testing (cbNIPT): an of fetal origin
alternative to chorionic villus sampling to confirm diagnosis of Fraction of fetal cells 20/20 1-6 2
unaffected fetuses following preimplantation genetic testing for receiving a diagnosis
monogenic disorders (PGT-M)
Agreement between fetal 7/7 - -
C.L.F. Toft1,2, H.J. Ingerslev3, U.S. Kesmodel2,3, L. Hatt4, R. Singh4,
cells, original PGT-M
K. Ravn4, B. Hestbek4, I.B. Christensen4, M. Koelvraa4,
analysis and CVS analysis
L.D. Jeppesen4, P. Schelde4, I. Vogel5, N. Uldbjerg6, R. Farlie7,
per case basis.
S. Sommer8, M.L.V. Østergaard9, A.N. Jensen10, H. Mogensen11,
B. Degn1, H. Okkels1, A. Ernst1, I.S. Pedersen1,2
1
Department of Molecular Diagnostics, Aalborg University Hospital, Denmark ; Limitations, reasons for caution: Measures are warranted in order to
2
Department of Clinical Medicine, Aalborg University, Aalborg, Denmark ; increase the diagnostic success rate per cell sample, and larger series are
3
Fertility Unit, Aalborg University Hospital, Denmark ; needed to validate robustness before cbNIPT can replace CVS in the clinical
4
ARCEDI Biotech ApS, , ; practice.
5
Clinical Genetic Department, Aarhus University Hospital, Aarhus, Denmark ; Wider implications of the findings: Since cbNIPT can be performed in
6
Department of Gynecology and Obstetrics, Aarhus University Hospital, Aarhus, early pregnancy, as early as gestational week four, results can be obtained in
Denmark ; time for CVS to be performed in case of a failed analysis. Hence, cbNIPT can
7
Department of Obstetrics and Gynecology, Viborg Regional Hospital, Viborg, be offered as a primary non-invasive option of prenatal testing.
Denmark ; Trial registration number: Not applicable

SELECTED ORAL COMMUNICATIONS Results from mediation analysis (MEDIATE macro; Preacher & Hayes, 2004)
SESSION 47: DOES EMOTIONAL BALANCE BEFORE BEING showed that, at T1, higher childwish (ß=.40,SE=.09) and lower ability to disen-
PARENTS AND AFTER EXIST? gage (ß=-.28,SE=.10) were associated with higher parenthood-engagement. At
T2, higher T1 parenhood-engagement (ß=.59,SE=.08) and childwish
07 July 2020 Parallel 5 15:15 - 16:35 (ß=.14,SE=.07) and lower subjective blockage (ß=-.14,SE=.07) and ability to
disengage (ß=-.18,SE=.07) were associated with higher parenthood-engagement.
There were significant associations between parenthood-engagement and
mental-health.
O-183 In-depth analysis of what influences whether patients’ At T1, childwish (ß=.35,SE=.08) was associated with higher treatment-en-
commit to achieve parenthood (parenthood-engagement) and gagement and this with lower anxiety (ß=-.33,SE=.12) and depression
undergo fertility treatment (treatment-engagement) before and (ß=-.51,SE=.11). At T2, T1 treatment-engagement (ß=.46,SE=.10), being preg-
after a treatment cycle. nant (ß=.58,SE=.16) and lower ability to reengage (ß=-.21,SE=.08) were asso-
S. Gameiro1, S. Mesquita da Silva2, U. Gordon3, G. Baccino4, ciated with higher treatment-engagement, and this with lower depression
J. Boivin5 (ß=-.25,SE=.09).
1
School of Psychology, United Kingdom, Cardiff, United Kingdom ; Limitations, reasons for caution: Response and attrition rates were similar
2
University of Oporto, Faculty of Sports, Oporto, Portugal ; to other questionnaire studies but worse-prognosis patients were more likely
3
Bristol Fertility Clinic, Bristol Fertility Clinic, Bristol, United Kingdom ; to attrite from the study. Engagement measures were self-reported but their
4
fivmadrid, fivmadrid, Madrid, Spain ; predictive power of undergoing another cycle (n=76 patients without preg-
5
Cardiff University, School of Psychology, Cardiff, United Kingdom nancy/childbirth) was as expected: parenthood-engagement did not predict
uptake, OR=.925,95%CI[.811,1.053], but treatment-engagement did,
Study question: Which factors are associated with patients’ commitment/ OR=1.261,95%CI[1.060,1.500].
effort to achieve parenthood (parenthood-engagement) and undergo fertility Wider implications of the findings: The forces associated with parenthood
treatment (treatment-engagement) before and after a treatment cycle? and treatment engagement change from before to after a treatment cycle. Before,
Summary answer: Before the cycle parenthood-engagement and treat- patients’ engagement with parenthood and treatment reflect their strong desire
ment-engagement are associated with stronger chidlwish, after the cycle parent- for children. After, they reflect a conflict between patients’ desire for children
hood-engagement is associated with stronger childwish and treatment-engagement and their immediate emotional reaction to the cycle outcome.
with achieving pregnancy. Trial registration number: not-applicable
What is known already: Infertile patients have to actively engage with their
parenthood goal (parenthood-engagement) if they want to achieve it, which
often means engaging with fertility treatment (treatment-engagement). O-184 Meditation and mindfulness reduce stress in women with
Engagement is the level of commitment/effort one puts towards achieving a recurrent pregnancy loss: A randomized controlled trial
goal, as opposed to stop pursuing it (disengagement) and pursuing other goals K. Kirchheiner Jensen1, M.C. Krog2, A.M. Kolte2, S. Hedegaard2,
(reengagement). Psychological theories predict that engaging is adaptive (i.e., M. Chonovitsch1, A.L. Lunøe2, E.C. Koert2, L. Schmidt3, H. Svarre
results in better mental-health) while goals are achievable, but when they become Nielsen1
totally blocked it is adaptive to disengage and reengage. Infertility research has 1
Amager Hvidovre University Hospital, Department of Obstetrics and Gynecology,
provided weak support to these predictions, maybe because of methodological Hvidovre, Denmark ;
limitations (e.g., poor blockage operationalization, neglect of other 2
Rigshospitalet Copenhagen University Hospital, Recurrent Pregnancy Loss Unit-
predictors). Fertility Clinic 4071, Copenhagen, Denmark ;
Study design, size, duration: Prospective study with two moments: before 3
University of Copenhagen, Department of Public Health, Copenhagen, Denmark
a treatment cycle (T1), 6 weeks after its outcome (T2). Determinants of engage-
ment were participants’ strength of childwish, objective (prognosis, cycle out- Study question: Does a 7-week meditation and mindfulness program reduce
come) and subjective (perceptions of ) blockage, and patient’s general ability to perceived stress among women with recurrent pregnancy loss (RPL) ?
disengage and reengage when facing blocked goals. Engagement was expected Summary answer: A 7-week meditation and mindfulness program significantly
to mediate associations between determinants and mental-health (anxiety, reduced perceived stress in women with RPL compared to a standard supportive
depression) and was conceptualized in terms of effort/commitment to achieve care program for women with RPL.
parenthood (parenthood-engagement) and to continuing with fertility treatment What is known already: Previous studies have shown that both perceived
(treatment-engagement). stress and moderate-severe depression is significantly more prevalent among
Participants/materials, setting, methods: The NHS-UK provided ethical women with RPL compared to other women trying to conceive. Despite the
approval. Patients starting an IVF, ICSI, IUI cycle using own or donated gametes severe mental impact on women experiencing RPL, evidence-based care for the
were consecutively recruited and completed T1 and T2 questionnaires. Childish psychological consequences is currently limited. Meditation and mindfulness
and subjective blockage were assessed with single-item questions. Objective interventions have shown to be beneficial in reducing stress and negative feelings
blockage was prognosis data (T1) and cycle outcome (T2), collected from med- as well as improving well-being for patients with a wide range of medical condi-
ical records. General ability to disengage and reengage were measured with the tions. However, to our knowledge, it has never been investigated if meditation
Goal Disengagement and Reengagement Scale, parenthood-engagement with and mindfulness can reduce stress among women with RPL.
the OPS-scale, treatment-engagement with the FertiQol-persistence, men- Study design, size, duration: A two-armed randomized controlled trial
tal-health with HADS. (RCT) designed to evaluate a meditation and mindfulness intervention for
Main results and the role of chance: The final sample comprised 117 women with RPL versus standard supportive care for women with RPL.
participants (33% response rate, 47% attrition rate). Average age was 35.81 Participants were included from November 2018 to April 2019. Of 163 invited,
(SD=4.14) years, 77 (66%) were women, 19 (16%) had children and 49 (42%) 76 were enrolled and randomly assigned to either supportive care (control group
achieved pregnancy on their treatment cycle. 38 participants) or to a 7-week meditation and mindfulness program lead by a
Mixed ANOVAs WS:Time(T1, T2) x BS:Outcome(not pregnant, pregnant) professional instructor in addition to supportive care (intervention group 38
were run to investigate changes from T1 to T2 on determinants, engagement participants).
and mental-health. Subjective blockage decreased for pregnant participants Participants/materials, setting, methods: Patients in a tertiary RPL unit
(η2=.223,p<.01) but remained stable for non-pregnant (η2=.036,p >.05). were invited to participate. The intervention group had three meditation and
Disengagement ability increased for all participants (η2=.045,p<.05). Parenthood- mindfulness workshops with two to three-week intervals. Additionally, the inter-
engagement decreased (η2=.104,p<.001) for all participants. Treatment- vention group meditated daily, supported by an online audio guide. The control
engagement decreased for non-pregnant participants (η2=.386,p<.001) but group was instructed not to meditate during the 7-week study period.
remained stable for pregnant (η2=.010,p>.05). Participants’ mental health was evaluated twice by the perceived stress scale,

the major depression inventory, COMPI fertility problem stress scales and (range=4.8%-59.4%; SD=16.9). Eighty-five percent of women over-estimated
COMPI marital benefit measure. their IVF-LBR (mean overestimation 33.66%; SD=20.02) and 47.8% even
Main results and the role of chance: There were no differences between expected their IVF-LBR to be more than double their calculated IVF-prognosis
the groups at baseline. The main outcome, perceived stress, decreased significantly (mean overestimation 46.47%; SD=16.10). Eighty-eight percent of men over-es-
both in the intervention group (mean difference 5, standard deviation 4, p-value timated their IVF-LBR (mean overestimation 38.81, SD=21.84) and 53.7% even
0.001) and in the control group (mean difference 2, standard deviation 4, p-value expected their IVF-LBR to be more than double their calculated IVF-prognosis
0.006). However, the perceived stress score in the intervention group decreased (mean overestimation 51.10%; SD=17.75). Men expected significantly higher
significantly more compared with the control group, p-value 0.027. The infertili- IVF-LBRs compared to their female partners (64.4% vs. 58.6%; Paired t-test;
ty-specific stress in the personal domain (COMPI Fertility Problem Stress Scales) p=0.028) and their degree of mis-estimation was also significantly higher (2.3 vs
decreased significantly in the intervention group (p-value 0.04), otherwise all other 1.8; Paired t-test; p=0.013). Male and female partners did not differ in level of
secondary outcomes showed no significant differences between the two groups. optimism (Paired t-test; p=0.074) and the correlation between the level optimism
Seven women left the intervention group after the first session, indicating that it and expected IVF-LBR was rather weak (Pearson correlation coefficient in
was too stressful to participate, whereas two women left the control group. women: 0.428; p=0.000 and in men: 0.254; p=0.038). The correlation between
Limitations, reasons for caution: It is unknown if participants in the control the IVF-prognosis and the level of optimism was also weak (women: Pearson
group meditated, despite being instructed not to. Likewise, we are unaware if correlation coefficient of 0.022; Men: Pearson correlation coefficient of -0.163).
the participants in the intervention group meditated as instructed. Meditation Limitations, reasons for caution: Recruitment is ongoing to end up with a
and mindfulness can be an overload for women experiencing RPL since seven larger scale prospective cohort study with follow-up until the studied IVF-cycle
participants left the intervention group. is completed, by achieving a live birth or a negative pregnancy test after the
Wider implications of the findings: This is the first RCT study investigating transfer of the last (fresh and frozen) embryo.
the effect of meditation and mindfulness on the mental health of women expe- Wider implications of the findings: This study documents (the extend of )
riencing RPL. Meditation and mindfulness reduced perceived stress significantly the interesting overestimation of IVF-LBRs by couples going through IVF.
more than our routine supportive care. Guided self-administered meditations Whether couple’s overestimation leads to distress and ultimately IVF-
could be a useful tool in the care for women experiencing RPL. discontinuation, as suggested by qualitative interviews (Peddie et al., 2005), and
Trial registration number: NCT03905395 is associated with actual LBRs will be followed up.
O-185 The realism of men and women’s expected IVF live birth
rates
This
K. abstract
Peeraer 1
has been
, J. Devroe 1
chosen2,for
, T. D’Hooghe the press
J. Boivin 3
, J. Vriens4, O-186 Sperm donor conception and disclosure to children aged
E. Dancet1 from <1 to 17 years old: a follow-up study of parents attitudes
programme and will not be made public until the
1 and children reactions to disclosure.
Leuven University Hospital, Departement of Development and Regeneration,
abstract
Leuven, is presented.
Belgium ; V. Drouineaud1, M. Benchaib2, E. Chapel-Lardic1, N. Mendes3,
2
KU Leuven, Departement of Development and Regeneration, Leuven, Belgium ; J.P. Wolf1, C. Chalas1, L. Ferreux1, D. Rivet-Danon1, C. Jean1,
3
Cardiff University, School of Psychology, Cardiff, United Kingdom ; C. Patrat1, O. Blagosklonov4, A. Condat3
4 1
Laboratory of Endometrium- Endometriosis & Reproductive Medicine, Assistance Publique – Hôpitaux de Paris AP- HP Centre- Université de Paris-,
Department of Development and Regeneration, Leuven, Belgium Service de Biologie de la Reproduction - CECOS, Paris, France ;
2
Hôpital Femme Mère Enfant- Hospices Civils de Lyon-, Service de médecine de la
Study question: Which live birth rates (LBR) do IVF-patients expect and are reproduction-, Lyon, France ;
they in line with their IVF-prognosis and affected by their gender and general 3
Assistance Publique-Hôpitaux de Paris AP-HP- Groupe Hospitalier de La
optimism? Pitié-Salpétrière-, Service de Psychiatrie de l’enfant et de l’adolescent-, Paris,
Summary answer: During IVF women and especially men expect unrealisti- France ;
cally high IVF-LBRs and the difference between men and women is not explained 4
CHU Minjoz, CECOS Franche-Comté - Service de Biologie et Medecine de la
by general level of optimism. Reproduction - Cryobiologie-, Besançon, France
What is known already: The general public is known to overestimate in vitro
fertilisation (IVF) success rates. Qualitative interviews showed that well informed Study question: When do heterosexual parents disclose their use of sperm
women cryopreserving their oocytes were unrealistically optimistic about their donation to their child according to the age?
chances on a live birth as they thought they and/or their gynaecologist would Summary answer: 45.1% of children aged from <1 to 17 years old were
perform better than average (de Groot et al., 2016). To the best of our knowl- informed the day of inclusion. 51.35% were informed between 0-2 and 40.54%
edge, the LBRs expected by patients during their IVF-cycle have yet to be studied between 3-5.
and compared to patient’s personalized IVF-prognosis. In addition, whether What is known already: In France, gamete donation is free, voluntary and
these expected LBRs are affected by factors like gender and level of optimism anonymous since 1994. The Bioethics Law does not give any indication concern-
is unknown. ing the attitude with regard to the disclosure to the child conceived with donor
Study design, size, duration: We prospectively surveyed the expected LBRs gamete. In the past, couples conceiving through gamete donation were adviced
and degree of general optimism of 69 couples having an oocyte aspiration in to maintain secrecy. For many years, centers for the study and preservation of
our clinic between March and December 2019. Men and women were asked to eggs and sperm (CECOS) invite couples before ART to disclose to their future
each fill out their own questionnaire. Additionally, couples’ personalized IVF- child about their origin ton ensure trust. A previous 10-year retrospective fol-
prognosis was calculated using the van Loendersloot prognostic model (van low-up French study reported that 38% of the couples disclosed to the child.
Loendersloot et al., 2013, Sarais et al.,2016) after validation and calibration on Study design, size, duration: A retrospective follow-up study was per-
our clinics data (c-statistic of 0.74). formed. Parents with 82 children conceived with donor sperm and aged from 1
Participants/materials, setting, methods: Eligible couples completed at to 17 years old were included.
least one 2nd-6th IVF-cycle with own gametes after a previous IVF-cycle with the Participants/materials, setting, methods: Parents of 82 children were
same partner in our clinic. The level of general optimism was assessed with the asked questions of quantitative and qualitative nature about decisions towards
reliable ‘LOT-R’ questionnaire (Scheier et al., 1994). The degree of mis-estima- their child and other people about their use of donated sperm, how they dis-
tion was calculated with the following formula: (expected IVF-LBR - prognosis)/ closed and children reactions to disclosure. Five groups were analysed according
prognosis. A positive sign shows overestimation, a negative sign shows under- to the children age: 0-2; 3-5; 6-10; 11-14 and 15-17 years old. The study was
estimation and the absolute value quantifies the extend of mis-estimation. set in two centers for the study and preservation of eggs and sperm (CECOS):
Main results and the role of chance: The 67 participating couples (partic- APHP, Cochin, Paris and Besançon.
ipation rate=97.10%) had a mean IVF-prognosis (i.e. calculated LBR per com- Main results and the role of chance: 45.1% of children aged from <1 to
pleted IVF-cycle, including fresh and frozen embryo transfers) of 31.8% 17 years old were informed the day of inclusion: 21.05% of the children aged

0-2 , 45.00% of those aged 3-5, 52.38% of those aged 6-10, 52.94% of those bond with their child, some parents (mainly fathers) had fears around the donor’s
aged 11-14 and 80.00% of those aged 15-17. role in the children’s lives and the consequences for the position of the non-ge-
51.35% of the children were informed between 0 and 2 years old, 40.54% netic father. A few parents (mainly mothers) saw genetics as relevant in terms
between 3-5, 8.11% between 6-10, 0% between 11-17. In 54.05% of the cases, of possible inherited illnesses from the donor.
both parents disclosed to the child while 2.70% did it separately. Most of the Limitations, reasons for caution: There is a possible self-selection bias, with
parents who informed their child had informed their family and/or friends about mostly people who are open about donor conception and non-genetic parent-
gamete donation before ART (94.59%) and 5.41% after the birth. Although hood in their own lives being represented in the study.
73.17% couples intended to inform their children about gamete donation before Wider implications of the findings: This study gives initial insights into how
conception, 58.33% disclosed to offspring after birth and 41.67% kept the origin heterosexual couples in the Netherlands diagnosed with male infertility experi-
secret. All parents disclosed to their child in a voluntary way. 35.13% used a book ence non-genetic parenthood. The findings show the importance of undertaking
and 40.54% told a story and used a book simultaneously. After disclosure, the more qualitative and quantitative studies into experiences of non-genetic par-
reactions of the child were different according to the age: indifference (50%) and enthood, both to improve guidance for donor conception and to increase knowl-
curiosity (45%) were the most observed in children aged 0-2 while 50% of the edge on the topic.
kids aged 3-5 showed indifference and/or accepted disclosure. Children aged Trial registration number: Not applicable
6-10 were often curious (66%).
Limitations, reasons for caution: Only parents were interviewed in this
study and it should be of great interest to query children conceived through O-188 Mental health in balance of infertility treatment and
sperm donation. Despite the high response rate, rare parents refused to partic- employment: Japan-Female Employment and Mental health in
ipate in the study, suggesting that they might not be inclined to disclose the donor Assisted reproductive technology (J-FEMA) Study
origin to their child. Y. Ikemoto1, K. Keiji1,2, E. Motoki3, A. Tanaka4, S. Rikikazu2,5,
Wider implications of the findings: Attitudes moved towards greater open- K. Nakagawa2, Y. Sato6,7, Y. Kuribayashi5, K. Tomooka3, Y. Imai3,
ness about the use of donated gametes, probably due to the evolution of men- A. Ochiai1, M. Kitade1, T. Tanigawa3, A. Itakura1, S. Takeda1
talities and awareness of the couples by CECOS centers. Psychological counselling 1
Juntendo University Faculty of Medicine, Obstetrics and Gynecology, Tokyo, Japan ;
of couples is of particular importance before treatment but also after the child 2
Sugiyama Clinic Shinjuku, Obstetrics and Gynecology, Tokyo, Japan ;
has been born. 3
Juntendo University Faculty of Medicine, Public Health, Tokyo, Japan ;
Trial registration number: NA 4
Saint Mother Obstetrics and Gynecology Clinic, Institute for ART, Fukuoka,
Japan ;
O-187 ‘Doing’ kinship: Experiences of non-genetic parenthood of 5
Sugiyama Clinic Marunouchi, Obstetrics & Gynecology, Tokyo, Japan ;
Dutch heterosexual couples using donor sperm treatment 6
Tatedebari Sato Hospital, Obstetrics & Gynecology, Gunma, Japan ;
M. Siermann1, M. Visser1, A. Schrijvers1, M. Mochtar1, T. Gerrits2 7
Takasaki Art Clinic, Obstetrics & Gynecology, Gunma, Japan
1
Academic Medical Centre Amsterdam, Centre for Reproductive Medicine,
Amsterdam, The Netherlands ; Study question: The aim of the present study was to identify the risk factors
2
University of Amsterdam, Faculty of Social and Behavioural Sciences, Amsterdam, associated with severe psychological stress in women undergoing infertility
The Netherlands treatments.
Summary answer: Severe psychological stressors in women undergoing infer-
Study question: How do Dutch heterosexual couples diagnosed with male tility treatments include long infertility duration, no childbirth history, low family
infertility who achieved parenthood through donor sperm treatment experience income, resignation from job, and infertility-related harassment.
non-genetic parenthood? What is known already: Infertility is one of the most stressful experiences
Summary answer: For participants genetics were both relevant and irrelevant: for couples who desire to have a child. In Japan, women are increasingly getting
genetics were irrelevant for parenthood, but highly relevant for creating similar- married at a later age after their prime reproductive age, thereby leading to
ities between father and child. declining birthrates and increasing proportions of infertile women. The ratio
What is known already: Male infertility can lead to emotional distress, stigma, of the employed women has also risen up to 44.5%, which includes more than
and questioning one’s sense of masculinity. Non-genetic parenthood can lead 70% of the reproductive-aged women. Therefore, the number of working
to perceived difficulties in parenthood and inequality between parents. women undergoing infertility treatments has increased. These women face
Furthermore, donor conception may challenge norms on the role of genetics psychological stress not only during their infertility treatments, but also in
for kinship. There is limited knowledge on how heterosexual couples experience their jobs.
male infertility, non-genetic parenthood and kinship. Study design, size, duration: This was a cross-sectional multi-center study
Study design, size, duration: We performed a qualitative in-depth interview that was performed from August 2018 to December 2018. We conducted a
study between September 2018 and January 2019 with both partners of thirteen survey by administering anonymous self-report questionnaires to 1727 subjects
Dutch heterosexual couples who conceived after donor sperm treatment. in four IVF clinics. This study was approved by the local ethics committee.
Participants were recruited through the participant list of a previous mixed-meth- Participants/materials, setting, methods: The questionnaire consisted
ods study of the University of Amsterdam and the Academic Medical Centre of questions on infertility treatments, work/life conditions, and included the
Amsterdam on guidance to donors, parents and children. Kessler Six-question Psychological Distress Scale (K6) score for the objective
Participants/materials, setting, methods: We held semi-structured indi- analysis of the psychological distress. Of the 1727 subjects, 1672 subjects were
vidual interviews with thirteen genetically related mothers and thirteen non-ge- recruited for the analysis; among the excluded 55 subjects, including 36 under
netically related fathers (Mage fathers = 43.4 years, range 36-64 years; Mage treatment for mental disorders and 19 with no data of the K6 scores. We also
mothers = 39.2 years, range 33-46 years). Their 26 children (average: 2 children focused on 1335 subjects who were working when they started the infertility
per couple) were between 1 and 13 years old (Mage = 5.4 years). Interview treatments.
questions were based on literature and clinical experiences of experts in the Main results and the role of chance: The mean age of the 1672 subjects
field of donor conception. Interviews were transcribed and analyzed using the- was 37.6 ± 4.8 years, and the mean K6 score (range: 0–24) was 4.8 ± 4.4; 103
matic analysis. women (6.2%) had K6 scores ≥ 13 (K6 high), and they were diagnosed with
Main results and the role of chance: All parents in the study were ‘doing’ severe psychological disorders. Multivariate logistic regression analysis showed
kinship: they negotiated the importance of nature versus nurture. Most parents that “K6 high” is strongly associated with long infertility duration of 2 years or
(men and women) saw genetics as irrelevant for experiencing parenthood, for more (odds ratio [OR] 1.874, 95% confidence interval [CI] 1.080–3.253), no
bonding with their children, and for the preferred role of the donor in their experience of childbirth (OR 2.037, 95% CI 1.046–3.969), and low family
future lives. Yet, most of them saw genetics of the donor as relevant for gener- income with 6.0 million yen (49,000 euro) or less (OR 1.887, 95% CI 1.041–
ating similarities between father and child. Having the same donor for all children 3.418). Among the 1335 subjects who were working when they started the
was seen as essential for nearly all of the parents. Based on the donor’s genetic infertility treatments, 266 women (19.9%) had to resign from their jobs for the

infertility treatment. Multiple regression analysis showed that “high K6” is generalized linear models were used with Poisson distribution and log link func-
strongly associated with low family income (OR 2.830, 95% CI 1.517–5.278), tion for adjusted oocyte counts, and a binomial distribution and log link function
the experience of resignation from their jobs (OR 2.081, 95% CI 1.045–4.144), for adjusted clinical outcomes.
infertility-related harassment at the workplace (OR 2.074, 95% CI 1.080–3.983), Main results and the role of chance: 110 women had a median serum
and difficulty in working during the infertility treatments (OR 2.938, 95% folate concentration of 20.55 ng/ml (interquartile range: 10.8, 32.9 ng/ml). Age,
CI 1.150–7.504). type of follitropin alpha, ovulation trigger, baseline level of estradiol and LH were
Limitations, reasons for caution: The pregnancy outcomes and the time- not associated with both early and clinical ART outcomes in univariate regression
dependent changes in these women could not be analyzed because we had only models. For each outcome, we fit a full multivariable model including all covari-
conducted the survey in the form of anonymous self-report questionnaires in ates with univariate p≤0.10 and reduced to a final model retaining covariates
this cross-sectional study. with p < 0.05. Women with higher serum folate had significantly lower total
Wider implications of the findings: Severe psychological stress is associated oocytes yield (p =0.08), lower number of matured oocytes (p=0.04) and lower
with increased risks of infertility and pregnancy loss. Meanwhile, a beneficial fertilized oocytes (p=0.02) after adjustment for BMI, level of AMH, international
effect of mental care on pregnancy loss has been reported. Therefore, a com- normalized ratio, duration of stimulation, dose of FSH, ratio serum calcium/
passionate mental care system should be established and women should be magnesium (additionally for male factor in model of fertilized oocytes). Women
supported for balancing their infertility treatment schedules with their jobs. in highest quartile of serum folate (>=33.0 ng/ml) had 0.23 odds ratio (95% CI:
Trial registration number: 1 0.04, 1.34; p=0.046) for biochemical pregnancy, 0.12 odds ratio (95% CI: 0.02,
0.79; p=0.02) for clinical pregnancy, and 0.1 odds ratio (95% CI: 0.01, 0.7;
p=0.03) for live birth rate compared with women in the lowest quartile (<10.8
ng/ml) in models adjusted for red blood cells, level of FSH, ratio serum calcium/
SELECTED ORAL COMMUNICATIONS magnesium (additionally adjusted for level of antithrombin in model of biochem-
SESSION 48: RELATING THE RELEVANCE OF BIOMARKERS ical pregnancy).
TO INFERTILITY Limitations, reasons for caution: Normogonadotropic non-obese study
women aged 20-35 years old were unrestricted in use of different supplements,
07 July 2020 Parallel 6 15:15 - 16:30
including folic acid and folate. Further pre-conception large-scale studies with
known folate status of both parents, including daily doses, serum folate, homo-
cysteine and hormone levels are needed.
Wider implications of the findings: We suggested that baseline higher
O-189 Does higher level of serum folate before ovarian serum folate concentration was associated with worse ART outcomes. Our
stimulation worsen outcomes of assisted reproductive findings might be useful for choose of safe dosage of folate supplementation for
technologies in normogonadotropic women? both fertile women and women undergoing infertility treatment.
M. Polzikov1, D. Blinov2, T. Ushakova1, Z. Barakhoeva3, Trial registration number: parental study - NCT03088137 (clinicaltrials.gov)
M. Ovchinnikova4, L. Vovk5, Y. Fetisova5, O. Sergeyev6
1
IVFarma LLC, n/a, Moscow, Russia C.I.S. ; O-190 The mechanism of action of oxytocin receptor antagonists
2
Institute for Preventive and Social Medicine, n/a, Moscow, Russia C.I.S. ; (OTRan) in ART – a study of nolasiban on biomarkers of uterine
3
“AltraVita” Human Reproduction Clinic, n/a, Moscow, Russia C.I.S. ; receptivity in healthy female volunteers
4
Group of companies “Mother and Child”, Hospital Lapino, Moscow, Russia C.I.S. ; O. Pohl1, L. Marchand1, P. Pierzyński2, C. Blockeel3, S. Mackens3,
5
Group of companies “Mother and Child”, Perinatal Medical Centre PMC, U. Lorch4, J.P. Gotteland1
Moscow, Russia C.I.S. ; 1
ObsEva SA, Research and Development, Plan-les-Ouates Geneva, Switzerland ;
6
Belozersky Institute of Physico-Chemical Biology- Lomonosov Moscow State 2
Uniwersytecki Szpital Kliniczny, Klinika Rozrodczości i Endokrynologii
University, Group of Epigenetic Epidemiology, Moscow, Russia C.I.S. Ginekologicznej, Białystok, Poland ;
3
Universitair Ziekenhuis Brussel, Centre for Reproductive Medicine, Brussels,
Study question: To investigate the association of serum folate levels before Belgium ;
in vitro fertilization with outcomes of ART in a single ovarian stimulation (OS) 4
St George’s University of London, Richmond Pharmacology Limited, London,
GnRH-antagonist protocol. United Kingdom
Summary answer: Using multivariable models we suggested that higher serum
folate concentrations in normogonadotropic patients was associated with worse Study question: How does nolasiban (an OTRan) effect markers of endome-
early and clinical ART outcomes. trial receptivity such as uterine contractions, and endometrial perfusion and gene
What is known already: Women of reproductive age are recommended to expression.
consume about 400 mg folic acid per day from fortified foods, supplements, or Summary answer: Administration of nolasiban resulted in decreased uterine
both 2–3 month before conception and during pregnancy. Folic acid supplements contractions, increased endometrial blood flow, and modulation of gene expres-
with different dosage are commonly used by infertile women which can lead to sion relevant for endometrial receptivity and implantation.
elevated folate concentration. Study by Murto et al. (2014) has shown that What is known already: The rate of uterine contractions during embryo
women with unexplained infertility had significantly higher median plasma folate transfer (ET) has been shown to be negatively correlated with pregnancy rates,
concentrations than fertile women. However Gaskins et al. (2015) found that and endometrial blood flow (perfusion) has been shown to be an important
higher serum concentration of folate before ART treatment was associated with predictive parameter for endometrial receptivity. Oxytocin receptors are
higher live birth among subjects of EARTH study using three treatment protocols. expressed during human peri-implantation phase in the myometrium, endome-
Study design, size, duration: A sub-analysis of data obtained from multicenter, trium and uterine blood vessels, and several clinical studies have shown that
randomized, embryologist-blinded, parallel-group, therapeutic equivalence study blocking activation of oxytocin receptors by an OTRan, has the potential to
of recombinant FSH. 110 women were enrolled using following inclusion criteria: decrease uterine contractions, increase endometrial perfusion and improve
women aged 20-35 years old, tubal and/or male causes of infertility factors, first pregnancy rates following ET. The highest dose of nolasiban tested in clinical
or second attempt at IVF/ICSI; 18 ≤ BMI ≤ 30 kg/m2; FSH <10 IU/l, estradiol trials is a single administration of 900mg.
level <50 pg/ml; AMH≥1.0 ng/ml. No restrictions for folic acid and other food Study design, size, duration: A randomized, double-blind, parallel-group
supplements/vitamins were applied before, during and after OS. study was conducted in 42 healthy, pre-menopausal women aged 18 to 37 years.
Participants/materials, setting, methods: All subjects underwent OS using Following hormonal pre-treatment, on the day corresponding to a Day 5 ET,
a GnRH-antagonist protocol and one ART cycle per women. Over the 5-day women received a single oral administration of 900mg or 1800mg nolasiban or
fixed-dose regimen, the women received 150 IU/day of follitropin alpha, fol- matching placebo. Pharmacodynamic assessments up to 24 hours after treatment
lowed by dose adaptation. Baseline serum folate was measured by using a che- included ultrasonographic measurements of uterine contraction frequency and
miluminescence assay by Architect system (Abbott, USA). Multivariable endometrial perfusion. Endometrial biopsies were collected at 24 hours.

Participants/materials, setting, methods: Subjects were pre-treated with transgenicm mice were used as animal models. The phenotypic observation
an identical hormonal preparation as that used for women undergoing fro- lasted for 20 weeks to clarify the role of Trim28 in the pathophysiological changes
zen-thawed ET (FET). Transvaginal echography at 0, 4, 8 and 24 hours was used of PCOS and explore the molecular mechanism involved.
to measure uterine contraction frequency, and endometrial perfusion parame- Main results and the role of chance: We found that the TRIM28
ters, including vascularity index (VI), flow index (FI) and vascularity flow index in the granulosa cells (GCs) of women with PCOS were significantly elevated
(VFI), were measured via 3D power Doppler. Endometrial tissue mRNA expres- compared with those in the control group (P<0.05). We modelled our clinical
sion was analyzed using Next Generation Sequencing with standard analytical findings by transgenic mice with ovarian-specific overexpression of Trim28 and
methodologies. followed a PCOS-like phenotype of metabolic abnormalities, including obesity,
Main results and the role of chance: Both 900mg and 1800mg doses of insulin resistance, hyperandrogenism and follicular dysplasia. Transcriptome
nolasiban had measurable and durable effects on uterine contractions, with a analysis of KGN cells revealed that a series of genes were significantly enriched
similar decrease in median contraction frequency at 24-hours after treatment. in the ovarian steroidogenesis pathway, and the combination analysis of
At 1800mg, effects were observed from the 4-hour time point onwards and in RNA and ChIP sequences revealed that the mRNA expression of PDE10A
post-hoc statistical analyses were at p<0.10 at the 4- and 8-hour time points was significantly upregulated via TRIM28 in GCs of PCOS women as well as
compared to placebo. in KGN cells with TRIM28 overexpression, compared with that in the control
Endometrial perfusion parameters showed marked and sustained median group (P<0.05). Since PDE10A hydrolyzes cyclic adenosine monophos-
increases and were comparable for both doses. The most noticeable increase phate (cAMP), the cAMP and PKA activation in TRIM28-overexpressing KGN
in VI was between the 8-hour and the 24-hour time points with p<0.10 at cells were inhibited, which further led to the inhibition of the MAPK-PKA-ERK
the 8-hour time point at 1800mg. FI also increased over time and post hoc signaling pathway and ultimately caused steroidogenesis. Our study thus sug-
statistics showed p<0.10 at the 24-hour time point for both doses. Similarly, gests that elevated TRIM28 levels reduce ERK phosphorylation by upregulating
an appreciable increase in endometrial VFI over time was seen at 900mg the expression of PDE10A, leading to ovarian steroidogenesis dysfunction in
and 1800mg compared to placebo; p<0.10 at the 8-hour time point at GCs and resulting in follicular arrest and hyperandrogenism, and further con-
1800mg. firms the critical role of TRIM28 in the pathophysiological changes of PCOS at
Within 24 hours of 1800mg nolasiban administration, 10 endometrial genes the in vivo level.
were found to be significantly differentially expressed (adjusted p<0.05). Of Limitations, reasons for caution: This study still has certain limitations. We
these, OLFM4, DPP4 and CXCL12 were regulated in the same direction as did not explore the reasons for the increased TRIM28 in GCs of PCOS patients,
Window-of-Implantation-associated genes. In addition, three genes (DPP4, for example, whether it was caused by genetic factors or epigenetic factors.
CXCL12 and IDO2) have previously been associated with decidualization and Wider implications of the findings: Our study first clarified the regulatory
endometrial receptivity. mechanism of TRIM28 for PDE10A and their role in the multiple phenotypic
Limitations, reasons for caution: This study was conducted in healthy interpretations of PCOS and successfully constructed the Trim28-ovarian specific
volunteers without a diagnosis of infertility but pre-treated as for FET. overexpression animal models. The above findings provide new basis and guid-
Extrapolation of these results to patients with infertility who are undergoing ance for proposing feasible and effective risk prediction programs and clinical
fresh embryo transfer should be made with caution. intervention measures.
Wider implications of the findings: These data expand the understanding Trial registration number: 2017YFC1001303
of the mechanism-of-action of OTRan in increasing live birth rates following ET.
These results also suggest faster, broader and more potent effects of the nola- O-192 Androgens increase accumulation of advanced glycation
siban 1800mg dose compared to the 900mg dose, supporting testing at higher end products (AGEs) in granulosa cells by activating endoplasmic
doses and potential alternate regimens in IVF patients. reticulum (ER) stress in polycystic ovary syndrome (PCOS)
Trial registration number: 2018-003702-36 M. Harada1, J. Azhary, MK1, C. Kunitomi1, A. Kusamoto1,
N. Takahashi1, E. Nose1, N. Oi1, O. Wada-Hiraike1, Y. Urata1,
O-191 TRIM28 induces follicular arrest and hyperandrogenism T. Fujii1, Y. Osuga1
Y. Liu1, X. Li1, J. Xu1, H. Huang1 1
Faculty of Medicine- University of Tokyo, Dept. of Obstetrics and Gynecology,
1
The International Peace Maternity and Child Health Hospital, Shanghai Jiao Tong Tokyo, Japan
University School of Medicine, Shanghai, China
Study question: What is the underlying mechanism regulating AGE – receptor
Study question: What is the expression pattern of TRIM28 in granulosa for AGEs (RAGE) system in granulosa cells from PCOS patients?
cells(GCs) and the role of that in the mechanism of polycystic ovary syndrome? Summary answer: Targeting AGE-RAGE system, that is upregulated in PCOS
Summary answer: The elevated TRIM28 levels reduce ERK phosphorylation ovary and induced by testosterone via activation of ER stress, improves repro-
by upregulating PDE10A, leading to ovarian steroidogenesis dysfunction in GCs ductive phenotype of PCOS.
and resulting in follicular arrest and hyperandrogenism. What is known already: PCOS is associated with hyperandrogenism.
What is known already: Polycystic ovary syndrome (PCOS) is a common Previously, we demonstrated that androgen activates ER stress in granulosa cells
endocrinopathy of unknown etiology that affects women of reproductive age. from PCOS patients and mice, and that this contributes to the pathology of
Previous studies have built a convincing argument that an abnormal AR is associated PCOS, including ovarian fibrosis and the growth arrest of antral follicles. In
with PCOS and have documented the unique roles of Androgen receptor(AR) addition, recent studies demonstrated the accumulation of AGEs in granulosa
Splicing variants(ASVs)) in GCs in the pathogenesis of PCOS. The AR ASV exhibited cells from PCOS patients contribute to the pathology by affecting a number of
a genome recruitment pattern differing from that of wild-type AR and dramatically cellular processes, such as steroidogenesis, glucose metabolism, and the pro-
induced the expression of androgen-related genes. Bioinformatics analysis sug- duction of proinflammatory cytokines.
gested that this abnormal genomic recruitment pattern of AR ASVs is highly similar Study design, size, duration: The expression of RAGE was compared
to that of multiple transcription factors, of which TRIM28 ranks first. between human granulosa-lutein cells (GLCs) obtained from 11 PCOS patients
Study design, size, duration: We compared the gene expression pattern of and ten control patients undergoing oocyte retrieval. Ovaries of three PCOS
TRIM28 in granulosa cells(GCs) from 139 PCOS women and 120 women with patients and three control patients, harvested at surgery, were evaluated immu-
tubal blockage undergoing IVF-ET between June 2014 and July 2018. We used nohistochemically for AGEs and RAGE. For in vitro experiments, GLCs were
a human granulosa-like tumor cell line(KGN) to explore the molecular mecha- obtained from pooled follicular fluids. For in vivo experiments, a dehydroepi-
nism at the in vitro level. We observed the phenotype of animal modes for androsterone (DHEA)-induced PCOS mouse model was used and forty mice
20 weeks. were allocated to four treatment groups.
Participants/materials, setting, methods: 139 PCOS women and 120 Participants/materials, setting, methods: Human GLCs were incubated
women with tubal blockage undergoing IVF-ET in our center were recruited. with testosterone and RAGE expression and AGEs accumulation were exam-
GCs and follicular fluids were collected during oocyte retrieval. KGN cell line ined, while GLCs were preincubated with an ER stress inhibitor to examine
was used for in vitro test. The ovarian specific overexpression of Trim28 the intermediary role of ER stress. RAGE expression and AGEs accumulation

in human GLCs and ovaries were compared between control and PCOS Summary answer: In our dataset, women with PCOS do not have shorter
patients. PCOS mice were treated with an ER stress or RAGE inhibitor, fol- LTL, but do show less telomere attrition between ages 31 and 46.
lowed by examination of vaginal smears and quantification of atretic antral What is known already: Telomeres are DNA-protein complexes that pro-
follicles. tect chromosome ends from DNA damage and act as a indicator of cellular
Main results and the role of chance: Treatment with testosterone increased ageing. In most somatic tissues, a small part of telomeric DNA is lost with
RAGE mRNA and protein expression in cultured human GLCs. Pre-incubation every cell division, causing telomere shortening. Current evidence, almost
with a clinically available ER stress inhibitor tauroursodeoxycholic acid (TUDCA) entirely from cross-sectional observations, supports negative associations
abrogated the stimulatory effect of testosterone on RAGE expression. Treatment between LTL and adverse lifestyle and cardio-metabolic risk factors. Women
with testosterone increased AGEs accumulation in GLCs, and this was reduced with PCOS are affected by co-morbid conditions that are also associated with
by pre-treatment with TUDCA, similar to the pattern for RAGE expression. shorter LTL, such as obesity and chronic inflammation. To date, few studies
When we knocked down C/EBP homologous protein (CHOP), a transcription have investigated the association between PCOS and LTL and/or LTL short-
factor activated during ER stress, by RNA interference, CHOP-deficient cells ening with age.
demonstrated significantly lower expression of RAGE expression following tes- Study design, size, duration: We used a community-based birth cohort
tosterone treatment, than control siRNA-treated cells, with a concomitant (N=5889 women) with clinical follow-up at ages 31 (N=3115) and 46 (N=3280)
reduction in AGEs accumulation. GLCs harvested from PCOS patients exhibited including measures of LTL at both ages (N=2906 and N=3233). We defined
higher expression of RAGE mRNA and protein than those from control partic- study groups as follows: age 31, both oligo/amenorrhea and hirsutism: “both
ipants. Immunohistochemical analysis revealed that RAGE expression and AGEs symptoms” (N=73); age 46, self-reported PCOS diagnosis (DG-PCOS, N=155);
accumulation were increased in granulosa cells of the ovaries from PCOS self-reported PCOS (both symptoms and/or DG-PCOS: srPCOS, N=207);
patients. Increased RAGE expression and AGEs accumulation in granulosa cells controls: women without PCOS-related symptoms or diagnosis (age 31:
of PCOS subjects were further confirmed in PCOS model mice. The adminis- N=1054; age 46: N=1324).
tration of a RAGE inhibitor (FPS-ZM1) or TUDCA to PCOS mice reduced the Participants/materials, setting, methods: Mean relative LTL was measured
expression of RAGE and the accumulation of AGEs in granulosa cells, and this by monochrome multiplex quantitative PCR. The amount of telomeric DNA
was accompanied by an improvement in estrous cycling and a reduction in the sequence was quantified relative to a single-copy gene and normalised using a
number of atretic antral follicles. common reference DNA sample. The possible association between LTL and
Limitations, reasons for caution: It is not determined whether the concen- PCOS was analyzed using linear regression models adjusted at both ages for
trations of testosterone usually utilized in human GLC culture studies are appro- BMI, smoking, alcohol consumption and socioeconomic status. Pearson correla-
priate as a model of in vivo hyperandrogenism. Furthermore, the concentrations tions between LTL and BMI, waist, hip, testosterone and sex hormone-binding
of testosterone in follicular fluid of antral follicles of PCOS women, not of globulin were performed at both ages.
preovulatory follicles, are still unknown. Main results and the role of chance: Since LTL was measured using a dif-
Wider implications of the findings: Our findings suggest that the hyperan- ferent reference DNA sample at ages 31 and 46, this was standardized [z score=
drogenism of PCOS increases the accumulation of AGEs in the ovary by acti- (LTL–mean (LTL))/SD(LTL)] separately for each measurement.
vating ER stress, and that targeting of the AGE–RAGE system, either by using a Unadjusted mean LTL did not differ between women with PCOS symptoms
RAGE inhibitor or a clinically available ER stress inhibitor, may provide a novel and controls at age 31 (0.01±(SD)1.10 vs. 0.07±0.99, P=0.632) or between
therapeutic approach for PCOS. DG-PCOS women and controls at age 46 (0.03±0.93 vs. 0.01±1.02, P=0.860).
Trial registration number: Grants-in-Aid for Scientific Research from the Similarly, LTL did not differ when comparing srPCOS with controls (at age 31
Japan Society for the Promotion of Science ( JSPS) (19k09749 , 19k24021., for srPCOS 0.02±0.99 vs. controls 0.07±0.99, P=0.593; at age 46 for srPCOS
19k24045, 19h00319 .), a grant from the Japan Agency for Medical Research -0.05±0.95 vs. controls 0.01±1.02, P=0.414). After respective adjustments
and Development (18gk0110014h003) for the aforementioned covariates at age 31 or 46, none of the results
changed.
Interestingly, we observed a greater mean difference (MD) of LTL z-scores
O-193 Polycystic ovary syndrome and leukocyte telomere length: between ages 46 and 31 in the control group (MD: -0.13±1.10, P<0.001) than
results of cross-sectional and longitudinal analyses in a birth in the srPCOS group (MD: -0.01±1.10, P=0.958). The observed difference
cohort study suggests that the longitudinal change in LTL is markedly less in women with PCOS
P. Pinola1, J. Pölönen1, J. Ronkainen2, A. Blakemore3, J. Buxton4, when compared to controls.
M.R. Järvelin5, J. Tapanainen6, S. Franks7, T. Piltonen1, S. Sebert2, In the srPCOS group there were only weak negative correlations between
L. Morin-Papunen1 LTL and BMI (r=-0.17, P=0.019), and LTL and waist circumference (r=-0.19,
1
Oulu University Hospital- University of Oulu, Department of Obstetrics and P=0.009) at age 31 whereas the correlations did not reach significance
Gynaecology- Medical Research Center and PEDEGO, Oulu, Finland ; at age 46.
2
University of Oulu, Center for Life Course Health Research and Biocenter Oulu, Limitations, reasons for caution: The symptoms and diagnosis of PCOS
Oulu, Finland ; were self-reported. The questionnaire at age 46 did not differentiate
3
College of Health and Life Sciences- Brunel University London and Imperial between women with polycystic ovaries and the syndrome. The study pop-
College London, Department of Life Sciences and Department of Medicine, ulation was relatively young to express significant telomere shortening. The
London, United Kingdom ; sample size of the PCOS women with telomere measurement data was
4
Kingston University London, Department of Biomolecular Sciences- School of Life relatively small.
Sciences- Pharmacy and Chemistry, London, United Kingdom ; Wider implications of the findings: We observed less LTL attrition in PCOS
5
Imperial College London and University of Oulu, MRC-PHE Centre for women than in controls. Analyses included correction for BMI and lifestyle fac-
Environment and Health- Department of Epidemiology and Biostatistics- School of tors, but before concluding whether there is a causal association between PCOS
Public Health and Center for Life Course Health Research- Faculty of Medicine, and LTL, further research is needed to clarify the underlying mechanisms and
London and Oulu, United Ki ; compare with data from other biomarkers of ageing.
6
University of Helsinki and Helsinki University Hospital, Department of Obstetrics Trial registration number: NA
and Gynecology, Helsinki, Finland ;
7
Imperial College London, Institute of Reproductive and Developmental Biology,
London, United Kingdom
Study question: Does mean relative leukocyte telomere length (LTL) or lon- SESSION 49: EMBRYO METABOLISM AND DEVELOPMENT
gitudinal change in LTL differ in women with PCOS symptoms or diagnosis in 07 July 2020 Parallel 1 17:00 - 18:00
comparison to controls?

O-194 Human blastocysts show distinct changes in their 3

Paris Descartes University, Laboratoire de Chimie et Biochimie Pharmacologiques
metabolism between day 5 and 6 et Toxicologiques- UMR 8601-CNRS- Université Paris Descartes- ComUE Sorbonne
D. Sakkas1, M. Venturas2, J.S. Shah1, T. Sanchez2, D.J. Needleman2 Paris Cité- 45 rue des Saint-Pères- 75006- Paris- France, Paris, France
1
Boston IVF, Research, Waltham- MA, U.S.A. ;
2 Study question: To evaluate the relation between metabolic follicular fluid
Harvard University, Molecular and Cellular Biology and School of Engineering and
(FF) composition and ART outcomes in terms of embryo quality and live birth
Applied Sciences, Cambridge, U.S.A.
occurrence in endometriosis patients.
Summary answer: Embryo quality and live birth occurrence seem to be
Study question: Do day 5 and 6 human blastocysts exhibit metabolic changes correlated to a specific metabolic signature in endometriosis patients.
that can be detected by non-invasive metabolic imaging? What is known already: Among different mechanisms explaining infertility
Summary answer: Imaging of NADH and FAD reveals metabolic differences occurrence in endometriosis, oocyte quality alteration has been raised. This last
between day 5 and day 6 discarded human blastocysts. point is still under debate despite of recent studies confirming the deleterious
What is known already: Based on their functional role in cell biology, intra- impact of endometriosis on the oocyte competence. Metabolomics studies are
cellular NADH and FAD have a diagnostic potential as natural biomarkers for increasingly being used to measure alterations of metabolic pathways associated
cellular redox reactions and energy metabolism. NADH and FAD are auto-flu- with complex disorders. Among the various sources of study samples in endo-
orescent, and their concentration and local environment can be measured using metriosis, FF is one of the most interesting to analyze in a context of infertility.
Fluorescence Lifetime Imaging Microscopy (FLIM). It has previously been shown Very few studies have focused on the metabolic signature of endometriosis FF
that FLIM can quantitatively measure distinct stage differences in the metabolic and its correlation with ART outcomes.
state of preimplantation mouse embryos over the course of development. Study design, size, duration: This is a prospective study started on February
Human embryos are known to undergo metabolic changes during the 8-16 cell 2018 and still recruiting (ClinicalTrials.gov NCT03241329). A total of 9 endo-
stage however less is understood of the metabolic idiosyncrasies during the later metriotic patients were included for this analysis.
stages of preimplantation. Participants/materials, setting, methods: Endometriotic patients enrolled in
Study design, size, duration: We performed a prospective observational an ART attempt without any male infertility associated factor. After controlled ovarian
study with 88 day 5 and 68 day 6 blastocysts (108 patients, mean age 35.3 years). stimulation, oocyte retrieval was performed by transvaginal aspiration under ultra-
Blastocysts were discarded and donated for research under an approved insti- sound guidance. Each follicle was individually aspirated and only unique oocyte deriv-
tutional review board protocol . All, but 2, embryos had a Gardner score of A ing from the same follicle was retained for the evaluation. FF was then centrifuged
and/or B for the inner cell mass and trophectoderm and were either full or and stored at -80°C before 1D-1H NMR-based metabolomics analysis. Data were
expanded blastocysts. The vitrified blastocysts were thawed and cultured for 2 compared to blastocyst quality assessment and to clinical outcome after transfer.
hours prior to imaging. Main results and the role of chance: Analysis was carried out on a total
Participants/materials, setting, methods: A FLIM measurement provides of 26 FF samples for which embryo quality was assessed and on 8 FF samples
9 metabolic parameters which can be used to analyze the metabolic signatures for which clinical outcome information was available. Univariate analysis revealed
of individual embryos. We imaged each blastocyst at three different focal planes, that the concentration of various statistically relevant metabolites could be asso-
enabling three metabolic measurements per embryo. We then compared those ciated with embryo quality. In particular, the metabolic profile of FF resulting in
metabolic signatures in relation to the day of vitrification of the embryo (day 5 better embryo quality was characterized by a decrease in glucose level associated
or 6), using multilevel models. to an increase in lactate level. Among other discriminant metabolites, citrate,
Main results and the role of chance: FLIM was used to compare the met- acetate and amino acids, such as tyrosine also appear as being particularly rele-
abolic differences between day 5 and 6 discarded human blastocysts. Of the vant with a lower concentration level observed for good embryos. Finally, live
nine metabolic parameters obtained from each blastocyst the 4 for NADH birth occurrence appears to be correlated to the same trend of variation in
showed no significant difference (P>0.05) when comparing the Day 5 and 6 glucose and lactate levels, but also to an increase in lipids levels such as HDL
blastocysts: NADH short (t1) and long (t2) fluorescence lifetime, fluorescence cholesterol, 2-octenoate and alkane.
intensity (I), and fraction of the molecule engaged with enzyme (F). In contrast, Limitations, reasons for caution: Even if the methodology of our study
all 4 FAD parameters showed highly significant differences (P<0.001) between was robust, the analysis was realized using a small number of samples. These
the Day 5 and 6 blastocysts: FAD-t1; FAD-t2, FAD-I and FAD-F. The redox ratio preliminary results need to be confirmed on larger series.
(Intensity of NADH)/(intensity of FAD), was also compared between Day 5 Wider implications of the findings: Embryo quality and live birth occur-
and 6 blastocysts and showed a substantial significant difference (P<0.001). rence seem to be correlated to a specific metabolic signature in endometriosis
Subsequent analysis comparing Day 5 and 6 blastocysts with similar expansion patients. Our results could be linked to the oocyte quality and could then rep-
rates and similar Gardner grade inner cell mass and/or trophectoderm showed resent a new and non-invasive tool for the prediction of succeeding chance in
the same pattern. pregnancy favorable outcomes.
Limitations, reasons for caution: The study was limited by the type of Trial registration number: not applicable
human blastocysts available as they were discarded and donated for research
without any knowledge of their clinical outcome. It would also be expected that
the blastocysts would be a mixture of euploid and aneuploid. O-196 Blastocyst quality modulates the generation of an
Wider implications of the findings: Metabolic imaging can non-inva- inflammatory microenvironment by decidualized cells
sively detect a significant shift in metabolism between Day 5 and 6 human L. Fernandez1, E. Grasso1, E. Soczewski1, M.S. Gori1, G. Calo1,
blastocysts. This analysis indicated that Day 5 and 6 blastocysts may be V. Hauk1, G. Martinez2, M. Irigoyen2, C. Perez Leiros1,
undergoing intricate changes in respiration as they ready for implantation. R. Ramhorst1
Further studies examining metabolic signature over time may provide more 1
CONICET. Universidad de Buenos Aires., Instituto de Química Biológica de la
information. Facultad de Ciencias Exactas y Naturales IQUIBICEN. Laboratorio de
Trial registration number: Not applicable Inmunofarmacología y Laboratorio de Inmunología Reproductiva de la Fertilidad.,
Buenos Aires, Argentina ;
2
Fertilis Medicina Reproductiva, Laboratorio de embriologia, San Isidro, Argentina
O-195 NMR metabolomics study of follicular fluids reveals specific
signature correlated to ART data in endometriosis patients. Study question: Could soluble factors secreted by blastocyst (blastocyst con-
K. Pocate1, P. Santulli2, F. Kateb3, M. Bourdon2, C. Maignien2, ditioned media or BCM) control the inflammatory response during the peri-im-
C. Patrat1, J.P. Wolf1, G. Bertho3, C. Chapron2 plantation period according to blastocyst quality?
1
Hopital Cochin, Department of Histology- Embryology- and Biology of Summary answer: Decidualized cells increased the expression/production
Reproduction, Paris, France ; of inflammatory cytokines as well as neutrophils migration and activation in
2
Hopital Cochin, Gyneacology-Obstetrics and Reproduction clinical unit, Paris, France ; response to BCM from impaired development (ID) blastocyst.

What is known already: The decidualization program in humans starts on each What is known already: Embryos examined on the first day after retrieval
menstrual cycle and implies not only phenotypical changes on the endometrial are usually classified as viable or atretic. However, little more can be established
stromal cells, but also in their secretory profile. This secretome includes pro-im- regarding their future development, based on routine morphological studies. All
plantatory factors as well as pro and anti-inflammatory cytokines. Despite blasto- AI algorithms developed so far for embryo selection are based on time-lapse
cyst implantation needs a sterile inflammatory response, it should be later video analysis of cleavage stage embryos. Oocyte and zygote stage visual assess-
controlled in favour of a tolerogenic microenvironment. In this sense, decidualized ment was not used so far for the prediction of embryo quality.
cells display the ability to change their secretome according to the quality of the Study design, size, duration: A retrospective analysis of the first 24 hours
blastocyst. of embryo development based on 1560 consecutive time-lapse videos from a
Study design, size, duration: Human endometrial stromal cell line (HESC) single IVF center.
was decidualized with medroxiprogesterona and dbcAMP during 8 days. Then, Participants/materials, setting, methods: Using computer vision algo-
decidualized HESC cells were stimulated with human blastocyst conditioned rithms, we identified unique features, some of which were never before recog-
media (BCM) obtained from developing blastocysts (normal development or nized, during the first 24 hours of embryonic development. We then developed
ND) or arrested ones (impaired development or ID). Non decidualized cells an automated AI algorithm to extract and measure these features from time-lapse
were used as control. videos. A deep neural network was developed based on the parameters recog-
Participants/materials, setting, methods: BCM were recovered from 5 nized by the computer vision module for each image. A classifier was used to
days individually cultured blastocyst obtained from IVF/ICSI and classified as nor- match these parameters to the endpoint, high-quality blastocysts.
mal or impaired development (ND/ID) according to Istanbul consensus. Main results and the role of chance: Our model was able to predict high-
Neutrophils were obtained from peripheral blood from healthy donors. Cytokine grade blastocysts on day 5 using only day-1 time-lapse data with an AUC of
expression/production was evaluated by RT-qPCR/flow cytometry/ELISA. 0.665 [95% CI 0.650-0.681] and 10-fold stratified cross-validation of the training
Caspase-1 activity was measured using Flica probe. Neutrophils migration towards set. Our test results showed that the AUC was reproducible. These results are
HESC supernatants was evaluated using a transwell system. ROS production was comparable to studies attempting to make similar predictions based on a much
assessed by CDFH-DA probe. MMPs activity was evaluated by gelatin zymography. longer observation period of cleavage stage embryos.
Main results and the role of chance: We observed that ID-BCM stimulation Limitations, reasons for caution: Future prospective validation of our AI
increased caspase-1 activation and IL-1β production by decidualized cells, while algorithm is required using different patient populations, although the number
ND-BCM reduced IL-1β production (p<0.05, ANOVA). When a cytokine profile of embryos analyzed is outstanding.
was evaluated, we observed that ID-BCM increased CCL2 and CXCL12 expres- Wider implications of the findings: Our results broaden the understanding
sion as well as IL-8 secretion, while the production of the anti inflammatory of the potential capabilities for IVF of newly introduced AI systems. We showed
cytokine IL-10 was reduced (p<0.05, ANOVA). In contrast, ND-BCM stimula- that a deep neural net can solve one of the most challenging tasks, distinguishing
tion did not modulate these cytokines. Since IL-8 and CXCL12 are associated between top-grade embryos, solely based on unique visual features identified
to neutrophils recruitment towards the inflammatory sites, we evaluated whether in the first 24 hours after fertilization.
the HESC supernatants were able to modulate neutrophils migration using a Trial registration number: not applicable
transwells system. In this sense, ID-BCM increased neutrophils recruitment.
Additionally, when neutrophils were stimulated with HESC supernatants,
ID-BCM treatment leaded to higher ROS production (p<0.05, Friedman test),
suggesting an increased activation of these immune cells. Since menstruation is SELECTED ORAL COMMUNICATIONS
accompanied by a neutrophils influx, we hypothesize that ID-BCM stimulation SESSION 50: ANDROGEN TREATMENT IN FERTILITY
induces a “menstruation like” pro-inflammatory microenvironment. Due to the MANAGEMENT
fact that MMPs are the final effectors of menstruation, we tested MMPs activity
on HESC conditioned media. In this sense, ID-BCM stimulation increased gelet- 07 July 2020 Parallel 2 17:00 - 18:00
inolitic activity of MMP9 (p<0.05, ANOVA).
Limitations, reasons for caution: The present results were obtained using
immortalized cell lines and in vitro models. Further studies are necessary to
elucidate whether the mechanisms operate similarly in vivo and rule out any O-198 A prospective study of testosterone gel treatment in poor
factor not contemplated in vitro. ovarian reserve in IVF-ICSI cycles
Wider implications of the findings: These results provide new clues about R. Singh1, M. Singh2
the embryo maternal crosstalk and the natural embryo selection. Decidualized 1
BHOPAL TEST TUBE BABY CENTRE, INFERTILITY, BHOPAL, India ;
cells respond to blastocyst derived factors according to their quality either con- 2
BTTB Centre, Infertility, Bhopal, India
trolling or inducing a pro-inflamatory microenvironment. These findings might
contribute to a better understanding of reproductive disorders such as in vitro Study question: Does transdermal testosterone gel pretreatment
implantation failures. improve the outcome in women with poor ovarian reserve under-
Trial registration number: not applicable going IVF-ICSI Cycles ?
Summary answer: The testosterone gel has a significant impact on
O-197 What can we learn from the first 24 hours of embryo the fertility rate in women with a poor response in the IVF cycles.
development? A fully automated AI-based algorithm for What is known already: Poor ovarian reserve to external gonado-
identifying high-quality blastocysts. tropin drugs is one of the problems with IVF-ICSI cycles which can
D. Gilboa1, M. Meseguer2, R. Maor1, R. Weiner1, L. Alon1, L. Bori2, lead to cycle stop, access to fewer oocytes and embryos, and finally
L. Alegre2, D.S. Seidman3 reduced pregnancy rates.
1 No effective approach has been found yet to treat poor response
AiVF, IVF Research and Development, Tel Aviv, Israel ;
2 to ovarian stimulation.
IVIRMA Global, IVF Unit, Valencia, Spain ;
3 However, there are possible methods affecting the performance
Sheba Medical Center, IVF Unit, Ramat Gan, Israel
of gonadotropins on the ovaries such as high-dose gonadotropins,
growth hormone, glucocorticoids, and low-dose aspirin. Another
Study question: Can an AI-based system that analyses the first 24 hours of treatment is the use of low-dose androgens to improve ovarian
embryonic development open, for the first time, a window to the development response to gonadotropins which acts by increasing the intrafollicular
of the embryo? androgen and the number of follicle-stimulating hormone (FSH)
Summary answer: Using AI algorithms we have shown that a combination receptors on granulosa cells.
of previously unrecognized features within 24 hours after fertilization can help Study design, size, duration: 52 patients from July 2017 to July 2019,
identify high-quality blastocyst were randomly divided into two groups , 26 patients treated with a

placebo (lubricant gel, control group) and 26 patients treated with What is known already: Despite the controversy about its effect and
testosterone gel (Study group). uncertainty about the exact mechanisms, DHEA is added as an adjuvant in
Inclusion criteria were : patients for IVF cycles, patients older than over a quarter of IVF treatment protocols in patients with poor ovarian
40 years, a cycle with previous poor response, i.e., to obtain 3 or response. DHEA stimulates primordial follicles initiation and preantral follicular
<3 oocytes of the cycles by normal stimulation, AFC <5–7, development in the gonadotrophin responsive stages. It is believed that DHEA
AMH <0.5–1.1 ng/ml, Fertility outcomes were compared. increase ovarian susceptibility to FSH stimulation at the later stage but there
Participants/materials, setting, methods: 52 patients were randomly has not been an established evidence to support at the follicular/molecu-
divided into two groups. 26 patients treated with a placebo gel and lar level.
26 patients treated with testosterone gel . Study design, size, duration: In order to determine the efficacy of DHEA both
Patients who met inclusion (Bologna) criteria were placed in the alone and combination with the COH protocol in the low and normalovarian reserve
antagonist cycle group. The patients were randomly divided into two experimental groups, 75 Spraque Dawley female rats were divided into 15 groups.
groups each included 26 participants treated with a placebo and tes- (Control group, VCD group, VCD+DHEA group, VCD+DHEA+KOH group,
tosterone gel . Fertility outcomes were compared between two groups. VCD+DHEA+SF group, VCD+DMSO group, VCD+DMSO+KOH group,
The two groups were not statistically different in terms of FSH , VCD+DMSO+SF group, DMSO group, DMSO+DHEA group, DMSO+DHEA+KOH
AFC, AMH, group, DMSO+DHEA+SF group, DMSO+DMSO group, DMSO+DMSO+KOH
Main results and the role of chance: The number of oocytes and group and DMSO+DMSO+SF group).
embryos in the study (testosterone gel ) group were significantly Participants/materials, setting, methods: The experimental animal model
higher than in the control group. of DOR was composed by 4-Vinylcyclohexene diepoxide (VCD) injections
The mean number of oocytes obtained was 3.12 ± 1.14 versus 1.27 (160mg/kg/day for 15 days). After VCD or DMSO injections, DHEA injections
± 1.03 and embryos was 2.10 ± 1.08 versus 0.39 ± 0.48. were administered (60mg/kg/day for 45 days). COH protocol was then applied
The clinical pregnancy rate was 15% ( 4/26) in the study (testos- to the subjects. In ovarian tissues obtained from sacrificed subjects, COX-2 gene
terone gel ) group ,were significantly higher versus than in the control expression levels were analyzed by RT-qPCR, morphological evaluation and
group 04 % ( 1/26). ovarian follicle count were performed by hematoxylin-eosin staining.
In conclusion, there is evidence from this study that the use of Main results and the role of chance: It was determined that VCD injections
transdermal testosterone prior to ovarian stimulation in women caused a decrease in the number of follicles at different stages of develop-
who are considered poor responders, and this treatment has shown ment (p<0.05) and an increase in the number of atretic follicles (p<0.05) and
to significantly improve live birth rates and reduce the doses of FSH COX-2 gene expression levels (p<0.05) in the ovarian tissues. While DHEA
required for ovarian stimulation. injections did not affect folliculogenesis and COX-2 gene expression levels in
Androgen receptors are expressed in granulosa cells at early stages subjects with normal ovarian reserve, it was determined that DHEA injections
of follicle maturation, it is surprising that such a short treatment up caused a decrease in atretic follicle counts (p<0.02) and an increase in COX-2
to 20 days of testosterone supplementation could achieve signifi- gene expression levels in subjects with low ovarian reserve (p<0.01). DHEA
cantly higher live birth rates. Hence, extending testosterone supple- injections before the COH protocol affected folliculogenesis positively in both
mentation for a longer period could enhance the pool of follicles low and normal ovarian reserve experimental groups, and thus enhanced the
sensitive to gonadotrophins and therefore increase the number of efficiency of the COH protocol. Also, DHEA decreased COX-2 gene expression
oocytes available for retrieval. levels in all subjects(p<0.001).
Limitations, reasons for caution: Transdermal-testosterone may Limitations, reasons for caution: The limitations of the study is that clinical
improve the clinical outcomes for poor-ovarian-reserve. outcomes have not been evaluated.
One limitation is the low number of participants and exact sub- Wider implications of the findings: While evidence of using DHEA in
group of poor-ovarian-reserve who would benefit from this treat- clinical setting is currently uncertain, this study shows that DHEA can modify the
ment still needs to be identified. ovarian microenvironment and modulate the systemic inflammatory response
Although trends in all parameters appear to favour testosterone by reducing COX-2 expression.DHEA treatment potentially may be useful clin-
supplementation, further investigations are needed to confirm these ically as a means to increase the number of gonadotropin-responsive follicles
findings. for ovarian stimulation.
Wider implications of the findings: According to the results of our Trial registration number: Not applicable
study, the testosterone gel has a positive impact on fertility rate in
patients with poor-ovarian-reserve.
The identification of poor responders that could especially benefit O-200 Endocrine and histological effects of androgen treatment in
from testosterone treatment should be addressed in further female-to-male transsexual patients: a study model of long-term
studies. high-dose ovarian androgenization
Large studies on larger populations are recommended to be G. Casals Soler1, A. Borrás Capó1, A. Goday Cibeira1,
conducted. M. Mora Porta2, S. Peralta Flores1, F. Fàbregues Gasol1,
Trial registration number: not applicable J.M. Calafell Pozo1, D. Manau Trullàs1, F. Carmona Herrero3
1
Hospital Clinic de Barcelona, Assisted reproduction unit, Barcelona, Spain ;
O-199 The efficacy and molecular mechanism of 2
Hospital Clinic de Barcelona, Endocrine department, Barcelona, Spain ;
dehydroepiandrosterone in diminished ovarian reserve 3
Hospital Clinic de Barcelona, Gynecology department, Barcelona, Spain
C. Çakır1, G. Kuspınar1, S. Işıklar1, K. Aslan2, I. Kasapoglu2,
G. Uncu2, B. Avcı1 Study question: Which are the hormonal and ovarian histological effects of
1
Uludag University School of Medicine, Histology and Embryology & Gynecology a long-term high-dose androgen treatment using female-to-male transsexual
and Obstetric ART Center, Bursa, Turkey ; patients (FTMTP) as a study model?
2
Uludag University School of Medicine, Gynecology and Obstetric ART Center, Summary answer: According to this study model, long-term high-dose andro-
Bursa, Turkey gen treatment is associated with sequential preoperative hormonal changes,
heterogeneous postoperative endocrine parameters and characteristic ovarian
Study question: What is the effectiveness and molecular mechanism of dehy- histological features.
droepiandrosterone(DHEA) added to the controlled ovarian hyperstimula- What is known already: FTMTP treated with high-doses of androgens for a
tion(COH) protocol in diminished ovarian reserve (DOR) in vivo rat model? long period of time constitutes an excellent endocrinological, metabolic and his-
Summary answer: DHEA improves ovarian respons by increasing folliculo- tological study model of ovarian androgenization. The aim of the present inves-
genesis and suppresses apoptotic process of follicles with modulation of the tigation is: a) to identify the sequence of endocrinological changes in these patients,
systemic inflammatory response by reducing COX-2 gene expression. from the baseline period prior to the start of androgenization to the postoperative

stage and beyond, with controls up to 6 months after surgery; b) histological between endogenous androgen and achievement of pregnancy, with conflicting,
ovarian assessment after a long period of treatment with androgens at high doses. however, results, that have not been yet systematically reviewed.
Study design, size, duration: Longitudinal observational prospective study Study design, size, duration: A systematic review and meta-analysis was
including 60 FTMTP patients treated with long-term high-dose of systemic andro- conducted after performing a literature search in MEDLINE, CENTRAL until
gens (testosterone undecanoate, Reandron® 1g every 3 months by intramuscular October 2019. The main outcome was the association between levels of tes-
injection). The study points were three: before starting androgen treatment tosterone / DHEAS and the probability of pregnancy. Meta-analysis was con-
(point 0); after two years of androgen treatment, performed the day before ducted by pooling data to calculate standardized mean differences (SMD) with
surgery, which consisted in a laparoscopic hysterectomy and bilateral adnexec- 95% confidence intervals (CI), using random effects model.
tomy (point 1); and 6 months after surgery (point 2). Participants/materials, setting, methods: Thirteen eligible studies, includ-
Participants/materials, setting, methods: We performed serum hormonal ing 3977 patients, published between 1992 and 2017 were identified. In these
tests including AMH, FSH, LH, estradiol, androgenic profile (androstenedione, studies, testosterone and DHEAS levels were assessed prior to initiation of
testosterone, SHBG) and fasting insulin in each point of the study. stimulation among women who after embryo transfer achieved or not pregnancy.
Ultrasonographic scans were performed in 32 patients at point 0 and 1 of study. Three out of the thirteen studies were prospective, eight were retrospective
The histological examination was performed in histological sections stained with and in two studies the design was unclear.
hematoxylin and eosin. The stromal structure, the thickness of the tunica Main results and the role of chance: Inhibition of premature luteinizing
albuginea and the developmental stages of the follicles were evaluated, counting hormone (LH) surge was performed using gonadotrophin-releasing-hormone
follicles of each category separately. (GnRH) agonists (n=9 studies), GnRH-antagonists (n=2 studies), either
Main results and the role of chance: The mean age (±SD) was 27.81±5.81 GnRH-agonists or antagonists (n=1 study), while this information was not
years old (range 20-34). Five patients presented previous polycystic ovary syndrome reported in one study. Ovarian stimulation was performed with recombi-
(PCOS) criteria, with a transvaginal ultrasonography with ≥20 follicles <10mm, nant-FSH (n=6 studies), urinary FSH (n=4 studies), both recombinant and
ovarian volume ≥10 mm3 and oligomenorrhea. All patients became amenorrheic urinary FSH (n=2 studies), while this information was not reported in
after starting the hormonal treatment. At point 1 of the study, all the patients one study.
showed hirsutism, no follicular activity by ultrasonography, high testosterone serum Significantly lower testosterone levels were present in patients who achieved
levels (781,5 ng/ml ± 325,9) and other serum hormonal parameters in the reference live birth compared to those who did not (SMD: -0.21, 95%CI: -0.38 to -0.04,
ranges: AMH (3,23±1,4 ng/ml), FSH (5,95±1,97U/L), LH (4,7±3,45 U/L) and p=0.02, one study, 731 patients).
estradiol (46,8±24,9 pg/ml). The histological follicular population study demon- Testosterone levels were not significantly different between patients who
strated 88% of primordial follicles, 3.1% of primary follicles, 0.7% of secondary achieved clinical pregnancy and those who did not (SMD: -0.17, 95%CI: -1.27
follicles, 4.9% of antral follicles and 3.4% of atretic follicles. We must highlight a high to +0.93, p=0.76, eight studies, 2016 patients) as well as between those who
presence of luteinization of stromal cells (68.5% of cases) and only two cases with had positive pregnancy test compared to those who did not (SMD: -0.16, 95%CI:
stromal hyperplasia (1.8%). The thickness of the tunica albuginea was heterogeneous -0.43 to +0.11, p=0.24, four studies, 1230 patients).
(mean=0.55±0.22 mm; range 0.15-1.45). These ovarian samples share some char- DHEAS levels were not significantly different between patients who achieved
acteristics with PCOS ovaries although they do not reproduce their complete his- clinical pregnancy and those who did not (SMD: +0.05, 95%CI: -0.09 to +0.18,
tological criteria (Hughesdon, 1982). Hormonal assessment showed high p=0.53, six studies, 942 patients) as well as between those who had positive
heterogeneity at point 3 of study (mean ± SD; range): FSH 52.0±36.0 U/L (0.4- pregnancy test compared to those who did not (SMD: -0.01, 95%CI: -0.73 to
102.4), LH 26.5±21.59 U/L (0.1-68.3) and estradiol 36.2±7.9 pg/ml (27-45). +0.71, p=0.98, one study, 120 patients).
Limitations, reasons for caution: The sample size is limited. We could not Limitations, reasons for caution: Heterogeneity was present regarding
include a control group of non-androgenized patients due to ethical reasons. patient population, as well as the type of ovarian stimulation protocol used,
Ovarian samples from surgery for other indications did not present adequate which could not be meaningfully explored by subgroup analysis, due to the limited
characteristics to be considered as controls due to histological changes secondary number of eligible studies.
to different associated pathologies. Wider implications of the findings: The current study does not suggest
Wider implications of the findings: We demonstrated that long-term high- that there is a positive association between testosterone / DHEAS levels at
dose systemic treatment with androgens is associated with: a) sequential hor- initiation of stimulation and pregnancy achievement, questioning the rationale
monal changes during the preoperative period and heterogeneous postoperative behind testosterone and DHEAS pre-treatment in patients undergoing ovarian
data, possibly due to a hypothalamus and/or pituitary gland inhibition in a stimulation for IVF.
selected group of patients; b) characteristic follicular and stromal changes in the Trial registration number: Not applicable
ovarian histological study.
Trial registration number: Not necessary
O-201 Association between levels of testosterone /DHEAS at SELECTED ORAL COMMUNICATIONS

initiation of ovarian stimulation for in vitro-fertilization and the SESSION 51: RM: NEW DIAGNOSTIC AND THERAPEUTIC
probability of pregnancy: a systematic review and meta-analysis ASPECTS
V. Christoforaki1, J. Bosdou1, G. Lainas1, T. Lainas2, 07 July 2020 Parallel 3 17:00 - 18:10
A. Kalpatsanidis1, A. Mouza1, G. Grimbizis1, E. Kolibianakis1
1
Aristotle University of Thessaloniki- Medical School, Unit for Human
Reproduction- 1st Department of Ob/Gyn, Thessaloniki, Greece ;
2 O-202 The transcriptomic profile of endometrial receptivity in
EUGONIA, Assisted Reproduction Unit, Athens, Greece
recurrent miscarriage
Study question: Is the probability of pregnancy associated with levels of tes- L. Craciunas1, O. Pickering1, J. Chu1, M. Choudhary2,
tosterone/ dehydroepiandrosterone-sulphate (DHEAS) at initiation of ovarian J. Zurauskiene3, A. Coomarasamy1
stimulation for in-vitro fertilization (IVF)? 1
University of Birmingham, Tommy’s National Centre for Miscarriage Research,
Summary answer: At initiation of ovarian stimulation, a significant negative Birmingham, United Kingdom ;
association exists between testosterone and live birth, whereas a non-significant 2
Royal Victoria Infirmary, Assisted Conception Unit, Newcastle, United Kingdom ;
association exists between DHEAS and clinical/biochemical pregnancy. 3
University of Birmingham, Centre for Computational Biology, Birmingham, United
What is known already: It has been shown that androgen action is required Kingdom
for normal follicular growth and progression beyond pre-antral stage. Androgens
exert direct autocrine and/or paracrine effects that regulate follicular develop- Study question: What is the transcriptomic profile of the endometrium in
ment, by augmenting follicle stimulating hormone (FSH) receptor expression in women who suffered unexplained recurrent miscarriage and can it predict the
granulosa cells. Until today, several studies have examined the association fate of the subsequent pregnancy?

Summary answer: Endometrium biopsied from women who suffered high Summary answer: This meta-analysis has demonstrated the importance of
order recurrent miscarriage has a distinct transcriptomic profile and the differ- screening for microbiota, including Bacterial Vaginosis (BV) and Chlamydia
ences may predict the fate of subsequent pregnancies. Trachomatis (CT).
What is known already: Embryo chromosomal abnormalities represent the What is known already: Approximately 15% of pregnancies end up in mis-
most common cause of first trimester miscarriage in addition to a broad range carriage, contributing to one of the most common adverse pregnancy outcomes.
of maternal health problems. Genomic studies based on the products of con- Most causes remain unknown, although a complex interplay of different factors
ception reported decreasing rates of chromosome abnormalities as the order are thought to be involved and the presence of infection plays a pivotal
of miscarriage increased, with a significant turning point after four or five mis- role. Microbiota affects all facets of reproduction including pregnancy loss; how-
carriages, suggesting the cause of miscarriage more likely to be non-chromo- ever, few studies have been conducted to quantify the prevalence of each micro-
somal in higher order miscarriages. Endometrial transcriptomics aim to describe biome following ART (assisted reproductive technology) and its role with
the full range of RNA transcripts that are produced at the level of the endome- recurrent miscarriage.
trium with an aim to develop a new test of endometrial receptivity. Study design, size, duration: Systematic review and meta-analysis of pub-
Study design, size, duration: This was a multicentre cohort study performed lished controlled studies. Searches were conducted from January 1999 to June
in the Tommy’s National Centre for Miscarriage Research in Birmingham, Saint 2019 on MEDLINE and EMBASE using the following search terms: miscarriage
Mary’s Hospital in Manchester and Royal Devon & Exeter Hospital, United OR recurrent miscarriage OR microbiome (bacterial vaginosis (BV), chlamydia,
Kingdom. The study was conducted between December 2017 and December herpes simplex virus (HSV), human papillomavirus (HPV), cytomegalovirus
2019. It included 24 women diagnosed with unexplained recurrent miscarriage (CMV) and toxoplasma or toxoplasmosis) AND IVF. Studies were limited to
based on strict inclusion criteria to rule out known causes and risk factors for those published in English.
miscarriage, while increasing the risk of an unidentified endometrial receptivity Participants/materials, setting, methods: A total of 225 full-text articles
problem. were assessed for eligibility from 310 records identified through database search-
Participants/materials, setting, methods: Endometrial biopsies were ing. 30 studies were included in the quantitative synthesis (meta-analysis).
obtained during implantation window from 18-35 years old women, not pregnant References were based on title and abstract and assessed utilising the Newcastle–
and regularly menstruating, diagnosed with unexplained recurrent miscarriage Ottowa Quality Assessment Scales.
by undertaking standard investigations according to the ESHRE guidelines. They Main results and the role of chance: For patients undergoing IVF treat-
were excluded for miscarriage risk factors such as smoking, obesity or hyper- ment, a significant association was observed with early spontaneous miscarriage
prolactinemia. The RNA transcripts abundances were quantified using Kallisto. for Bacterial Vaginosis (BV) (RR 1.56, 95% CI 1.14–2.21), which supports existing
R packages tximport and DESeq2 were used to summarize count estimates at literature. Chlamydia Trachomatis (CT) had two studies which limited clinical
the gene level and to analyse the differential gene expression. significance and Herpes Simplex Virus (HSV) did not show any statistical signif-
Main results and the role of chance: When compared to controls (two or icance for miscarriage following IVF (RR 1.28, 95% CI 0.85-1.92). There were
three miscarriages), women who suffered four or more miscarriages had 19 no studies for the other microbiota. There was no significant relationship
differently expressed genes after adjustment for multiple comparisons. They were between microbiota infection and recurrent miscarriage; however, meta-analysis
related to biological processes such as immunity (HLA-DMA, CCR8, ALOX5), was carried out for the association between CT and Cytomegalovirus (CMV)
energy production (ATP12A), hormone secretion (CGA), adhesion (CHAD, with recurrent miscarriage (OR 1.30, 95% CI 0.61-2.78 and OR 2.58, 95% CI
ADGRF2, AQP5, TBCD, CTNND1, NKD2) and cell proliferation (NCCRP1). 0.84-7.90 respectfully). There were insufficient studies for the remaining micro-
Important differences in the transcriptomic profiles were identified when biota to perform a meta-analysis.
women who suffered a subsequent miscarriage were compared to women who Limitations, reasons for caution: There is a paucity of data for specific
had a live birth following the endometrial biopsy. The number of significantly micriobiota. Some sample sizes were too small to find any significant association
differently expressed genes was 421. Their gene ontology mapped to processes with miscarriage and further study is necessary.
relevant to embryo implantation and early stages of pregnancy such as biosyn- Wider implications of the findings: This study has shown the importance
thetic process (129 genes), signal transduction (124 genes), response to stress of screening before ART treatment and for women with recurrent miscarriages.
(113 genes), immune system process (84 genes), cell differentiation (75 genes), There may be a role for specific commensals including Lactobacillus that may
catabolic process (67 genes), cell death (66 genes), cell proliferation (59 genes), improve the intrauterine environment to aid with implantation. Being the first
homeostatic process (41 genes), cytoskeleton organization (40 genes), cell adhe- meta-analysis within this field is valuable for patient counselling.
sion (38 genes) or reproduction (26 genes). Trial registration number: not applicable
Limitations, reasons for caution: This was a first discovery study to eval-
uate endometrial receptivity using transcriptomics in a relatively small selected O-204 The metabolomic profile of endometrial receptivity in
population of women who suffered unexplained recurrent miscarriage. The recurrent miscarriage
findings reported here have not been validated in an independent sample L. Craciunas1, J. Chu1, O. Pickering1, L. Mohiyiddeen2, W. Dunn3,
set yet. A. Coomarasamy1
Wider implications of the findings: Abnormal transcriptomic profiles may 1
University of Birmingham, Tommy’s National Centre for Miscarriage Research,
explain variations in endometrial receptivity and build up on the evidence to Birmingham, United Kingdom ;
support non-chromosomal causes for higher order miscarriages. Validation of 2
Central Manchester University Hospitals, Saint Mary’s Hospital, Manchester,
the biological changes identified prior to a subsequent miscarriage or live birth United Kingdom ;
may aid in developing a prediction test for use in clinical practice. 3
University of Birmingham, School of Biosciences, Birmingham, United Kingdom
Study question: What is the metabolomic profile of the endometrium in
women who suffered unexplained recurrent miscarriage and can it predict the
O-203 Systematic review and meta-analysis of prevalence of fate of the subsequent pregnancy?
microbiota in ART cycles and recurrent pregnancy loss Summary answer: Endometrium biopsied from women who suffered high
S. Robati1, E. Chronopoulou1, E. Theodorou1, P. Serhal2, W. Saab2, order recurrent miscarriage has a distinct metabolomic profile and the differences
V. Seshadri2 may predict the fate of subsequent pregnancies.
1
Institute for Women’s Health- University College London, Faculty of Population What is known already: Embryo chromosomal abnormalities represent the
Health Sciences, L0ndon, United Kingdom ; most common cause of first trimester miscarriage in addition to a broad range
2
The Centre for Reproductive & Genetic Health CRGH, Population Science and of maternal health problems. Genomic studies based on the products of con-
Women’s Health, London, United Kingdom ception reported decreasing rates of chromosome abnormalities as the order
of miscarriage increased, with a significant turning point after four or five mis-
Study question: Does microbiota influence miscarriage rates in ART cycles carriages, suggesting the cause of miscarriage more likely to be non-chromo-
and contribute to recurrent miscarriage? somal in higher order miscarriages. Endometrial metabolomics aim to describe

the full range of metabolites identified in a tissue sample obtained from the a specific gene expression profile suitable for endometrial function evaluation.
endometrium with an aim to develop a new test of endometrial receptivity. ER Map® is a molecular diagnostic tool able to accurately predict endometrial
Study design, size, duration: This was a multicentre cohort study performed receptivity status by analysing the gene expression profile of an endometrial
in the Tommy’s National Centre for Miscarriage Research in Birmingham, Saint biopsy by high-throughput RT-qPCR. In this study, the experience of the clinical
Mary’s Hospital in Manchester and Royal Devon & Exeter Hospital, United application of ER Map® for WOI identification and personalised scheduling of
Kingdom. The study was conducted between December 2017 and December embryo transfer in subfertile couples is presented in detail.
2019. It included 24 women diagnosed with unexplained recurrent miscarriage Study design, size, duration: This is a retrospective study analysing endo-
based on strict inclusion criteria to rule out known causes and risk factors for metrial function and ART outcomes of 2256 patients undergoing endometrial
miscarriage increasing the risk of an unidentified endometrial receptivity receptivity assessment by ER Map® between March 2016 and September 2019.
problem. Results obtained when single embryo transfers were scheduled either on the
Participants/materials, setting, methods: Endometrial biopsies were moment of endometrial receptivity (WOI timeframe) as recommended by ER
obtained during implantation window from 18-35 years old women, not preg- Map®, or deviating from this recommendation are analysed and compared.
nant and regularly menstruating, diagnosed with unexplained recurrent miscar- Participants/materials, setting, methods: Patients referred for ER Map®
riage by undertaking standard investigations according to the ESHRE guidelines. analysis were included in the study. Endometrial biopsy samples obtained in a
They were excluded for miscarriage risk factors such as smoking, obesity or HRT cycle at P4+5.5 were ER Map® tested and classified into ‘Receptive”, ‘Pre-
hyperprolactinemia. The metabolite composition and relative concentrations receptive” or ‘Post-receptive”. Based on ER Map® results, a recommendation
of samples were analysed applying ultra-high performance liquid chromatogra- for embryo transfer was performed. Single blastocyst transfers (sET) were per-
phy-mass spectrometry. Raw data were processed applying XCMS and statistical formed in all cases. Clinical outcome measures were: positive β-HCG rate [βR],
analysis was applied using the software MetaboAnalyst. clinical pregnancy rate [CPR] and pregnancy loss rate [PLR].
Main results and the role of chance: There were distinct metabolomic Main results and the role of chance: ER Map® analysis of endometrial
profiles between the endometrial samples obtained from women with higher receptivity status showed that 771 out of 2256 patients (34.2%) studied had a
order miscarriage defined as more than four recurrent miscarriages and more displaced WOI.
than five recurrent miscarriages, respectively. More biological differences were Analysis of ART outcomes showed a significantly higher clinical pregnancy
identified when the cut-off for the extreme group was set at five miscarriages rate in sETs scheduled within the WOI predicted by ER Map® compared to
and the analysis showed groups of metabolites from the same metabolite class, transfers that deviated more than 12h from ER Map® recommendation (βR
supporting the class-specific changes with more confidence. These groups of 56.09% vs 41.54%, X2 test p=0.02; CPR 44.35% vs 23.08%, X2 Test p <0.001).
metabolites related to cardiolipins, ceramides and glycerophospholipids. Pregnancy rates were even lower when the deviation from the WOI identified
Cardiolipins are mitochondrial cell membrane lipids and indicate mitochondrial by ER Map® exceeded 24h (βR 56.09% vs 26.92%, X2 test p=0.012; CPR
stress and morphological changes. Ceramides are generally believed to be 44.35% vs 19.23%; X2 test p=0.004). The deviation of embryo transfers from
lipotoxic, while glycerophosphoinositols are considered to be signalling the WOI predicted by ER Map® had also an impact on the progression of
molecules. pregnancy. A significant increase in pregnancy loss (~2-fold) was detected in
Important biological changes were identified in the metabolomic profiles of the group of transfers that deviated from ER Map® recommendation compared
women who suffered a subsequent miscarriage compared to those who had a to transfers performed within the ER Map® WOI (PLR 41.4% vs 21.05%, X2
live birth following the endometrial biopsy. The most relevant metabolite classes Test p <0.022).
were triacylglycerides (fatty acid storage molecules), purine and pyrimidine This study provides strong evidence that ER Map® endometrial receptivity
metabolism and ether glycerophospholipids. evaluation can reliably identify the WOI and improve clinical outcomes.
Limitations, reasons for caution: This was a first discovery study to evaluate Personalisation of progesterone duration pre-treatment before transfer renders
endometrial receptivity using metabolomics in a relatively small selected popu- significantly improved ART results, increasing the likelihood of pregnancy and
lation of women who suffered recurrent miscarriage. The findings reported here reducing the risk of miscarriage.
have not been validated in an independent sample set yet. Limitations, reasons for caution: ER Map® test can serve as a valuable tool
Wider implications of the findings: Abnormal metabolomic profiles may to improve ART results, however, to determine the true extent of any clinical
explain variations in endometrial receptivity and build up on the evidence to benefits, other types of investigations, such as non-selection studies and ran-
support non-chromosomal causes for higher order miscarriages. Validation of domized controlled trials, will also be necessary.
the biological changes identified prior to a subsequent miscarriage or live birth Wider implications of the findings: The application of ER Map® for the
may aid in developing a prediction test for use in clinical practice. identification of cases of WOI displacement and personalised embryo transfer
Trial registration number: NCT03442335 scheduling is an effective strategy for improving ART outcomes. Not only patients
suffering from implantation failure but also couples experiencing recurrent mis-
carriage can benefit from the accurate identification of the WOI.
O-205 The precise identification of the window of implantation Trial registration number: Not applicable
using the molecular tool ER Map® in ART cycles significantly
improves clinical outcomes O-206 Recurrent pregnancy loss: diagnostic workup after two or
M. Enciso1, J. Aizpurua2, B. Rodriguez-Estrada1, I. Jurado1, three pregnancy losses? A systematic review of the literature and
M. Ferrandez-Rives1, E. Rodriguez3, E. Pérez-Larrea3, meta-analysis.
A.B. Climent4, K. Marron5, J. Kennedy5, J. Sarasa1 M. Van Dijk1, A. Kolte2, J. Limpens3, E. Kirk4, S. Quenby5,
1
iGLS, Research and Development, Alicante, Spain ; M. Van Wely1, M. Goddijn1
1
2
IVF-Spain, Gynaecology, Alicante, Spain ; Amsterdam UMC- University of Amsterdam, Center for Reproductive Medicine,
3
IVF Donostia, Gynaecology, San Sebastian, Spain ; Amsterdam, The Netherlands ;
2
4
IVF-Spain, Statistics, Alicante, Spain ; Recurrent Pregnancy Loss Unit- Fertility Clinic 4071- University Hospital Copenhagen-
5
Sims IVF Clinic, IVF Unit, Dublin, Ireland Rigshospitalet-, Recurrent Pregnancy Loss Unit, Copenhagen, Denmark ;
3
Amsterdam UMC- University of Amsterdam, Medical library, Amsterdam, The
Study question: Does the identification of the window of implantation using Netherlands ;
the endometrial receptivity test ER Map® improve ART outcomes? 4
Royal Free Hospital, Obstetrics and Gynaecology, London, United Kingdom ;
Summary answer: The use of ER Map® test for endometrial receptivity 5
University Hospital Coventry- Warwick Medical School., Division of Reproductive
evaluation and personalised scheduling of embryo transfer significantly improves Health, Warwick, United Kingdom
pregnancy rates.
What is known already: The endometrium reaches a receptive status for Study question: Do abnormal test results for definite or probable risk factors
embryonic implantation around days 19-21 of the menstrual cycle. During this for RPL occur with equal frequency in women with two versus women with
period, known as the window of implantation (WOI), the endometrium shows three or more pregnancy losses?

Summary answer: No difference in prevalence in uterine abnormalities and Summary answer: The weight of individual parameters is not static but
APS was found, a difference in prevalence of chromosomal abnormalities, throm- evolves at each step of IVF process, contributing to higher accuracies in live birth
bophilia and thyroid disorders cannot be excluded. prediction (C-index).
What is known already: Recurrent pregnancy loss (RPL) occurs in 1 – 3% What is known already: Several predictive models have been developed to
of all couples trying to conceive. No consensus exists when to perform testing help ART clinicians in counselling patients and improve decision-making process.
for risk factors in couples with RPL. Some guidelines recommend testing if a The first validated models, proposed by Templeton et al. in 1996, then improved
patient has had two pregnancy losses whereas others advise to test after three by Nelson et al. in 2011, integrated demographic and clinical parameters before
losses. Before trying to conceive, couples and clinicians attempt to find an expla- starting IVF. In 2016, Mc Lernon et al. proposed a new predictive tool based on
nation for their pregnancy losses and a treatment that will prevent a recurrence, additional information collected during the first IVF attempt providing an indi-
especially in cases with modifiable risk factors, such as thyroid disorders and vidualized estimate of a couple’s cumulative chances of live birth.
APS. Is it necessary to wait before the next pregnancy loss will occur? To our knowledge, no predictive model exists that dynamically adjusts the
Study design, size, duration: A systematic review of literature and meta-anal- LBP throughout the IVF process, as new data become available.
ysis was performed. Study design, size, duration: We designed our study on retrospective data
Participants/materials, setting, methods: Relevant studies were identified of 9587 IVF cycles between 2016-2017 from four IVF centers. Only cycles with
by a systematic search in OVID Medline and EMBASE from inception. A search verified, consistent and complete data up to final outcome were included.
for recurrent pregnancy loss was combined with a broad search for terms indic- Parameters were split into four categories following the IVF process: (1) demo-
ative of number of pregnancy losses, screening/testing for pregnancy loss or graphic (patient baseline characteristics), (2) ovarian, (3) laboratory and (4)
the prevalence of known risk factors. Meta-analyses were performed in case of transfer data.
adequate clinical and statistical homogeneity. The quality of the studies was Participants/materials, setting, methods: We implemented a univariate
assessed using the Newcastle-Ottawa scale. and multivariate evolutive logistic regression, validated with Random Forest,
Main results and the role of chance: From a total of 1985 identified pub- with iterative parameter selection based on 5% significance level. No parameters
lications, 21 were included in this systematic review and 19 were suitable for were included or excluded by default. Validation was internally based on a 70%-
meta-analyses. For uterine abnormalities (7 studies, OR 1.00, 95%CI 0.79 – 1.27, 30% split. Results are reported as odd ratios and C-index.
I2=0%) and for antiphospholipid syndrome (3 studies, OR 1.04, 95%CI 0.86 – LBP were re-calculated by fitting a new model at the addition of each of the four
1.15, I2=0%) we found low quality evidence for a lack of a difference in preva- categories, finally yielding probabilities for each fresh or frozen embryo transfer.
lence of abnormal test results between couples with two versus three or more Main results and the role of chance: Accuracy of the model in predicting
pregnancy losses. We found insufficient evidence of a difference in prevalence live birth progressively increased from 0.64 to 0.72 as ovarian stimulation, lab-
of abnormal test results between couples with two versus three or more preg- oratory and embryo transfer parameters were respectively added.
nancy losses for chromosomal abnormalities (10 studies, OR 0.78, 95%CI 0.55 Female age remains strongly predictive of LBP at all steps. After fresh embryo
– 1.10), inherited thrombophilia (5 studies) and thyroid disorders (2 studies, OR transfers, LBP decreases linearly with increasing female age, whereas its impact
0.52, 95%CI: 0.06 – 4.56). becomes significant beyond 37 years for frozen transfers (OR=0.59; 95%CI=0.4-0.85).
Limitations, reasons for caution: A methodological limitation of this study Female BMI influence is constant throughout all steps (OR=0.98;
is the definition of the study groups. On average, 15-20% of women with two 95%CI=0.97-0.99).
losses will experience a loss in the next pregnancy some of the women, some Among infertility causes, endometriosis showed the worst impact on LBP.
will be in the other group if evaluated at a different time point. AFC and AMH are significant predictors when only baseline characteristics
Wider implications of the findings: The results of this study may support are included in the model, but their impact is not statistically significant anymore
investigations after two pregnancy losses in couples with RPL, it should be when the number of retrieved oocytes becomes available.
stressed that additional studies of the prognostic value of investigations used High gonadotrophin dose negatively impacts LBP in fresh embryo transfers
in RPL are urgently needed. An evidenced-based treatment is currently not avail- (OR=0.52; CI=0.42-0.64).
able in the majority of cases. Endometrial thickness >7mm positively impacts LBP at the transfer step
Trial registration number: Not applicable (OR=1.07; CI=1.04-1.10).
As an example, a 38 year-old woman whose baseline characteristics give her
22% LBP for her first fresh transfer if AMH is 2.5ng/mL, has 22% recalculated
SELECTED ORAL COMMUNICATIONS chances if 3 oocytes are retrieved and 28% if 8 are collected. If that same woman
SESSION 52: AI. A NEW TOOL TO ASSESS ART OUTCOMES has an AMH of 1.5ng/ml, her LBP drops to 17%, but increases to 21% and 27%
AND HELP PATIENTS? if 3 and 8 oocytes are retrieved, respectively.
Limitations, reasons for caution: Our models need to be further evaluated
07 July 2020 Parallel 4 17:00 - 18:00
by performing an external validation: infertile population in France might be
slightly different than in other countries (ie: BMI, ethnicity, etc). The limited
O-207 Patient-oriented counselling: predicting live birth number of cycles results from deliberate stringent quality control process,
probability (LBP) at each step of the In Vitro Fertilization (IVF) whereby only verified, consistent, and complete cycle data were kept.
process via machine learning. Wider implications of the findings: The prospect of this work is to develop
V. Grzegorczyk Martin1, T. Fréour2, C. Avril1, A. De Bantel - Finet1, a practical and evolutive tool to help clinicians accompany their patients by always
P. Barrière2, J.L. Pouly3, M. Grynberg4, I. Parneix5, J. Roset1, providing the latest personalized estimates of LBP, before and throughout the
T. Grzegorczyk6 IVF intervention, at each fresh and/or frozen embryo transfer.
1
Clinique Mathilde, Centre d’Assistance Médicale à la Procréation, rouen, France ; Trial registration number: not-applicable
2
CHU de Nantes, Service de Biologie et Médecine de la Reproduction, Nantes, France ;
3 O-208 Is IVF making more male embryos and does maternal
CHU Clermont Ferrand, Unité de Fécondation In Vitro Département de
AMH play a role in this?
Gynécologie Obstétrique et Reproduction Humaine Polyclinique de l’ Hotel-Dieu,
Clermont Ferrand, France ; K. Hammer1, C. Cherston1, S. Vagios1, C. Sacha1, D. Pepin2,
4
Hôpital Antoine-Béclère, Médecine de la reproduction et préservation de la C. Bormann1, M. Kumar Kanakasabapathy3, P. Thirumalaraju4,
fertilité, Clamart, France ; H. Shafiee5, M. Morris-Sabatini1
1
5
Polyclinique Jean Villar, Centre d’AMP - IFREARES Bordeaux, Bruges, France ; Massachusetts General Hospital, OBGYN-Reproductive Endocrinology & Infertility,
6
Teranalytics, Data management, Boston, U.S.A. Boston, U.S.A. ;
2
Massachusetts General Hospital, Pediatric Surgical Research Laboratories-
Study question: How does a machine learning tool for live birth prediction Department of Surgery, Boston, U.S.A. ;
accuracy evolve to counsel couples as demographic, ovarian stimulation, labo- 3
Harvard Medical School- Brigham and Women’s Hospital, Department of
ratory and transfer data become available? Medicine, Boston, U.S.A. ;

4 3
Harvard Medical School, Harvard-MIT Division of Health Sciences and UCL Institute for Women’s Health, Department of Reproductive Health, London,
Technology, Boston, U.S.A. ; United Kingdom
5
Harvard Medical School, Department of Medicine, Boston, U.S.A.
Study question: What is the time to pregnancy for users of the Natural Cycles
Study question: Does anti-mullerian hormone (AMH) predict neonatal sex if fertility awareness based mobile application to identify the fertile window?
undergoing in vitro fertilization? Summary answer: Women <35 years, with no anovulation, regular cycles
Summary answer: Increasing AMH levels correlate with an increased trend and who regularly logged sex (52% of cohort) had a 12-month pregnancy prob-
for male births, likely related to rate of developmental progression of male ability of 96%.
embryos by day 5. What is known already: Women trying to get pregnant are encouraged to
What is known already: A sex skew toward male neonates after IVF has identify their fertile window using methods such as their menstrual cycle dates,
been described, but it is unclear why this skew exists. This phenomenon may basal body temperature, changes in cervical mucus and lutenising hormone surge
be related to speed at which a male embryo develops to the blastocyst stage identification. But these methods can become time consuming and stressful.
leading to an increase in their selection for transfer. We propose that maternal Using an app which can calculate the fertile window using an algorithm may be
AMH also plays a role in early embryonic development. AMH is known for its easier and more accurate. This is one of the first studies to examine the time to
role in sexual differentiation and as a marker for ovarian reserve. AMH receptor pregnancy for women using an app that identifies the fertile window.
has been heavily identified in male placentae and fetal membranes and likely Study design, size, duration: This observational study included 5376 women
impacts early embryonic development. with an age range of 18 to 45 years. They used the app in Plan pregnancy mode
Study design, size, duration: Retrospective data analysis of 658 IVF cycles between September 31st 2017 and August 31st 2018 to allow at least 12 months
from January 2015 to December 2019 at an academic institution in the United of potential usage up to the cutoff date of September 1st 2019.
States. Data include SART primary outcomes, PGT outcomes, artificial intelli- Participants/materials, setting, methods: The time to pregnancy was
gence (AI) data on embryo selection and sex by either PGT or neonatal outcome. analysed related to age, gravidity, BMI, cycle length variation, average cycle length,
We hypothesize that a higher AMH level will lead to more male high-quality and behavioural characteristics such as frequency of sexual intercourse via hazard
embryos on day 5 of embryo culture. ratios (HRs) using Cox regression adjusted for fixed covariates.
Participants/materials, setting, methods: SART data included women Main results and the role of chance: 3166 women achieved pregnancy
ages 27-44 with AMH levels drawn within 1 year of IVF. PGT cycles additionally within 13 cycles, while 570 women were still trying to get pregnant at the end
included EmbryoScope data using an AI algorithm trained to select the highest of the observation period. Overall the one-year pregnancy probability was 83%
quality embryo by morphology for transfer on day 3 and day 5 of development. (95% CI: 82%-84%). The mean time to pregnancy for the whole cohort was 3.3
Results were analyzed by AMH level within quartiles for the following endpoints: cycles (95% CI: 3.2-3.4). Women <35 years, with no anovulation, regular cycles
ratio male:female neonate sex, ratio male:female PGT result, sex of day 3 and and regularly log sex (52% of cohort) had a 12 month pregnancy probability of
day 5 embryo selected using AI. 96% and a mean time to pregnancy of 2.4 cycles (95% CI: 2.2-2.6). Lower
Main results and the role of chance: SART contained 658 singleton new- pregnancy probability was associated with the following exposures: ages 35 to
borns, 53.8% (n=354) were male and 46.2% (n=304) female. AMH correlated 45 (HR 0.69, 95% CI: 0.61-0.78), high cycle length variation (HR 0.89, 95% CI:
with male neonate; for every 1ng/mL increase in AMH the odds for a male 0.77-1.01) and sexual intercourse logged on less than 10% of days (HR 0.84,
neonate increased 5.4%, p=0.017. Quartile analysis demonstrated that at or 95% CI: 0.71-1.01). Anovulatory cycles were also associated with a worse out-
above the seventy-fifth percentile, AMH=5.11ng/mL, was associated with male come with a one-year pregnancy probability of 72%.
sex; male to female 60.7% (n=99) vs 39.3% (n=64). AMH less than the seven- Limitations, reasons for caution: Users had higher educational level and
ty-fifth percentile had a male to female ratio 51.5% (n=255) vs 48.5% (n=240), lower BMI than would be expected in the general population. Close to 50% had
p=0.04. characteristics associated with sub-fertility such as age > 35 years, highly variable
PGT-A cycles, n=209, had more female than male embryos, 52,1% (368/706 cycle length or anovulatory cycles. Sex logging data was incomplete which limited
embryos) female vs 47.9% (368/706 embryos) male. AI analysis of the highest the conclusions that could be made.
quality embryo demonstrated a difference in sex ratio between day 3 and day Wider implications of the findings: Fertility awareness apps have an import-
5. Day 3 cohort favored female embryos at higher AMH levels than male ant role in educating women and their partners about fertility and facilitating
embryos, female (median 2.3, IQR 1.4-3.8) vs male (median 1.3-3.0), p=0.042. discussions around the topic. In the future smart algorithms may facilitate early
Day 3 embryos and AMH above the seventy-fifth percentile demonstrated a identification of couples who may benefit from infertility assessment.
trend towards female sex, male 35.7% (n=10) vs female 64.3% (n=18), p=0.136. Trial registration number: 2016/2037-31
Day 5 embryos had comparable AMH values for both sexes, male median 2.1
(IQR 1.4-3.7) and female median 2.1 (IQR 1.3-2.9), p=0.45. AMH above the O-210 Interactive information provision during IVF/ICSI
seventy-fifth percentile had an increased trend toward male embryo selection treatment by using an app: A randomized controlled trial
67.9% (n=19) male vs 32.1%(n=9) female, p=0.23. M. Keijsers1, T. Timmers2, I. Van Rooij1, J.M.J. Smeenk1
Limitations, reasons for caution: This is a retrospective review of SART 1
Elisabeth Twee Steden Hospital, Obstetrics & Gynecology, Tilburg, The
data combined with data from a verified artificial intelligence algorithm to identify
Netherlands ;
the highest quality embryos during development. More work is needed to elu- 2
Interactive Studios, software development, Rosmalen, The Netherlands
cidate how AMH exposure of the oocyte may differently impact the develop-
ment of male vs female embryos. Study question: An application (app) has been developed providing patients
Wider implications of the findings: AMH levels in the follicle may have with information in an interactive way; will the use of this tool result in a change
implications for early embryo development kinetics. Providers must account for of knowledge, self-reliance and care consumption?
sex differences in embryo development so as not to artificially skew the sex ratio Summary answer: Patients in the app group achieved a higher level of
when selecting embryos for transfer. knowledge and were more satisfied with treatment compared to the con-
Trial registration number: not applicable trol group.
What is known already: Patients undergoing an ivf/icsi treatment experience
a lack of knowledge, which causes (unnecessary) stress during their treatment.
O-209 Time to pregnancy for women using a fertility awareness
Furthermore, they receive a lot of information in a limited timeframe in the clinic
based mobile application to plan a pregnancy
often resulting in an insufficient uptake of relevant details. Patients would like to
J. Pearson1, S. Rowland1, C. Favaro1, E. Berglund-Scherwitzl1, see an improvement in providing information. Routinely providing comprehen-
R. Scherwitzl1, K. Gemzell-Danielsson2, J. Harper3 sible and structured information, in particular about the practical aspect of the
1
Natural Cycles Nordic AB, Luntmakagatan 26, Stockholm, Sweden ; treatment, in a patient controlled way could be a solution.
2
Karolinska Institute, Head Department of Women´s and Children´s Health, Study design, size, duration: A single center randomized controlled trial
Stockholm, Sweden ; was conducted. The app group had access to the informative and personal app,

including informative videos and quiz questions.The control group received the and Peters. It involved five key stages: 1) identifying the research question; 2)
current/normal form of information provision. Each patient received four ques- identifying relevant studies; 3) study selection; 4) charting the data; and 5) col-
tionnaires at fixed timepoints prior to, during and after treatment. The queston- lating, summarising and reporting results.
naires focussed respectively on information provision, self-reliance, care Participants/materials, setting, methods: Articles were included if they
consumption, level of knowledge and finally the satisfaction of care during treat- addressed one or more of the following topics: 1. What are the factors that
ment was evaluated. shape the commercial organisation of ART? 2. How are commercial influences
Participants/materials, setting, methods: In the period from April 2018 to currently identified, appraised, regulated and managed? 3. How should commer-
September 2019, 51 patients were included into the study. Native patients, so cial influences be addressed, by whom, and with what guidance? The search
without any experience with the use of gonadotrophins and with a medical indi- strategy was developed based on keywords and index terms related to com-
cation for IVF/ICSI treatment were invited to participate in a randomized con- mercialisation of ART in PubMed based on Medical Subject Headings
trolled trial prior to their intake regarding the IVF/ICSI treatment. Twenty-two (MeSH) terms.
patients were included in the control group and twenty-nine patients in the Main results and the role of chance: Commercial influences on ART are
app group. a frequent topic of discussion and critique in the academic literature, however
Main results and the role of chance: The groups were found not to differ these discussions are widely dispersed and often implicit in discussions of 1. The
significantly on baseline characteristics and outcome parameters. The patients pricing of ART services, and variability in these costs 2. The number and timing
in the app group scored higher on the QPP-IVF questionnaire (indicating satis- of interventions, particularly when it comes to the use of more invasive (and
faction) than the patients in the control group, although this did not reach sig- expensive) interventions in patients who might benefit from less invasive treat-
nificancy (52,89 versus 47,92, p=0,13). Two days after the ivf intake in the clinic ments and patients in whom success is unlikely 3. The expansion into non evi-
both the control group (from 17,33 to 23,25, p=0,22) and the app group (from dence-based interventions 4. The ways in which interventions are marketed and
20,74 to 26,58, p=0,02) scored higher in the relative level of knowledge com- justified to patients 5. The workings of international markets and 6. Other com-
pared to baseline findings. Only the increase in the app group was found to be mercial interests, such as joint ownership of diagnostic, counselling and hospital
significant. Five days after the ovum pick up, a significant increase in the level of services and relationships with other health-related industries.
perceived knowledge is also observed in the app group (from 20,74 to 26,89, Limitations, reasons for caution: A scoping review is a semi-systematic
p=0,05), but not in the control group (from 17,33 to 26,00, p=0,48). No sig- literature review that aims for thematic saturation rather than aiming to collect
nificant differences were found on self-reliance and care consumption (p>0.05). the full range of articles on a topic. In cannot, therefore, draw quantitative
Limitations, reasons for caution: The limited sample size of the pilot study conclusions about the literature. Our search was also limited to articles published
is a reason for caution. The observed differences need to be replicated in a larger in English.
study and in a multicentred design. Wider implications of the findings: The implications of the commerciali-
Wider implications of the findings: Knowledge on the use of a patient sation of ART cannot be understood without systematically considering a wide
journey app is limited within IVF/ICSI. The preliminary results indicate that the range of ethical, organisational, political, economic, scientific and clinical issues,
app could help patients in the uptake of information at their own pace, resulting across a diverse range of literatures. We provide a novel “map” of these issues
in more knowledge and satisfaction. This could also have a beneficial effect on that can help to guide debate on this highly contentious topic.
care consumption and costs. Trial registration number: Not applicable
Trial registration number: NA
O-212 Expanded carrier screening in medically assisted
reproduction: An ethical exploration of a new screening option in
the light of professional and parental responsibilities
SELECTED ORAL COMMUNICATIONS S. Van der Hout1, W. Dondorp1, G. De Wert1
SESSION 53: CONTROVERSIES IN ART 1
Maastricht University, Health- Ethics & Society, Maastricht, The Netherlands
07 July 2020 Parallel 5 17:00 - 18:00
Study question: May professionals recommend that applicants of assisted
reproduction undergo expanded carrier screening, and that identified carrier
couples apply for pre-implantation genetic testing to avoid the birth of an
affected child?
O-211 Commercial influences on assisted reproductive Summary answer: While an offer of ECS should be non-directive, carrier
technology: a scoping review couples of very serious disorders should have access to MAR only if they apply
W. Lipworth1, M. Wiersma1, N. Ghinea1, A. Newson1, I. Kerridge1, for PGT.
C. Waldby2, W. Ledger3, R. Norman4 What is known already: Expanded carrier screening (ECS) entails a screening
1
University of Sydney, Sydney Health Ethics, University of Sydney, Australia ; offer for multiple recessive disorders simultaneously, and allows testing of cou-
2
Australian National University, Research School of Social Sciences, Canberra, ples regardless of ancestry. A growing number of European fertility clinics offer
Australia ; ECS on a routine basis. However, little attention has been given to the ethics
3
University of New South Wales, Obstetrics & Gynaecology, Sydney, Australia ; of ECS in the context of medically assisted reproduction (MAR). MAR-
4
University of Adelaide, Robinson Research Institute, Adelaide, Australia professionals have indicated a need for ethical guidance, as an offer of ECS
addressing MAR-patients raises ethical questions that do not arise in other
Study question: How are commercial influences on assisted reproductive tech- carrier screening contexts: the involvement of MAR-professionals in creating
nology (ART) understood, debated and studied in the academic literature? the child implies that both professional and parental responsibilities should be
Summary answer: Commercial influences raise a numerous ethical, organi- taken into account.
sational, economic and clinical issues. These are discussed in diverse contexts, Study design, size, duration: In this ethics study we explore the morally
not limited to direct discussions of commercialisation relevant aspects of the scenario of offering ECS to MAR-patients, focusing on
What is known already: While advances in technologies have increased the possible tensions between considerations of reproductive autonomy and paren-
range of ART options available to patients, concerns exist about the steps taken tal/professional responsibility. This study is part of a larger project in which we
by ART providers to ensure that their businesses are successful, profitable and will use the findings to conduct an exploration of relevant stakeholder views.
sustainable (henceforth “commercial influences” on ART). Distinguishing the key The overall aim is to contribute to a robust normative framework that can guide
issues associated with these commercial influences is complicated by the diversity the practice of ECS in MAR.
of ways and contexts in which they are discussed, and there have been no sys- Participants/materials, setting, methods: This is a desk research study
tematic attempts to draw these together. bringing together the scientific literature relevant to the possible (wider) intro-
Study design, size, duration: This scoping review used the methodological duction of ECS in MAR and debates conducted in philosophical and applied
framework first proposed by Arksey and O’Malley, and later refined by Levac ethics literature on the nature and scope of relevant principles including

‘reproductive autonomy’, ‘professional responsibility’ and ‘responsible parent- Participants/materials, setting, methods: One of the objectives of the
hood’. We use the method of ‘wide reflective equilibrium’ as a framework for focus group was to deliberate about the ethical justifications that support the
our analysis. This method has a proven value for the normative analysis of right to know the genetic origins, mainly based on the protection of health and
bioethical problems. the right to identity, but also taking into account other values that affect the
Main results and the role of chance: MAR-professionals have a respon- different participants in the donation For this, we used the deliberative method
sibility to take account of the welfare of the child that they are causally and to deal with conflicts, as it seeks to protect the values that may be at stake.
intentionally involved in creating. ESHRE has recommended that they should Main results and the role of chance: Spanish Society of Fertility has pro-
refrain from providing assistance when there is a high risk that the child will duced an Anonymity in Donations Framework Document, divided into five
have a seriously diminished quality of life. What follows from this norm with distinct parts: demographic, clinical, psychological, social, ethical and legal aspects.
regard to offering ECS to MAR-patients? May (or should) professionals recom- From an ethical perspective, the following arguments regarding gamete donation
mend, or even insist, that patients have ECS, and make use of pre-implantation anonymity were suggested:
genetic testing (PGT) or other preventive options when both partners are Health of children: Spanish policy, despite its requirement of anonymity, explic-
identified as carriers? We recommend that this depends on a) the level of itly indicates that donor-conceived individuals should have access to non-iden-
genetic risk; and b) the severity of the disease under consideration. If MAR- tifiable information about the donor, including medical information, so, the
patients have an a priori low carrier risk, ECS should be offered in a non-di- anonymous donation is not incompatible with providing relevant medical
rective way and facilitate autonomous reproductive decision-making. However, information.
if both partners are identified as carriers of a serious genetic disorder, profes- Sense of identity: there is no solid empirical evidence to show that children
sionals may well be justified in urging them to actively consider PGT, or even conceived by the donor in general suffer “genealogical confusion”.
make access to MAR conditional on the patients’ use of preventive options. Other values examined were: autonomy of the parents and the right to pri-
We conclude that the preventive options created by new genomic testing vacy, genetic inheritance, donor confidentiality, justice, responsibility, quality and
possibilities are not morally indifferent, but may bring along new professional economic value.
and parental responsibilities. Following the bioethical deliberation mentioned above, and after having exam-
Limitations, reasons for caution: This is an ethical exploration aimed at ined the main values that become part of the two extreme courses (anonymity
contributing to further debate about the responsible introduction of ECS in MAR. of the donation against its suppression), the best intermediate course would be
Wider implications of the findings: Our exploration of the relationship the one that tried to safeguard the greater number of values involved, namely,
between respect for autonomy and acknowledging professional and parental donor-conceived individuals are morally entitled to access general (non-identi-
responsibilities for the welfare of the future child has a wider relevance for the fying) information about their origin.
ethics of (shared) decision-making in the highly dynamic field of MAR as Limitations, reasons for caution: Although there is no evidence of any
a whole. conflict in Spain for the rule of anonymity, the opinion of the patients, donors
Trial registration number: Not applicable and offspring, would be lacking.
Wider implications of the findings: Before introducing mandatory disclo-
sure of the donor´s identity, the commitment should be to ensure that this
O-213 Ethical justifications of Spanish Fertility Society to preserve change entails advantages for those involved, which have not been demonstrated
anonymity of gamete donation so far. Strategies should be adopted to implement education programs that do
R. Nunez - Calonge1, M. Muñoz2, F. Abellan3, I. Cuevas4, L. Feito5, not stigmatize the fact of being born through donor gametes.
A. De la Fuente6, L. Martínez7, D. Mataro8, M. Roca9 Trial registration number: Not applicable
1
URA, Ethics and Reproduction, Madrid, Spain ;
2
IVIRMA, Medical Director, Alicante, Spain ; O-214 First sperm and egg bank in Latin America with open
3
Sociedad Española de Fertilidad, Legal, Madrid, Spain ; identity donors: a five-year study on patients’ and donors’
4
Hospital General, IVF Laboratory, Valencia, Spain ; preferences between anonymous and open-identity donation.
5
Faculty of Medicine- Complutense University, Public Health & Maternal and Child P. Regalado1, V. Rawe1
Health, Madrid, Spain ; 1
REPROBANK, Research, Buenos Aires, Argentina
6
Instituto Europeo de Fertilidad, Medical Director, Madrid, Spain ;
7
Hospital Virgen de las Nieves, Reproduction, Granada, Spain ; Study question: Would gamete donors and patients choose open-identity
8
Eugin Clinic, Reproductive Endocrinology and Infertility, Barcelona, Spain ; given the chance? Several countries worldwide have laws regulating anonymity
9
Barcelona University, Psychology, Barcelona, Spain in gamete donation, mostly European and North American with opposing stand-
points between them.
Study question: What is the view of Spanish Society of Fertility (SEF) regarding Summary answer: Ours results show many patients demanding open identity
ethical aspects of the removal of anonymity in gamete donation? and half of gamete donors willingly donating this way, concurring with interna-
Summary answer: SEF recommends disclosure to the donor-conceived chil- tional tendency to unveiling donor identity.
dren their biological origin preserving the anonymity since there are no convincing What is known already: In Latin America few countries have laws regulating
ethical arguments for removal. Assisted Reproductive Techniques (ART) and anonymity donation is set by
What is known already: Although most gamete donations in the world are default. The lack of empirical data is alarming: having non-official statistics,
made anonymously, a growing number of countries have questioned the morality national surveys or publications addressing open identity vs anonymous donation.
of preserving anonymity, and laws are being enacted that allow children born by A tendency toward unveiling donor identity to offspring is increasing in Europe
donation to identify donors. and North America. However, in Latin America few countries have laws regu-
The main criticisms of the anonymity of gamete donation focus on the primary lating ART and anonymity donation is set by default. Our study is rich and pioneer
interest of donor-conceived children, and more specifically on health and interest for Latin American, it shows our cultures demand to allow access to genetic
in forming their own identity. On the other hand, those who support anonymity origins by lifting anonymity and leave the choice to offspring.
argue that its practice respects the interests of the donor as well as the parents’ Study design, size, duration: Report of 5 years of experience of the first
wishes to determine the best interest of offspring. open identity program (PIA) in Argentina, offering both anonymous and open
Study design, size, duration: Reflecting concern about the anonymity of identity donor sperm samples and later extended to eggs. We intended to
gamete donation, the Spanish Society of Fertility convened a focus group with discover donors’ and patients’ choice on open vs anonymous donation, reasons
key figures in ethics, law and reproductive medicine to identify scientific, psy- and motivations for choosing either. We gathered information from all donors
chological, legal and ethical arguments supporting the anonymous gamete dona- and patients that participated or went through ART.
tions. The project was set-up as a qualitative study to call in to question the Participants/materials, setting, methods: The sample includes 192 donors
justifications that are often thought to ground a right to know one’s genetic (132 sperm and 60 egg), 2391 ART patients and over 150 parents of open
origins and took place between May and October 2019. identity donors´ offspring. Descriptive, correlational and inferential statistics were

applied with parametrical and non-parametrical analysis. Also, we surveyed the The women were applied intervention or control procedure in four face-to-face
sample with two ad hoc questionnaires regarding reasons or setbacks for choos- session (first treatment day, second control day, OPU, embryo transfer) and one
ing open or anonymous donations, opinions on current legislation on the matter, mobile session (from embryo transfer day to pregnancy test day) throughout
future plans on identity revelation to offspring, among other queries. IVF treatment cycle.
Main results and the role of chance: Statistics show that open identity Participants/materials, setting, methods: Interventions based on
donation was chosen by 55% of egg donors, 48.5% of sperm donors and 29% Hypnofertility including affirmations, visualization, imagination and relaxation
of ART patients. Regarding donors, more egg than sperm donors chose open were applied to women undergoing with IVF treatment from the first day of
identity, however the difference was not statistically significant. Regarding treatment to the day of pregnancy test. The standart IVF center follow up pro-
patients, results showed statistically significant differences based on type of family cess was provided to control group. The cortisol level was evaluated trough
structure: open identity donation is chosen in 1 out of 10 heterosexual couples, saliva sample (four times), for determining preparedness of women the Fertility
3 out of 10 lesbian couples, and 4 out of 10 single women. More in depth results Preparedness Scale (three times) was applied and pregnancy result was evaluated
are discussed in relation to patients’ and donors’ general reasons, attitudes and by blood beta HCG test.
motivations toward open identity and anonymity. Also, post-treatment parents Main results and the role of chance: The group equivalence on baseline
with open identity offspring past choice satisfaction is addressed. socio-demographic and infertility characteristics was shown in similar (X2: 0.007,
An international wide controversy has existed since sperm donation became p: 0.06; X2: 4.419, p: 0.93). There was no significant difference between the two
a part of ART. Anonymity continues to depend on judicial regulations. Empirical groups in terms of fertility preparedness evaluated at baseline (t=0.930, p=.35).
research needs to guide laws based on ethical and social-need grounds. The Although there was higher improvement in fertility preparedness in intervention
past decade has shown an international tendency towards removing anonymity group (second session p=0.04, third session p=0.01), the results of repeated
or allowing access to genetic origins. Anonymity in many countries continues measures, showed no difference in total in terms of group*time (F=2.177,
to be defended while social movements and organizations of donor offspring p=.13). Saliva samples collection time of both groups were similar (p=.84; p=.41;
continue to demand the human right to access genetic origins. Research on p=.68; p=.50, respectively). When the cortisol levels of the women in the
open identity vs. anonymity mostly reflects European and North American intervention and control groups were evaluated, a statistically significant differ-
cultures. ence was found in terms of group*time interaction (F=4.187, p=.00). It was
Limitations, reasons for caution: Samples were gathered from a single observed that the interventions decreased the cortisol levels of the women in
gamete bank because it is the only one in the country offering an open identity the intervention group compared to the control group (p=.00). Cortisol levels
program. Samples don’t represent argentine ART patients. However, it does of the intervention group gradually decreased during the treatment period
show a tendency in favor of what has been found in other countries. (p=.01). The last measurement has the lowest results in spite of the most stress-
Wider implications of the findings: Our findings are rich and pioneer for ful days of treatment (the embryo transfer day). Even though interventions
Latin American, evidencing a cultural demand to address anonymity. Few coun- relieved the women during treatment process, there was no the expected effect
tries in Latin America have ART laws. Research is fundamental for future on pregnancy outcomes (X2: 0.098, p=.75).
legislations. Limitations, reasons for caution: The main limitation of this randomized
Trial registration number: not applicable controlled trial was the limited number of samples. Since the cost of each mea-
surement of cortisol level in saliva measured in the study was high, and the
support obtained from University Scientific Research Project Coordination Unit
was limited.
SELECTED ORAL COMMUNICATIONS Wider implications of the findings: Although interventions based on
SESSION 54: MODERN TECHNIQUES PROMOTE VARIETY Hypnofertility not effect pregnancy clinical outcomes, they reduced the cortisol
IN FERTILITY NURSING RESEARCH levels of women. Also the fertility preparedness was improved in clinically mean-
07 July 2020 Parallel 6 17:00 - 18:00 ingful level.
O-216 Assessment of a French web-based patient decision aid

(ptDA) of ertility preservation (FP) for women with breast cancer
O-215 The Effect of Hypnofertility on Cortisol Levels, Fertility
A.C. Benoit1, G. Moutel2, R. Morello3, G. Grandazzi2, A. Mayeur1,
Preparedness and Pregnancy Rates in Women Undergoing
M. Grynberg1
InVitroFertilization: A Randomized Controlled Study
1
Antoine Beclere University Hospital, Reproductive Medicine and Fertility
S. Fata1, M. Aluş Tokat1
Preservation, Clamart, France ;
1
Dokuz Eylul University, Obstetric and Gynecological Nursing, Izmir, Turkey 2
Normandie University, Inserm U1086- ANTICIPE, Caen, France ;
3
University Hospital- Caen, Biostatistics and Clinical Research Unit, Caen, France
Study question: Does Hypnofertility have remarkable effect on cortisol levels,
fertility preparedness and pregnancy rates? Study question: Does a French web-based ptDA in addition to standard
Summary answer: Hypnofertility is a powerful and effective mind-body pro- counselling by a specialist in reproductive medicine improve informed choice of
gram and aims to improve relaxatation, regulate hormone secretion and increase women with breast cancer?
the pregnancy rates. Summary answer: The web-based ptDA we have developed improves
What is known already: It is found that there is a negatively correlation patients’ knowledge and informed choice without influencing their attitude
between stress levels and fertilization rates, pregnancy rates, total births, live towards FP and decrease the level of decisional conflict.
births. There is information in the literature that controling of cortisol levels is What is known already: Chemotherapy may cause infertility in young survi-
important and can give idea about stress experienced before starting assisted vors of breast cancer, impairing their quality of life in the long term. Various FP
reproductive treatment. Health professionals can be enterprise some applica- techniques increase the likelihood of survivors becoming genetic mothers. The
tions to reduce the stress and cortisol levels. One of the methods that can reduce complex emotional situation, the need to make the right decision in a short space
stress is Hypnofertility. Hypnofertility is a powerful and effective mind-body of time and conflicting ideas about life and death may mean that the decision
program with aim to relax couples, improve the preparedness to treatment about FP is a difficult one to take. Weighing up the risks and benefits of FP options
process and therewithall increase the pregnancy rates. may seem a mammoth task. Use of a ptDA could be increase patients’ knowledge
Study design, size, duration: It was randomized controlled trial conducted of treatment options and congruency between informed values and care.
in Dokuz Eylül University InVitroFertilization (IVF) Center in Turkey between Study design, size, duration: The study was a single-center randomized
December 2017-March 2019. The 61 women with unexplained infertility (30 controlled trial conducted in the FP department of an academic hospital in France
women for intervention group and 31 women for control group) were reached. from September 2018 to November 2019. After being referred by oncologic

centers to specialists in reproductive medicine for fertility counselling, women Participants were not amenable to alternative medical structures (i.e. group
were randomly assigned to the control group with usual care or to the inter- sessions for emotional support), but were interested in anonymous digital plat-
vention group with use of the web-based ptDA before and during counselling. forms to meet psychosocial needs. In response to these data, as well as previous
Participants/materials, setting, methods: A total of 125 women aged 18 study team data, our team developed an internationally-focused and non-com-
to 40, with a primary diagnosis of breast cancer, who have not yet started any mercial peer-support platform (https://all-about-fertility.com/) to allow men
cancer treatment and able to read, write and speak French were included and with MFI to be able to disclose to others with MFI and seek anonymous emo-
randomized to the control group (n=65) or the intervention group (n=60). We tional support online, whilst also receiving accurate and unbiased medical infor-
have compared the decision-making process between control group and inter- mation to support their fertility journey and decision-making. Contributions from
vention group by quantitative data collected about fertility-knowledge, attitudes global Reproductive Health/Science experts provides men with confidence when
towards FP, and decisional conflict after the FP consultation. seeking health information.
Main results and the role of chance: Better knowledge was found in the Limitations, reasons for caution: As the results are based on qualitative
intervention group (8.6/10 (±1.34)) compared to the control group (6.49/10 data from 19 men with MFI, the results cannot readily be generalized to larger
(±1.89)). In both groups, patients had a positive attitude towards FP (96%). The populations. Implementation of the peer-support platform is still being optimized
final decision was not influenced by membership in one group or another. Thus, and its usefulness and impact to meet stated goals will be evaluated/reported
at the end of the FP consultation, 73.6% (92/125), i.e. 69.2% (45/65) of patients in the coming years.
in the control group and 78.3% (47/60) of patients in the intervention group, Wider implications of the findings: Men with MFI have unmet psychosocial
chose to keep their oocytes, embryos and/or ovarian tissue. The proportion and informational needs. Development of a peer-support platform is a positive
of informed choice was statistically higher in the intervention group than in the step in addressing some of these needs and has the potential to have a wide
control group (respectively 75% versus 38.5%, p<0.001). In addition, the level reach into areas with limited support (geographically, economically).
of decision-making conflict among patients in the intervention group was lower Trial registration number: n/a
than in the control group (respectively 14 (±12.5) versus 21.9 (±15.7), p<0.01).
Limitations, reasons for caution: Investigators were not be blinded to the
intervention, which represents a limitation. Furthermore, the study was specif- O-218 A preparatory information movie for the first oocyte
ically focused on women with breast cancer. Therefore, the results are not aspiration in addition to care as usual: protocol for the POAM
applicated to other malignancies. randomized controlled trial
Wider implications of the findings: The present study led to the development L. Dias1, P. De Loecker2, T. D’Hooghe1, K. Peeraer3, E. Dancet1
of the first French language ptDA of FP. Our results suggest that a French web- 1
KU Leuven, Regeneration and Development, Leuven, Belgium ;
based ptDA has the potential to help supporting women with breast cancer to 2
GZA Ziekenhuizen campus Sint-Augustinus, Gynaecologie - Verloskunde -
make informed fertility-related decisions and might decrease decisional regrets. Fertiliteit, Wilrijk, Belgium ;
UZ Leuven, Regeneration and development, Leuven, Belgium
O-217 Addressing the psychosocial and informational needs of Study question: Can a preparatory information movie on oocyte aspiration
men with severe male factor infertility (MFI) with a peer-support (POAM) decrease the anxiety of women on the day of their first oocyte aspi-
platform ration? A protocol for a randomized controlled trial.
E. Lowndes Stevenson1, K. Baker2, K. McEleny3 Summary answer: This RCT protocol provides a framework to examine the
1
Duke University Medical Center, School of Nursing, Durham NC, U.S.A. ; effect of a preparatory information movie on oocyte aspiration on women’s
2
Duke University Medical Center, Department of Urology, Durham, U.S.A. ; anxiety and other secondary outcomes.
3
Newcastle upon Tyne Hospitals NHS Trust, Newcastle Fertility Center, Newcastle- What is known already: Limited success rates of the fertility clinic trajectory
upon-Tyne, United Kingdom could be increased by preventing discontinuation, a couple’s decision to stop
treatment despite financial stability and ongoing wish for a child. The main reason
Study question: What do men with MFI desire in order to address their for discontinuation is distress, which increases during an IVF cycle, with anxiety
psychosocial and educational needs during the fertility process? peaks on the days of oocyte aspiration and pregnancy test. Moreover, pre-treat-
Summary answer: Men indicate desiring digital health communication and ment anxiety predicts discontinuation from especially the first IVF cycle. Detailing
anonymous digital platforms to meet their psychosocial and informational needs. procedural steps of out-patient clinic invasive procedures decreases patient’s
What is known already: Men with MFI experience higher levels of psycho- anxiety. Information leaflets however often increase anxiety because they can
logical challenges such as depression and sexual dysfunction than fertile men. be very complicated. An educational movie simply showing what will happen
Men respond differently than their partners to the infertility diagnosis. Our pre- can overcome this problem.
vious research found that men employed avoidance strategies/did not disclose Study design, size, duration: In the POAM-RCT 190 women are randomized
infertility status to social networks, experienced affective symptoms (sadness, (1:1 allocation ratio; computerized randomization) during a recruitment period
shock, disbelief, denial), and lacked access to evidenced-based information in of 19 months to the parallel groups of ‘care as usual’ (a one-hour face-to-face
part because of their margination within the fertility process, secondary to the information session 1-3 months prior to the start of treatment on all procedural
female-facing nature of fertility healthcare. steps of an IVF cycle, also summarized on a take-home leaflet) or ‘care as usual
Study design, size, duration: This IRB-approved prospective qualitative study supplemented with the Preparatory information on Oocyte Aspiration Movie
recruited men seeking a MFI consultation with a urologist between 2018 and (POAM)’.
2019. Men over 18 years of age with primary or secondary MFI were phone Participants/materials, setting, methods: The setting for this monocentric
interviewed after providing informed consent. Each digitally recorded interview interventional single-masked RCT is the fertility clinic of the GZA Ziekenhuizen,
lasted between 15 and 40 minutes and was transcribed verbatim. Nineteen a private hospital in Belgium. Dutch speaking women with Dutch speaking male
participants were recruited (until data saturation reached). partners who are about to start their first IVF cycle (with or without ICSI or
Participants/materials, setting, methods: Inclusion criteria were men (≥ PGT) are eligible. Women who already experienced an IVF cycle and therefore
18 years old) seen for a new patient MFI consultation having received a MFI know the patient journey on the day of the oocyte aspiration are not eligible.
diagnosis during the preceding 12 months and could read/write English. Exclusion Main results and the role of chance: In case of randomization to the
criteria were a history of vasectomy or stated absence of a fertility concern intervention group, couples receive the day prior to their first oocyte aspiration,
despite a fertility related diagnosis. Nineteen men were recruited from a fertility in addition to ‘care as usual’, a secured link to the POAM (5-minute movie
urology clinic in a large academic medical center. Interviews were conducted by visualizing the patient journey on the day of oocyte aspiration detailing which
phone. Data were analyzed using content analysis. procedures will happen when, in which room, in the presence of which persons)
Main results and the role of chance: Mean age was 35.3 years. Results by email.
indicated that participants prefer technology based communication to receive The primary outcome of this RCT is women’s state anxiety assessed with the
health information (i.e. ‘mychart’) because of the ease, access, and efficiency. reliable STAI-state questionnaire. Secondary patient-related outcomes are men’s

state anxiety and also women’s and men’s infertility-specific distress on the day Limitations, reasons for caution: A potential limitation of this RCT is selec-
of the oocyte aspiration, assessed with the Infertility-Distress Scale. Previous tion bias, as women who are more anxious or stressed will be less likely to
research in the field of reproductive medicine showed that men are significantly participate. Randomization aims to adjust for this effect. Another potential lim-
less distressed if they receive an information leaflet before a diagnostic sperm itation is the single-masked character of the RCT as only the statistician will be
test. Finally, women’s and men’s intervention evaluation are assessed by ques- masked during data processing.
tioning how often they watched the movie and whether or not they would Wider implications of the findings: With this RCT, we expect to demon-
recommend it. strate the value of additional information using a preparatory movie in couples
Secondary clinical outcomes that will be followed-up in the electronic medical undergoing their first oocyte aspiration. If these couples are positively affected,
records are clinical pregnancy, miscarriage, IVF delay, IVF discontinuation and this technique should be implemented in each fertility clinic. We also want to
cumulative pregnancy. The evidence on whether women’s anxiety affects (cumu- show the value of publishing a study protocol.
lative) clinical pregnancy rates and miscarriage rates is conflicting. Trial registration number: NCT03717805

human
reproduction

professionals in the UK. The HFEA is continuing to work on a range of initiatives
designed to set standards and inform patients about the use of treatment add-ons
INVITED SESSION in the UK. The HFEA has published information for patients on some of the
add-ons being offered in the UK based on an annual review by our independent
SESSION 55: COCHRANE SESSION - BETTER EVIDENCE,
Scientific and Clinical Advances Advisory Committee of relevant published
BETTER POLICIES
research. The HFEA has tightened its guidance for clinics to require that any
08 July 2020 Parallel 1 08:30 - 09:30 information provided to patients by clinics on the safety and effectiveness of any
add-on explicitly references the HFEA website. In future we envisage that new
treatments or technologies under development will only be offered to patients
O-219 Better RCTs in reproductive medicine outside of a research setting once safety and effectiveness have been demon-
M.Van Wely1 strated. More high-quality research including Randomised Controlled Trials,
1
meta-analyses, and follow-up of patients is needed. We want to move towards
Academic Medical Center, Center for Reproductive, Amsterdam, The Netherlands
a more consistent and transparent approach to the use of treatment add-ons in
O-220 Where do we go after the RCTs fertility services.
S. Cheshire1, P. Thompson2, C. Ettinghausen3, D. Halai4
1
HFEA, Chair of the Board, London, United Kingdom
2
HFEA, Chief Executive, London, United Kingdom INVITED SESSION
3
HFEA, Director of Strategy and Corporate Affairs, London, United Kingdom
4 SESSION 56: FRONTIERS IN ANDROLOGY
HFEA, Head of Scientific Policy, London, United Kingdom
08 July 2020 Parallel 2 08:30 - 09:30
Abstract text
ART is a fast moving and innovative area of medicine. And while innovation can
be a force for good, offering patients improved chances of success, too often O-221 Capturing the sperm flagellum: Mechanistic andrology
new drugs, treatments or equipment are introduced with little robust evidence
D.Smith1
that they will improve outcome. For regulators this presents a difficulty: how
1
best to encourage innovation while protecting the patient? University of Birmingham, School of Mathematics, Birmingham, United Kingdom
More high-quality research including Randomised Controlled Trials, meta-anal- O-222 When will we start transplanting prepubertal testis tissue?
yses, and follow-up of patients is needed. But where do we go if these are not
C. Wyns1
readily available now?
1
The UK’s Human Fertilisation and Embryology Authority (HFEA) is ~30 years n/a, n/a, BRUSSEL, Belgium
old and has led the way in regulation in this sector. Our experience of licensing
mitochondrial donation suggested a different way of thinking about regulation Abstract text
and innovation, where good regulation provides the conditions that encourage Testicular tissue transplantation in humans was already performed back in 1889
innovation while ensuring every patient has access to the right information, at by Brown-Sequard to treat hypogonadism. However, grafting the tissue to
the right time. restore the reproductive potential has never been reported.
We in the fertility sector in the UK are concerned that many patients are being Efforts to preserve the fertility of prepubertal boys facing fertility-threatening
frequently offered, and charged for, optional extras to their treatment which therapies by cryopreserving their immature testicular tissue have motivated
claim to improve their chances of having a healthy baby. These additional ther- researchers to develop methods to give these young patients a hope to father
apies and techniques are collectively known as treatment ‘add-ons’. They cover their own genetic child. In this regard, auto-transplantation of cryostored tissue
a range of interventions including genetic tests, drugs, surgery and equipment. can be considered if there is no risk of cancer cell contamination of the tissue
The problems posed by treatment add-ons are well known but crucially, the i.e. not in case of haematological cancers or metastasizing tumours.
evidence base for most add-ons is not robust. Over recent years, there has been The proof of principle of the technique was recently obtained with the birth
a significant increase the number of clinics, both in the UK and elsewhere, offering of GRADY in macaques but many research questions remain before pilot trials
additional treatments – in 2019, 77% [DH1] of clinics in England offered at least can be considered in humans where most often only small amounts of tissue are
one add-on according to their websites. available for future use.
This talk will focus on the actions the HFEA is taking to encourage more Xenotransplantation experiments with human prepubertal testicular tissue in
responsible innovation in the way in which clinics offer treatment add-ons, and nude mice showed an important loss of spermatogonia in grafts, regardless of
the way in which patients can understand the risks and benefits involved. The prior cryopreservation, justifying optimization of the avascular transplantation
ultimate aim has always been to balance scientific and medical advancement with procedure.
individual rights and needs. Our add-ons work touches on the quality of evidence, High DNA fragmentation by TUNEL was observed in the first days following
patient information and informed consent. grafting suggesting that cell damage is likely due to hypoxia before vascular supply
Given our limited powers on the introduction of new treatments, we cannot of oxygen to the grafted tissue is in place. In this regard, finding the most appro-
simply set conditions of the use of add-ons. In January 2019, the HFEA published priate graft size to reduce tissue necrosis and cell loss is a first matter of concern.
a consensus statement signed by an additional 10 signatory bodies which sets Only one study compared graft outcome for different tissue sizes and no differ-
out the principles of responsible innovation which we believe should guide ences were observed up to 16mm³. Attempts to shorten the ischemic period
BK-OUP-ESHRE2020_ABSTRACTS-200150-Wednesday.indd 99 22/06/20 8:43 PM

before revascularization of the graft were also made by culturing the tissue lastly , proximal surgery keeps good indication after sterilization when the
fragment before grafting in media containing vascular endothelial growth factor patients wishes to restaure her fertility. datas suggest that microsurgical reca-
(VEGF) as this method increased the number of seminiferous tubules with elon- nalization of the tube through robot or microsurgery gives better outcome in
gating spermatids in bovine. Unfortunately, this could so far not be confirmed term of pregnancy than IVF/ICSI.
for human prepubertal tissue although a potential benefit was suggested in a Therefore we consider that tubal surgery, in the hands of trained team, has
report of two cases. a role to play ,providing that a good diagnosis is established, a careful selection
However, controlled local drug delivery of VEGF covering the time period of patients is made and a complete information is delivered to patients .
needed for the stabilization of the neo-vasculature could be more efficient. Nevertheless, nowadays, few surgeons are trained in tubal surgery, except in
Therefore, nanoparticles (NPs) containing growth factors e.g. VEGF, platelet-de- Far East countries(such as India and China) where tubal surgery is still very
rived growth factor (PDGF) and necrosis inhibitors have been developed as well popular.
as tissue embedding matrices allowing proper migration of endothelial cells. We should probably think to re-promote the teaching of tubal surgery in the
Higher recovery rates of undifferentiated spermatogonia in autografts were initial training of young surgeons, in order not to compete with IVF /ICSI but
reached in an alginate hydrogel and further improvement was achieved with NPs to provide an additional tool which may be beneficial in some infertile patients
controlled drug delivery.
Incomplete and abnormal differentiation of spermatogonia in xenografts of
human prepubertal testicular tissue was also observed, most likely due the phy-
logenetic distance between mice and human, but not only. Indeed, graft devel- SELECTED ORAL COMMUNICATIONS
opment seems also influenced by its hormonal environment as autografting SESSION 58: IMPROVING SPERM CRYSOPRESERVATION
experiments in marmosets showed less advanced germ cell development in OUTCOMES
hemi-castrated than castrated animals. While it was reported that human pre- 08 July 2020 Parallel 4 08:30 - 09:30
pubertal Leydig cells show maturation features based on the presence of key
enzymes of steroidogenesis and ultrastructural modifications after xenografting,
only few studies have investigated the impact of the hormonal environment on
the spermatogonial stem cell niche in grafts. Some questions still need to be O-225 Does cryopreservation affect live birth outcome in
addressed: what would be the real impact of the post-chemotherapeutic hyper- surgically retrieved sperm: – an analysis of a decade of nationwide
gonadotropic hypogonadic state of candidate patients for autotransplantation data
on transplant outcome, and would hormone requirements of the transplanted J. Lewin1, E. Yasmin1
tissue depend on the age of the patient at cryopreservation? Furthermore, and 1
University College London Hospital, Gynaecology, London, United Kingdom
so far, the absence of a valid preclinical model for human prepubertal testicular
tissue transplantation precludes to answer the question of the time needed to Study question: Is there a difference in live birth rate after ICSI between
achieve complete spermatogenesis in grafts and thus the optimal moment for cryopreserved and fresh surgically-retrieved sperm?
sperm retrieval. Summary answer: There was no difference in live birth rate between cryo-
preserved and fresh sperm used in ICSI over 10 years analysed from all UK
fertility centres
What is known already: Multiple small studies have been conducted com-
INVITED SESSION paring success rates from fresh and cryopreserved surgically-retrieved sperm
SESSION 57: EVIDENCE-BASED SURGICAL for use in ICSI. A 2014, meta-analysis showed no significant difference in live
INTERVENTIONS birth rate. Most included studies were small and subsequent studies have demon-
08 July 2020 Parallel 3 08:30 - 09:30 strated conflicting results. Concerns are raised about the effect of cryo-pro-
tectants and freezing/thawing techniques on function and structure of sperm
such as DNA fragmentation and acrosomal reaction.
Study design, size, duration: Annual data pertaining to ICSI cycles from all
IVF centres in the UK were obtained from the UK Human Fertilisation and
O-223 Isthmocele management in infertile patients
Embryology Authority (HFEA). The time period studied included all years from
O. Donnez1 2007 to 2017 inclusive. Data from 235,296 ICSI cycles were therefore obtained.
1
Elsan, Gynecologic and oncologic surgeon, polyclinique Urbain V, MD, PhD, France Participants/materials, setting, methods: By law all clinics in the UK offer-
ing assisted reproductive technology treatments must collect accurate data from
O-224 Indications for tubal surgery
all cycles including pregnancy outcome, which is then submitted to the HFEA.
A. Watrelot1 We obtained anonymised data from the HFEA on live births per cycle of treat-
1
Hospital Natecia, gynecologic surgery, Lyon, France ment according to sperm and egg source from 2007-2017. We compared live
birth outcomes between cryopreserved and non-cryopreserved sperm from
Abstract text different sources using Chi square, ANOVA and Pearson correlation coefficient
Interest in tubal surgery has decreased progressively in the last decades due to where appropriate.
IVF/ICSI results which have dramatically increased in the same time.However Main results and the role of chance: A total of 235,296 ICSI cycles were
IVF access is not easy in many countries and more and more patients wish , if performed in the UK between 2007 and 2017 inclusive. Data obtained from
possible, to conceive naturally. Therefore once may ask what is the place for these cycles included the number of cycles and live births/ per cycle of treatment
tubal surgery in 2020. for each year categorised by source of egg and sperm (donor or partner) and
First of all it is important to consider the tubal pathology: the most common further divided by method of sperm retrieval (ejaculate, epididymal or testicular)
pathology is the one affecting the distal part of the tube . Everybody knows and whether the used sperm were fresh or cryopreserved. Any sub-group with
today the detrimental effect of hydrosalpinx on fertility and IVF results. <5 cycles/live births were listed as <5 to protect patient anonymity, so these
In this case, salpingectomy is often proposed but with a careful selection, a were excluded from analysis, including all data from 2007 and from donor gamete
conservative treatment by salpingoplasty may produce very good results in term cycles. 197,836 ICSI cycles were therefore included, which resulted in 61,344
of spontaneous pregnancy. live births over the 10-year period. Of these cycles 184,246 used ejaculated
A “new” chapter in distal tubal pathology represented by the subtle tubal sperm, 5249 epididymal and 8341 testicular, which had live birth rates of 30.93%,
abnormalities is raising (such as paratubal cysts, sacculation,accessory tubes),and 32.94% and 31.60% respectively. There was no difference in the live birth rate
we have more and more evidence that these minor abnormalities should be between fresh and frozen ejaculated (30.94% vs 29.78%, p=0.208), epididymal
diagnosed and treated allowing spontaneous pregnancy in more than 50% (32.59% vs 33.20%, p=0.64) or testicular (31.59% vs 31.64%, p = 0.97) sperm.
of cases. There was no difference between any of the groups when analysed by one-way

ANOVA (p=0.089), and no statistically significant correlation between the year Wider implications of the findings: Our findings suggest that Cryotop and
and the live birth rate for any group. Cell Sleeper methods would be suitable and clinically useful for men with severe
Limitations, reasons for caution: ICSI cycles using donor oocytes were male infertility who hope to have biological children. Furthermore, our effective
excluded from analysis due to low numbers. The primary diagnosis (obstructive sperm storage methods contributed to reduce the time-consuming and exhaust-
or non-obstructive azoospermia) was not known in patients undergoing surgical ing search for individual sperm in the laboratory.
retrieval of testicular sperm and a sub-group analysis between live outcome in Trial registration number: not applicable
obstructive and non-obstructive azoospermia would have provided important
clinically relevant information. O-227 comparison of permeable cryoprotectant free vitrification
Wider implications of the findings: The lack of difference in live birth by droplet method versus rapid freeze using glycerol in abnormal
outcomes in different sources of partner sperm allays some of the concerns semen sample
about using frozen sperm for ICSI especially in surgically retrieved sperm. This S. Ranganathan1, S. Reddy1, M. Daniel1, S. Srinivasan1
analysis can serve as evidence to counsel patients undergoing surgical retrieval 1
Sri Ramachandra Institute of Higher education and Research, Reproductive
of sperm. Medicine, Chennai, India
Study question: Is permeable cryoprotectant free vitrification using droplet
O-226 Novel Technologies for Single-Sperm Vitrification with method of abnormal semen sample a good alternative to conventional rapid
Cryotop and Cell Sleeper: A Follow-Up Study freeze using glycerol?
Y. Endo1, S. Mitsuhata1, M. Hayashi1, Y. Fujii1, H. Motoyama1 Summary answer: The permeable cryoprotectant free sperm vitrification
1
Kurashiki Medical Clinic, IVF Center, Kurashiki, Japan protocol tested in this study renders considerably better recovery rates of abnor-
mal semen sample compared to rapid freezing.
Study question: Do the novel sperm vitrification devices (Cryotop and Cell What is known already: Semen cryopreservation in contemporary practice
Sleeper) efficiently vitrify small numbers of spermatozoa? is conventional rapid freezing; however, more recently, vitrification has also been
Summary answer: Single-sperm recovery was successful with both devices; used. The rapid freeze method has been associated with a decline in sperm
fertilization and embryo cleavage occurred normally, embryo transfer was suc- quality with negative effects on both structural and functional sperm features.
cessful, and four babies were liveborn. One of the recently emerged technology within the field of cryobiology is sper-
What is known already: We developed the effective single-sperm vitrification matozoa vitrification.This method is based on rapid cooling of cells by immersion
methods by using the nonbiological carriers Cryotop (Kitazato, Tokyo, Japan) and into liquid nitrogen and, thereby, is the key to reducing the chance of forming
Cell Sleeper (Nipro, Osaka, Japan) and were the first to report the successful big ice crystals. Vitrification is still yet to be explored, since only limited studies
delivery of infants from the individually vitrified sperm from a man with nonob- shows its efficacy in abnormal semen sample.
structive azoospermia (Endo et al., 2011, 2012). Since the publication of our Study design, size, duration: This study included 80 oligozoospermic sperm
reports, however, there have been few reports of live birth after the use of sperm samples from patients seeking ART treatment between 2018-2019. The effects
from men with extremely severe male infertility. Because the population of such of permeable cryoprotectant free vitrification protocol on functional sperm
men is quite limited, insemination with donor sperm is a common option in quality parameters in comparison to fresh and rapid freeze using glycerol samples
such cases. were assessed.
Study design, size, duration: This was a retrospective study. From January Participants/materials, setting, methods: All samples were divided into
2010 to December 2018, vitrification of small numbers of spermatozoa was three aliquots : fresh(F), vitrification, warming(V), rapid freeze(R). Sperm total
performed for 9 men with azoospermia after testicular biopsy and for 3 men motility, progressive motility, vitality and spontaneous acrosome reaction were
with cryoptozoospermic semen. The ICSI treatments were performed in 16 assessed and compared between both the groups.
cycles for sperm from Cryotops and 14 cycles for those from Cell Sleepers. Main results and the role of chance: Results showed better preservation
Participants/materials, setting, methods: Vitirification: All motile spec- of sperm features after vitrification compared to conventional rapid freeze.
imens were vitrified before the day of oocyte pickup. Two to 14 spermatozoa Permeable cryoprotectant free vitrification using droplet method presented a
were loaded into cryoprotectant solution in each device (Cryotop and Cell significantly higher percentage of total motile, progressively motile and live sper-
Sleeper), which was placed in liquid nitrogen vapor. Subsequently, they were matozoa compared to rapid freezing. The percentage of sperms undergoing
stored in a cryogenic tank. Warming: On the day of oocyte retrieval, premature capacitation was lower in frozen thawed vitrified samples compared
Cryotops and Cell Sleepers were taken out of the tank. Sperm were retrieved to rapid freeze samples. The total motility in V group was 37.79±11.98 and R
with intracytoplasmic sperm injection (ICSI) needles and used in ICSI group was 36.28±11.90. Progressive motility in V group was 22.01±8.45 and R
procedures. group was 18.31±8.18. The vitality in V group was 51.56±12.31 and R was
Main results and the role of chance: A total of 475 sperm in 55 Cryotops 49.95±12.22. Acrosome reacted sperms were significantly high in R group with
and 171 sperm in 18 Cell Sleepers were vitrified. During ICSI, 30 Cryotops and 43.29±9.96 compared to V group 31.93±8.72. Acrosome intact sperms were
16 Cell Sleepers were warmed, and the sperm recovery rate was 83% significantly high in V group with 65.21±8.86 compared to R group 54.09±9.44.
(218/264) and 91% (135/148), respectively (P < 0.05). After warming, 34% The analysis of sperm quality parameters studies revealed that the V group
(75/218) of sperm from Cryotops and 15% (20/135) from Cell Sleepers were preserved the sperm quality much better than the R group, indicating that sam-
motile, and the difference was significant (P < 0.01). A total of 87 Cryotop ples are better preserved through vitrification than rapid freezing.
sperm and 74 Cell Sleeper sperm were used in ICSI and injected into oocytes Limitations, reasons for caution: This study has been done only on oligo-
individually. The normal fertilization rate was significantly higher in the Cryotop zoospermic sperm samples. It is necessary to compare these results in surgically
group than in the Cell Sleeper group (48% [42/87] vs. 32% [24/74]; P < 0.05), retrieved sperm samples to evaluate the influence of the application of this
but the rate of embryo cleavage did not differ (88% [37/42] vs. 96% [23/24]). methodology in routine clinical practice.
Fresh and frozen embryos from both groups underwent 16 and 10 transfer Wider implications of the findings: The sperm vitrification protocol
cycles, respectively, and 16 embryos from the Cryotops group and 11 from described here guarantees better maintenance of sperm quality parameters than
the Cell Sleeper group were transferred to patients. The rates of pregnancy conventional rapid freeze and can prove to be a better and economical alterna-
were 13% (2/16) with Cryotop group and 20% (2/10) with Cell Sleeper group. tive to preserve sperm samples from patients seeking ART treatment. Further
Finally, 2 healthy babies from Cryotop group and 2 from Cell Sleeper group it opens new doors in IVF and ICSI.
were born. Trial registration number: not applicable
Limitations, reasons for caution: Fewer than 0.7% (12/1837) of patients
undergoing oocyte pickup required the novel technologies of single-sperm vit- O-228 Sperm banking before gonadotoxic treatment: is it worth
rification during the experimental period in Kurashiki Medical Clinic (Okayama, the effort?
Japan). Further clinical follow-up studies will provide more information about E. Reiser1, K. Vomstein1, T. Kriesche1, B. Böttcher1, G. Pinggera2,
which device has more clinical benefit for single-sperm vitrification. B. Toth1

1
Medical University Innsbruck, Department of Gynecological Endocrinology and Summary answer: A model based on laboratory-specific morphokinetics was
Reproductive Medicine-, Innsbruck, Austria ; found to be complementary to the general models and an important tool for
2
Medical University Innsbruck, Department of Urology, Innsbruck, Austria improving single embryo selection.
What is known already: Generally applicable algorithms for selection and
Study question: Do cancer patients already show impaired sperm quality prior mainly deselection of embryos from a cohort are routinely used in IVF labs. It
to gonadotoxic treatment? was shown that clinical and laboratory conditions influences embryo morphoki-
Summary answer: Sperm quality was already impaired prior to gonadotoxic netics. In order to avoid the differences between labs, the general algorithms
treatment. were developed by using large range of cells division timing. It is recommended
What is known already: Cryopreservation of semen is a well-established that laboratories develop a model that uses the specific embryo division timing
method as chemotherapy, surgery and radiotherapy have deleterious effects on obtained under local conditions. However, it was not reported whether a specific
spermatogenesis. However, patients suffering from malign, as well as benign model can be developed in a lab that embryos were already selected for transfer
diseases show impaired semen characteristics already prior to gonadotoxic treat- by using the general models.
ment. Therefore, this study aims to compare semen quality between different Study design, size, duration: During 2013-2018, 12,944 embryos were
cancer types and benign diseases. Furthermore, the usage rate of cryopreserved incubated in our EmbryoScope (Vitrolife), using the general models for embryo
semen is reported to be as low as 8% for assisted reproductive treatments (ART) selection. There were 1879 KID (known implantation data) embryos, of which,
and was also analyzed in our study cohort. 425 were positive KIDs. For the outcome, we set three endpoints for KID
Study design, size, duration: Within this retrospective study, the semen quality definition: gestational sacs (GS), clinical pregnancy (CP), and live birth (LB).
and the utilization of cryopreserved semen, was assessed in a total of 264 patients Comparison between positive and negative KID embryos for cell division timings
suffering from cancer and benign diseases prior to any gonadotoxic therapies. Patients was analyzed separately for ICSI and IVF, at patient’s age 18-41 years old.
were referred to the Department of gynecological endocrinology and reproductive Participants/materials, setting, methods: Implanted and non-implanted
medicine, Innsbruck, Austria between 01/2008 and 07/2018. Pre-treatment semen embryos were analyze for statistical differences in cell division timing and cell
analyses were studied and compared in accordance with the WHO guidelines. In cycles intervals. We used EmbryoScope Stats software for model building. The
addition, the usage of cryopreserved semen for ART was evaluated. timing parameters were tested for their contribution to the scoring in the model.
Participants/materials, setting, methods: Pre-treatment semen analyses The algorithms were tested for the area under the receiver operating charac-
were studied and compared in accordance with the WHO guidelines in 264 teristic curve (AUC) in KID embryos for developing Day 2,3 and 5 models. The
patients with testicular cancer, hematological malignancies and benign diseases stability of the algorithms structure was verified by performing calibration-vali-
prior to any gonadotoxic therapies. In addition, the usage of cryopreserved dation procedures.
semen for ART was evaluated. Main results and the role of chance: Since significant differences in cell division
Main results and the role of chance: Patients with testicular cancer (TM) timings were found between implanted and non-implanted embryos, we were
showed a lower sperm concentration (12 *106/ml) compared to hematological able to develop a laboratory-adapted model. The algorithms were developed for
malignancies (HM) (28.5 *106/ml, p=0.0029) and benign diseases (Ben) (29 *106/ selection of Day 2, 3 and 5 ICSI embryos. AUC at most cases were higher than
ml, p=0.0008). Within the TM group, the group of seminomas (8.5 *106/ml) 0.65 which indicates that these models are valid in our laboratory. In addition,
presented the lowest sperm counts. Although most of the individual mean values these values for AUC were obtained across all GS, CP and LB KID embryos
were within the WHO reference limits, only 38 % of TM and 45 % of sarcoma databases tested. An increase in the predictability of the models was observed
patients had a normozoospermia. In the other groups (HM, Ben) only 60 % of the from Day 2-3 to Day 5 models. AUC test results ranged around 0.658, 0.671 for
patients showed a normozoospermia. No correlation between sperm quality and Day 2, Day 3 respectively and 0.872 for Day 5 model. Our results show that
Ann- Arbor stadium, TSH- values or BMI was present. Only 5 (1.9 %) patients although embryos were selected for transfer using the general models, specific
used their frozen semen for a total of 9 ART cycles, which resulted in 5 live births. laboratory model is contributory for better selection of embryos. It may improve
Limitations, reasons for caution: The low use rate might be explained by embryo selection and increase the number of single embryo transfers.
the short follow up time after cryopreservation and patients’ age. The storage Developing a laboratory-specific model requires many stages of sorting and
of semen for fertility preservation is currently not covered by health insurance characterization. It demands accurate annotation and statistical analysis for
in Austria compared to other countries, where storage costs play a minor role. every parameter alone and in combination, to evaluate which parameter should
Wider implications of the findings: Depending on the underlying disease, be included in the model. Many conclusions can be drawn about the mod-
patients show different sperm quality, with TM and HM patients having the lowest el-building process, which may facilitate and improve the process in other
sperm concentrations. Due to the low usage rate of cryopreserved semen, a control laboratories.
sperm test should be part of the follow-up cancer care - to avoid costly storage. Limitations, reasons for caution: Laboratory specific in house model is an
Trial registration number: x important tool that helps selecting embryos for transfer, but should be used with
caution and in combination with other selection method such as general models
and embryo classical morphology evaluation by the embryologists.
Wider implications of the findings: We recommend developing in-house
SELECTED ORAL COMMUNICATIONS model for embryo selection in addition to the general models. This may enable
SESSION 59: NEW MORPHOKINETIC INSIGHTS OF EMBRYO single embryo transfers with more confidence, reducing multiple pregnancies
DEVELOPMENT and the number of transfers required to achieve pregnancy. The experience we
08 July 2020 Parallel 1 10:00 - 11:45 gained in the process may be of assistance to other laboratories.
Trial registration number: Institutional study registration number:
0135-18-CMC
O-229 Improving embryo selection by development of laboratory- O-230 Trophectoderm specification: compaction, polarisation
adapted Time-lapse model and inner and outer cells occur simultaneously in the human
I. Blais1, S. Lahav-Baratz1, M. Koifman1, I. Feferkorn1, M. Dirnfeld1,2 preimplantation embryo
1
Lady Davis Carmel Medical Center, Division of Reproductive Endocrinology and A. Demtschenko1, W. Essahib1, G. Verheyen2, K. Sermon3, H.
IVF- Department of Obstetrics and Gynecology, Haifa, Israel ; Tournaye2, H. Van de Velde1
2
Technion – Israel Institute of Technology, Ruth and Bruch Rappaport Faculty of 1
VUB, Reim, Brussels, Belgium ;
Medicine, Haifa, Israel 2
Uz Brussel, crg, Brussels, Belgium ;
3
VUB, Rege, Brussels, Belgium
Study question: Is it applicable to develop morphokineric algorithm for a
specific IVF laboratory, can it provide more information for embryo selection in Study question: How is the trophectoderm lineage specified in the human
addition to general models? preimplantation embryo?

Summary answer: Outer cells at the fully compacted embryo stage display euploid and aneuploid embryos. Moreover, there are live births reported from
characteristics of trophectoderm cells such as apical polar distribution of p-ERM multinucleated embryos. One of the theories about these discrepancies is the
and nuclear GATA3/YAP1/TEAD4 protein expression. location of the multinucleated cells once the blastocyst is formed. In this way, these
What is known already: In mice, the first indication of inner cell mass (ICM) cells may be part of the inner cell mass, the trophectoderm or be excluded.
and trophectoderm (TE) segregation occurs three days post fertilisation (dpf3). Study design, size, duration: Retrospective study involving 20,779 embryos
After compaction, asymmetric divisions give rise to inner and outer blastomeres from 5,621 cycles performed between 2014 and 2019. Two main groups were
that ubiquitously express transcription factor TEAD4 and sequester polarity considered: Control Group (CG;N=16,897), embryos without multinucleation
markers (p-ERM), and TE lineage specifiers (YAP1, GATA3) to outer blasto- and, Multinucleation Group (MNC;N=3,879), embryos showing at least one
meres. This is considered the onset of TE lineage segregation in the mammalian multinucleated blastomere. Embryos transferred belonging to MNC Group
embryo. Studies regarding this event in the human embryo are absent and extrap- (N=307) were subdivided according to the multinucleated cell location after
olations from the mouse model cannot be a surrogate for studies in the blastulation: MNC-1 (N=142), no cells excluded; MNC-2 (N=73), mononucle-
human embryo. ated cells excluded; MNC-3 (N=46), multinucleated cells excluded. All groups
Study design, size, duration: Human embryos used in this study, were were homogeneous.
donated to research as a surplus of IVF-ICSI treatment at our IVF-center after Participants/materials, setting, methods: All the embryos were cultured
the cryo-storage expiration date of 5 years with informed consent. The project until blastocyst stage using one-step culture media. We used the time-lapse
was approved by the institutional ethical committee and the federal committee technology to follow up the location of the multinucleated cells and we had to
for research on human embryos. exclude from the study those transferred embryos that could not be monitored
Participants/materials, setting, methods: Cryopreserved human 8-cell (N=46). Single embryo transfer was performed to all patients and the clinical
stage embryos, donated to research, or zygotes created for research after outcome rates were compared between groups performing the statistical analysis
informed consent were warmed (Vit Kit -Thaw, Irvine Scientific, USA) and cul- Chi-square test.
tured until dpf5 according to standard laboratory procedures of the associated Main results and the role of chance: Our results showed that despite
IVF-clinic. Time course immunofluorescence was performed on fixed embryos presenting a significantly lower rate of blastocyst formation (MNC=20.0%;
at predefined stages for TEAD4, YAP1, GATA3, p-ERM as well as F-actin and CG=58.0%; p<0.05), the transfer of multinucleated embryos that were able to
Hoechst for imaging (LSM800, ZEISS) and the manual estimation of individual reach the blastocyst stage at D5/6 had remarkable reproductive success.
blastomere counts per embryo (ImageJ). Blastocysts that were able to exclude multinucleated cells (MNC-3) achieved
Main results and the role of chance: The compaction process of the human higher clinical outcomes respect those including multinucleated cells (MNC-1
embryo is interrupted by cleavage divisions this results in a fully compacted and MNC-2). Pregnancy rate in MNC-3 (63.0%) was significantly higher than in
embryo at dpf4.These fully compacted embryos are comprised of a total of the other groups (MNC-1=50.7%; MNC-2=41.1%; p<0.05). The clinical preg-
16±3 cells, n=21, of which 76%, n=16/21 contain 1±1 inner blastomere in their nancy rate in MNC-3 (56.5%) was higher than MNC-1 (41.5%) and MNC-2
core excluded from the exterior. The nuclei of the outer blastomeres are positive (32.9%), being statistically significant with MNC-2 (p<0.05).
for TE-specifier YAP1 (69±22%, n=8) and begin to co-express TE-transcription Miscarriage rate of MNC-3 was lower than the other groups but the differ-
factor GATA3 (15±22%, n=8). The apical membranes of the outer blastomeres ences were not significant (MNC-1=16.2%; MNC-2=9.6%, MNC-3=6.5%).
appear polarised by p-ERM staining (100%, n=8) and TEAD4 is expressed ubiq- It was in the live birth rates when MNC-3 (48.9%) showed the highest signif-
uitously (100%, n=8). While inner cells also express TEAD4 they remain apolar icant differences regarding the other two groups (MNC-1=24.8%, MNC-
and rarely express YAP1. This inner/outer pattern is maintained in the blastocyst 2=22.2%; p<0.05).
ICM and TE. All nuclei of TE cells of dpf5 blastocysts are positive for YAP1/ Moreover, we found that embryos excluding multinucleated cells (MNC-3)
GATA3 and TEAD4 and apical membranes of TE cells show polar distribution reached equivalent clinical outcome results to CG (with 4210 blastocysts trans-
of p-ERM. Whereas the ICM remains apolar, TEAD4 positive, GATA3 negative, ferred): pregnancy rate: 63.0% vs. 60.8%; clinical pregnancy rate: 56.5% vs. 50.6%;
and very rarely contains YAP1 positive cells. live birth rate: 48.9% vs. 34.4%.
Limitations, reasons for caution: In spite of available functional data on the Limitations, reasons for caution: There is a wide difference on sample size
interaction between YAP1 and TEAD4 and their effect on GATA3 expression between MNC and CG groups. 14.7% of the MNC transferred embryos had
and embryo development in the mouse and the bovine model, functional exper- to be excluded from the study because they could not be well monitored under
iments in the human embryo still need to confirm the link between polarity, time-lapse viewer.
nuclear TEAD4/YAP1/GATA3, TE specification and blastocyst formation. Wider implications of the findings: We consider that MNC should be
Wider implications of the findings: The same molecular determinants are classified according to their ability to locate multinucleated cells. The exclusion
involved in the TE lineage segregation of mice and humans, but their expression of multinucleated cells could be part of some error detection mechanism.
differs according to the morphological development of the respective species. Embryos that are able to discard them while reaching the blastocyst stage have
In contrast to the stepwise model of the mouse, compaction, polarisation and the same reproductive potential as blastocyst without multinucleation.
inner and outer cells occur gradually in the human embryo. Trial registration number: Doesn’t apply
O-232 Deep learning using embryo preimplantation videos
O-231 Embryos excluding multinucleated cells during blastocyst can automatically predict the potential of human embryos to
formation increase their reprodutive potential blastulate and implant
A. MUNUERA PUIGVERT1, S. Novo1, L. Almenara1, A. García- Y. Ken-Tor1, N. Zabari1, A. Szeskin1, A. Tamar1, D. Richter2, Y. Or3,
Faura1, B. Marquès1, F. García1, C. Castelló1, M. López-Teijón1 Z. Shoham3, A. Hurwitz4, I. Har-Vardi2, M. Gavish1, A. Ben-Meir4,
1
Institut Marquès, Reproductive Medicine Service, Barcelona, Spain A. Buxboim5
1
Hebrew university, Computer science, Tel Aviv, Israel ;
Study question: Has the inclusion or exclusion of blastomeres in multinucle- 2
Soroka University Medical center, Fertility and IVF Unit- Department of Obstetrics
ated embryos any effect on embryo development and clinical outcomes on IVF and Gynecology, Beer Sheeva, Israel ;
treatments? 3
Kaplan Medical Center, Fertility and IVF Unit, Rehovot, Israel ;
Summary answer: Embryos excluding multinucleated cells during blastulation 4
Hebrew University Hadassah Medical Center, Fertility and IVF Unit- Department
appear to have the capacity for self-correction that allows them to keep intact of Obstetrics and Gynecology, Jerusalem, Israel ;
their ability to become healthy babies. 5
Hebrew university, The Alexender Grass Center for Bioengineering- School of
What is known already: The effects of multinucleation on embryo development computer Science and Engineering, Tel Aviv, Israel
and on clinical outcomes are unclear. Some studies have correlated multinucleation
with low embryonic development, low implantation potential and aneuploidies Study question: Does the morphodynamic data in preimplantation human
increase. However, there is still a debate about transferring multinucleated embryos, embryo videos can be analyzed using deep learning algorithms to predict the
as some other studies have observed that multinucleation is equally present in potential to reach blatulation and implantation.

Summary answer: Blastulation prediction on test-set embryos, reached area Summary answer: The results for embryos that present DC and develop
under the ROC curve 0.73 at 72 hour. prediction on of day-5 transferred test-set into good quality blastocysts (GQB) are equivalent to those that do not
embryos, outperformed manual KIDScore™-D5. present DC.
What is known already: In IVF treatments, early identification of embryos What is known already: After the appearance of time-Lapse technology,
with high implantation potential is required for shortening time to pregnancy various studies based on morphokinetics have identified the presence of direct
while avoiding clinical complications caused by multiple embryo pregnancy. cleavage in 14% - 26.1% of all evaluated embryos. These embryos present a
Current classification tools are based on manually annotated morphological and significant reduction in blastocyst formation, euploidy rate and implantation
morphokinetic parameters. However, manual annotation introduces inter-ob- potential compared to control groups without DC. However, depending on the
server and intra-observer variability and provides a discrete representation of cycle where DC is observed, the impact may not be so negative.
preimplantation development while ignoring dynamic features that may be asso- Study design, size, duration: A retrospective study including 11168 embryos
ciated with embryo quality. Deep learning AI, which offers a powerful toolbox from 3175 cycles between January 2018 to October 2019. Two groups were
for carrying out automated and standardized classification tasks involving expan- defined: a DC-negative group (DC-) containing 10051 embryos without DC in
sive datasets, is gradually incorporated into the health care system worldwide any of their division cycles and a DC-postive group (DC+) including 1117
and IVF clinics specifically. embryos with at least one DC in any of their first three cycles. DC+ group was
Study design, size, duration: 16,000 3D videos of preimplantation subdivided into DC+1 (n=493), DC+2 (n=476) and DC+3 (n=148) based on
embryos were annotated for morphological and morphokinetic parameters. the cycle in which DC was observed.
6,200 were blastulation-labelled and > 5,500 were implantation-labelled. Each Participants/materials, setting, methods: For all cycles intracytoplasmatic
embryo video file was associated with maternal information and clinical meta- sperm injection technique was performed and the resulting embryos were cul-
data. The classification was evaluated using fivefold cross-validation and using tured in time-lapse incubators to blastocyst stage using one-step culture media.
an uncontaminated test set consisting of 20% of the labeled embryos. Blastocyst, good quality blastocyst (GQB), multinucleation and ploidy rates were
Generality was verified via leave-one-clinic-out and leave-one-age-group out compared between both groups, with and without DC. Furthermore, groups
cross-validation. DC+1, DC+2 and DC+3 were also compared among each other. The chi-square
Participants/materials, setting, methods: Embryos were cultured in nine test was used for statistics.
time-lapse incubators during the past six years in four medical centers across Main results and the role of chance: At least one DC event was observed
Israel. We developed fully automated and standardized classifiers by training in 10% of embryos on culture day 2 or 3. This rate was lower compared to values
time point sensitive deep neural networks directly on video frames. Using published previously by others laboratories. The in vitro development rate of DC+
machine learning we combined the frame scores into a single embryo score for embryos up to day 5/6 (42.4%) was significantly lower than the DC- embryos
blastulation or implantation prediction. Using SHAP values we identified key time (73,6%) (p<0.05). Presence of multinucleated cells was more frequent in DC+
points for prediction and impactful video frames. embryos (487/1117; 43.6%) than DC- embryos (1411/10051; 14%) (p<0.05).
Main results and the role of chance: Blastulation prediction on test set GQB rate was significantly lower the earlier DC happened (DC+1 46/493; 9.3%,
embryos increased monotonically with the time of prediction measured from DC+2 106/476; 22.3%, DC+3 66/148; 44.6%; p<0.05). It is noteworthy that
ICSI, reaching area under the ROC curve (AUC) 0.73 at 72 hours, 0.88 at 96 DC+3 GQB rate was similar to the control group (5298/10051; 52.7%)
hours and 0.94 at 110 hours. Blastulation prediction was further demonstrated (p=0.0601). When only considering embryos biopsied on day 3, DC+ embryos
using high-quality embryos that reached 8 cells. had lower euploidy rates (8/84; 9.5%) than DC- ones (316/1036; 30.5%) (p<0.05).
KID predictive strength increased with time of prediction as evaluated for the However, on day 5/6 biopsy, the euploidy rate was equivalent (DC+ 30/82; 36.6%
same cohort of day-5 transferred test-set embryos. AUC increases slowly from vs DC- 497/1300; 37.9%; p=0.8569), as was pregnancy rate (DC+ 7/12; 58.3%
48 to 84 hours and more rapidly from 84 hours onward. Our deep learning vs DC- 228/325; 70.2%; p=0.5786) and ongoing pregnancy rate (DC+ 5/12;
prediction is as accurate as KIDScore-D3 on day-3 and more accurate than 41.7% vs DC- 185/325; 56.9% p=0.4531). Differences in mosaicism rates were
KIDScore-D5 on day-5 as evaluated for the same test-set embryos. not significant (DC+ 10/82; 12.2% vs DC- 237/1300; 18,2%; p=0.2168).
Both blastulation and implantation results were consistent with fivefold Limitations, reasons for caution: Due to their decreased potential depend-
cross-validation, leave one-clinic-out and leave one-age-group out from the ing on the moment of DC, the number of DC+ embryos biopsied and/or
training session, supporting generality. transferred was lower compared to DC-. A larger sample in the study group is
Using SHAP analysis we identified key temporal points that can direct blastu- necessary to confirm the above results.
lation and implantation prediction without including the rest of the video. we Wider implications of the findings: Embryos with early DC+ should never
trained again the embryo-learning blastulation and implantation classifiers using be the first option for transfer. From our results, in cases where PGT is indicated,
only the key temporal points. The new models reached comparable AUC for biopsy must be performed on day 5/6. If an embryo is capable of developing
both goals supporting the importance of a few time points. into a GQB, events that are linked to bad prognosis may have been resolved.
Limitations, reasons for caution: Prediction accuracy is limited due to Trial registration number: not applicable
lacking critical information about endometrial receptivity and using a homoge-
nous dataset of embryos that were retrospectively preselected for transfer O-234 Application of the embryo-uterus statistical model for
according to established morphological and/or morphokientic criteria. prediction of implantation after day 3 embryo transfer by using
Wider implications of the findings: Deep learning provides full automation time-lapse morphokinetics and female age
and standardization of embryo classification and improves accuracy. Our classi- E. Van Marion1, E. Baart1, E. Van Santbrink2, M. Avo Santos2,
fiers mark the first step towards the development of a decision support tool. J. Laven1, M. Eijkemans3
This framework opens the door for clinical implementation of deep learning 1
Erasmus University Medical Centre Rotterdam, Division of Reproductive
classification tools that will improve conception rates while shortening time to Endocrinology and Infertility- Department of Obstetrics and Gynaecology,
pregnancy. Rotterdam, The Netherlands ;
Trial registration number: not applicable 2
Reinier de Graaf Groep, Division of Infertility- Department of Obstetrics and
Gynaecology, Voorburg, The Netherlands ;
O-233 Direct cleavage in late stages of embryo progession does 3
University Medical Centre Utrecht, Biostatistics and Research Support, Utrecht,
not affect in vitro development, ploidy nor the reproductive The Netherlands
potential of human embryos
J. Masso Hernaez1, S. Novo1, V. Moens1, À. García-Faura1, B. Study question: Can we develop a robust and clinically applicable time-lapse
Marquès1, F. García1, C. Castelló1, M. López-Teijón1 morphokinetic model, which predicts embryo implantation potential after single
1
Instituto Marques, Reproductive Medicine Service, Barcelona, Spain embryo transfer (SET)?
Summary answer: A prediction model based on the time interval between
Study question: What are the effects of direct cleavage (DC) on embryo the four- and five-cell stage and female age showed adequate performance on
development and preimplantation genetic testing (PGT) results? an independent data set.

What is known already: Over the past years, time-lapse embryo culture is injection (ICSI) demonstrated an improvement in fertilization, cleavage and blas-
increasingly used as a semi-quantitative tool to research the timing of embryo tulation rates in couples with fertilization failures or severe male factor infertility.
development and the correlation with implantation. Studies with different sample However, there is no data available on the morphokinetic evaluation of embryos
sizes and statistical approaches have led to either centre specific or generally developed following activation.
applicable models to support embryo selection. The predictive capability of Study design, size, duration: This retrospective cohort study included 798
time-lapse selection algorithms may be influenced by patient characteristics, type embryos from 484 couples undergoing ICSI from 2018-2020. We used propen-
of data included in the analysis and the used statistical methods. Earlier studies sity score matching to compare the morphokinetic parameters of embryos that
excluded double embryo transfer (DET) cycles of which only one embryo were developed following ICSI with or without oocyte activation via ready to
implanted, introducing bias in the data. use calcium ionophore.
Study design, size, duration: This is a retrospective study of couples (n= Participants/materials, setting, methods: The indication for oocyte acti-
707) undergoing an in vitro fertilization (IVF) cycle with or without intracytoplas- vation was either severe male factor infertility or previously failed/low fertiliza-
mic sperm injection (ICSI) at the Erasmus University Medical Centre between tion(≤ 25%). After excluding women ≥40-year of age, the study group comprised
January 2012 and June 2019. Embryo transfer was either SET or DET. This 121 women and the control group 363 . Groups were re-analysed according to
resulted in 785 transferred embryos that reached at least the five-cell stage. female age and ICSI indication . All embryos were cultured in a timelapse incu-
Embryo selection was not guided by time-lapse morphokinetics but was per- bator in the same single step medium and annotated at t2, t3, t4, t5, t8 and tB
formed on day 3 according to classic morphological criteria. time points by a single embryologist.
Participants/materials, setting, methods: Embryos were cultured in the Main results and the role of chance: The mean female age was 33.7 years
EmbryoScopeTM and retrospectively annotated for developmental time points (19-40). Morphokinetic measurements showed normal distributions. The mean
up to the 8-cell stage. The final prediction model was established by using the annotation time points for “t2, t3, t4, t5, t8 and tB” in the control versus study
Embryo-Uterus model (EU) (Roberts, Stat Med 2007:p156) with the number group were in the following order: 26.8 vs 26.9, 35,0 vs 35.2, 38.4 vs 38.3, 46.6
of gestational sacs as the outcome variable. Correlations between embryos that vs 47.0, 56.1 vs 55.5, 104.9 vs 103.8 hrs (p>0.05 for all). When adjusted accord-
implanted simultaneously were included in the model. The model was externally ing to female age and ICSI indication, no difference was detected between two
validated on time-lapse data from 1269 fresh embryo transfers performed at groups for any time points.
the Reinier de Graaf hospital. Limitations, reasons for caution: This was a single center retrospective
Main results and the role of chance: We performed manual backward selec- study reporting on a limited number of ICSI cycles in which calcium ionophore
tion using P < 0.3 for inclusion in a multivariable EU-regression analysis with the treatment was deemed necessary.
following parameters: female age, time to three-cell stage (t3), time-interval Wider implications of the findings: Calcium ionophore treatment of
between the two- and the three-cell stage (t3-t2), time to four-cell stage (t4), oocytes to improve fertilization and/or clinical outcome in selected patients
time-interval between the four- and the five-cell stage (t5-t4), and the ratio of the does not have a significant impact on embryo kinetics.
time-interval between the three- and the five-cell stage and the two- and the five- Trial registration number: Not Applicable
cell stage (t5-t3/t5-t2). As the relationship between these parameters and gesta-
tional sacs was non-linear, they were included using cubic splines. All variables were
entered in the Embryo part of the model. This resulted in a final model including
female age and t5-t4, yielding the probability of clinical pregnancy after SET, with SELECTED ORAL COMMUNICATIONS
the optimal timing of t5-t4 between 10-14 hours. An embryo that needs less time SESSION 60: LONG TERM HEALTH, OBSTETRICS AND NEO-
between t5-t4 shows a steeper decrease in implantation potential than if it needs NATAL OUTCOMES RELATING TO INFERTILITY TREATMENT
longer. This was consistent for all female ages. The area under the receiver oper- 08 July 2020 Parallel 2 10:00 - 11:45
ating characteristic curve (AUC) of the final model was 0.624. The model was
capable of predicting clinical pregnancy in the external dataset with an AUC of
0.639 and showed adequate calibration with a slope of 1.235.
Limitations, reasons for caution: Although the model shows good calibra- O-236 Pregnancy, Delivery, and Neonatal Outcomes Among
tion and validation, prospective validation is needed to establish clinical applica- Women with Congenital Adrenal Hyperplasia: A population based
bility. Embryo transfer (ET) was performed on day 3. For clinics performing day study on 9.1 million pregnancies
5 ETs the model can be relevant for poor responders or patients that do not A. Badeghiesh@gmail.com1, H. Baghlaf2, E. Suarthana1, M. Dahan1
want to culture until day 5. 1
Mcgill University, Obstetric and Gynecology department, Montreal, Canada ;
Wider implications of the findings: We were able to minimize selection 2
University of Toronto, Obstetrics & Gynecology, Toronto, Canada
bias by analysing both SETs and DETs irrespective of the number of implanted
embryos and selecting embryos for transfer by morphology alone. This model
Study question: What are the pregnancy, delivery and neonatal outcomes in
can be used to counsel couples on their pregnancy chances before embryo
congenital adrenal hyperplasia (CAH) patients?
transfer, and as a decision tool for cryopreservation.
Summary answer: CAH patients have a two-fold higher risk of; chorioamni-
onitis, maternal infection and cesarean section also fetal increases in small for
gestational age and congenital malformation
O-235 Does calcium ionophore treatment have any effect on What is known already: Congenital adrenal hyperplasia (CAH) is a cluster
embryo kinetics? of inherited enzymatic defects of adrenal steroid biosynthesis. Deficiencies of
I. Keles1, B. Balaban2, A. Isıklar2, S. Ertaş2, B. Urman2, B. Ata1, K. each enzyme required in the steroid biosynthesis pathway are well known, and
Yakin3 these deficiencies are all inherited as autosomal recessive disorders. Women
1
Koc University Hospital, Assisted Reproduction Unit, ISTANBUL, Turkey ; with CAH have decreased fertility because of oligo-ovulation. Conception
2
American Hospital of Istanbul, Assisted Reproduction Unit, Istanbul, Turkey ; requires a combination of proper therapeutic compliance, careful endocrine
3
Koc University, Obstetrics and Gynecology, Istanbul, Turkey monitoring, and often ovulation induction. There are significant gaps about preg-
Study question: Is oocyte activation with calcium ionophore associated with nancy, delivery and neonatal outcomes among CAH patients. The purpose of
morphokinetic parameters in cleavage and blastocyst stage embryos? this study is to investigate these outcomes.
Summary answer: Embryos that were grown from activated oocytes did not Study design, size, duration: We conducted a retrospective popula-
show any difference in terms of morphokinetic parameters. tion-based cohort study utilizing data from the Health Care Cost and Utilization
What is known already: Alterations in calcium signalling may be one of Project-Nationwide Inpatient Sample database (HCUP-NIS) over 11 years from
the underlying reasons for defects in cell growth and clevage. It has been shown 2004 to 2014. We created a cohort of all deliveries between 2004 and 2014
in humans that calcium fluctuations were detected with a peak shortly before inclusively. Within this group, all deliveries to women with CAH were identified
cell division and these calcium oscillations disappeared in arrested embryos. as part of the study group (n=299), and the remaining deliveries were catego-
Clinical use of calcium ionophore treatment after intracytoplasmic sperm rized as non-CAH births and comprised the reference group (n=9,096,489).

Participants/materials, setting, methods: Analysis was performed to in our institution between March 2013 and December 2015. Maternal serum
identify the prevalence of pregnant women with CAH over the study duration. and cord blood vitamin D levels were analysed at the day of delivery.
Demographic and clinical characteristics were compared between women with Main results and the role of chance: Maternal vitamin D levels did not
and without CAH using Chi-square test. All confounding variables were adjusted differ significantly in women with PCOS and without PCOS (p=0.998), nor did
for using multivariate logistic regression, based on any significant differences the vitamin D levels of their respective offspring (p=0.692). Vitamin D deficiency
between the two groups generating adjusted odds ratios (aOR). (< 20 ng/mL) was found in 26.9% and 22.5% of women with and without PCOS
Main results and the role of chance: There were 9,094,499 deliveries (p=0.430). There was a strong positive correlation between maternal and neo-
during the study period. 299 pregnant women were found to have CAH. natal vitamin D levels in both investigated groups. Linear regression estimates
Compared to the control group, CAH subjects were likely to be: older of cord blood vitamin D levels are about 77% of serum vitamin D concentrations
(p<0.001), white (p<0.001), have higher incomes (p<0.001), have private of the mother. Compared to healthy controls, the risk for maternal complications
medical insurance (P<0.001), be obese (p<0.001), have a previous cesarean was increased in PCOS women (48% versus 65%; p=0.009), while there was no
section (p<0.001), chronic hypertensive (7.4% versus 1.8%, p<0.001), pre-ges- significant difference in neonatal complications (22% versus 22%; p=1.0).
tational diabetics (5% versus 0.9%, p<0.001), and have thyroid disease (11.7% However, vitamin D levels were similar between mothers and infants with and
versus 2.5%, p<0.001). Chorioamnionitis was higher in CAH compared to without perinatal complications.
controls (aOR 2.67, 95% CI 1,17 -6.06). The rate of caesarian section (aOR Limitations, reasons for caution: Fetal cord blood was used for the analysis
2.10, 95% CI 1,44 – 3.07) and maternal infection (aOR 2.63, 95% CI 1,22 of fetal hormonal levels.
-5.63) were higher in CAH. Rates of pregnancy induced HTN, preeclampsia, Wider implications of the findings: Although the share of women and
eclampsia, gestational diabetes, preterm delivery, preterm premature rupture infants with vitamin D deficiency was high in women with and without PCOS, it
of membranes and postpartum hemorrhage did not differ between CAH group seems that the incidence of adverse perinatal outcome was not affected. The
and the control group. long-term consequences for mothers and infants with a vitamin D deficiency
At birth, 8% and 2.2% of the neonates were found to be small for gestational have to be investigated in future studies.
age (SGA) in CAH and control groups respectively (aOR 3.37 95% CI 1.86 - Trial registration number: NCT02106676
6.11). Congenital anomalies were encountered in 2.7% of the CAH group com-
pared to the control group (0.4%) (aOR 5.24 95% CI 2.31 - 11.90) O-238 Early ovarian ageing and long-term health consequences: Is
Limitations, reasons for caution: This retrospective analysis utilizes an number of oocytes harvested in ART associated to an earlier and
administrative database, with its inherent limitations. Significant medical history increased risk of age-related diseases?
or adverse pregnancy outcomes may be more often reported in patients with
more significant conditions or outcomes This
M.W. abstract
Christensenhas
1
been
, U.S. chosen
Kesmodel 2
forKirkegaard
, K. the press3
programme
, H.J. Ingerslev2
and
Århuswill notHospital,
be made public until the Aarhus
abstract is ;
1
Wider implications of the findings: CAH patients are at risk of: chorioam- University Obstetrics and Gyneacology, N, Denmark
2
Aalborg University Hospital, Fertility Unit, Aalborg, Denmark ;
nionitis, maternal infection and cesarean section. CAH is associated with small presented.
3
Aarhus University Hospital, Obstetrics and Gyneacology, Aarhus N, Denmark
gestational age infants possibly related to chronic maternal steroid use. The rate
of congenital malformations likely related to elevated androgens in female off-
Study question: Do young women with early ovarian ageing defined as unex-
spring and chronic glucocorticoid used in pregnancy was about 2.7%.
plained, repeatedly few oocytes harvested in ART have an increased risk of
age-related diseases?
Summary answer: At follow-up young women with idiopathic early ovarian
O-237 Vitamin D concentration at term in newborn-mother pairs ageing had an increased risk of age related diseases compared to young women
with and without polycystic ovary syndrome: association with with normal ovarian ageing.
perinatal outcome What is known already: Early and premature menopause is associated with
M. Kollmann1, B. Obermayer-Pietsch2, G. Pregartner3, E. an increased risk of cardiovascular diseases (CVD), osteoporosis and death.
Lerchbaum4, S. Feigl1, P. Klaritsch5 Identifying women at risk may thus allow early preventive health initiatives.
Repeatedly few oocytes harvested in well stimulated assisted reproductive tech-
1
Division of Obstetrics and Maternal Fetal Medicine, Department of Obstetrics nology (ART) cycles is a likely predictor of advanced menopausal age when seen
and Gynecology- Medical University of Graz, Graz, Austria ; in young women and may thus serve as an early marker of accelerated general
2
Endocrinology Lab Platform- Department of Obstetrics and Gynecology and ageing. Oocyte harvest in ART as a measure of ovarian ageing and thus as a risk
Division of Endocrinology and Metabolism, Department of Internal Medicine- predictor of age-related morbidity and mortality has not been investigated
Medical Uniersity of Graz, Graz, Austria ; previously.
3
Institute for Medical Informatics- Statistics and Documentation IMI, Medical Study design, size, duration: A register-based national historical cohort
University of Graz, Graz, Austria ; study. Young women (≤ 37 years) having their first ART-treatment in a Danish
4
Division of Endocrinology and Metabolism, Department of Internal Medicine- fertility clinic (public or private) during the period 1995-2014 was divided into
Medical University of Graz, Graz, Austria ; two groups dependent on ovarian reserve status: early ovarian ageing(EOA)
5
Division of Obstetrics and Maternal Fetal Medicine, Department of Obstetrics (n=1,234) and normal ovarian ageing(NOA) (n=18,614). Number of oocytes
and Gynecology, Graz, Austria harvested in first and subsequent cycles was used as a marker of ovarian reserve.
Several national registers were applied to assess morbidity and mortality.
Study question: Do mothers with/without PCOS have different vitamin D Participants/materials, setting, methods: EOA was defined as ≤ 5 oocytes
levels at term, how are vitamin levels reflected in their offspring, and are vitamin in minimum two well-stimulated cycles and NOA as ≥8 oocytes in minimum 1
levels associated with an adverse perinatal outcome? cycle. Known causes influencing the ovarian reserve (endometriosis, surgery,
Summary answer: Vitamin D levels did not differ significantly in women with/ chemotherapy etc.) was reason for exclusion. Primary outcome was overall-
without PCOS and their respective offspring. Vitamin D deficiency was not disease risk defined as either: CVD, osteoporosis, type-2 diabetes, cancer, all-
associated with adverse perinatal outcome. cause death, Charlson Comorbidity index, cataract, Alzheimer’s or Parkinson’s
What is known already: Studies suggest that non pregnant women with disease or early retirement benefit. Cox regression models were used to assess
PCOS may be at elevated risk of vitamin D deficiency. Furthermore, there is the disease risk after first ART-cycle.
evidence suggesting that vitamin D may also play an important role during preg- Main results and the role of chance: Median follow-up time from first
nancy. Data regarding vitamin D deficiency during pregnancy in PCOS patients ART-cycle to first disease event was 6.1 years (10/90 percentile 1.0/12.9) and
and its association with perinatal outcome is scarce. 6.7 years( 10/90 percentile 1.0/14.1) in the EOA -and NOA group respec-
Study design, size, duration: Prospective cross-sectional study tively. Women with EOA had an increased risk of overall- disease when com-
Participants/materials, setting, methods: We included 79 women with pared to women with a normal oocyte yield (Adjusted HR 1.26, 95 % CI
PCOS according to the ESHRE/ASRM 2003 definition and 354 women without 1.10;1.43). Stratifying on diseases categories, the EOA group had a significantly
PCOS and an ongoing pregnancy ≥ 37 + 0 weeks of gestation who gave birth increased risk for cardiovascular diseases (adjusted HR 1.39, 95 % CI 1.15; 1.67),

osteoporosis (adjusted HR 2.36, 95 % CI 1.48;3.74), Charlson comorbidity index deficiency in our studied population of 34 healthy women on reproductive age
(Adjusted HR 1.28, 95 % CI 1.06;1.54) and early retirement benefit ( adjusted when LC-MS/MS is utilized, and most of them were in the sufficiency range of
HR 1.52, 95 % CI 1.06;2.19). >30 ng/ml (n=23; 67.65%). In contrast, this population could be classified in
Limitations, reasons for caution: Due to register limitations we were unable the insufficiency range according to mean values obtained via ELISA method,
to identify the reason why no oocytes had been collected in case of cancelled most of them showing concentrations bellow <20ng/ml (n=23; 67.65%).
cycles and we may have missed women with the most severe forms of EOA. Limitations, reasons for caution: The study population was a rather homog-
Neither did we have information on the total doses of gonadotropin given in enous group of young, healthy women. These findings need to be confirmed in
each cycle. a larger, more diverse patient population.
Wider implications of the findings: These findings indicate that oocyte Wider implications of the findings: Utilizing ELISA to measure serum vita-
yield may serve as marker of later accelerated ageing when unexpected, repeat- min D levels results in an overestimation of vitamin D deficiency and may explain
edly few oocytes are harvested in young women. Counselling on life-style factors the increased prevalence in the population. Thus, LC-MS/MS should be consid-
as a phrophylactic effort against cardiovascular and other age related diseases ered as a more reliable procedure to measure Vitamin D in research and clinical
may be essential for this group of women. practice.
Trial registration number: The study was approved by the Danish Data Trial registration number: not applicable
Protection Agency ( J.nr 1-16-02-319-14)
O-240 Polycystic Ovary Syndrome as an independent risk factor
O-239 The worldwide epidemic of Vitamin D deficiency: are we for gestational diabetes and hypertensive disorders of pregnancy:
contributing by using inaccurate and unreliable measurement A population-based study on 9.1 million pregnancies
methods? G. Mills1, A. Badeghiesh2, E. Suarthana2, H. Baghlaf2, M. Dahan3
E.E. Lara Molina1, J.M. Franasiak2, A. Devesa-Peiro3, M. López- 1
McGill University, Ob GYN, Montreal, Canada ;
Nogueroles4, M. Florensa5, M. Martin5, D. Amoros6, A. 2
McGill University, Ob Gyn, Montreal- QC, Canada ;
Ballesteros7, A. Pellicer8, P. Diaz-Gimeno9 3
McGill University, McGill University Health Center Reproductive Center,
1
IVI RMA Global Barcelona, Egg Donation, Barcelona, Spain ; Montreal- QC, Canada
2
IVI RMA Global New Jersey, Chief Medical Officer of IVIRMA America, New
Jersey, U.S.A. ; Study question: Does polycystic ovary syndrome (PCOS) confer an indepen-
3
Valencia University, Gynecology and Obstetrics, Valencia, Spain ; dent risk for the development of gestational diabetes (GDM), gestational
4
Biomedical Research Institute La Fe, Analytical Unit, Valencia, Spain ; Hypertension (GHTN), and pre-eclampsia (PEC)?
5
IVI RMA Global Barcelona, IVF laboratory, Barcelona, Spain ; Summary answer: PCOS confers a two-fold higher risk of developing GDM,
6
IVI RMA Global Barcelona, General laboratory, Barcelona, Spain ; a 50% increased risk for of developing GHTN, and a 30% increased risk of
7
IVI RMA Global Barcelona, Reproductive Unit, Barcelona, Spain ; developing PEC.
8
IVI RMA Global Rome, IVI RMA Global President, Rome, Italy ; What is known already: Despite significant evidence of an increased prev-
9
Biomedical Research Institute La Fe, IVI Foundation IVI RMA Global, Valencia, alence of maternal pregnancy complications in women with PCOS, there remain
Spain significant gaps in understanding how PCOS affects the development of GDM,
GHTN, and PEC. This is most likely due to the complex, multifactorial etiology
Study question: Is Vitamin D deficiency diagnosis biased by which measure- of PCOS, its range of potential confounders for pregnancy complications, and
ment technique is utilized? the variable methodology of studies that have been conducted. To date, the
Summary answer: 25-hydroxyvitamin D (25OHD) serum concentrations are largest meta-analysis on this subject includes 11, 565 women with PCOS analyzed
significantly lower when measured via Enzyme-Linked Immunosorbent Assay for their risk of GDM and 5896 patients analyzed for their risk of PEC.
(ELISA) compared to Liquid Chromatography-Tandem Mass Spectrometry Study design, size, duration: We conducted a retrospective popula-
(LC-MS/MS). tion-based cohort study utilizing data from the Health Care Cost and Utilization
What is known already: Vitamin D deficiency is widely reported in general Project-Nationwide Inpatient Sample database (HCUP-NIS) over 11 years from
population and in women undergoing ART, but lack of accuracy when measuring 2004 to 2014. We created a cohort of all deliveries between 2004 and 2014
its metabolites remains as an unresolved issue. 25OHD is the most abundant inclusively. Within this group, all deliveries to women with PCOS were identified
vitamin D metabolite in the circulation, and is proposed to be the best indicator as part of the study group (n=14,882), and the remaining deliveries were cate-
of vitamin D status. Although LC-MS/MS for serum 25OHD measurement is gorized as non-PCOS births and comprised the reference group (n=9,081906).
theoretically a more accurate and reliable technique than immunoassay-based Participants/materials, setting, methods: The analysis was performed to
methods, these are faster and less laborious, and are more commonly used for identify the prevalence of pregnant women with PCOS over the study duration.
Vitamin D assessment in the general practice. Baseline clinical and demographic characteristics between both groups were
Study design, size, duration: 34 healthy women participating in our egg compared. Logistical regression analyses were conducted to explore associations
donation program were included during four months in this prospective, non-in- between PCOS and maternal metabolic outcomes through the estimation of
terventional cohort study. Serum samples were collected for quantification of odds ratio (OR) and 95% confidence intervals (CI). The regression models were
25OHD concentrations with the use of LC-MS/MS procedure and with ELISA. adjusted for the potential confounding effects of maternal demographic, preex-
25OHD levels according to IOM guidelines (<20, 20-30, and >30 ng/mL) were isting clinical characteristics, and concurrently occurring characteristics.
evaluated according each method results. Main results and the role of chance: At baseline, more pregnant women
Participants/materials, setting, methods: Serum was obtained in each with PCOS had obesity (22.3% vs. 3.5% p<0.001), chronic hypertension (8.4%
subject, and then separated into two samples. 25OHD concentrations in one vs. 1.8% p<0.001), pre-gestational diabetes (4.1% vs. 0.9% p<0.001), and thyroid
of the samples were measured via LC-MS/MS using a UPLC-TQ-S Xevo Waters disease (12.6% vs. 2.4% p<0.001). Women with PCOS were more likely to
system with a Waters Acquity BEH C18 (1,7μm 2,1 x100mm) column. A Vitamin have undergone IVF treatment (2.4% vs. 0.1% p<0.001), have multi-gestation
D Enzyme-Linked Immunosorbent Assay (ELISA) kit (ab213966) Abcam for the pregnancies (5.9% vs. 1.5% p<0.001), and more multiple gestations in the PCOS
quantitative determination of 25OHD was used for the other sample. A paired cohort were the result of IVF treatment than the Non-PCOS cohort (12.3% vs.
Wilcoxon test was performed for contrasting the mean differences between 2.3% p<0.001).
both techniques. In singleton pregnancies, women with PCOS were more likely to develop
Main results and the role of chance: All the cases were included and studied gestational diabetes (aOR 2.17, 95% CI 1.99-2.35), pregnancy-associated HTN
during autumn and winter months. None of them had taken vitamin D oral (aOR 1.41 95% CI 1.29-1.54 p<0.001), gestational hypertension (aOR 1.48,
supplements during the last six months before sampling. Mean value for 25OHD 95% CI 1.31-1.66), pre-eclampsia (aOR 1.31, 95% CI 1.15-1.50), and superim-
concentrations in serum was 36.96±15.78 ng/ml when measuring via LC-MS/ posed pre-eclampsia (aOR 1.34, 95% CI 1.07-1.67) after controlling for con-
MS, and significantly lower when ELISA method was used (20.74 ± 21.73 ng/ founding effects (age, race, income level, insurance type, obesity, IVF use,
ml, p-value=1.255e-05). According to IOM guidelines, there was no Vitamin D previous C/S, chronic HTN, pre-gestational diabetes, thyroid disease, smoking,

and recreational drug use). In multiple gestation pregnancies, PCOS only con- Limitations, reasons for caution: As a retrospective cohort study, there
ferred an increased risk of developing GDM (aOR 2.33 95% CI 1.92-2.83 may be some amount of selection and misinformation bias that cannot be
p<0.001). However, there was a non-significant trend towards an increased risk accounted for. This study has a relatively small sample size, suggesting the pos-
for developing pregnancy-associated HTN (aOR 1.92 95% CI 0.99-1.42 p=0.058) sibility of a type 1 error. However, given the levels of significance between group
in the multiple gestations group. outcomes, however, this risk remains low.
Limitations, reasons for caution: This retrospective analysis utilizes an Wider implications of the findings: Addition of GnRH antagonist for 7-10
administrative database, relies on the accuracy of individuals reporting and coding days to routine cabergoline treatment after an hCG trigger in agonist cycles may
data. Remote diagnoses of PCOS or mild phenotypes may not have been reduce the risk of developing moderate and severe OHSS compared to caber-
reported. Significant medical history and/or adverse pregnancy outcomes may goline alone. Further studies are warranted to determine if GnRH-antagonists
be more often reported in patients with more significant conditions and can be used as a treatment for OHSS once started.
outcomes. Trial registration number: Not Applicable
Wider implications of the findings: PCOS is an independent risk factor for
the development of GDM, GHTN, and PEC. It is important to consider the risk
O-242 Artificially prepared frozen embryo transfer cycles are
of all other co-existing metabolic conditions in women with PCOS, as these risks
associated with an increased risk of preeclampsia.
are additive and can significantly increase the risk of adverse complications in
pregnancy. C. Roelens1, A. Racca1, S. Mackens1, L. Van Landuyt1, L.
Trial registration number: not required Gucciardo2, M. De Vos1, H. Tournaye1, C. Blockeel1
1
UZ Brussel, Centre for Reproductive Medicine, Jette, Belgium ;
O-241 GNRH antagonist administration to decrease risk of OHSS 2
UZ Brussel, Department of Obstetrics and Prenatal Medicine, Jette, Belgium
in GNRH agonist cycles triggered with HCG
G. Mills1, S.Y. Dahan1, M.H. Dahan1 Study question: Is the development of preeclampsia (PE) following frozen
1
McGill University Health Centre, Obstetrics and Gynecology, Montreal, Canada embryo transfer (FET) related to the method of endometrial preparation?
Summary answer: Hormonal replacement therapy (HRT) FET cycles are
Study question: Does administration of a gonadotropin-releasing hormone associated with a significantly higher risk of developing PE when compared with
antagonist after HCG trigger in GnRH agonist cycles with significant stimulation natural cycle (NC) FET.
reduce the risk of developing ovarian hyperstimulation syndrome? What is known already: PE is a multisystem disorder encountered in approx-
Summary answer: Addition of GnRH antagonist for 7-10 days to cabergoline imately 2-5% of pregnancies and a leading cause of maternal and perinatal mor-
treatment after hCG-trigger in agonist cycles reduces the risk of moderate/ tality and morbidity. Although assisted reproductive technology (ART) is an
severe OHSS compared to cabergoline alone established risk factor for the development of PE, the exact underlying mecha-
What is known already: In ART cycles that cannot or have not undergone nism has not been elucidated. Recent studies have shown a correlation between
a GnRH agonist trigger, such as in GnRH agonist cycles or when OHSS signs and endometrial preparation before FET and the development of PE during a sub-
symptoms appear in low risk cycles after administration of hCG, secondary risk sequent pregnancy. The impact of the absence of a corpus luteum on the hor-
reduction strategies can be employed. Currently, the two main secondary OHSS monal environment and vascular adaptations in early pregnancy has been
risk reduction strategies are to avoid fresh embryo transfer to prevent endoge- hypothesized as a possible contributing factor in the development of PE after
nous production of hCG, and administration of the dopamine agonist, caber- HRT-FET cycles.
goline. However, when using these strategies, the risk of moderate and severe Study design, size, duration: We performed a retrospective cohort study
OHSS remains present. at a tertiary university-based hospital encompassing 537 unique patients who
Study design, size, duration: A retrospective cohort study of 171 IVF had a pregnancy following FET between 2010 and 2019 and delivered in the
patients at the McGill University Health Centre treated with GnRh-agonist pro- same institution. 324 patients underwent FET in a NC while 213 performed a
tocols and with unexpected exuberant response to stimulation from 2011-2019 HRT cycle. A sample size calculation was performed and showed that 396
was performed. Women received triggering with 5000IU urinary hCG or patients were needed to detect with 90% power a difference of 8.9% between
250mcg recombinant hCG, and were converted to freeze all cycles. study and control group.
Participants/materials, setting, methods: Patients were allocated to one Participants/materials, setting, methods: The primary outcome was the
of two groups. The NO-ANT group (n=123) received cabergoline (Pfizer, incidence of PE defined as the development of hypertension after 20 weeks of
Montreal, Canada) 0.5 mg daily, and the ANT group (n=48) received cabergoline gestation associated with one or more new-onset conditions: proteinuria, mater-
treatment and GnRH antagonist (ganirelix, Orgalutran® 0.25 mg/0.5 ml, Merck, nal or uteroplacental dysfunctions. Multivariable regression analysis was per-
Canada). Treatment in both groups lasted for 7 days starting on the oocyte formed to account for confounding factors knowing to affect the occurrence of
retrieval day. If moderate or severe OHSS occurred, GnRH antagonist was PE, including: NC versus HRT, body mass index (BMI), ethnicity, previous history
continued for 10 days. No fresh embryo transfers were performed. of hypertension during pregnancy and mean arterial pressure (MAP) at the first
Main results and the role of chance: 171 patients were identified as being prenatal consultation.
at risk for developing OHSS after receiving an hCG trigger as part of an agonist Main results and the role of chance: No difference was found in the following
cycle. There were no differences between the two groups (No-ANT vs. ANT, baseline demographic and clinical characteristics between NC and HRT FET cycles:
respectively) in terms of; age (36.2±2.8 vs. 35.8±3.2 years p=0.42), basal serum maternal age, BMI, nulliparity, smoking habits, associated medical conditions, part-
FSH levels (8.1±2.3 vs. 7.9±2.3 IU/L p=0.61), or AFC (12.2±2.4 vs. 11.6±1.9, ners age, developmental stage of the embryo, multiple pregnancy, term at delivery
p=0.12). The ANT group had more oocytes collected (18.6 vs. 17.1 p=0.03), and birth weight. Patients suffering from PCOS and ovulation disorders were more
more 2PN embryos (14.3 vs. 12.2 p<0.001), and more frozen blastocysts commonly represented in the HRT group as compared with the NC group (41.3%
(4.4 vs. 3.9 p=0.04). versus 6.2%, p<0.001). African ethnicity was more frequent in the HRT group
The NO-ANT group had more cases of moderate and severe OHSS (52% (15.9% versus 8.9% for HRT and NC respectively, p=0.021). Patients undergoing
vs 25% p = 0.001, and 25% vs 10% p = 0.03, respectively), more occurrences a HRT cycle delivered more often by caesarean section as compared with the NC
of free pelvic fluid (74% vs. 35% p<0.001), and more bloating and discomfort group (42.7% versus 29%, p=0.001). MAP at the first prenatal consultation was
(91% vs. 65% p<0.001) than patients in the ANT group. The number of peri- significantly higher in HRT FET cycles (87.9 mmHg ± 17.8 and 82.9 mmHg ± 14.9
toneal ascites drainages/cases of severe OHSS were lower in ANT group for HRT and NC respectively, p=0.001).
(0.4 vs. 1.2 p=0.01). The incidence of PE was significantly lower in NC FET cycles (3.7% versus
The ANT group had lower serum hemoglobin (14.2±1.4 g/dl vs. 15.1±1.3 11.3% for respectively NC and HRT FET cycles, p=0.001). Univariate and mul-
g/dl p<0.001), higher serum albumin (29.4±3.4g/L vs. 23.6±2.9g/L p<0.001), tivariate logistic regression analyses were performed in order to account for
higher serum sodium (132.9±2.6 vs. 132.0±2.2 mEq/L p=0.02), and lower relevant confounding factors. After confounder adjustment the incidence of PE
serum potassium levels (4.6±0.7 vs. 5.2±1.0 mEq p<0.001) than the was significantly lower in the NC FET group (NC versus HRT: aOR 0.35, 95%
NO-ANT group. CI 0.17-0.74, p=0.006).

Limitations, reasons for caution: Although the study adjusted for potential
confounding factors, the results remain limited by the retrospective nature of
the study and its potentially associated bias. SELECTED ORAL COMMUNICATIONS
Wider implications of the findings: The higher incidence of PE in HRT SESSION 61: UNDERSTANDING SPERMATOGENESIS
versus NC FET cycles found in our population adds further weight to the existing BEYOND HISTOLOGY
data on this topic. These significant findings should urge practitioners to prefer-
08 July 2020 Parallel 3 09:50 - 11:55
entially perform FET in a natural cycle instead of a HRT cycle in ovulatory
patients.
O-244 Spermiation in mouse is controlled by RARA-mediated
O-243 High anti-Müllerian hormone levels are associated with gene repression
preterm delivery in patients with polycystic ovary syndrome M. Teletin1,2, N. Vernet3, B. Feret3, M. Klopfenstein3, W. Bourguet4,
K. Hu1, F.T. Liu2, R. Li2 P. Germain4, P. Weber5, M. Mark1,2, N.B. Ghyselinck3
1
1
Peking University Third Hospital, Center for Reproductive Medicine- Department 1Institut de Génétique et Biologie Moléculaire et Cellulaire IGBMC- CNRS
of Obstetrics and Gynecology, Beijing, China ; UMR7104 - INSERM U1258- 1 rue Laurent Fries- BP 10142- F-67404 Illkirch
2
Peking University Third Hospital, Center for Reproductive Medicine, Beijing, China CEDEX, Functional genomics and cancer, Strasbourg, France ;
2
Hôpitaux Universitaires de Strasbourg HUS LBDR CECOS- 1 place de l’Hopital-
Study question: Is the serum AMH levels associated with the risk of preterm Strasbourg-, Cecos-Laboratoire de biologie de la reproduction, Strasbourg, France ;
3
delivery in PCOS patients? Institut de Génétique et Biologie Moléculaire et Cellulaire IGBMC- CNRS
Summary answer: High anti-Müllerian hormone levels are associated with UMR7104 - INSERM U1258- 1 rue Laurent Fries- BP 10142- F-67404 Illkirch
preterm delivery in patients with polycystic ovary syndrome. CEDEX, Functional genomics and cancer dept, Illkirch, France ;
4
What is known already: Anti-Müllerian hormone (AMH) levels are higher in Centre de Biochimie Structurale CBS- CNRS UMR 5048 - INSERM U1054- 29
patients with polycystic ovary syndrome. Accumulating evidence indicates that rue de Navacelles- F-34090, Biochimie Structurale, Montpellier, France ;
5
AMH has an impact on the physiology of the female reproductive system. Increased Laboratoire de Biométrie et Biologie Evolutive LBBE- CNRS UMR5558- UCB Lyon
AMH levels in PCOS patients are associated with PCOS severity and greater ovar- 1- Bâtiment Grégor Mendel- 43 Boulevard du 11 novembre 1918- F-69622-
ian stimulation during IVF treatment. During pregnancy, serum AMH levels are also Villeurbanne, Biologie Structurale, Lyon, France
higher in PCOS than non-PCOS patients, and the elevated AMH levels observed
in PCOS patients have an impact on the endocrine system of the fetus. Study question: What is the role of nuclear receptor (NRs) and their inter-
Study design, size, duration: This is a retrospective cohort study. A total action with cofactors in mouse spermiation?
of 25165 in vitro fertilization (IVF) cycles with AMH data performed from January Summary answer: Our data establish that gene repression mediated in Sertoli
2017 to July 2018 in Peking University Third Hospital were available for analysis cells by NCoR-bound RARA/RXRB heterodimers is instrumental to spermiation
in this study.. in mouse.
Participants/materials, setting, methods: Among the 25165 cycles, 10718 What is known already: The activity of nuclear receptors is thought to be
were fresh embryo transfer (ET) cycles, and 14447 were frozen-thawed embryo mediated by interaction with coactivators or corepressors. In the absence of
transfer (FET) cycles. There were 136 preterm deliveries and 1777 term deliv- their cognate ligands, it is admitted that NRs bound to the DNA response ele-
eries in the fresh ET group and 145 preterm deliveries and 1685 term deliveries ments located in their target genes recruit corepressors and inhibit mRNA
in the FET group. In PCOS patients, 423 term deliveries and 45 preterm expression. The physiological impact of NR-mediated gene repression is poorly
deliveries. characterized. We have shown previously that loss-of-function mutants of RARA
Main results and the role of chance: Serum AMH levels were not different or RXRB display spermiation defects.
between the term delivery and preterm delivery groups in the entire cohort Study design, size, duration: To address this question, we have generated
(3.8 vs 4.1 ng/mL, P >0.05). In patients diagnosed with PCOS, those with and analyzed the phenotypes of mice expressing a mutant form of all-trans
preterm delivery had higher AMH levels than were found in those with term retinoic acid (ATRA) receptor alpha (RARA) which specifically disrupts core-
delivery (9.3 vs 6.9 ng/mL, P <0.01). Preterm deliveries predominated in PCOS pressor binding without affecting agonist-dependent coactivator binding (RARA
patients with AMH levels above the 75th percentile (9.75 ng/ml) (adjusted P I396E mutation). We have additionally generated mice in which NCor1 and NCor2
<0.0001, adjusted OR=4.0 (95%CI 1.94, 8.08), adjusted for age, BMI, indicated were knocked out specifically in Sertoli cells.
delivery associated complications, embryo transfer number, cesarean or vaginal Participants/materials, setting, methods: Histological analysis of testes
delivery, fresh ET or FET, gravidity times, parity times, and sex of the infant) from RARA-I396E expressing mice and also of testes from NCor1 and NCor2
and frozen-thawed embryo transfer (FET) patients with AMH levels higher compound double mutants in Sertoli cells were performed together with in situ
than the 90th percentile (10.10 ng/ml) (adjusted P <0.05, adjusted OR=2.0 hybridization techniques, immunofluorescence and transcriptome analyses.
(95%CI 1.16, 3.36), adjusted for age, BMI, indicated delivery associated com- Main results and the role of chance: We demonstrate here that this specific
plications, diagnosis (PCOS or non-PCOS), embryo transfer number, cesarean mutations impair spermiation (i.e., the release of mature spermatids by the
or vaginal delivery, fresh ET or FET, gravidity times, parity times, and sex of seminiferous epithelium (SE) of the testis), a hallmark of the phenotype observed
the infant). Indicated-delivery associated complications and male infants were upon ablation of RARA or of RXRB in Sertoli cells. Moreover, the combined,
risk factors for preterm delivery (adjusted P =0.01, adjusted OR=3.0 (95%CI Sertoli cell-specific, ablation of the two genes encoding the corepressors NCor1
1.29, 6.87), adjusted P =0.02, adjusted OR=2.3 (95%CI 1.18, 4.58)). Serum and NCor2 mimics the spermiation defect observed in RARA-I396E expressing
AMH levels were not significantly associated with gestational diabetes or mice. Transcriptome analysis revealed deregulation of a specific set of genes,
pre-eclampsia. some of which may control spermatid release. This strongly suggests that gene
Limitations, reasons for caution: The main limitation of this study is its repression mediated in Sertoli cells by NCoR-bound RARA/RXRB heterodimers
retrospective study design and lack of sample size calculation. Some patients is controlling spermiation. It is likely that this repressing activity occurs when the
with repeated IVF failure may get pregnant more than one year after AMH level of ATRA available in the SE is low, which corresponds to stages I to VI of
measurement, which may be different from the exact AMH levels during the the SE cycle, just prior to spermiation.
cycle of successful pregnancy. Limitations, reasons for caution: Since our present results were obtained
Wider implications of the findings: Our results will guide clinicians to in mice, their relevance for human spermatogenesis remains to be
better manage the process of pregnancy in these patients. It will be interesting demonstrated.
for further studies to investigate the potential mechanisms underlying these Wider implications of the findings: Pharmacological studies strongly sup-
effects. port a role of the ATRA signaling pathway in human spermatogenesis. The
Trial registration number: not applicable evidence provided in this work might suggest that changing RARA activity by

pharmacological approaches in humans might be used to help patients with genes are associated with oligoasthenozoospermia and male infertility, probably
spermiation defects and to design new male contraceptive strategies. through influencing sperm count, motility, and morphology. This study lay the
Trial registration number: Not Applicable groundwork for future studies focused on investigating therapies for male
infertility.
Trial registration number: Ha195/11
O-245 MicroRNA-targeting in spermatogenesis: Over-
expressions of microRNA-23a/b-3p and its affected targeting O-246 Relevance of spermiogenic maturation of the male gamete
of the genes ODF2 and UBQLN3 in sperm of patients with through the epididymal journey
oligoasthenozoospermia
A. Melnick1, A. Parrella1, M.S. Wang1, D. Tavares1, M. Haddad1,
M. Abu-Halima1, B. Ayesh2, L. Becker1, M. Hammadeh3, A. Keller4, Z. Rosenwaks1, G.D. Palermo1
M. Eckart5 1
1
Saarland University, Institute of Human Genetics, Homburg/Saar, Germany ; Reproductive Medicine, New York, U.S.A.
2
al-Aqsa University, Department of Laboratory Medical Sciences, Gaza, Palestine ;
3
Saarland University, Department of Obstetrics and Gynecology- IVF and Andrology Study question: Which area of the epididymis yields spermatozoa with the
Laboratory, Homburg/Saar, Germany ; highest capacity for supporting embryonic development?
4
Saarland Univsersity, Chair for Clinical Bioinformatics, Saarbrücken, Germany ; Summary answer: The cauda epididymis yields spermatozoa with optimal
5
University of Saarland, Institute of Human Genetics, Homburg/Saar, Germany spermiogenic maturity, resulting in superior fertilization, embryo implantation,
and delivery rates.
Study question: To investigate whether microRNA-23a/b-3p targets the genes What is known already: Epididymal sampling is the preferred surgical treat-
ODF2 and UBQLN3 and whether this targeting impacts expression levels of ODF2 ment for men with obstructive azoospermia. The preferred site of retrieval is
and UBQLN3 in patients with oligoasthenozoospermia. the caput, which is believed to yield the largest number of spermatozoa with
Summary answer: The over-expression of microRNA-23a/b-3p and low- superior kinetic characteristics. Only a few studies have described the fertilization
er-expression of ODF2 and UBQLN3 genes are associated with male competence of spermatozoa retrieved from the caput, corpus, and cauda regions
subfertility. of the epididymis.
What is known already: Spermatogenesis is the process of male germ cell Study design, size, duration: Between 2011 and 2019, 46 men diagnosed
proliferation and differentiation within the testes. In this complex and highly with obstructive azoospermia (OA) underwent surgical retrieval of spermatozoa
regulated process, many genes are involved, the expression levels of which are from different areas of the epididymis. Spermatozoa were subsequently used
strongly or partially coordinated by microRNAs (miRNAs). MiRNAs are small, for ICSI cycles with the patients’ female partners (≤37 yrs old). Clinical outcome
non-coding RNAs that are involved in the post-transcriptional regulation of gene was compared among the different sources of spermatozoa.
expression. Transcriptome analysis shows that hundreds of genes are expressed Participants/materials, setting, methods: A total of 36 specimens from
exclusively or predominantly in male germ cells including ODF2 and UBQLN3, caput, 7 from corpus, and 3 from cauda were retrieved. Semen parameters were
which play a crucial role during spermatogenesis and/or sperm function. compared among the three sources, and spermatozoa were injected by ICSI in
However, the expression regulation of these two genes is still unclear. oocytes from the female partners. Fertilization, implantation, and delivery rates
Study design, size, duration: A total of 86 men were included in the study, were assessed and compared for all the three epididymal locations. Unpaired t
including 43 oligoasthenozoospermic men who attended the IVF center for and Fisher’s Exact tests were used to compare the outcomes. P values <0.05
infertility treatment at Saarland University, Germany and 43 age-matched nor- were considered statistically significant.
mozoospermic volunteers served as controls. Reverse transcription-quantitative Main results and the role of chance: A total of 46 men were divided in
PCR (RT-qPCR), Northern blot, and dual luciferase assay were used to validate three groups according to the source of spermatozoa used: caput, corpus, and
the over-expression of microRNA-23a/b-3p and lower-expression of ODF2 cauda. Maternal age was comparable among the three groups (32.0 ± 4, 32.1
and UBQLN3 genes. The study was conducted between 2019 − 2020, at the ± 4, and 35.0 ± 1, respectively). Average and standard deviation of spermatozoa
Institute of Human Genetics. concentration was 35.1± 39 in the caput, 28.7±29 in the corpus, and 30.2± 59
Participants/materials, setting, methods: Total RNA, including miRNA x106/ml in the cauda.
was isolated from sperm of oligoasthenozoospermic (n=43) and normozoosper- The mean of motility was 15.8±15% in the caput, rising to 40.1±29.4% in the
mic men (n=43). RT-qPCR was used to detect the expression levels of microR- corpus (P<0.01) and decreasing to 7±11.8% in the cauda (P<0.05). The mean
NA-23a/b-3p and ODF2 and UBQLN3 genes. In silico prediction and numbers of injected oocytes were 12.7, 11, and 12.5, respectively (P=NS). The
dual-luciferase assays were performed to confirm the potential links between fertilization rate was 65.8% with caput, 77.6% with corpus (P<0.05), and 88%
the over-expression of microRNA-23a/b-3p and lower-expression of ODF2 (P<0.05) with cauda spermatozoa.
and UBQLN3 genes. Correlation analyses of the miRNA and mRNA expression The implantation rate also progressively increased distally through the regions
levels were done for clinical sperm parameters. of the epididymis. In the caput, the implantation rate was 34.8%, rising to 44.4%
Main results and the role of chance: The expression levels of microRNA- and 57.1% in the corpus and cauda, respectively. The same trend was observed
23a/b-3p were significantly up-regulated and ODF2 and UBQLN3 genes were for delivery rates. In cycles using caput spermatozoa, the delivery rate was 48.4%
significantly down-regulated in oligoasthenozoospermic men compared with with 5.8% pregnancy loss.
age-matched normozoospermic men as determined by RT-qPCR. Using dual- In cycles using corpus spermatozoa, the delivery rate was 75%, with no preg-
luciferase assays, ODF2 and UBQLN3 genes were identified as direct targets of nancy loss. Ultimately, the cycles with cauda had the highest delivery rate,
microRNA-23a/b-3p. Mutations in the microRNA-23a/b-3p binding site within with 100%.
the 3′UTRs (3′untranslated regions) of ODF2 and UBQLN3 genes resulted in Limitations, reasons for caution: This novel study was performed on a
abrogated responsiveness to microRNA-23a/b-3p. Correlation analysis high- small number of subjects. While we limited female age to ≤37 years, it is not
lighted that sperm count, motility, and morphology was negatively correlated possible to exclude all confounding factors with certainty. In men with OA, it is
with microRNA-23a/b-3p and positively correlated with the lower expression not always possible to choose the site of surgical sampling
level of UBQLN3, while ODF lower expression level was positively correlated Wider implications of the findings: We have found evidence that the
with sperm motility. epididymis retains an important role in spermiogenesis. This has been confirmed
Limitations, reasons for caution: Despite the negative correlation between by the higher fertilization competence of spermatozoa retrieved from the cauda
the over-expression of microRNA-23a/b-3p and the lower-expression of ODF2 epididymis. Confirmation of these findings in a larger study population may serve
and UBQLN3 genes, further validation of these results is needed by an increased as a guide for epididymal sperm retrieval.
number of studied men and by a confirmation of the protein expression by Trial registration number: not applicable
Western blotting.
Wider implications of the findings: Findings suggest that the over-expres- O-247 Effect of malignancy on semen quality: An analysis of 3371
sion of microRNA-23a/b-3p or the lower-expression of ODF2 and UBQLN3 patients

E. Yasmin1, T. Lukaszewski1, E. Williamson2 2

Ben-Gurion University of the Negev, The Shraga Segal Dep. of Microbiology-
1
University College London Hospital, Obstetrics and Gynaecology, London, United Immunology and Genetics- The Center of Advanced Research and Education in
Kingdom ; Reproduction CARER- Faculty of Health Sciences-, Beer-Sheva, Israel ;
3
2
University College London Hospital, Reproductive Medicine, London, United Soroka Medical Center and Faculty of Health Sciences- Ben-Gurion University of
Kingdom the Negev, Dep. OB/GYN and The Center of Advanced Research and Education in
Reproduction CARER-, Beer-Sheva, Israel
Study question: How does malignancy and different diagnoses affect semen
quality as compared with World Health Organisation (WHO) thresholds of Study question: Do Sertoli cells secrete factors that may induce proliferation
semen analysis in fertile men? and differentiation of mouse spermatogonial cells to meiotic and post-meiotic
Summary answer: A wide range of malignancies adversely affect semen qual- stages in vitro?
ity led by testicular cancer and followed by bowel cancer, myeloma and leukaemia. Summary answer: Addition of Sertoli cells conditioned media to spermatogo-
What is known already: The WHO (2010) published a set of reference nial cells in vitro, using methylcellulose culture system induced their development
values for semen analyses based on the assessment of fertile men with the to cells that express acrosin and protamine.
threshold determined by fifth centiles. The WHO guideline is widely used to What is known already: Sertoli cells (SCs) are active in the process of
determine normality of semen analysis. Observational studies have shown the spermatogenesis and are in a direct contact with spermatogonial cells during
deleterious effect of malignancy on semen quality. Most studies have small their development from spermatogonial stem cells to sperm. Endocrine factors
cohorts and larger studies are limited to testicular cancer and haematological such as follicle stimulating hormone induce SCs to produce factors crucial for
malignancies. One of the larger analysis of 4480 cases included both cancer and spermatogenesis. In addition, SPGCs are involved in the regulation of SC func-
benign systemic diseases. This study demonstrated poorer sperm quality in dif- tion. SCs produce different factors that affect the proliferation of the sper-
ferent cancers but did not include lung, bowel, myeloma and prostate cancer. matogonial cells such as glial-derived nerve growth factor (GDNF), stem cell
Study design, size, duration: This is a retrospective observational study of factor (SCF), leukemia inhibitory factor (LIF) and others. However, the factors
single semen analysis prior to oncology treatment in 3371 patients. The data that are involved in the differentiation of the spermatogonial cell are not
spans a 35 year period (1984-2019). All the samples were collected in a single yet clear.
centre. Only major groups of diagnoses were included in the analysis. Very small Study design, size, duration: Seven-day-old mice were used to isolate cells
groups of heterogenous malignancies were excluded from the study. from the seminiferous tubules by 2-step enzymatic digestion. Sertoli cell cultures
Participants/materials, setting, methods: Semen analysis was performed were prepared by culturing the STCs for 3 days in 37 0C in 5% CO2 incubator.
in patients referred for fertility preservation and after they consented to sperm Thereafter, the cells were treated with hypotonic shock to eliminate the residual
cryo-preservation. All patients were post pubertal. Semen analysis was based germ cells, and re-culture in DMEM media containing 10% KSR. After over-night
on the standardised protocol of the centre. The diagnoses, semen parameters the media were replaced by fresh which was collected after 8 hours and stored
and frozen samples were entered into a secure database. The groups were at -80 0C.
based on diagnosis. Distribution of the semen parameters at 5th, 10th, 25th, Participants/materials, setting, methods: Isolate cells from the seminif-
50th, 75th and 95th was carried out and compared with the WHO references. erous tubules (STCs) which contain spermatogonial cells were cultured in vitro
Main results and the role of chance: Of the 3371 patients, the majority (66%) in methylcellulose culture system which contained GDNF, LIF, SCF with or with-
were 20-39 yrs with 15.5% under 20yrs. Nearly a third of patients had testicular out conditioned media (CM) which was collected from SC cultures. The effect
cancer (33%) and over a quarter had lymphoma (28%). Sarcoma was the third of this CM on the development of spermatogenesis in vitro was evaluated by
largest group (15%) followed by leukaemia (9%), prostate cancer (6.5%), brain examining the presence/expression of markers of pre-meiotic (VASA, GFR-
tumour (4%), bowel cancer (3.5%), myeloma (1.5%), lung cancer (0.5%) and mel- alpha, SALL4), meiotic (BOULE) and post-meiotic (ACROSIN, PROTAMINE)
anoma (0.5%). Oligozoospermia (<15mill/ml) was observed in nearly half (46.5%) stages by immunostaining or by qPCR analysis.
of patients with testicular cancer followed by bowel cancer (43%), myeloma (41%) Main results and the role of chance: Our results showed that isolated cells
and leukaemia (40%). Azoospermia was noted in 4-12% of patients across all from seminiferous tubules of 7-day-old mice contained only pre-meiotic cells
diagnoses bar melanoma. The centile distribution of sperm count, sperm concen- (VASA presence in around 8% of cells representing spermatogonial cells), but
tration, motility and volume revealed normal parameters (as per WHO threshold not meiotic or post-meiotic cells. These spermatogonial cells proliferated in vitro,
at 5th centile) only at 50th centile in testicular, brain and bowel cancer, leukaemia in methylcellulose culture system, in the presence of GDNF, LIF and SCF to
and melanoma. Normal values were seen at 25th centile in lymphoma, lung cancer pre-meiotic (VASA presence in around 18% of cells), meiotic (BOULE in around
and sarcoma. The threshold of normal was noted at 10th centile in the melanoma 12% of cells) and post-meiotic cells (ACROSIN in around 8% of cells) after 4-5
group. The study highlights that semen quality is significantly diminished across all weeks of culture (control system). Addition of 10% or 40% (v/v) of SC condi-
diagnoses. The centile distribution of the patients did not match the distribution tioned media to the spermatogonial cells in the culture did not significantly affect
in fertile men on which the WHO thresholds are based in any diagnosis. Semen their proliferation of premeiotic cells compared to the control system. However,
quality was significantly affected in bowel, lung and prostate cancer. addition of these SC conditioned media to the spermatogonial cells in vitro
Limitations, reasons for caution: A single sample of semen was used for significantly increased their development to the presence of acrosin in around
analysis and therefore variation was not accounted for. Period of abstinence 20% of the cells and around 1.5 fold of increase in the RNA expression.
could not be standardised as the samples had to be collected at short notice in Furthermore, an increase in the RNA expression of protamine around 3 fold
the context of fertility preservation. was found compared to the control system.
Wider implications of the findings: The findings of the study can be used Limitations, reasons for caution: The study was performed in mice system
to inform clinicians and counsel patients presenting for fertility preservation. The and the relevant to human need to be confirmed.
study extends information about semen quality across wider diagnoses. The Wider implications of the findings: This is the first demonstration that SCs
study also provided baseline information for comparison with semen analyses produce factors that could induce development of spermatogonial cells to
after treatment and in remission. post-meiotic cells in vitro. These differentiating factors present in SC conditioned
Trial registration number: Not applicable media need to be identified. Our results may assist in using the in vitro system
for male fertility preservation in the future.
Trial registration number: Hub Reproduction, Faculty of Health Sciences,
O-248 Conditioned media of Sertoli cell cultures induced Ben-Gurion University
prepubertal mice spermatogonial cells differentiation in vitro,
using methylcellulose system, to express high levels of acrosin and O-249 Appropriate Response of Fallopian Tube Cell Line to
protamine. Sperms with DNA Fragmentation through Modulating Specific
M. Huleihel1, S. Naor2, E. Lunenfeld3 Cytokines
1
Ben-Gurion University of the Negev, The Shraga Segal Dept. of Microbiology & S.O. Mousavi1, R. Mohammadi2, F. Amjadi3, S. Aghajanpour2,
Immunology, Beer - Sheva, Israel ; M. Eslami1, R. Aflatoonian2

1
Faculty of Advanced Science and Technology- Tehran Medical Sciences- Islamic Study question: How does DNA methylation co-vary between matched sam-
Azad University., Department of Genetics, Tehran, Iran ; ples of human whole blood and spermatozoa, and does obesity influence this
2
Reproductive Biomedicine Research Center- Royan institute for Reproductive DNA methylation covariation?
Biomedicine- Tehran- Iran., Department of Endocrinology and Female infertility-, Summary answer: DNA methylation in blood and spermatozoa is clearly
tehran, Iran ; distinct, with little evidence of covariation between the tissues. We identified
3
Iran University of Medical Science- Tehran- Iran., Department of Anatomical one cross-tissue obesity-associated DNA methylation site.
Science-, tehran, Iran ; What is known already: Paternal obesity and T2DM have been associated
4
Faculty of Advanced Science and Technology- Tehran Medical Sciences- Islamic with an increased risk of fathering low birth weight offspring. Human obesity
Azad University- Tehran- Iran., Department of Genetics-, tehran, Iran and T2DM are associated with altered DNA methylation in blood. Animal studies
suggest that obesity and insulin resistance are associated with DNA methylation
Study question: How sperms with DNA fragmentation can cause inflammation changes in spermatozoa, and that these could mediate intergenerational effects.
and influence the host defense? Such findings are lacking in humans. The largest previous study of DNA meth-
Summary answer: Some investigations suggest that sperms with DNA frag- ylation in matched human sperm and blood had only 8 participants.
mentation can change fallopian tube response with alteration in specific cytokines Study design, size, duration: Genomic DNA was extracted from matched
expression. samples of semen and blood from 68 lean and 22 obese males. DNA methylation
What is known already: Normal physiological homeostasis in the female at ~850,000 CpG sites across the genome was analysed using the Illumina
reproductive tract needs appropriate regulation of inflammation through cyto- MethylationEPIC array to test for 1) interindividual covariation in DNA meth-
kines expression. Cytokines are fundamental compartments of immune system. ylation between blood and sperm, 2) consistent obesity-associated DNA meth-
Interaction of sperms with the fallopian tube in the female reproductive tract ylation differences across both tissues, 3) obesity altered tissue-covariation, and
has an important role in different stages of pregnancy from fertilization to 4) to compare spermatozoal DNA methylation to that of ~6,000 somatic tissue
successful delivery. Investigations has shown that specific cytokines such as samples.
SPP1, MIF and C5 have relevance in host homeostasis and defense against Participants/materials, setting, methods: Males of proven fertility were
pathogens. recruited from the University College London Hospital antenatal clinic. All par-
Study design, size, duration: 30 patients who considered unexplained infer- ticipants provided informed consent and were phenotyped with regards to car-
tile were selected as donors of semen samples with normal features. After diovascular health.
washing sperms, they were categorized to two groups of normal and abnormal DNA methylation was measured using the Illumina MethylationEPIC array at
DFI by TUNEL assay. UCL Genomics and DNA methylation data was analysed using the wateRmelon
We also cultured fallopian tube epithelial cell into the culture flasks containing package in R.
DMEM/F12 with 10% FBS medium up to 70% of confluence. DNA methylation data from ~6,000 somatic tissue samples was obtained
Participants/materials, setting, methods: Fallopian tube epithelial cells from the Gene Expression omnibus and analysed using the bigmelon package in R.
were co-incubated with sperms for 24h. after washing cells and extracting the Main results and the role of chance: There were significant mean DNA
RNA, we synthetized the cDNA. The control group was fallopian tube epithelial methylation differences between blood and sperm at the majority (64%) of the
cells without sperm. Finally, the mRNA expression levels of the cytokines were interrogated 704,356 probes (P < 9 ´ 10-8). At 62% of these, sperm was relatively
evaluated by PCR Array and compared between 3 groups (control, normal DFI, hypermethylated. Sperm displayed a more polarised DNA methylation distri-
abnormal DFI, n=3×10). bution to blood; both low (<20%) and high (>80%) levels of methylation were
Main results and the role of chance: Our data are agreeing with previous more common.
researches which show that cytokines play critical role in sperm interaction with In comparing the spermatozoal DNA methylome to those of ~6,000 somatic
female reproductive tract. tissue samples, we identified 156,654 CpG sperm-specific hyper- and hypometh-
C5 expression in the abnormal DFI group was significantly higher than the ylated sites. Six gene ontology terms were enriched among sperm-specific sites,
control, however there was no significant difference between other groups. all of which related to transcriptional regulation.
Furthermore, SPP1 expression was higher in normal DFI group than the con- When interrogating DNA methylation covariation between blood and sperm,
trol, although it was significantly less than the abnormal DFI group. Finally, it is analyses were restricted to sites meeting minimum variability criteria; range of
notable that MIF had no significant changes between groups (P<0.05). middle 80% of samples ≥ 5% in blood and sperm. Out of 155,269 such sites,
Limitations, reasons for caution: Obtaining semen samples due to ethical 1% showed a significant correlation of DNA methylation between blood and
and logistical issues was the major limitation of this study. Furthermore, in vitro sperm (P < 9´ 10-8). Using the human dbSNP database, we identified SNPs in
co-incubation of fallopian tube epithelial cells with sperms may not directly mimic the probe sequences of most (>99%) of these correlated sites.
the in vivo interaction. We found no evidence that obesity impacts on DNA methylation covariation
Wider implications of the findings: According to our data, DNA- between blood and sperm. However, DNA methylation at one site (cg19357369)
fragmented sperms seem to be established as pathogens in female reproductive was significantly associated with obesity status in blood and sperm (P = 8.95 ´ 10-8).
system and significantly change these cytokines in fallopian tube which prevent Limitations, reasons for caution: A small obesity cohort (n = 22) limited
fertilization events in advance to other reproductive process. our ability to identify modest effects.
Trial registration number: not applicable Lacking the genetic sequence of our subjects meant we could only speculate
as to how SNPs influenced our results, not verify this directly.
O-250 A lack of DNA methylation covariation between human The finding of an obesity-associated CpG site in blood and sperm requires
blood and sperm make it unlikely to mediate intergenerational replication.
inheritance of acquired traits Wider implications of the findings: The lack of DNA methylation covari-
F. Asenius1, S.J. Marzi2, T.J. Gorrie-Stone3, A. Brew4, Y. Panchbaya5, ation between blood and sperm means that blood should not be used as a
E. Williamson6, L. Schalkwyk3, V. Rakyan4, D.J. Williams1 surrogate tissue for spermatozoa.
1
University College London, UCL EGA Institute for Women’s Health, London, Obesity has only a marginal influence on DNA methylation in human
United Kingdom ; spermatozoa.
2
Imperial College London, Department of Brain Sciences, London, It is unlikely that DNA methylation is mechanistically involved in intergener-
United Kingdom ; ational inheritance of acquired traits.
3
University of Essex, School of Biological Sciences, Colchester, United Kingdom ; Trial registration number: Not applicable
4
Queen Mary University of London, The Blizard Institute, London,
United Kingdom ; O-251 Fallopian tube response to DNA fragmented sperms: the
5
University College London, UCL Genomics, London, United Kingdom ; role of Tumor necrosis factor receptors
6
University College London Hospital, Fertility & Reproductive Medicine Laboratory, R. Mohammadi1, S.O. Mousavi2, M. Sabbaghian3, S. Aghajanpour4,
London, United Kingdom Z. Zandieh5, T. Maadani4, R. Aflatoonian6

1
Royan institute for Reproductive Biomedicine., Department of Endocrinology and Summary answer: The expression of piR-32678 in seminal plasma was iden-
Female infertility- Reproductive Biomedicine Research Center-, Tehran, Iran ; tified as a noninvasive evaluation of the spermatogenic function in testis.
2
Faculty of Advanced Science and Technology- Tehran Medical Sciences- Islamic What is known already: microRNAs (miRNAs) have been reported to be
Azad University, Department of Genetics, Tehran, Iran ; involved in spermatogenesis, and may serve as noninvasive biomarkers for eval-
3
Reproductive Biomedicine Research Center- Royan institute for Reproductive uating male fertility. Unlike ubiquitous miRNAs which are secreted from a variety
Biomedicine, Department of Andrology, Tehran, Iran ; of tissues and organs, piRNAs are mainly expressed in germ cell. In this concern,
4
Reproductive Biomedicine Research Center- Royan institute for Reproductive piRNAs in seminal plasma seems to more reliable to develop as candidate pre-
Biomedicine, Department of Endocrinology and Female infertility, Tehran, Iran ; dictor for male fertility.
5
Iran University of Medical sciences, Department of Anatomical science, Tehran, Study design, size, duration: This retrospective cohort study was conducted
Iran ; from January to April 2018. A total of 31 fertile controls with normal semen
6
Reproductive Biomedicine Research Center- Royan institute for Reproductive parameters, 18 oligospermia, 12 non-obstructive azoospermia (NOA) and 11
Biomedicine, Department of Endocrinology and Female infertility-, Tehran, Iran obstructive azoospermia (OA) patients were enrolled in this study. Semen analysis
Study question: Can DNA fragmentation of sperm induce significant changes was determined according to 2010 WHO reference criteria. Whilst, the seminal
in immunological response of fallopian tube? plasma was separated from the spermatozoa by high-speed centrifugation.
Summary answer: The results show that sperm DNA fragmentation may Participants/materials, setting, methods: Seminal plasma RNAs were
alter immunological response of fallopian tube. extracted and quantified. Samples from 5 cases of NOA and 5 cases of controls
What is known already: The interaction between the male and female gam- were subjected to an Arraystar HG19 4×44K piRNA microarray. Among differ-
etes and embryos in the female reproductive system plays an important role in ential expressed piRNAs, piR-32678 was selected for verification on 26 controls,
fertility, embryonic development, and implantation. Active immune systems 18 oligospermia, 7 NOA and 11 OA patients using RT-qPCR. The diagnostic
throughout the female genitalia are against viral pathogens and bacterial agents value of piR-32678 for the spermatogenic failure was evaluated by receiver
which cause sexually transmitted diseases. TNF receptors super family members operating characteristic (ROC) curve.
are one of the most important variables of immune cells that provide effective Main results and the role of chance: Firstly, there were totally 3652 piRNAs
host protection. During fertilization sperms carry proteins which are allogenic differentially exhibited in seminal plasma from patients with NOA compared with
to the female immune system. Therefore, TNF receptors play an important role those from fertile controls (fold change > 2, P < 0.05), including 1532 up-regulated
in the fallopian tube specially in the presence of sperm. piRNAs and 2120 down-regulated piRNAs. Secondly, the qRT-PCR result showed
Study design, size, duration: Fallopian tube epithelial cells were cultured that the relative expression of seminal plasma piR-32678 in oligospermia group
into the tissue culture flasks containing DMEM/F12 with 10% FBS medium. Sperm [3.22 (2.55, 6.34), U = 23.50, P < 0.001] and in the NOA group [ 2.31 (1.65, 3.02),
samples from 20 donors with normal features were collected. The extent of U = 24.00, P = 0.003] was significantly higher than that in the control group [1
sperm DNA fragmentation was measured by the TUNEL assay. Afterward, sam- (0.83. 1.42)]. However, no statistically difference was noticed between OA group
ples classified to two groups of normal and abnormal DFI. The third group was and the controls [ 0.92 (0.54, 2.55), U = 142.00, P = 0.973]. Thirdly, ROC analysis
fallopian tube cells without sperm. demonstrated that the area under the curve (AUC) of piR-32678 was 0.891 (95%
Participants/materials, setting, methods: Different sperms were co-in- CI: 0.812 - 0.970, P < 0.001) for diagnosing spermatogenic failure. When the cut
cubated with fallopian tube cells for 24h. RNA extraction from cells was then off value of the piR-32678 relative expression was set as 1.53, the sensitivity and
fallowed by cDNA synthetized. Finally, PCR array was performed to evaluate specificity were 0.960 and 0.757, respectively.
TNF receptors genes expression profiling. In addition, this data was validated Limitations, reasons for caution: Small sample size is the first limitation for
by q-PCR. this study. The findings require to be further validated in a large sample. The
Main results and the role of chance: The results of the data analysis indi- biological function of piR-32678 needs to be further investigated in the
cated that the expression of some TNF receptors in the vicinity of sperm sig- future study.
nificantly changes. Sperm DFI was assumed to be effective in different expression Wider implications of the findings: Seminal plasma piRNAs can be con-
of TNF receptors. However, The results has shown that the expression of TNF sidered as new innovative noninvasive molecular makers for evaluating male
receptors in cells exposed to abnormal-DFI sperms compared to the cells fertility. Furthermore, piRNAs might provide a new clue for studying the etiology
exposed to sperms with normal DFI had no significant changes. The present of spermatogenic disorders.
study on the effect of spermatozoa on TNF receptors production from the Trial registration number: Not applicable.
fallopian tube epithelial cells revealed that the expression of cytokines altered
between different groups. In conclusion, TNF receptors super family members
including TNF, TNFSF10, TNFSF11 and TNFSF11B had the highest expression
in the group without spermatozoa. SELECTED ORAL COMMUNICATIONS
Limitations, reasons for caution: The major limitation of this study was SESSION 62: PATHOPHYSIOLOGIC ASPECTS OF
obtaining semen samples due to ethical and logistical issues. On the other hand, IMPLANTATION
in vitro culture system may not directly mimic of in vivo environment. 08 July 2020 Parallel 4 10:00 - 11:45
Wider implications of the findings: This study indicate that abnormal DFI
can change the expression of cytokines which have essential roles in sperm
preservation and fertilization. Therefore, this promising novel outcome might
be true that alteration in immunological responses of fallopian tube can disrupt O-253 Endometrial gene expression profiling of recurrent
fertilization events. implantation failure after in vitro fertilization
Trial registration number: not applicable İ.G. Albayrak1,2, F.A. Zarnagh3, B. Karamustafaoğlu Balcı4,
E. Baştu5, T. Günel2
O-252 piR-32678 in the seminal plasma is a potential predictor for 1
Uskudar University, Molecular Biology and Genetics, Istanbul, Turkey ;
spermatogenic function in testis 2
Istanbul University, Molecular Biology and Genetics, Istanbul, Turkey ;
T. Wu1,2,3, L. Li1,2,3, M. Song1, C. Chen1,3, Y. Chen1,3, Y. Zeng1,2,3 3
Istanbul Medical Faculty, Molecular Medicine, Istanbul, Turkey ;
1 4
Shenzhen Zhongshan Urology Hospital, Fertility Center, Shenzhen, China ; Istanbul Medical Faculty, Department of Obstetrics and Gynaecology, Istanbul,
2
Shenzhen Zhongshan Urology Hospital, Shenzhen Key Laboratory of Reproductive Turkey ;
5
Immunology for Peri-implantation, Shenzhen, China ; Acibadem Fulya Hospital, Department of Obstetrics and Gynaecology, Istanbul,
3
Shenzhen Zhongshan Urology Hospital, Shenzhen Zhongshan Institute for Turkey
Reproductive Medicine and Genetics, Shenzhen, China
Study question: Has the expression of endometrial mRNAs in implantation
Study question: Is there any seminal plasma piwi-interacting RNA (piRNA) phase of menstrual cycle significantly altered between RIF (Recurrent Implantation
to predict male fertility? Failure) and the control group?

Summary answer: The study results showed that three selected mRNAs pregnancy and suspected trophoblastic disease were excluded from the study.
were significantly dysregulated in between RIF and control group samples. The study also excluded women who had endometriosis, adnexal masses, exog-
What is known already: Successful embryo implantation depends on a enous hormones, diabetes, women less than 16 years of age and different-
well-functioning endometrium as well as a normal healthy embryo. This process ly-abled women.
might be blocked if either of these variables is defective. Recurrent implantation Main results and the role of chance: Out of 278 women recruited, 15
failure (RIF) is diagnosed when good-quality embryos repeatedly fail to implant women miscarried (5.34%). Comparison of the miscarried women and those
after transfer in several in vitro fertilization (IVF) treatment cycles. Expression who continued pregnancy in women presenting with threatened miscarriage had
differences in maternal mRNAs could be reffering to so many diseases including shown that the two groups of women were different in the age ( P value 0.05)
recurrent implantation failure. A better understanding of the underlying mech- and an objectively assessed bleeding score (P-value 0.03). The two groups were
anisms of RIF will give better treatment opportunities and better outcomes in IVF. also different in the biomarkers hCG (P-value 0.04), progesterone (P-value 0.03),
Study design, size, duration: In this study, there are two groups; RIF and inhibin A (P value 0.02) and ultrasound markers MGSD (P-value 0.04), CRL
normal fertility samples which are collected as endometrial biopsies in implan- (P-value 0.03) and FHR (P-value 0.01). On stepwise multivariate regression
tation phase of menstrual cycle. In the first step, total RNAs isolated from endo- analysis, a regression model composed of the variables of age, hCG, inhibin and
metrium tissue. Then Droplet Digital PCR (ddPCR) performed for expression FHR gave the best sensitivity and specificity to predict miscarriage (P-value
analysis of target mRNAs (mRNAs of TNC, WWC1, MME genes) in samples. 0.0003). The ROC curve for the above regression model showed a sensitivity
And bioinformatic analysis of ddPCR results was performed. Study duration of 58% and specificity of 96%. The model gave a diagnostic odds ratio (95% CI)
lasted from November 2018 until December 2019. of 1.01 (1.01 – 1.02) to predict miscarriage.
Participants/materials, setting, methods: Endometrial biopsy samples Limitations, reasons for caution: Due to fewer numbers of miscarriages,
was collected from 35 RIF samples (age <40) and 35 fertile control samples (age the study was not able to establish a gestation age-specific cut off value for
<40). Samples were collected in the secretory phase of menstrual cylce (cycle predictive markers. Failure to eliminate those miscarriages due to chromosomal
day 20-24). In this study, mRNA expressions of three selected mRNAs were problems might have negatively contributed to the results of the study.
mesuared by ddPCR technique which is a highly sensitive method for measuring Wider implications of the findings: The proposed prediction model needs
expression levels of targeting mRNAs in biological sample. to be tested and validated in a prospective research setting with a different study
Main results and the role of chance: The significant feature of this study population. Contrary to the results of the previous systematic review, the current
is the analysis of mRNA expression level in healthy and RIF endometrial biopsy study established that ca 125 is not a useful marker to predict miscarriage in
samples which collected in implantation phase. Four target genes (MME, WWC1, women presenting with threatened miscarriage.
TNC) were chosen by PANOGA database regarding to their possible relation Trial registration number: T 15-29
with implantation process. Study results showed that, MME and WWC1 genes
expression levels are significantly (P<0,05) up-regulated and TNC gene expres- O-255 Does the outcome of fresh embryo transfer affect the
sion level is significantly (P<0,05) down-regulated in RIF samples comparing to outcome of the subsequent frozen-thawed embryo transfer
the control group. originating from the same cohort of retrieved oocytes
Limitations, reasons for caution: In this study, one of the challenges was G. Oron1,2, M. Ronen1,2, A. Wertheimer1,2, E. Shlush1,2, A.
endometrial biopsy step; sample obtaining from normal fertile women was Hochberg1,2, A. Ben-Haroush1,2, O. Sapir1,2, Y. Shufaro1,2
difficult. 1
Beilinson, Infertility and IVF Unit- Helen Schneider Hospital for Women- Rabin
Wider implications of the findings: The importance of endometrial gene
Medical Center- Beilinson Hospital, Petach - Tikva, Israel ;
expression analysis studies lies in the impact of any identification of one or more 2
Tel-Aviv University, Sackler Faculty of Medicine-, Tel-Aviv, Israel
markers specific for receptivity. In addition to identification of differentially
expressed genes, identifying the related pathways is important for uncovering
Study question: Does the outcome of the fresh transfer affect the subsequent
molecular processes in RIF and understanding the mechanisms underlying the
frozen-thawed transfer originating from the same cohort of oocytes
disease.
Summary answer: The clinical pregnancy rate of the first frozen-thawed
transfer is higher if a clinical pregnancy was attained in the fresh transfer regard-
less of live birth
O-254 Prediction of miscarriage in women presenting with What is known already: Numerous factors including maternal and treatment
threatened miscarriage in their first trimester of pregnancy using characteristics affect the outcome of an IVF treatment cycle. Current data is
biomarkers, ultrasound markers and demographic variables indecisive to whether the outcome of a fresh embryo transfer has an impact on
R. Pillai1, D. Tincello1, N. Potdar1 the outcome of the subsequent frozen-thawed transfer of embryos originating
1
University of Leicester, Department of health sciences, Leicester, United Kingdom from the same cohort of retrieved oocytes.
Study design, size, duration: The study included all oocyte collection cycles
Study question: Can we predict miscarriage in women presenting with threat- between January 2009 and September 2019 that had a fresh embryo transfer
ened miscarriage in the first trimester of pregnancy using biomarkers, ultrasound and at least one frozen-thawed embryo transfer cycle, in a tertiary medi-
markers and demographic variables? cal center.
Summary answer: A log regression model using age, hCG, inhibin A and fetal Participants/materials, setting, methods: Frozen-thawed cycles after a
heart rate can predict miscarriage with a sensitivity of 58% and specificity of 96%. fresh cycle with a clinical pregnancy were compared to frozen-thawed cycles
What is known already: A systematic review and meta-analysis on biomark- after a fresh cycle without a pregnancy. Logistic regression analysis was utilized
ers to predict miscarriage has shown ca 125 as the best marker in predicting to adjust for potential confounders including maternal age, treatment protocol,
miscarriage (Pillai et al., 2016). Another systematic review conducted on ultra- cause of infertility, treatment cycle number, number of oocytes retrieved, num-
sound markers to predict miscarriage has demonstrated fetal heart rate (FHR) ber of frozen embryos, day of fresh embryo transfer (cleavage vs. blastocyst)
as the most predictive marker for miscarriage with a cut off value of 110 bpm and outcome of the fresh embryo transfer
(Pillai et al., 2018). Maternal age is a well-established risk factor for miscarriage. Main results and the role of chance: During the study period 1,758 oocyte
Studies focussing on the combination of markers with FHR has shown promising retrieval cycles had a fresh embryo transfer and a subsequent frozen-thawed
results. However, existing evidence was limited by the quality of the studies and transfer of embryos originating from the same cohort of retrieved oocytes. The
inconsistencies in the reporting formats. clinical pregnancy rate from the fresh embryo transfers in the study group was
Study design, size, duration: A prospective cohort study was conducted 28.9% (508/1758). The clinical pregnancy rate in the first frozen- thawed embryo
from December 2015 till September 2017, recruiting 278 women. transfer was 30.1% (153/508) among patients who achieved a pregnancy in the
Participants/materials, setting, methods: The study included women pre- fresh transfer and 23.9% (299/1250) of those who did not (p=0.007). The clinical
senting with bleeding +/- pain into the early pregnancy assessment unit and had pregnancy rate from the subsequent frozen-thawed transfer was similar if a live
a confirmed fetal heartbeat on the scan from 6+0 to 11+6 weeks of gestation. birth was attained from the fresh embryo transfer 28.9% (69/239) or not 25.2%
Women with uterine anomalies/ myomas, multiple pregnancies, extrauterine (383/1519); p=0.229. The live birth rate from the subsequent frozen-thawed

transfer was similar if a clinical pregnancy or live birth were attained from the O-257 The effect of vaginal microbiota on the outcome of assisted
fresh embryo transfer or not. On a multivariate regression analysis, pregnancy reproductive technologies
in the fresh transfer was a significant independent predictor for a pregnancy in A. Karaer1, B. Doğan2, G. Tuncay3, S. Gunal4, A.N. Tecellioğlu5
the subsequent frozen-thawed embryo transfer p=0.043. Other significant pre- 1
Inonu University School of Medicine, Obstetrics and Gynecology, Malatya, Turkey ;
dictors were maternal age and the treatment protocol. 2
School of Engineering- Inonu University-, Department of Biomedical Engineering,
Limitations, reasons for caution: The limitation of the study is in the ret- Malatya-, Turkey ;
rospective nature of the study 3
Inonu University- School of Medicine, Department of Obstetrics and Gynecology-
Wider implications of the findings: Identifying predictive factors for the Division of Reproductive Endocrinology and Infertility-, Malatya, Turkey ;
success of frozen-thawed embryo transfers is important to maintain acceptable 4
School of Pharmacy- Inonu University, Medical Microbiology, Malatya, Turkey ;
pregnancy rates while reducing multifetal pregnancies. These can help physicians 5
Inonu University, Reproductive Sciences & Advanced Bioinformatics Application &
while counseling patients regarding the number of embryos to transfer taking Research Center, Malatya, Turkey
into consideration the outcome of the previous fresh cycle.
Trial registration number: not applicable Study question: Is there any impact of the vaginal microbiota on the pregnancy
rate in women undergoing assisted reproductive techniques (ART)?
O-256 Conception after early IVF pregnancy loss – Should Summary answer: Although the pregnancy rate is lower in women with
we wait? community site type (CST)-IV and CST-V, there was no significant association
M. Sharon-Weiner1, H. Gluska1, S. Farladansky-Gershenabel1, between CSTs and pregnancy rate.
H. Schreiber1, A. Wiser1, A. Shulman1, A. Hershko-Klement1 What is known already: Bacteria in the human body account for 2-3% of
1
Meir Medical Center, Obstetrics and Gynecology, Kfar Saba, Israel total body mass and urogenital tract accounts for around 9% of the total human
microbiota. In literature, microbiome studies conducted on female reproductive
Study question: Does the interval between early IVF pregnancy loss and the tract usually investigate the obtained vaginal microbiota data under five different
next IVF cycle predict the cycle’s outcome? community state types (CSTs). Where, CST-I, II, III and V forms the groups in
Summary answer: Shorter intervals between early IVF pregnancy loss and which Lactobacillus crispatus, Lactobacillus gasseri, Lactobacillus iners, and
reinitiating IVF treatment are related to the likelihood of live birth in the subse- Lactobacillus jensenii are dominated (relative abundance > 50%), respectively.
quent pregnancy. CST-IV forms the group in which none of the above Lactobacillus species are
What is known already: Approximately 5% of IVF cycles end in early preg- dominated. Growing evidence demonstrates the importance of vaginal micro-
nancy loss, representing 15% of pregnancies resulting from IVF. There is no biota in human reproductive functions.
consensus regarding the optimal interval between a miscarriage and the next Study design, size, duration: A cohort of 223 infertile patients undergoing
attempt to conceive. Short inter-pregnancy intervals (IPIs) are associated with ART in an university based ART center between May 2016 and May 2019 were
poor obstetric outcomes in the general population. Despite the abundance of enrolled in this study.
studies examining IPI in the general obstetric population, there is a paucity of Participants/materials, setting, methods: The vaginal swab were collected
studies specifically evaluating IPI in the IVF population. from posterior vaginal fornix immediately before embryo transfer. Genomic
Study design, size, duration: This retrospective cohort study included 289 DNA was extracted from vaginal samples and sequenced by the V3-V4 region
women who experienced first trimester IVF pregnancy loss, January 2014– of 16S ribosomal RNA (rRNA) gene. The vaginal microbiota profiles were
January 2018 and reinitiation of IVF treatment. assigned to one of the five CSTs based on the dominant bacterial species. The
Participants/materials, setting, methods: Women who experienced biochemical pregnancy rate (positive hCG test result day 14) was investigated.
first-trimester pregnancy loss (including chemical pregnancy and miscarriage) Main results and the role of chance: In total the vaginal microbiota of 223
following IVF were included. Treatments and follow-up occurred in a tertiary, women who underwent a fresh embryo transfer were analyzed. The vaginal micro-
university-affiliated, medical center. Miscarriages were classified as spontaneous/ biata of 123 (55.2%) women dominated by Lactobacillus species, 100 (44.8%)
medical/surgical termination. We calculated intervals between miscarriage and women had bacteria related with bacterial vaginosis predominantly. The overall
the next cycle, interval to the subsequent pregnancy and number of cycles pregnancy rate was 48.4% (108/223 ). The pregnancy rate was 53.8% (21/39)
required. Primary outcome measure was the result of the IVF cycle following in women with CST-I, 66.7% (10/15) in women with CST-II, 50% (32/64) in
the miscarriage. Secondary measure was the interval between miscarriage and women with CST-III, 44% (44/100) in women with CST-IV, and 20% (1/5) in
the next IVF pregnancy. women with CST-V. There was no statistically significant difference between CSTs
Main results and the role of chance: Among 289 women diagnosed with regarding to pregnancy rate. CST-IV was mostly dominated by Gardnerella vaginalis,
first trimester pregnancy loss following an IVF cycle, interval to the subsequent Streptococcus agalactiae and Streptococcus anginosus. A linear discriminant analysis
IVF treatment was not associated with the chance of achieving pregnancy. (LDA) of the vaginal microbiota data concluded that, the Streptococcaceae (family)
Patients after chemical pregnancy or spontaneous miscarriage experienced has the highest contribution (|log10 LDA score| > 4) in the separation of women
shorter intervals to next cycle, as compared with miscarriages managed medically with positive and negative pregnancy test result.
or surgically (Kaplan-Meier survival curve, P=0.01) and higher pregnancy rates Limitations, reasons for caution: The major limitation of sequencing are
(P=0.009). that it does not give information about the viability of the organism and it does
In a multivariable logistic model, type of miscarriage mattered: compared to not provide information about its biological function. These results are limited
chemical pregnancy, odds ratio (OR) for achieving pregnancy after a spontaneous to the studied ART population and do not reflect the microbiota profile of a
miscarriage was 0.7 (P=0.5); OR=0.3 (P=0.002) for a pregnancy after a medical general population.
termination, and OR=0.3 (P=0.001) for a pregnancy following surgical termina- Wider implications of the findings: Our data demonstrated that the abnormal
tion. However, the time interval was not a significant factor (P=0.8). vaginal microbiota is not associated with poor pregnancy rate for ART treatment.
When pregnancy was achieved in the first post-miscarriage cycle, the chance Trial registration number: Not applicable
of a live birth was higher with shorter intervals (median 57.5 days), while another
miscarriage was significantly related to a longer interval (median 82.5 days) O-258 First trimester pregnancy outcomes of subchorionic
between miscarriage and the next IVF cycle (P=0.03). haematomas in a recurrent pregnancy loss population at a tertiary
Limitations, reasons for caution: This was a non-randomized, retrospective London teaching hospital.
study. Time intervals between cycles was not planned or randomized. C. Brooks1, S. Webb1, Y. Mulji1, R. Rai1
Wider implications of the findings: Following miscarriage, the subsequent 1
St Mary’s Hospital Imperial College Healthcare NHS Trust, Obstetrics and
IVF cycle should not be postponed, as shorter intervals are associated with Gynaecology, London, United Kingdom
greater likelihood of a live birth. We cautiously state that IVF cycles following
chemical pregnancy or spontaneous miscarriage, rather than medically or sur- Study question: Do subchorionic haematomas increase the likelihood of mis-
gically managed miscarriage result in higher pregnancy rates. carriage in recurrent pregnancy loss population; are they related to use of aspirin
Trial registration number: Not applicable and low molecular heparin?

Summary answer: Subchorionic haematomas do not increase the likelihood Participants/materials, setting, methods: All women had ³3 consecutive
of miscarriage in the recurrent pregnancy loss population. pregnancy losses and a regular menstrual cycle, while none had significant chro-
What is known already: There is conflicting evidence on the role of subcho- mosomal or uterine abnormalities. Frequencies of different subgroups of p-MBL
rionic haematomas (SCH) on pregnancy outcome and specifically in the recurrent levels in RPL patients were compared to MBL levels in the control group.
pregnancy loss (RPL) population. Some studies report an association with mis- Frequencies of adverse perinatal outcomes between subgroups defined by
carriage and increase of SCH incidence with the use of Aspirin. This is an import- p-MBLlevels were compared. A multivariate analysis was performed identifying
ant finding to confirm as aspirin and low molecular weight heparin (LMWH) are risk factors including low p-MBL level for adverse obstetrical outcomes in the
both established treatment regimens for conditions that can cause recurrent patients.
pregnancy loss. Main results and the role of chance: Significantly more RPL patients had
Study design, size, duration: A retrospective observational cohort study low p-MBL levels (<500 mg/l) (odds ratio [OR] = 2.45, 95% CI 1.61-3.71; p
on all new patients referred to the early pregnancy clinic at St Mary’s Hospital, <0.001) and less had high MBL levels (>3000 mg/l) (OR = 0.51; 95% CI 0.33-
London, was conducted between January 2017 and June 2019. 0.80; p=0.003) compared to controls, while comparing the intermediate p-MBL
Participants/materials, setting, methods: The patients seen in the early levels showed no differences (OR = 0.73, 95% CI 0.49-1.07; p = 0.11). A pre-
pregnancy unit have a history of RPL and are referred by their GP. They are seen vious moderate (>500 ml) or severe peripartum haemorrhage (>1000 ml)
in clinic at 6 weeks gestation and have a transvaginal ultrasound scan performed. (p<0.001 for both) and previous birth of a boy (p<0.001) were associated with
Patient data was collected from both Cerner © electronic records and care- secondary RPL (sRPL). Smoking, increasing age, BMI and number of consecutive
stream PACsweb imaging. miscarriages but not low p-MBL levels were significantly associated with no
Main results and the role of chance: In this pilot study of 226 patients the pregnancy or pregnancy loss after admission. No clear association was observed
median age of the RPL women were 36 (range 24-45) and had an median number between maternal p-MBL levels and birthweight or gestational age, neither
of miscarriages per couple of 4 (range 1-15). There was a miscarriage rate before nor after RPL.
of 28%. Limitations, reasons for caution: Only 93 (37.5 %) women gave birth after
There was a subchorionic haematoma incidence of 17%. RPL in the follow-up period, which were too few to find any clear associations
Comparing the RPL patients on medication with those on no medication, there between p-MBL levels and adverse perinatal outcomes after RPL.
was no increased likelihood of developing a SCH; aspirin alone (OR 1.6 CI 0.55- Wider implications of the findings: While low p-MBL levels are strongly
4.73 p=0.39); LMWH alone (OR 2.1CI 0.66-6.89 p= 0.2) or LMWH and aspirin associated with RPL, high levels may play a protective role. A previous large
combined (OR1.3 CI 0.5-3.37 p=0.5). peripartum haemorrhage, previous delivery of a boy and low maternal p-MBL
Women who were on medication for an identified cause of RPL were less likely may predispose to a large feto-maternal transfer of fetal cells triggering an abnor-
to miscarry than those who were on no medication. (OR 0.38 CI0.19-0.76 mal maternal immunization against fetal or trophoblast cells.
p=0.006). Therefore the likelihood of having a miscarriage is higher in women Trial registration number: NCT04017754
on no medication where no cause was found for the RPL.
Having a SCH did not increase the likelihood of having a miscarriage in this
population (OR 0.65 P 0.31)
Limitations, reasons for caution: It was a retrospective study and therefore SELECTED ORAL COMMUNICATIONS
relies on accurate record keeping. There will be inter-operator bias, as different SESSION 63: PROTECTING GAMETE QUALITY
sonographers scanned during the clinic. 08 July 2020 Parallel 5 10:00 - 11:45
Wider implications of the findings: In the RPL population we can reassure
that having a SCH does not increase the risk of miscarriage in the first trimester.
Aspirin and LMWH do not increase the risk of developing a SCH and therefore
medication should not be stopped if they develop a SCH. O-260 LH prevents follicular damage and preserves the meiotic
Trial registration number: Not Applicable potential of oocytes exposed to chemotherapy at the primordial
stage
O-259 Low mannose binding lectin level in plasma is a risk factor L.M. Castillo1,2, M.J. Soriano1, J. Martinez1,2, A. Pellicer1,3, S.
for recurrent pregnancy loss Herraiz1
C. Nørgaard-Pedersen1, R. Steffensen2, O.B. Christiansen1 1
IVI Foundation - IIS La Fe, Reproductive Medicine, Valencia, Spain ;
2
1
Aalborg University hospital, Obstetrics and gynaecology, Aalborg, Denmark ; University of Valencia, Department of Pediatrics- Obstetrics and Gynecology,
2
Aalborg University Hospital, Bioanalytic department, Aalborg, Denmark Valencia, Spain ;
3
IVI-RMA Rome, Reproductive Medicine, Rome, Italy
Study question: Are low or high plasma mannose binding lectin (p-MBL) levels
associated with recurrent pregnancy loss (RPL) and the perinatal outcome before Study question: Does luteinizing hormone (LH) treatment protect follicle
and after RPL diagnosis? viability and meiotic potential of chemotherapy-exposed primordial follicles?
Summary answer: While low p-MBL levels are significantly more frequent in Summary answer: LH improves the meiotic potential of murine metaphase
RPL patients, high levels are significantly less frequent. No association with II oocytes (MII-oocytes), avoiding the alkylating agents´ gonadotoxic effects, by
adverse perinatal outcomes was found. the promotion of follicular DNA repair mechanisms.
What is known already: Low p-MBL levels have been associated with What is known already: High-dose chemotherapy with alkylating drugs
RPL, while relations to high levels have been poorly studied. Reports con- induces detrimental changes on ovaries. Follicle viability is severely affected by
cerning association between RPL and perinatal outcomes including birth DNA damage and apoptosis of oocytes and granulosa cells (GCs), leading to
weight and gestational age are conflicting. Some but not all studies suggest impaired follicular development and depletion. Early activation of DNA repair
an association between low maternal p-MBL levels and reduced birthweight mechanisms, like homologous recombination through the ataxia-telangiecta-
and gestational age, which could be an important factor for the higher fre- sia-mutated (ATM) pathway, is crucial for cell survival after cytotoxic events.
quency of preterm birth and lower birth weight often reported in pregnancies Previous results suggested LH treatment as an alternative for fertility preservation
after RPL. based on its protective role on the ovarian reserve against chemotherapy.
Study design, size, duration: A combined case-control and cohort study in Therefore, we aimed to assess the follicular protective mechanisms of LH and
248 RPL women admitted to the RPL Centre of Western Denmark from January the meiotic potential of chemotherapy-exposed MII-oocytes in a mouse model.
2016 to December 2019. At first consultation blood samples including p-MBL Study design, size, duration: Experimental study where twenty-four 7-week
measurement and perinatal data from previous births were collected. Women old CD-1 female mice were exposed to three experimental conditions (n=8/
were followed until birth or end of study. P-MBL levels were compared to those group): Control, chemotherapy (ChT) and ChT+LH. The ChT-treated groups
of 185 female blood donors, while perinatal outcomes were compared to all were intraperitoneally injected with 12mg/Kg-busulfan and 120mg/kg-cyclo-
3354 singleton births at our hospital in 2017. phosphamide. The LH-treated animals received a pre-treatment dose with 1IU,

24 hours before ChT, followed by a second 1IU-dose administered with che- Study question: What is de fertilization capacity of In Vitro Matured Oocytes
motherapy. Control-mice received saline. Ovaries from 6 animals/group were found during oVArian tissues preparation(IViMOVA), subsequent embryo devel-
collected at 12 and 24 hours, while the remaining mice were maintained for opment and are these embryos euploid or do they harbor chromosomal
30 days. abnormalities?
Participants/materials, setting, methods: The ATM-pathway, by Rad51 Summary answer: IViMOVA oocytes show lower fertilization rates, impaired
gene expression, was evaluated by RT-qPCR, while apoptotic (cleaved caspase-3) embryo development characterized by irregular cleavage patterns and early
and anti-apoptotic (Bcl2) proteins were quantified by western-blot, on ovarian embryo arrest apparently not related to chromosomal abnormalities.
samples at 12 and 24h. Additionally, the 12h samples were screened for follicle What is known already: The collection of COCs during preparation of OT
DNA damage and apoptosis by γH2AX-staining and TUNEL-assay, respectively. has been described as the maximization of fertility preservation. This technique
The remaining animals were superovulated (10IU-PMSG + 10IU-hCG, 18 hours is sometimes mistaken with IVM in Polycystic Ovary Syndrome(PCOS) patients
later) for MII-oocyte collection. Thus, spindle formation and chromosome dis- where mild stimulation or primed ovarian punction show maturation rates of
position, referred to equatorial plate, were analyzed by confocal microscopy. 70% and successful treatment outcomes. On the contrary, IViMOVA studies
Main results and the role of chance: LH treatment increased the expression in literature show an average maturation rate of around 30% and the descrip-
of the DNA repair gene Rad51 at the 12h (Control: 1, ChT: 2.0±0.5; ChT+LH: tion of the developmental capacity of these oocytes is largely absent. IViMOVA
2.7±1.3; p=0.020 and p=0.019, respectively) and 24h timepoints (Control: 1, reports in literature are mostly case reports and to our knowledge, so far 3
ChT: 0.5±0.5, ChT+LH: 1.4±0.3). The activation of the DNA repair signalling live births have been described (Segers I.,2015, Uzelac P.,2015, Prasath
led to a rise of Bcl2/cleaved caspase-3 protein ratio, decreased by chemotherapy, EB.,2014).
enhancing cell survival on 24h-ovaries (Control: 2.5±0.9, ChT: 0.8±0.3, Study design, size, duration: During OT preparation, COCs were collected
ChT+LH: 2.0±1.3). from surplus medulla, in vitro matured (MediCult IVM medium,Origio) for
Furthermore, LH treatment reduced the significant increase in the number of 44-48h and MII were vitrified(Irvine Scientific). Warmed intact MII were injected
γH2AX-positive oocytes induced by chemotherapy (Control: 19.6±0.8%, ChT: after 2h and cultured in sequential medium(Cook) in the Embryoscope®(Vitrolife).
64.6±3.1%, ChT+LH: 42.6±4.1%; p=0.034 and p=0.021, respectively). For genetic analysis, cells were lysed and DNA was amplified using the Sureplex
Moreover, these positive effects were also observed in the GC integrity, by Amplification system(Illumina), followed by shallow whole genome sequencing
reducing the amount of follicles with >20% of TUNEL-positive GCs (Control: (sWGS) (average genome coverage of 0.1-1x), for the detection of aneuploidy
1.6±0.8%, ChT: 8.5±0.8%, ChT+LH: 5.1±0.9%; p=0.034 and p=0.043), during or subchromosomal aberrations (resolution ~5Mb).
the first 12 hours after treatment. Participants/materials, setting, methods: From Nov.2015-Jan.2019, in
The meiotic potential, referring to MII-oocytes derived from follicles at the total 93 persons (median age 20.7y) underwent hysterectomy with bilateral
primordial stage during chemotherapy administration, was seriously affected in oophorectomy in the context of gender affirming surgery after a median duration
the ChT group, with a 34.5% decrease in spindle area (Control: 127.7±20.5µm2, of testosterone treatment of 75.3weeks. At the time of hysterectomy serum
ChT: 83.7±7.7µm2; p=0.019). Nevertheless, LH treatment was able to avoid was analyzed for AMH-FSH-LH-E2-Progesterone-Testosterone-SHBG. A neg-
this effect, preserving control-like values (ChT+LH: 119.7±2.7µm2; p=0.034). ative binomial regression model for counts was applied (SPSS) to find biochemical
Furthermore, LH diminished the number of MII-oocytes with at least one mis- parameters associated with MII vitrified. Frozen thawed sperm form 1 donor
aligned chromosome compared to ChT group (Control: 12.5%, ChT: 83.3%, (30y, proven fertile) was used for all ICSI procedures.
ChT+LHx1: 58.3%). Main results and the role of chance: For the developmental capacity study,
Limitations, reasons for caution: Although these findings represent the 1765 COCs of 80 patients were in vitro matured in 5%CO2, 6%O2 at 37°C
first steps of a new strategy for fertility preservation in cancer patients, this is which resulted in 390MII oocytes (22%maturation rate) being vitrified. After
an animal model study developed in mouse ovarian samples. Therefore, these correction of demographic variables (duration of testosterone treatment, age
results should be validated in order to properly identify the repairing mechanisms and BMI), AMH was significantly (P=0.0058) associated with the number of MII
in a preclinical approach with human samples. vitrified. 238MII oocytes were donated for this study of which 208 were warmed
Wider implications of the findings: LH treatment minimizes the deleterious and 151 (72.6%) intact. ICSI was performed in 92%(139/151) of the intact
effects induced by alkylating agents on follicular viability. The enhancement of warmed oocytes. After ICSI, 35% of the oocytes showed normal fertilisation
DNA repair systems seems to be one of the main protective mechanisms pro- (48/139); 12%(17/139) were 1pn, 19%(27/139) were 0pn and 15%(21/139)
moted by LH. This improvement would contribute to produce MII-oocytes with were 3pn or more. After ICSI, 19%(26/139) were degenerated. Only 2pn were
increased potential to properly complete the meiosis II. further cultured and embryo development was monitored. On day 3, 52% of
Trial registration number: not applicable the 2pn (25/48) developed up the cleavage stage of which 44%(11/25) were 6
cell or more. Out of 139 injected oocytes and 48 normally fertilized, only 1 (2%)
blastocyst (4BB) was obtained on day 5. Timelapse imaging showed direct or
O-261 The developmental capacity of In Vitro Matured Oocytes reversed cleavage, no extrusion of the 2nd polar body and arrest in the cleavage
originating from cumulus oocyte complexes (COCs) found during stage. For genetic analysis, 32 embryos were available and was successful in
oVArian tissues (OT) preparation is compromised 30/32 embryos. Interpretable results showed normal chromosomal patterns in
K. Tilleman1, S. Lierman2, A. Tolpe3, I. De Croo4, S. De Gheselle5, 43% (13/30) of the embryos.
J. Defreyne6, G. T’Sjoen6, M. Baetens7, A. Dheedene7, B. Menten7, Limitations, reasons for caution: This descriptive cohort study used
P. De Sutter8 tissue from transgender patients undergoing men with testosterone treat-
1
Ghent University Hospital, Department for Reproductive Medicine- Ghent Fertility ment, hence it is possible that this could affect the developmental capacity
And Stem cell Team G-FAST, Gent, Belgium ; of the oocytes although previous work from our group has shown that the
2
Ghent university Hospital, Department for Reproductive Medicine- Ghent Fertility oocytes look morphologically normal and display normal spindles (Lierman
And Stem cell Team G-FAST, Gent, Belgium ; S., 2017).
3
Ghent University Hospital, Department for Reproductive Medicine- Ghent Fertility Wider implications of the findings: IViMOVA should be considered an
And Stem cell Team G-FAST, Ghent, Belgium ; innovative technique (Provoost V. 2014). As this technique is often described as
4
Ghent University Hospital, Department for Reproductive Medicine- Ghent Fertility a maximization of fertility preservation for oncology patients, information should
And Stem cell Team G-FAST, Ghent, Belgium ; be given on the uncertainty concerning the developmental capacities of these
5
Ghent University Hospital, Department for Reproductive Medicine- Ghent Fertility oocytes and the lack of data in scientific literature.
And Stem cell Team G-FAST, Ghent, Belgium ; Trial registration number: This study was funded by FWO TBM T001016N
6
Ghent University Hospital, Department of Endocrinology- Center for Sexology and and was approved by the Ghent University Hospital Ethical Committee (UZ
Gender, Ghent, Belgium ; Ghent Reference: 2015/0124, Belgian registration number B670201523543)
7
Ghent University Hospital, Center for Medical Genetics, Ghent, Belgium ; and the injection of the in vitro matured oocytes and creation of embryos for
8
Ghent University Hospital, Department for Reproductive Medicine- Ghent Fertility this study was approved by the Belgian Federal Ethics Committee (reference
And Stem cell Team G-FAST, Ghent, Belgium Adv_056).

O-262 Effects of cancer on biology of granulosa cells, follicular Summary answer: Cisplatin exposure causes acute loss of germ cells in the imma-
fluid and quality of oocytes ture human testis and this effect was not rescued by short-term exposure to GCSF.
G. Younes1, W. Buckett1, Q. Yang2, W.Y. Son1, A. Volodarsky- What is known already: Long-term survival rates for children with cancer
perel1, T. Tulandi1, H.J. Clarke2 are more than 80%. However, childhood cancer treatment may result in subse-
1 quent infertility. Cisplatin is one of the most commonly used drugs for childhood
Mcgill University Health Centre, reproduction centre, montreal, Canada ;
2 cancers. Knowledge of the effects of Cisplatin exposure in childhood is still
Mcgill University Health Centre, research institute, montreal, Canada
limited. Moreover, there are currently no clinical agents to protect spermatogo-
Study question: Do factors associated with cancer damage the granulosa cells nial stem cells from chemotherapy-induced damage. Animal studies suggest that
of the ovarian follicle or alter the follicular fluid, leading to abnormal development GCSF may protect germ cells in males and females, however, there is no infor-
of the egg? mation whether GCSF could prevent loss of germ cells in human testes.
Summary answer: Inhibitors of Wnt signaling pathway and regulators of cell Study design, size, duration: In-vitro culture of human fetal testis tissues was
growth and differentiation, cell-cell interactions and adhesion are downregulated utilised as an established model for pre-pubertal testis development. Tissues
in cancer patients. were exposed to: 1) 0.5 μg/ml Cisplatin or vehicle control on day 4 for 24hrs
What is known already: The adverse effects of chemotherapy and radio- and kept in culture until 72 and 240 hours post-exposure; 2) combined treatment
therapy on fertility are well known. Cryopreservation of oocytes and embryos of Cisplatin with GCSF (5 or 20 ng/ml) for first 7 days (3 days prior, during and
are considered the standard of care for preserving fertility in reproduc- 3 days after Cisplatin exposure).
tive-age cancer patients before starting treatments. Participants/materials, setting, methods: Second trimester human fetal
Whether the malignancy itself has a negative effect on the reproductive system testis tissue fragments (14-22 gestational weeks; n=3-14) were cultured in a
remains unclear. Some clinical studies have shown apparent adverse effects of ‘hanging drop’ system. Tissue was cultured for 7 and 14 days in total which
malignant disease on the response to ovarian stimulation and on the quality and reflected 72 and 240 hours post-exposure to Cisplatin.
performance of oocytes, whereas others have shown no difference when com- Quantification of germ cell populations (cells per tubular area (mm2)) was
pared to healthy controls. performed on sections stained for AP2γ (gonocytes) and MAGE-A4 (pre-sper-
Study design, size, duration: This prospective pilot study included 20 matogonia). Statistical analysis was performed using two-way ANOVA to
patients treated at our academic reproductive centre. Patients could be divided account for inter-individual variation between fetuses.
into two groups: a study group that consisted of 10 cancer patients treated for Main results and the role of chance: Cisplatin exposure significantly
fertility preservation before chemotherapy or radiotherapy and a control group reduced the number of gonocytes (478.7±85.30 vs 379.7±92.29 cells/mm²;
of age-matched 10 healthy fertile patients treated with IVF due to male factor p<0.01) at 72 hours post-exposure compared with vehicle-exposed controls,
or social fertility preservation. Research and Ethics Board of McGill University whereas the number of pre-spermatogonia was unchanged. After 240 hours,
Health Centre approved the study. exposure to Cisplatin resulted in significant reduction in both gonocyte
Participants/materials, setting, methods: Cumulus cells and follicular fluid (327.1±45.13 vs 193.5±37.94 cells/mm²; <0.0001) and pre-spermatogonial
were collected from all patients. Total RNA of ten samples was extracted from the (393.0±36.73 vs 324.0±60.45 cells/mm²; p<0.01) numbers.
cumulus cells. We evaluated mRNA expression profiles of cumulus cells using next In combined treatment experiments, addition of GCSF from 3 days prior to
generation sequencing by Illumina Novaseq 6000. The follicular fluid of all twenty 3 days after Cisplatin exposure did not protect against the reduction of germ
samples was tested by automatic immunoassay for C -reactive protein (CRP) as an cell numbers 72 or 240 hours post-Cisplatin exposure.
inflammatory marker, cortisol as a stress marker, and insulin like growth factor. Limitations, reasons for caution: Human pre-pubertal testis tissue is of
Main results and the role of chance: All patients were treated with the antag- limited availability, thus, a validated in-vitro system using human fetal testis was
onist protocol. The number of total oocytes retrieved, and the number of mature utilised. These tissues contain similar germ cell sub-populations, which includes
or immature oocytes in the 2 groups were comparable. Three samples from the gonocytes (present in infancy) and pre-spermatogonia (present throughout
study group and three others from the control group were further analyzed by pre-puberty). ‘Hanging drop’ culture might not recapitulate all in-vivo aspects of
mRNA sequencing. Compared with controls, 85 known mRNAs were differentially immature testis microenvironment.
expressed in cancer patients’ cumulus cells, including 45 mRNAs upregulated and Wider implications of the findings: This study shows that Cisplatin expo-
40 mRNAs downregulated. The potential regulating roles of mRNA and gene sure causes early loss of gonocytes in the immature human testis, whereas the
annotation showed downregulation of genes that act as regulators of cell growth effect on (pre)spermatogonial number is delayed. This germ cell reduction could
and differentiation, cell-cell interaction and adhesion (GPC4, ID3, CGC20, IGSF-9, not be protected by concurrent GCSF treatment. Alternative regimens of GCSF
CPXM1). We also found that two strongly downregulated genes are inhibitors of supplementation are currently under investigation and will be discussed.
the canonical Wnt signaling pathway (SFRP4, WIF1). The concentrations of CRP, Trial registration number: Not applicable
cortisol, or insulin like growth factor in the two groups were comparable.
Limitations, reasons for caution: This pilot study has a small sample size. O-264 In vitro fragmentation of ovarian tissue activates
Two patients were treated with letrozole which may have an effect on the results. primordial follicles through the Hippo pathway
Wider implications of the findings: Our data suggest that cellular regulators C. De Roo1, S. Lierman1, K. Tilleman1, P. De Sutter1
and inhibitors of the Wnt signaling pathway may be downregulated in cancer. 1
University Hospital Ghent UZ, Reproductive Medicine- Department of
These changes may have negative effects on cumulus oocyte expansion, ovu- Gynaecology, Ghent, Belgium
lation, and luteinization.
Our results provide novel candidates for molecular targets in the research of Study question: What is the role of the Hippo and PI3K/Akt pathway in
the effect of cancer on female fertility. follicles during ovarian tissue culture in tissue derived from oncological patients
Trial registration number: none and transgender men?
Summary answer: Results highlight a Hippo pathway driven primordial follicle
activation in vitro, predominantly from day 0 to day 4
O-263 Pre-treatment with Granulocyte-Colony Stimulating What is known already: In vitro OT culture aims at activating and maturing
Factor does not protect germ cell populations in the immature primordial follicles for fertility restoration in patients with a threatened ovarian
human testis from Cisplatin-induced germ cell loss reserve. Not all patients are eligible for ovarian cortex transplantation and there-
G. Matilionyte1, R.A. Anderson1, R.T. Mitchell1 fore several groups attempt to culture ovarian tissue in vitro.
1
The University of Edinburgh, Centre for Reproductive Health, Edinburgh, United Cortex fragmentation disrupts the Hippo pathway, leading to increased
Kingdom expression of downstream growth factors and follicle growth. The PI3K/Akt
pathway is considered the intracellular pathway to where different extracellular
Study question: Does Cisplatin affect germ cell populations in the immature factors involved in primordial follicle activation in vivo converge to. In order to
testis and can this effect be rescued by treatment with Granulocyte-Colony optimize current ovarian tissue culture models, information on progression of
Stimulating Factor (GCSF)? these pathways during tissue culture is mandatory.

Study design, size, duration: The first step of a multistep cortex culture shown complete elimination of rhabdomyosarcoma cells by efficient ex vivo
system was performed using 144 ovarian cortex pieces of in total 6 patients. Per purging of ovarian cortex fragments is possible, allowing treatment of all cortex
patient, 24 cortical strips were cultured for 6 days and 6 pieces per patient were fragments without compromising ovarian tissue integrity.
collected for downstream analysis of follicle development and Hippo and PI3K/ Study design, size, duration: Human ovarian cortex tissue with experimen-
Akt pathway targets every second day. tally induced tumour foci of CML and AML were exposed to a 24 h treatment
Participants/materials, setting, methods: OT was obtained from onco- with an AURKB/C inhibitor to eliminate malignant cells by invoking mitotic
logical (N=3; 28.67+/-4.51years) and transgender (N=3; 23.33+/-1.53 ears) catastrophe. After treatment the inhibitor was removed followed by an additional
patients. Follicles were analysed using hematoxylin-eosin staining and pathways culture period of 6 days to allow any remaining tumour cells to form new foci.
were studied using immunohistochemistry and precise follicle excision by laser Ovarian tissue integrity was analysed by 4 different assays. Appropriate controls
capture microdissection for RT-qPCR analysis. MIQE guidelines for RT-qPCR were included in all experiments.
were pursued. Reference gene selection (GAPDH, RPL3A, 18s rRNA) was Participants/materials, setting, methods: Tumour foci of CML and
performed using GeNorm Reference Gene Selection Kit. Statistical analysis was AML cell lines were induced in ovarian cortex from transgender men under-
conducted with IBM SPSS Statistics 23 (Poisson regression, negative binomial going oophorectomy. Presence of any remaining cancer cells after ex vivo
regression, ANOVA and paired t-test). treatment was analysed by (immuno)histochemistry of serial sections from
Main results and the role of chance: Immunohistochemical analysis con- entire tissue fragments. Effect of the AURKB/C inhibitor exposure on the
firmed a Hippo pathway driven primordial follicle activation by mechanical manip- viability of ovarian cortex tissue and follicles was determined by histology,
ulation of the cortical strips. Ovarian tissue preparation and culture induced the glucose uptake assay, follicular viability assay and an assay for the in vitro growth
inhibitory pYAP to disappear in granulosa cells of primordial follicles on day 2. of small follicles.
The stimulatory YAP on the contrary appeared in primordial granulosa cells over Main results and the role of chance: Foci of metastasized CML and AML
increasing culture days. Looking at the YAP target CTGF, a significantly up reg- cells in ovarian cortex tissue were severely affected by a 24 h ex vivo treatment
ulated CTGF was noted in primordial follicles when comparing day 2 and day 4 with an AURKB/C inhibitor, leading to the formation of multi-nuclear syncytia
(ratio day 2/4 = 0.082; p<0.05), clearly showing an effect on the Hippo pathway and large scale apoptosis. Only the megakaryocytic CML cell line MEG-01, known
in primordial follicles during tissue culture. for its aberrant AURKB expression, was not affected. Ovarian tissue morphology
Follicle classification objectified a significant drop in estimated primordial follicle and viability was not compromised by the treatment, as no significant difference
counts in the oncological cohort (-78%; p=0.021) on day 2 and in transgender was observed regarding the percentage of morphologically normal follicles, fol-
cohort on day 4 (-634%; p=0.008). Intermediate follicle counts showed a non-sig- licular viability, glucose uptake or in vitro growth of small follicles between the
nificant trend to increase during culture and this follicle recruitment and growth ovarian cortex treated with the AURKB/C inhibitor and the control.
resulted in a significant raise in estimated primary follicle counts on day 6 in onco- Limitations, reasons for caution: The persistence of CML and AML met-
logical patients (170%; p=0.025) and although limited in absolute numbers, a sig- abolically-active multinuclear syncytia after treatment precludes the use of
nificant increase in secondary follicles on day 4 (367%; p=0.021) in the transgender molecular techniques to verify the complete absence of viable tumour cells. The
cohort. Subsequent antral follicle development could not be observed. functional integrity of the ovarian cortex tissue after ex vivo treatment requires
Limitations, reasons for caution: A limitation is the small sample size, inher- further investigation in vivo.
ent to this study subject, especially as a large amount of tissue was needed per Wider implications of the findings: Purging of CML/AML metastases in
patient to reduce inter patient variation in different downstream analysis tech- ovarian cortex is possible by targeting the Mitotic Catastrophe Signalling Pathway
niques. A particular and specific weakness of this study is the inability to include using an AURKB/C inhibitor without harming ovarian tissue. This provides a
an age-matched control group. therapeutic strategy to prevent reintroduction of leukaemia and enhances safety
Wider implications of the findings: These findings support an adapted of autotransplantation in leukaemia patients currently considered at high risk for
tissue preparation for Hippo pathway disruption and a shorter first phase of ovarian involvement.
tissue culture. This work may also have a reflection on transplantation of cryo- Trial registration number: not applicable
preserved tissue as larger strips (and thus slower burnout because of less Hippo
pathway disruption) could be a benefit. O-266 The role of F-actin and MYO10 distribution in quality
Trial registration number: This research was financially supported by the assessment of fresh and frozen-thawed human growing follicles
Foundation Against Cancer (Stichting tegen Kanker) and Flemish Foundation of S. Granados Aparici1, T. Tulandi1, W. Buckett1, W.Y. Son1, G.
Scientific Research (FWO Vlaanderen), Belgium Younes1, J.T. Chung1, S. Jin1, H. Clarke1, A. Volodarsky-Perel1
1
McGill University Health Centre, Obstetrics and Gynecology, Montreal, Canada
O-265 Purging human ovarian cortex of contaminating leukaemic
cells by targeting the Mitotic Catastrophe Signalling Pathway Study question: Does the distribution of F-actin and MYO10 protein differ in
L. Eijkenboom1, C.L. Mulder2, B.A. Van der Reijden3, N.M. Van human preantral follicles from fresh and frozen human ovarian tissue?
Mello4, J. Van Leersum1, D.D.M. Braat1, C.C.M. Beerendonk1, Summary answer: Cryopreservation and the freezing method have an impact
R. Peek1 on the distribution pattern of F-actin and MYO10 protein in granulosa cells and
1
Radboudumc, Obstetrics and Gynaecology, Nijmegen, The Netherlands ; oocytes.
2
Amsterdam UMC, Reproductive Biology, Amsterdam, The Netherlands ; What is known already: Cryopreservation of ovarian tissue has emerged as
3
Radboudumc, Laboratory Medicine, Nijmegen, The Netherlands ; an option for fertility preservation for oncological patients. Slow freezing and
4
Amsterdam UMC, Obstetrics and Gynaecology, Amsterdam, The Netherlands vitrification, the two major methods of ovarian tissue cryopreservation, may
disturb essential communication between oocyte and granulosa cells. Granulosa
Study question: Is it possible to eliminate metastasized chronic myeloid leu- cells communicate with the oocyte via specialized actin-rich filopodia termed
kaemia (CML) and acute myeloid leukaemia (AML) from ovarian cortex by inhi- transzonal projections (TZPs) that enable the oocyte to be supplied with key
bition of Aurora B/C kinases (AURKB/C)? nutrients and regulatory signals. MYO10 protein regulates filopodial growth and/
Summary answer: Inhibition of AURKB/C ex vivo effectively eradicated or function and may work similarly in TZPs. However, the effects of freeze-
experimentally induced CML and AML tumour foci and viability of ovarian tissue thawing on the expression and distribution of MYO10 and F-actin in TZPs remain
was not compromised by this treatment. unclear.
What is known already: Despite the success of ovarian tissue cryopreser- Study design, size, duration: Frozen ovarian tissue was donated by patients
vation, autotransplantation is not without risk since malignant cells may be pres- who underwent ovarian tissue cryopreservation due to malignant disease. Fresh
ent in the graft. Safety procedures to detect minimal disseminated disease render ovarian tissue was prospectively collected from patients who underwent ovarian
the fragment useless for autotransplantation and remaining fragments could still surgery during the period from 2017 to 2019. Only patients with normal ovaries
harbour malignant cells. Strategies to separate follicles from possibly contami- were included in this analysis. Ovarian follicles at primordial to secondary stages
nated stromal cell compartment followed by in vitro maturation or grafting to an were retrieved from patients who donated fresh ovarian tissue (n=4), underwent
artificial ovary are actively pursued, but are experimental. We have recently vitrification (n=3), and slow freezing (n=5) of ovarian tissue.

Participants/materials, setting, methods: Fresh and thawed samples were spermatogenesis genes was measured by qRT-PCR. The presence of DNA
immediately delivered for follicle harvesting. Follicles comprising a morphologi- strand breaks was detected by gamma-H2AX and cell proliferation by PCNA.
cally normal oocyte and non-pyknotic granulosa cells were stained using phal- Sperm cells were isolated from the epididymis, counted and analyzed for motility
loidin and anti-MYO10. Confocal microscopy was used to image equatorial and morphology.
optical sections of each follicle. Parameters measured were the number of Main results and the role of chance: TERT expression and activity in the
MYO10 foci and intensity of F-actin in the granulosa cells and oocytes. The mouse testis increased following a single AGS dose. The expression of sper-
correlation between the oocyte diameter and the structures associated with matogenesis markers (VASA and CREM) increased followed by a significant
TZP development (MYO10 foci and F-actin) was evaluated. enhancement in the sperm count. Exposure of mice to X-ray radiation (2.5 Gy)
Main results and the role of chance: A total of 180 good-quality follicles damaged the testis tissue and decreased sperm count and sperm parameters
with a mean oocyte diameter of 36.6±9.1 µm from women of 22-37 years of (motility, normal morphology). Treatment of the X-ray irradiated mice with a
age were included in the analysis. A positive correlation between oocyte diam- single dose of AGS, significantly restored testis tissue morphology, the expression
eter and MYO10 distribution in granulosa cells (correlation estimate (CE) 0.63; of spermatogenesis markers and decreased the number of seminiferous tubules
95% confidence interval (95%CI) 0.43-0.77; P=0.001) and in oocytes (CE 0.43; that exhibited DNA strand breaks. AGS treatment of irradiated mice restored
95%CI 0.19-0.63; P=0.001) was demonstrated in fresh follicles. In frozen-thawed the sperm count and sperm motility and reduced the percentage of sperm cells
follicles, a correlation was observed for MYO10 distribution in granulosa cells with altered morphology. A higher sperm count and an increase in testis tissue
(CE 0.22; 95%CI 0.04-0.38; P=0.02) but not in oocytes. When vitrified follicles regeneration, following X- ray radiation, was also detected in AGS treated mice
were compared to those subjected to slow freezing, the correlation between 21 and 30 days after irradiation and treatment.
oocyte diameter and MYO10 distribution in granulosa cells was revealed only Limitations, reasons for caution: We could not use an X- ray dose that
in the slow freezing group (CE 0.38; 95%CI 0.17-0.55; P=0.0005). Strikingly, completely abolished sperm production. Therefore, under these conditions no
large MYO10 aggregates within the oocyte were significantly more common significant differences in the ability to produce offspring, between X-ray irradiated
after slow freezing compared to vitrification [25(30.5%) vs. 4(9.1%); P=0.007]. and X- ray irradiated + AGS treated mice, were observed
Correlation between oocyte diameter and F-actin intensity was observed in the Wider implications of the findings: This study suggests the ability of telo-
slow freezing group (CE 0.31; 95%CI 0.09-0.49; P=0.006) but not in the vitrified merase increasing compounds to restore the normal tissue morphology, the
follicles. The different patterns of MYO10 and F-actin distribution in growing spermatogenesis process and the sperm count in damaged testes. Therefore,
fresh and thawed follicles suggests a significant impact of cryopreservation and these compounds may be used as possible therapy in conditions that reduced
method of freezing on the structures mediating physical communication between male fertility.
oocyte and granulosa cells. Trial registration number: NA
Limitations, reasons for caution: Only follicles from patients without ovar-
ian pathology were included. The effect of ovarian pathology on the structures
associated with TZP development in fresh and frozen-thawed ovarian tissue is
a matter for further studies. SELECTED ORAL COMMUNICATIONS
Wider implications of the findings: Our study suggests that MYO10 dis- SESSION 64: PROSPECTIVE CARRIER SCREENING OF ART
tribution and F-actin intensity in granulosa cells as well as the presence of MYO10 COUPLES
aggregates in oocytes can be used for quality assessment of growing follicles in 08 July 2020 Parallel 6 10:00 - 11:45
fresh and frozen-thawed ovarian tissue.
O-267 Protection of the mouse testis tissue and sperm production O-268 Insights from the largest study on the genetics of sporadic
from X-ray induced damages by pharmaceutical compounds that and recurrent miscarriage
increase telomerase T. Laisk1
H. Elul1, E. Priel1 1
Institute of Genomics- University of Tartu, Estonian Genome Center, Tartu,
1
Ben-Gurion University of the Negev, The Shraga Segal Dept. Immunology- Estonia
Microbiology & Genetics, Beer-Sheva, Israel Study question: Can we discover and map the maternal genetic susceptibility
and underlying biology of sporadic and recurrent miscarriage using large-scale
Study question: Do pharmaceutical compounds (AGS) that increase telomer- biobank data?
ase protect the mouse testes from damages induced by X-ray? Summary answer: Our results confirm miscarriage as a complex partly her-
Summary answer: A single AGS treatment increased telomerase in the mouse itable phenotype and implicate novel biology through regulation of genes
testes and protected the testis and sperm production from the damages induced involved in placental biology.
by X-ray radiation. What is known already: Miscarriage is a common complex trait that affects
What is known already: The telomerase reverse transcriptase (TERT) is 10-25% of all clinically confirmed pregnancies. Miscarriage is associated with
expressed and active in the testes and is important for the spermatogenesis excessive bleeding, infection, depression, infertility and an increased lifetime risk
process. Short telomeres were identified in men with oligozoospermia and of cardiovascular disease. A variety of predisposing factors have been associated
idiopathic infertility. Shortened telomeres in spermatozoa are markers for abnor- with increased miscarriage risk, and it has a genetic component that thus far has
mal spermatogenesis. In addition to its role in the re-elongation of telomeres, remained poorly characterized.
TERT possesses non-canonical functions: protects cells from apoptosis, partic- Study design, size, duration: Genome wide association study (GWAS)
ipates in the repair of DNA strand breaks and regulates the expression of genes. meta-analysis of up to 69,118 sporadic miscarriage cases from seven different
We synthesized novel compounds that transiently increased TERT expression ancestries, 750 recurrent miscarriage cases of European ancestry, and up to
and activity in various human and mouse cells and tissues. The compounds 359,469 female controls from biobanks all over the world.
protected cells from damages induced by oxidative stress. Participants/materials, setting, methods: We investigated the genetic
Study design, size, duration: Adult mice were divided to various treatment architecture of miscarriage with biobank-scale GWAS, Mendelian randomization,
groups (>10 mice per group): with/ without exposure to X-ray radiation and heritability, genetic correlation and functional annotation analyses.
with/without AGS treatment or vehicle treatment. At various intervals, post Main results and the role of chance: We identify one genome-wide signif-
irradiation and treatment, the mice were sacrificed and their testes including the icant association (rs146350366, minor allele frequency (MAF) 1.2%, P=3.2×10-8,
epididymis were removed and subjected to further analysis. odds ratio (OR)=1.4) for sporadic miscarriage in our European ancestry
Participants/materials, setting, methods: Testis slices were stained with meta-analysis and three genome-wide significant associations for recurrent mis-
hematoxylin- eosin for tissue morphology. To identify the various cell types in carriage (rs7859844, MAF=6.4%, P=1.3×10-8, OR=1.7; rs143445068,
the seminiferous tubules we used specific markers (VASA, CREM, acrosin) apply- MAF=0.8%, P=5.2×10-9, OR=3.4; rs183453668, MAF=0.5%, P=2.8×10-8,
ing the immunohistochemistry procedures. The expression of TERT and specific OR=3.8). Following functional annotation linked these associations with genes

related to placental biology. We found a heritability of 29% (95%CI 20%-38%) Atrophy (SMA) (n=3, 0.5%), Fragile-X Syndrome (n=3, 0.5%), Smith-Lemli-Opitz
for miscarriage. Consistent with observational associations, we found significant Syndrome (n=1, 0.2%) and Duchenne/Becker Dystrophy (n=1, 0.2%). Notably,
genetic correlations between sporadic miscarriage and number of children 3 of these ARCs were identified among couples trying to conceive by natural
(p=7.210-9). The Mendelian randomization analyses suggest that smoking may conception. Moreover, the identification of a patient carrying a pathogenic variant
causally increase the risk of sporadic miscarriage. Finally, our analysis of health on the SMN1 gene led to cascade testing of his relatives, revealing another ARC
outcomes associated with miscarriage confirms previously observed observa- for SMA in the family. At present, among ARCs that were followed up, all of
tions and identifies several novel ones. them (N=10) pursued actions to reduce their reproductive risk by undergoing
Limitations, reasons for caution: Only maternal genetic data was analysed. IVF with PGT for monogenic conditions (PGT-M). To date, 4/10 (40.0%) couples
Further studies are needed to assess the functional impact of the associated variants. completed their PGT-M cycle with euploid/healthy embryos. Two of these
Wider implications of the findings: Our study shows the potential involve- couples have already achieved a healthy pregnancy after embryo transfer.
ment of genes regulating placental function in the etiopathogenesis of miscarriage Limitations, reasons for caution: The main limitation of this study involves
and highlights the utility of large population-based biobank data for understanding the lack of ethnic diversity in our cohort (mainly Caucasians). Also, the use of
these understudied pregnancy complications. This research is presented on a limited panel of core gene-disease pairs can underestimate the rate of carriers
behalf of the International Miscarriage Genetics Consortium. and ARCs detectable in this cohort of individuals.
Trial registration number: NA Wider implications of the findings: Extensive application of ECS detects
a remarkable proportion of ARCs, improving couples’ reproductive autonomy
O-269 High detection rates in both IVF and general population and providing opportunities for cascade genetic analysis. Continuous reduction
confirm the clinical utility of Expanded Carrier Screening for the in sequencing costs and improvement in variant interpretation and genotype-phe-
management of reproductive genetic risk notype association are predicted to widen the diagnostic scope of ECS, leading
M. Fabiani1, S. Caroselli1, L. Girardi1, C. Patassini1, M. Poli1, C. to increasingly higher detection rates.
Simon1,2,3,4, A. Capalbo1,2 Trial registration number: not applicable
1
Igenomix Italia, Reproductive Genetics, Marostica VI, Italy ;
2 O-270 A novel long-range DNA sequencing approach improves
Igenomix Foundation, Reproductive Genetics, Valencia, Spain ;
3 the design of new protocols for preimplantation genetic testing of
Harvard School of Medicine- Harvard University-, Department of Obstetrics and
monogenic disease (PGT-M).
Gynecology, Boston MA, U.S.A. ;
4
Valencia University- and INCLIVA, Department of Obstetrics and Gynecology, M. Leaver1, N. Kubikova1, X. Tao2, C. Jalas2, D. Wells3
1
Valencia, Spain University of Oxford, Nuffield Department of Women’s and Reproductive Health,
Oxford, United Kingdom ;
Study question: What is the Detection Rate (DR) and the clinical utility of 2
The Foundation for Embryonic Competence, New Jersey, New Jersey, U.S.A. ;
Expanded Carrier Screening (ECS) application on the management of patients 3
Juno Genetics, Oxford, Oxford, United Kingdom
trying to conceive?
Summary answer: The high DR of At-Risk Couples (ARCs) and the remark- Study question: Can long-range next-generation sequencing assist in the
able proportion opting for IVF/Preimplantation Genetic Testing (PGT) treatment work-up of PGT-M cases, with potential to increase patient access, improve
demonstrate ECS clinical utility in reproductive contexts. accuracy and reduce costs?
What is known already: Around 1-2% of couples are at risk of conceiving a Summary answer: Long-range DNA sequencing provides a powerful, low-
child with an autosomal recessive or X-linked disorder. Carrier screening (CS) cost method that reveals the informative SNPs closest to gene mutations and
identifies ARCs improving their reproductive autonomy. Previously, CS was eliminates the need for additional family samples.
employed to test single diseases (e.g., Cystic Fibrosis). Current genetic technol- What is known already: To avoid misdiagnoses caused by allele dropout
ogies allow simultaneous/parallel testing of multiple disorders. However, the (failure to amplify one of the two alleles in an heterozygous cell), PGT-M strat-
previous low cost-effectiveness, suboptimal construction of test panels and egies usually involve parallel analysis of several diagnostically relevant sites (e.g.
inexperience with such genetic analysis across medical professionals are still mutations+linked polymorphisms). Polymorphisms need to be as close as pos-
undermining mainstream application of ECS. Here, a minimal ECS gene-panel sible to mutation sites because of the possibility of recombination. To determine
was routinely applied on a large population of reproductive patients providing which polymorphic alleles are associated with the disease, DNA from additional
evidence of its DR, clinical utility and cascade testing opportunity family members is usually analysed. However, relatives are not always available,
Study design, size, duration: A total of 2,013 individuals without family as patients may carry de novomutations, relatives may be untested or deceased,
history of genetic disorders were enrolled at affiliated clinics in different Italian or patients may not wish to disclose their PGT-M treatment to others.
regions between March 2017 and December 2019. Of these, 1,172 individuals Study design, size, duration: A novel work-up method for PGT-M was
were from couples trying to conceive (902 through In Vitro Fertilization (IVF) evaluated. DNA was obtained from 13 couples undergoing PGT for different
and 270 by natural conception). Additionally, 181 individuals undergoing homol- monogenic disorders. For each mutation, two primer sets (A and B) were
ogous IVF, 636 heterologous IVF and 24 natural conception were enrolled designed, allowing amplification of the mutation plus an additional ~10kb
through One-Member Screening Strategy. Data were collected and elaborated upstream (A) or downstream (B). Amplicons were sequenced as single contig-
anonymously following IRB approval. uous reads using the MinION (Oxford Nanopore). This identified informative
Participants/materials, setting, methods: Monogenic disorders included single nucleotide polymorphisms (SNPs) and revealed which alleles exist on the
in our panel (n=10) were selected based on ACMG-recommended criteria same chromosome as mutations.
(prevalence, carrier rate, severity) and unequivocally associated with highly pen- Participants/materials, setting, methods: 10kb regions flanking (and
etrant childhood conditions. Analytical methods employed include qPCR/ encompassing) mutation sites were sequenced from the 26 patients. Traditionally,
Taqman assays for SNPs/indel variants, triplet-primed PCR and melting curve candidate linked polymorphisms are identified from databases, but many turn
analysis for expansions and qPCR-based CNV analysis for exons/introns dele- out to be uninformative. Alternatively, parental samples can be analysed by
tions. Couples were defined at-risk when both partners carried an autosomal microarray, simultaneously assessing many SNPs, but only evaluating a fraction
recessive pathogenic variants on the same gene or when woman was a carrier of the variations in the genome. The SNPs found to be informative are some-
of X-Linked disorder. times relatively distant from the affected gene, increasing the chances of recom-
Main results and the role of chance: A total of 174 patients (8.6%) were bination between the SNP and mutation.
identified as carriers of one of tested pathogenic variants: CFTR (n=59, 2.9%); Main results and the role of chance: All 18 mutations in the 13 couples
SMN1 (n=46, 2.3%); DHCR7 (n=23, 1.1%); PMM2 (22, 1.1%); ACADM (n=15, were successfully detected using long-range sequencing. Additionally, between
0.7%); HADHA (n=4, 0.2%); FMR1 (n=3, 0.4%); ARSA (n=1, 0.05%) and DMD 2 and 83 (average 18) informative SNPs were found in the 10kb flanking regions.
(n=1, 0.1%). Remarkably, across the 586 couples tested, 13 (2.2%) were found The average distance from mutation sites to the nearest informative SNPs was
to be at increased risk of having an affected child (ARCs). In particular, the 2529bp.This compares to an average distance of 32,179bp when microarrays
identified ARCs were carriers of Cystic Fibrosis (n=5, 0.8%), Spinal Muscular were used to identify suitable SNPs. The extremely close proximity of

polymorphisms identified by long-range sequencing means that diagnostic chal- and analyzed to identify diet-modulated changes in enrichment which could be
lenges due to separation of SNPs from the disease-causing mutation by meiotic implicated in offspring phenotypes.
recombination can be virtually ruled out. In 28% of patients the closest infor- Main results and the role of chance: Sires (F0) fed a high-fat diet became
mative polymorphism had a minor allele frequency <0.1. Such SNPs are rarely obese, glucose intolerant and insulin insensitive, with increased adiposity, irre-
heterozygous and are therefore unlikely to be included in SNP-microarrays and spective of their genotype. Intergenerational effects of paternal diet were
are also unlikely to be chosen from databases as a candidate marker for PGT-M observed in male offspring only, while metabolic functions in female descendants
work-up.Additionally, novel informative intragenic SNPs, not present in any were not impacted by paternal high-fat diet. Males sired by obese transgenics
database, were identified in three couples. Because mutations and SNP alleles had enhanced metabolic phenotypes compared to wildtype obese descendants.
were contained within the same sequencing ‘read’, phasing was successfully Interestingly, transgenerational effects of high-fat diet were only observed in
accomplished in all cases, without any need for samples from additional family transgenic descendants, suggesting that paternal exposure to a multitude of
members. This will be of great value for couples who have no relatives suitable environmental stressors may exacerbate descendants’ risk for obesity and met-
for phasing of polymorphisms (approximately one quarter of all PGT-M couples). abolic syndrome. Sex-specific effects were further observed in the F2 generation,
Limitations, reasons for caution: Unlike generic PGT-M methods (e.g. suggesting males may be more susceptible to paternal and grand-paternal diets.
haplarithmisis/karyomapping), this strategy requires a customised protocol for Sperm chromatin profiling revealed diet-induced alterations in enrichment.
each couple. The use of direct mutation testing combined with analysis of the Differentially enriched regions occurred at genes with functions corresponding
closest possible SNPs flanking the mutation site is an approach that is unsur- to the observed offspring phenotypes, including genes involved in placenta devel-
passed in accuracy, but requires primer design and therefore slightly more opment, as well as glucose and lipid metabolism.
work-up. Limitations, reasons for caution: Further studies are required to better
Wider implications of the findings: By providing a simple, inexpensive, rapid understand the molecular mechanisms underlying the links between diet-induced
method of identifying the closest informative polymorphisms to parental muta- obesity, an aberrant sperm epigenome and offspring metabolic functions.
tions, long-range sequencing potentially improves PGT-M accuracy, reduces costs Wider implications of the findings: This is the first report linking high-fat
of customised protocols and accelerates test development. Furthermore, this feeding and alterations in the sperm epigenome at the level of a histone modi-
approach removes the need to obtain DNA from any family members other fication. These findings shed light on the potential contribution of chromatin in
than the couple undergoing PGT. sperm in paternal transmission of complex diseases.
Trial registration number: N/A Trial registration number: not applicable
O-271 KDM1A mediates transgenerational metabolic O-272 Germline Characterization of Genes Associated with
disturbances in a sex-specific manner and is linked to diet-induced Spermatogenesis and Embryonic Developmental Competence in
altered sperm chromatin signatures Azoospermic Men
A.S. Pepin1, C. Lafleur2, V. Dumeaux3, D. Sloboda4, S. Kimmins2 S. Cheung1, Z. Rosenwaks1, G.D. Palermo1
1 1
McGill University, Department of Pharmacology and Therapeutics, Montreal, Weill Cornell Medicine, The Ronald O. Perelman and Claudia Cohen Center for
Canada ; Reproductive Medicine, New York, U.S.A.
2
McGill University, Department of Animal Science, Montreal, Canada ;
3
Concordia University, PERFORM Centre, Montreal, Canada ; Study question: Can DNA sequencing of spermatozoa from azoospermic
4
McMaster University, Department of Biochemistry and Biomedical Sciences, men identify germline mutations related to the etiology of their infertility and
Hamilton, Canada ability to support a pregnancy?
Summary answer: Men with secretory azoospermia have mutated key genes
Study question: Our objectives were to investigate whether epigenome-en- that impair sperm production and affect the ability of their spermatozoa to
vironment interactions can lead to enhanced metabolic phenotypes transgener- support embryonic development.
ationally, and whether transmission of phenotypes are linked to sperm chromatin What is known already: Azoospermia accounts for about 15% of male factor
signatures. infertility cases. Although it can be caused by pre-testicular factors, the most
Summary answer: Combining a genetic model of epigenetic inheritance with recognized forms are testicular and post-testicular. Post-testicular azoospermia,
a diet-induced obesity model led to sex-specific transgenerational metabolic the most severe form, is characterized by scattered functional germinal epithelia
disturbances and aberrant sperm chromatin profiles. that strive to support the meiotic process during gamete development. To shed
What is known already: Obesity occurs in 650 million people worldwide light on the etiology of this condition, genetic studies have been performed,
(World Health Organization, 2016). Known factors contributing to obesity risks exclusively on peripheral blood. We performed a genetic assessment of the
include genetics, lifestyle and maternal factors. Epidemiological studies and animal spermatozoa to preferentially detect germline mutations that may be passed
model data indicate that paternal diet also affects offspring risks to develop onto offspring.
adult-onset metabolic disorders. However, the mechanisms underlying this Study design, size, duration: During the last 18 months, we performed
non-genetic inheritance of complex metabolic disease remain elusive. Sperm DNAseq on epididymal and testicular spermatozoa from men with acquired
histone methylation has been implicated in transgenerational epigenetic inheri- azoospermia (OA) (n=17) and nonobstructive azoospermia (NOA) (n=10),
tance and offspring health (Siklenka et al. 2015). This was previously demon- respectively, as well as on the ejaculated gametes from 3 fertile donor controls.
strated using a transgenic mouse model that overexpresses the histone Gene mutations related to azoospermia origin were categorized and compared.
demethylase KDM1A specifically in the germline, giving rise to males with an Gene mutation profiles of the OA and NOA men were then assessed in relation
abnormal sperm epigenome. to their ability to generate a pregnancy (fertile) or not (infertile).
Study design, size, duration: KDM1A transgenic males with a pre-existing Participants/materials, setting, methods: DNA was extracted and ampli-
compromised sperm epigenome and C57BL6NCrl wildtype sires, were exposed fied from at least 500 spermatozoa (DNA concentration, 705±562 ng/ul; quality,
to either a low- or high-fat diet (10% or 60% kcal fat, respectively) for two 1.7±0.1 nm). Following NGS, gene mutations, duplications, and deletions were
spermatogenic cycles (10-12 weeks). The next generations were generated by detected using the CLC Genomic Server 9.0. Genes were considered duplicated
mating males with chow-fed C57BL6NCrl females. All animals were subjected or deleted when the read depth was >1.5 or <0.5 times the median read depth
to a series of metabolic tests at 4 months of age (F0: n=15-25 animals per group, in the control. Common gene mutations from the OA and NOA cohorts were
F1: n=28-49, F2: n=8-21) and sacrificed at 5 months. assessed according to the couples’ clinical outcome.
Participants/materials, setting, methods: Metabolic assessment was per- Main results and the role of chance: Of the 27 couples (paternal age,
formed using glucose and insulin tolerance tests and baseline blood glucose 41.3±5yrs) included in this study, 17 OA men underwent surgical sperm retrieval,
levels. At necropsy, mesenteric adipose depots were weighted to assess adipos- with an average concentration of 1.3±3x106/ml and 7±14% motility. Ten NOA
ity. Liver RNA was extracted (n=4-6 per sex per group per generation) for men yielded spermatozoa with a concentration of 0.03±0.2x106/ml and 0.5±1%
differential gene expression. Sperm of F0 sires (n=5 per group) was subjected motility. NGS assessment did not show a significant difference in overall sperm
to chromatin immunoprecipitation sequencing targeting histone H3 methylation aneuploidy between the two groups (OA, 1.7%; NOA, 1.9%).

In the OA group overall, only 3 housekeeping genes were mutated (ATP4A, Main results and the role of chance: Our sample (n=387) consists of
SLC17A7, and OR1D4). In the NOA patients, however, 5 genes involved in RNA 265 females (68.5%) and 122 males (31.5%). Most of the participants were
transcription (POLR2L), apoptosis (AP5M1), and basic spermiogenic function below 34 years old (72.9%), didn’t have children (68.6%) and were currently
(AP1S2, AP1G2, and APOE) were deleted. in a relationship (69.1%). Of those who were in a relationship, 52.1% had a
The OA patients were treated in 17 ICSI cycles (maternal age, 34.8±3yrs), future child wish. More than half of the participants (53.6%) estimated their
resulting in a pregnancy and delivery rate of 47.1% (8/17). The fertile and chance of being a carrier for a recessive disorder (very) low. Likewise 64.8%
infertile OA cohorts had only 1 mutated gene, ZNF749 and PRB1, respectively. of participants estimated their chance of conceiving a child with a recessive
Both were unrelated to spermatogenesis or embryo developmental disorder (very) low. Offering ECS to couples with a child wish was found
competence. acceptable by 86% of participants. However, fewer participants would consider
When NOA men were treated in 10 ICSI cycles (maternal age, 38.2±2yrs), ECS for themselves in the future (61%). Only 19 (4.9%) participants answered
the pregnancy rate was 70% (7/10). While the fertile cohort displayed 1 mutated all knowledge questions correctly. Half of the participants (50.9%) preferred
gene (MPIG6B) related to stem cell lineage differentiation, the infertile NOA the disclosure of individual results, while 35.2% preferred couple-based results.
cohort had deleted genes involved in spermatogenesis (n=6), apoptosis (n=4), Most participants indicated that ECS should be offered through the gynecologist
acrosomal function (n=2), and early embryonic development (n=8). (81.1%), followed by the GP (71.5%) and the Center for Human Genetics
Limitations, reasons for caution: This is a novel study with a limited number (64.8%). About 68.9% of participants were willing to pay out-of-pocket for an
of observations, and it cannot demonstrate genetic association with obstructive ECS test. If participants were willing to pay themselves, they indicated that the
azoospermia independently of reproductive performance. In addition, although test should have a maximum cost of 150 euros (45.3%) or between 150 – 300
female partner age was controlled for, confounding factors cannot be excluded euros (40%).
with certainty. Limitations, reasons for caution: Our study employed convenience sam-
Wider implications of the findings: This novel DNAseq study aims to pling to recruit participants, therefore our reported results should be interpreted
identify germline mutations. The reproductive performance of OA men was not with caution. Another limitation of our study is that we didn’t offer an actual
associated with any genetic mutations. In the NOA cohort, fertile men had only ECS-test to participants so some results are only hypothetical. Actual participa-
one deleted gene involved in sperm production, while infertile men had several tion might differ from the intention to do a behavior.
deleted genes involved in spermatogenesis and embryonic development. Wider implications of the findings: Our study reports on the views of the
Trial registration number: not applicable potential users of ECS. These results can guide health care providers and policy
makers when implementing ECS in the future to ensure that couples makes
informed reproductive decisions based on accurate knowledge and consistent
O-273 Knowledge, attitudes and preferences regarding expanded with the values of the couple.
carrier screening among reproductive-aged men and women in Trial registration number: Not applicable
Belgium
E. Van Steijvoort1, I. Geysen1, S. Van Epperzeel1, H. Devolder1, H. O-274 Single nucleotide polymorphisms (SNPs) in FSHR/FSHβ
Peeters2, K. Peeraer3, G. Matthijs2, P. Borry1 genes do not modify ovarian response to stimulation with rFSH. A
1 prospective multicentre study in Europe and Asia
KU Leuven, Department of Public Health and Primary Care, Leuven, Belgium ;
2
KU Leuven, Department of Human Genetics, Leuven, Belgium ; N.P. Polyzos1, C. Blockeel2, C. Spits3, B. Alvaro Mercadal1,
3
KU Leuven, Department of Development and Regeneration, Leuven, Belgium S. Garcia4, P. Ma5, L. Le5, M. Ho5, G. Duqué3, J. Mertens3, D.
Stoop6, P. Drakopoulos2, H. Tournaye2, F. Martinez1, N. Vuong7
Study question: What is the knowledge, attitudes and preferences regarding 1
Dexeus University Hospital, Reproductive Medicine Department, Barcelona, Spain ;
expanded carrier screening among reproductive-aged men and women in 2
Universitair Ziekenhuis Brussel-Vrije Universiteit Brussel, Centre for Reproductive
Belgium? Medicine, Brussels, Belgium ;
Summary answer: Health care providers and policy makers should take the 3
Vrije Universiteit Brussel, Research Group Reproduction and Genetics, Brussels,
current knowledge level, attitudes and preferences of potential users in account Belgium ;
when implementing ECS. 4
Dexeus University Hospital, Statistics Department, Barcelona, Spain ;
What is known already: Through carrier screening couples at-risk of con- 5
My Duc Hospital, IvfMD, Ho Chi Minh City, Vietnam ;
ceiving a child with an autosomal recessive or X-linked condition can be identified 6
Ghent University Hospital, Centre for Reproductive Medicine, Ghent, Belgium ;
prior to conception, allowing prospective parents to make informed reproductive 7
University of Medicine and Pharmacy, Medical Statistics Department, Ho Chi
decisions when planning for a family. In the last decade, expanded carrier screen- Minh City, Vietnam
ing has become available to (prospective) parents. Following recommendations
made by the Superior Health Council (SHC) of Belgium, a Belgian Genetic Study question: Does the presence of SNPs in FSHR/FSHβ influence oocyte
Carrier Screening test (BeGECS) was launched in October 2019. However, little yield and Follicular output rate (FORT) in predicted normal responders treated
is known about the interest, knowledge level, attitudes and preferences of poten- with rFSH?
tials users. More insights are need to ensure responsible implementation of the Summary answer: The presence of SNPs in FSHR/FSHβ (rs6165, rs6166,
BeGECS offer. rs1394205, rs10835638) does not influence ovarian response in predicted nor-
Study design, size, duration: A cross-sectional survey was conducted using mal-responders treated with a fixed-dose of 150IU rFSH
convenience sampling. Individuals of reproductive age visiting their public phar- What is known already: Ovarian reserve markers have been a breakthrough
macist were invited to answer a self-administered questionnaire assessing knowl- in response prediction following ovarian stimulation. However, a significant per-
edge, attitudes and preferences regarding expanded carrier screening (ECS). centage of patients show a disproportionate lower ovarian response, as compared
Prior to filling in the questionnaire, participants were asked to read an information with their actual ovarian reserve. Studies on pharmacogenetics demonstrated a
letter explaining some key concepts. Based on our sample size calculation we relationship between FSHR/FSHβ genotyping and drug response, suggesting a
aimed to collect 385 completed questionnaires. Data collection was carried out potential effect of individual genetic variability on ovarian stimulation. However,
between September 2019 and December 2019. evidence from these studies is inconsistent, due to inclusion patients with variable
Participants/materials, setting, methods: The study population consisted ovarian reserve, use of different starting gonadotropin doses and allowance for
of reproductive aged (18-49 years) men and women. Participants were recruited dose adjustments during treatment. This highlights the necessity of a well-controlled
through five public pharmacies in Flanders (Belgium). Potential participants were prospective study, in homogenous population treated with an identical protocol.
approached about the study by researchers present in the pharmacies and were Study design, size, duration: We conducted a multicenter multinational
asked to fill in the anonymous questionnaire on the spot after reading an infor- prospective study, including 368 patients from Vietnam, Belgium and Spain (168
mation sheet explaining some key concepts. The questionnaire contained ques- from Europe and 200 from Vietnam), from November 2016 till June 2019. All
tions on socio-demographic characteristics, risk perception, intention to have patients underwent ovarian stimulation followed by oocyte retrieval in an antag-
ECS, knowledge, attitudes, preferences, etc. onist protocol with fixed daily dose of 150IU of rFSH until triggering. Blood

sampling and DNA extraction was performed prior to oocyte retrieval, followed 95% of cases The prevalence of EP has doubled since 1960 and accounts for
by genotyping of 4 SNPs from FSHR/FSHβ (rs6165, rs6166, rs1394205, about 2% of the pregnancies in the first trimester. In recent years, its incidence
rs10835638). has increased due to the increase in incidence of pelvic inflammatory diseases,
Participants/materials, setting, methods: Eligible were predicted normal use of fertility drugs, and pelvic surgery. Transvaginal ultrasound and serial mea-
responder women <38 years old undergoing their 1st/2nd ovarian stimulation surement of serum beta-hCG levels are the most common diagnostic methods
cycle. Laboratory staff and clinicians were blinded to the clinical results and for EP. Despite the use of transvaginal ultrasound and measurement of beta-hCG
genotyping respectively. The number of oocytes and the FORT (number of levels, about 40% to 50% of the initial cases of the disease are not diagnosed.
follicles on trigger day/AFCx100) were compared between different FSHR/ Therefore, the need for new diagnostic tools, preferably serum biomarkers, is
FSHβ genotypes. Results were analysed with x2 or oneway ANOVA and were essential.
corrected for multiple comparisons as appropriate. Several diagnostic biomarkers for EP have been proposed over the past
Main results and the role of chance: The prevalence of homozygous SNPs decades. Progesterone along with beta-hCG are the first and widely used bio-
in the FSHR was: rs6166(c.2039 A>G) 15.8%, rs6165(c.919 A>G) 34.8% and markers. Progesterone however, although represents a good indicator of early
rs1394205(c.-29G>A) 14.1% with significant differences between European and pregnancy viability, is a poor predictor of pregnancy location. To produce the
Asian women. FSHβ rs10835638(c.-211 G>T) was very rare: 0.5%. optimal biomarker for early EP detection, several obstacles must be overcome.
Comparison between different SNPs and genotypes, wild-type (WT), hetero- Firstly the behaviour of an EP and the substances produced during the first few
zygous(HT) or homozygous(HZ), did not reveal significant differences in terms days following implantation, might vary considerably in cases where the preg-
of ovarian response. nancy is a result of assisted reproduction technology, the site of implantation
The number of oocytes was comparable between the 3 genotypes (WT vs.HT and whether the EP is intact ruptured or aborted. Investigators studied several
vs.HZ) for all 3 variants in FSHR : biomarkers produced by the fallopian tube, the early embryo, the endometrium,
the ovary, or peritoneal inflammatory factors. So far none of these substances
1. a. rs6166 (mean(SD)): 14.81(7.07) vs. 13.49(7.30) vs. 13.69(5.82), have been widely used in clinical practice.
p=0.066 Cancer antigen 125 has been reported as a marker for pregnancies “likely to
2. b. rs6165: 14.39(7.08), 13.59(7.54), 14.25(6.30), p=0.095 continue”. Inhibin A, activin A, and high sensitivity C-reactive protein (hsCRP)
3. c. rs1394205: 15.06(8.10), 13.24(5.89), 13.61 (6.95), (p=0.088) were not significantly different in women with successful and failed expectant
management. On the other hand, inhibin A may be useful for predicting sponta-
neous resolution of pregnancy of unknown location (PUL) but is not as good as
Similarly, no differences were observed on the FORT for rs6166 (p=0.814),
progesterone. Recent attention has focused on metalloprotease 12 (ADAM 12)
rs6165 (p=0.974) and rs1394205 (p=0.081). Linear regression analysis to
and fibronectin as potentially promising candidate markers in PUL pregnancies.
predict the number of oocytes in relation to genetic variants (rs6166,
Serum Interleukin-15 (IL-15) and anti-C1q antibodies although showed promising
rs1394205, rs6165) adjusted for age, AMH, continent and duration of stimu-
results in differentiating between an abortion and EP, they had no further clinical
lation, did not show differences between WT, HT, HZ for rs6166 and rs6165.
application. Similar results presented when placental growth factor (PlGF) and the
Although rs1394205 HZ had statistically significant lower oocyte yield com-
soluble Flt-1 (sFlt-1) receptor of vascular endothelial growth where investigated.
pared with WT, this was of minimal clinical significance, 0.84 95%CI [0.71;0.98]
Several studies outline the utility of creatine phosphokinase (CPK), which is
oocytes less.
an intracellular enzyme found in the fallopian tube, as a marker for early diagnosis
Finally, the proportion of women with optimal response (>10 oocytes) was
of EP. The theory behind the idea is that since CPK is an intracellular enzyme,
also comparable between the 3 groups (WT vs. HT vs. HZ) for rs6165, rs6166
lysis of trophoblast cells leads to an increase in plasma CPK levels. Therefore,
and rs1394205
the level of this enzyme can be used for evaluation of tubal ectopic pregnancy
Limitations, reasons for caution: The study was performed in relatively
because increased CPK can be associated with trophoblastic invasion and tro-
young women with normal ovarian reserve to eliminate biases related to age-re-
phoblast mass. However different studies produced conflicting results.
lated fertility decline; thus, caution is needed when extrapolating results to other
Metabolomics and epigenomics represent two rather new fields of research
populations. In addition, no analysis was performed for FSHβ rs10835638 due
in medicine. Metabolomic markers have been produced promising results in
to the very low prevalence of homozygotes (n=2).
recent studies. Epigenetic studies are mainly focused on detecting micro-RNAs
Wider implications of the findings: Based on our results, genotyping SNPs
that regulate expression of EP-associated genes such as VEGFA, EGFR, ESR1
rs6165, rs 6166, rs1394205, rs10835638 prior to initiating an ovarian stimulation
and immune response-related genes. Most of these studies have a limited number
with rFSH in predicted normal responders should not be recommended since their
of subjects, therefore, their results need to be confirmed by future studies.
presence does not modify ovarian response in these women. Future research may
The ideal diagnostic tool for an EP would be a single serum marker to replace
focus on other genes related to folliculogenesis or steroidogenesis
ultrasound and serial biochemistry. This implies earlies management and the
opportunity for conservative management in most cases. To date this ideal serum
biomarker is under investigation.
Trial registration number: -
Study funding: -
INVITED SESSION Funding source: -
SESSION 65: BIOMARKERS OF FAILED PREGNANCY
O-276 Developing biomarkers for non-viable pregnancies: the
08 July 2020 Parallel 1 12:00 - 13:00
laboratory expert perspective
A. Horne1
1
University of Edinburgh, MRC Centre for Reproductive Health, Edinburgh,
United Kingdom
O-275 Biomarkers of failed pregnancy (miscarriage ectopic). The
clinician’s perspective Abstract text
A. Daponte1, O. Koukoura2 The development of biomarkers for prompt diagnosis, and safe timely treatment,
1
University of Thessaly- School of Health Sciences- Faculty of Medicine, Obstetrics of non-viable pregnancies is essential. A blood-based biomarker that accurately
and Gynaecology, Larissa, Greece identifies an ectopic pregnancy could be used to offer early diagnostic certainty
2
University Hospital of Larissa, Obstetrics and Gynaecology, Larissa, Greece in cases where ultrasound cannot determine the location of the embryo (‘a
pregnancy of unknown location’). Here, I review the literature on the develop-
Abstract text ment of molecularly targeted diagnostics for ectopic pregnancy, miscarriage and
Ectopic pregnancy (EP) occurs when a blastocyst abnormally implants outside viable pregnancies. Molecules examined so far can be broadly grouped into
the endometrium The fallopian tube is the commonest location in more than biological themes of relevance to reproduction: (i) Fallopian tube (dys)function,

(ii) embryo/trophoblast growth, (iii) corpus luteum function, (iv) inflammation, O-279 Impact of COVID-19 pandemic on the psychological status
(v) uterine function and (vi) angiogenesis. While a sensitive and specific bio- of infertile patients who had in vitro fertilization treatment
marker for ectopic pregnancy has yet to be identified, it is possible that improve- interrupted or postponed
ments in platform technologies or a multi-modal biomarker approach may yield S. Ferrero1, C. Scala2, M. Altieri1, F. Barra1
an accurate diagnostic biomarker test. Furthermore, with the advent of better 1
IRCCS Ospedale Policlinico San Martino- University of Genoa,
imaging technology, the need for a blood-based biomarker test may be super- Academic Unit of Obstetrics and Gynecology, Genoa, Italy
seded by improvements in ultrasound or magnetic resonance imaging technology. 2
Gaslini Hospital, Unit of Obstetrics, Genoa, Italy
Study question: To understand the changes in the psychological status of

infertile patients who had in vitro fertilization (IVF) treatment interrupted or
INVITED SESSION
postponed because of the COVID-19 pandemic.
SESSION 66: SYNTHETIC EMBRYOLOGY: MYTH OR REALITY? Summary answer: Infertile women who had IVF treatment postponed
08 July 2020 Parallel 2 12:00 - 13:00 because of COVID-19 pandemic have psychological symptoms. IVF centers must
strengthen psychological counseling to improve patients’ mental health.
What is known already: Infertility is a severely distressing experience for
many couples. Depression and anxiety are psychological disorders associated
O-277 Modelling blastocyst formation and in utero implantation
with infertility, and they may worsen during infertility treatments.
solely from stem cells
Study design, size, duration: This study was conducted in April 2020. A
N. Rivron1 survey’s link was e-mailed to the partecipants. A reminder e-mail was sent every
1
IMBA- Austrian academy of science, Laboratory for synthetic development, Vienna, Austria week for one month.
Participants/materials, setting, methods: This study included all the
Abstract text patients attending our institution who had IVF treatment interrupted or delayed
The pre-implantation conceptus (blastocyst) forms all embryonic and extra-em- because of the COVID-19 pandemic. The severity of anxiety and depression
bryonic tissues. It consists of a spherical thin-walled layer, the trophectoderm, was assessed using the Generalized Anxiety Disorder-7 (GAD-7) and the Patient
that surrounds a fluid-filled cavity sheltering the embryonic and primitive endo- Health Questionnaire-9 (PHQ-9). Subjects with pre-existing psychiatric disorders
derm cells. From mouse blastocysts, both trophoblast and embryonic stem cell were excluded.
lines can be derived, which are in vitro analogues of the trophectoderm and Main results and the role of chance: 503 patients (294 women and 209
embryonic compartments, respectively. Our lab showed that trophoblast and men) completed the questionnaire (response rate: 80.9%; n = 622). The mean
embryonic stem cells self-organize in vitro to form structures that morphologi- (± SD) age of the women included in the study was 37.3 (± 4.6) years; the mean
cally and transcriptionally resemble blastocysts (blastoids). Blastoids form prim- age of men was 38.4 (±6.2). The median number of previous failed IVF cycles
itive endoderm-like cells, the second extra-embryonic lineage, and implant was 2 (range, 0-8). The prevalence of anxiety or depression, or both was 37.0%
upon in utero transfer. Like blastocysts, blastoids form via inductive signals (186 of 503). The prevalence of anxiety or depression or both was significantly
originating from the inner embryonic cells and driving outer trophectoderm higher in women (41.5%; 95% C.I. 35.8%-47.4%) than in men (30.6%; 95% C.I.,
development. However, the nature and function of these molecular signals are 24.4%-37.4%; p = 0.012). The median total score on GAD-7 was 11 (interquar-
largely unexplored. Genetically and physically uncoupling the embryonic and tile range, IQR: 9-14). The median total score on PHQ-9 was 10 (IQR: 9-14).
trophectoderm compartments, along with single cell transcriptomics, revealed The occurrence of anxiety and/or depression was significantly associated with
an extensive list of inductive signals. We show that the embryonic cells maintain a diagnosis of poor ovarian reserve (odds ratio, OR=2.13), diagnosis of endo-
trophoblast proliferation and self-renewal, while fine-tuning trophoblast epithelial metriosis (OR=1.54), diagnosis of uterine fibroids (OR=1.34) and time spent
morphogenesis. Altogether, these embryonic inductions are paramount to form on COVID-19 related news per day (> 1 hour per day, OR=1.47). Notably, 187
a trophectoderm state that robustly implants and triggers a genuine patients (37.2%; 32.9%-41.6%) declared that they would like to undergo the IVF
decidualization in utero. Thus, at this stage, the nascent embryo fuels the treatment despite the COVID-19 pandemic. 384 patients (76.3%; 72.4%-80.0%)
development and implantation of the future placenta. Altogether, the blastoid is declared that they would be reassured if they knew when they could restart the
a powerful embryo model that can be reproducibly generated in large numbers, IVF treatment.
finely tuned, contains all the cell types to form the conceptus, and implants in utero. Limitations, reasons for caution: A longitudinal follow-up would be helpful
References: in tracking the changes in anxiety and depression levels at different stages of the
Rivron NC [corresponding author] et al. Blastocyst-like structures generated epidemic.
solely from stem cells. Nature (2018). doi:10.1038/s41586-018-0051-0. Wider implications of the findings: This study reveals that the COVID-19
Rivron NC. Formation of blastoids from mouse embryonic and trophoblast epidemic caused a sharp increase in the prevalence of anxiety and depression
stem cells. Protocol Exchange (2018) doi:10.1038/protex.2018.051 among infertile patients undergoing IVF. IVF center must strengthen the psycho-
Frias-Aldeguer J. et al. Embryonic signals perpetuate polar-like trophoblast stem logical counseling for these patients to improve their sleep quality and men-
cells and pattern the blastocyst axis. BioRxiv 510362; doi: https://doi. tal health.
org/10.1101/510362
Vrij E. et al. Chemically-defined induction of a primitive endoderm and epi-
blast-like niche supports post-implantation progression from blastoids.
BioRxiv 510396; doi: https://doi.org/10.1101/510396. O-280 Patient experiences of fertility clinic closure during the
COVID-19 pandemic: Appraisals, coping and emotions
O-278 Stem cells in reproductive biology: myth or reality? J. Boivin1, C. Harrison1, R. Mathur2, G. Burns3, A. Pericleous-
G P. Schatten1 Smith4, S. Gameiro1
1
1
University of Pittsburgh, Department of OB/GYN & Reproductive Sciences & Cell Cardiff University, School of Psychology, Cardiff, United Kingdom
2
Biology & of Bioengineering; UP Cancer Institute, U.S.A Manchester University NHS Foundation Trust and Manchester Academic Health
Sciences Centr, OB-Gyn, Manchester, United Kingdom
3
Fertility Network UK, Director, London, United Kingdom
4
British Infertility Counselling Association, Chair, York, United Kingdom
INVITED SESSION
Study question: What are appraisals, coping strategies and emotional reactions
SESSION 67: COVID-19 - PSYCHOSOCIAL IMPACT OF
to COVID-19 fertility clinic closure?
DELAYED TREATMENT
Summary answer: Clinic closure was appraised as stressful due to uncertainty
08 July 2020 Parallel 3 12:00 - 13:00 and threat to attainability of parenthood goal but patients were able to cope
using accommodative strategies.

What is known already: According to stress and coping theory, imbalance Summary answer: The implications of COVID-19 on individuals has been
between threat and coping resources leads to stress reactions. People facing drastic and include uncertainty, delay in care and anxiety about pregnancy out-
disasters generally experience more stress than usual, but remarkably most are comes during the pandemic.
able to cope and recover, with some eventually seeing benefits from the situation. What is known already: COVID-19 has been declared a pandemic during
Research to date on experiences of COVID-19 in the general population indi- which fertility services in the United Kingdom were temporarily suspended.
cates more anxiety and depression among respondents than historical norms, Recently, services have started the process of resumption, but there remains
worry about becoming mentally unwell due to uncertainty and loss of control significant uncertainty in the way fertility care will be delivered in the United
but nevertheless ability to cope. Reports of reactions to COVID-19 in infertility Kingdom.
patients shows these are moderately to extremely upset about COVID-19 treat- Study design, size, duration: The study involved two phases; an online
ment cancellation. questionnaire designed to obtain an overview of participants perceptions of
Study design, size, duration: Cross-sectional design. Mixed-methods, Covid-19, impact on fertility care and perceived barriers to care from 422
English, anonymous, online survey posted from April 09 to April 21 to social participants. In phase two, pertinent themes from the questionnaire findings
media. Eligibility criteria was being affected by COVID-19 fertility clinic closures, were explored further in greater depth and detail in 15 individual semi-struc-
18 years of age or older and able to complete survey in English. In total 946 tured interviews through purposive sampling in May 2020. The study was
people clicked on the survey link, 76 did not consent, 420 started the survey but designed with patient and public engagement, as well as a clinicians and
did not complete it, and 450 completed the survey (48% completion, 446 academics.
women, 4 men). Participants/materials, setting, methods: The study was advertised via
Participants/materials, setting, methods: On average participants were online platforms throughout the UK. Eligible participants included anyone who
33.6 years (SD=4.4) and had been trying to achieve pregnancy for 3.5 years has experience of fertility services within the past one year. The ‘open-ended’
(SD=2.22). The survey comprised quantitative questions about intensity of questionnaire results and transcribed semi-structured interview findings were
threat, emotions, and ability to cope with clinic closure. Open-text questions analysed through thematic analysis (familiarisation with the data, creating initial
covered understanding of COVID-19, fears, concerns and perceived benefits of codes, generating and reviewing subsequent themes, as well as refining themes
clinic closure, and desired information about closure and future re-opening. in the context of the research question). Percentages were also used to com-
Inferential statistics were used on quantitative data and thematic qualitative anal- pliment the qualitative findings.
ysis (inductive coding) performed on open text data. Main results and the role of chance: 718 participants replied to the ques-
Main results and the role of chance: Overall 82.2% (n=367) had tests/ tionnaire with the majority identifying as White British (82%) and 5% as male.
treatments postponed. Patients appraised fertility clinic closure as significantly Over 60% had used fertility services within the last one year. 92% of individuals
more negative than positive [t(447)=45.2, p < .001], and to be very or explained that COVID-19 had a ‘negative impact’ on their fertility treatment,
extremely uncontrollable [t(445)=38.99, p < .001] and stressful namely ‘delay in care’. 62% of participants discussed concerns about the ‘uncer-
[t(445)=27.44, p < .001]. Most reported below average ability to cope with tainty’ they felt about fertility services; these included the ‘unknown impact of
the closure (11.9% not at all able) [t(445)=5.57, p < .001]. Question by COVID-19 on pregnancy outcomes’, the ‘unknown impact on general gynae-
question textual analysis revealed 33 broad themes, grouped into four meta- cology services’ and the ‘unknown impact of COVID-19’ on ‘fertility success’.
themes across questions. First, COVID-19 effects were unknown and clinic The influence of the media and its focus on COVID-19 at present was a per-
closure perceived as precautionary or as unfair relative to advice about getting ceived barrier to care as participants felt fertility care was ‘less prioritised by the
pregnant given to the public. Second, closure was appraised as a threat to government’.
attainability of the parenthood goal largely based on uncertainty of effects Through semi-structured interviews with fifteen participants, we learned about
of delay and intensification of pre-existing hardships of fertility problems. the concerns they had about the extension of the 10-year storage limit, partic-
Third, threat emotions and uncertainty taxed personal coping resources but ularly in relation to embryos. Interestingly, participants from a Black and Minority
most managed the situation using diverse strategies (e.g., thought-manage- Ethnic (BAME) background discussed the ‘cultural pressures’ they faced during
ment, getting mentally and physically fit for next treatments, strengthening COVID-19 and the implications on their wider social circumstances. Participants
their social network, and keeping up-to-date with clinics. Finally, almost all were mindful about the ‘pressures on the service’ when re-opening, and there-
participants reported significant stress, worry and frustration at the situation fore ‘advancing maternal age’, ‘socio-economic background’ and ‘previous unsuc-
with a minority reporting intense feelings of hopelessness. More information cessful fertility treatment’ were the main factors individuals considered important
about effects of delay and eventual wait-list prioritisation at re-opening was when ‘prioritising’ fertility care.
desired. Limitations, reasons for caution: The questionnaire findings are represen-
Limitations, reasons for caution: The survey captured reactions at a specific tative mainly from individuals in the UK and from those identifying mainly as
point in time (during lockdown before clinics announced re-opening) but results White British; subsequently, cultural influences that may affect participants views
offered future directions for handling of information in COVID-19 conditions. may not be representative. Those individuals without online connectivity may
Participants were self-selected, 25% non-UK countries and only 4 men all of have had valuable insight but we were not able to capture this.
which may affect generalisability. Wider implications of the findings: This study has truly enabled participants
Wider implications of the findings: Fertility stakeholders (e.g., clinics, voices to be heard. The findings from this study can be used by fertility service
patient groups, regulators, professional societies) need to work together pro- providers to appreciate the patient perspective when considering the re-opening
actively to address the continuing impact of COVID-19 on patients. Transparent of fertility services nationally and internationally and be mindful of patient
processes for COVID-19 and sign-posting to information and coping support concerns.
resources will bolster patients existing coping resources and support staff deliver
the new normal for clinics.
O-318 Covid-19 does not stop fertility preservation: The Italian
situation during the pandemic emergency.
O-317 A study to explore participants experiences of fertility A. Anastasi1,8, L. Sosa Fernandez2,8, D. Cimadomo3,8, F.G.
services during the Covid-19 lockdown; a biphasic mixed methods Klinger4,8, E. Licata5,8, C. Scarica6,8, L. De Santis7,8
study 1
Hospital “del Delta”, Physiopathology of Human Reproduction Center,
B. Karavadra1, A. Balen2, E. Morris1 Lagosanto- Ferrara., Italy
2
1
Norfolk & Norwich University Hospital, Obstetrics & Gynaecology, Norwich, Embryos Fertility Center, Reproductive Medicine, Battipaglia, Italy
3
United Kingdom Clinica Valle Giulia - Genera Center, Reproductive Medicine, Rome, Italy
4
2
Leeds Teaching Hospital, Obstetrics and Gynaecology, Leeds, United Kingdom University of Rome “Tor Vergata”, Department of Biomedicine and Prevention-
Section of Histology and Embryology, Rome, Italy
Study question: What is the impact of Covid-19 on individuals using fertility 5
Sandro Pertini Hospital, Physiopathology of Reproduction and Andrology Unit,
services in the United Kingdom? Rome, Italy

6
Casa di Cura Villa Salaria in partnership with Institut Marques, Reproductive O-281 Complex mosaic embryos after preimplantation genetic
Medicine, Rome, Italy testing: go for a second biopsy?
7
San Raffaele Scientific Institute- University Vita-Salute San Raffaele, Department Z. Shuang1, X. Pingyuan2, H. Liang3, T. Yueqiu3, L. Ge4
of Obstetrics & Gynecology- IVF Unit, Milan, Italy 1
8
Hunan Guangxiu Hospital, Laboratory, Changsha- Hunan, China ;
On behalf of the Italian Society of Embryology, Reproduction and Research (SIERR) 2
Hunan Normal University, School of Basic Medicine, Changsha- Hunan, China ;
3
Central South University, Basic Medical College, Changsha- Hunan, China ;
Study question: How Coronavirus disease (Covid-19) influenced fertility 4
Reproductive and Genetic Hospital of Citic-Xiangya, Research Department,
preservation cycles in Italy? A SIERR (Italian Society of Embryology, Reproduction
Changsha- Hunan, China
and Research) survey.
Summary answer: The survey carried out by SIERR shows how in Italy the Study question: What is the incidence of complex mosaic in pre-implantation
pandemic influenced in a limited fashion fertility preservation treatments. genetic testing (PGT) embryos and how to manage these embryos in clinical
What is known already: At the beginning of March 2020 Italy was severely practice?
hit by SARS-CoV-2, whose rapid spreading has forced the government to impose Summary answer: The incidence of complex mosaic blastocysts in PGT was
the total lockdown of the country. Therefore, all ART centres in Italy decided 1.9%(253/13451) and it can be safe re-biopsy, implant and result in live births
to postpone their activities and reorganize their schedule to assure the safety What is known already: Some studies have reported that the births of babies
for both patients and staff members. However, given the known reproductive after mosaic embryo transfer though the pregnancy rates were lower and the
risks of cancer therapies, there has been a special attention to fertility preser- miscarriage rates were higher. The PGDIS and COGEN were given the practice
vation. In fact, during the pandemic, most Italian ART centres guaranteed the recommendations on chromosome mosaicism, they were suggested if non-mo-
execution of fertility preservation treatments. Currently, ART treatments are saic euploid embryos are not available, a non-complex, low-level mosaic embryo
gradually restarting for any clinical indication. may be considered for transfer. But the complex mosaicism (mosaicism observed
Study design, size, duration: The aim of this longitudinal survey is to verify three or more chromosomes) were not recommended to transfer. The incidence
whether the pandemic has reduced the number of male and female fertility of complex mosaicism appears to be lower, however, for the poor prognosis
preservation procedures. The survey questionnaire consisting of fifteen questions patients with only complex mosaic embryo, the clinical management becomes
(six multiple choice/nine brief answers), has been sent to all SIERR members very complicated.
on May 12th. On May 17th we no longer accepted responses for the first call. Study design, size, duration: Trophectoderm (TE) biopsy and NGS for genetic
We will send a recall on May 22nd trying to collect a greater amount of data.
testing was performed on blastocysts generated from 13451 embryos between
Participants/materials, setting, methods: In this highly stressful and 2018 to 2019 in the IVF centre of the Reproductive and Genetic Hospital of
demanding period, 46 SIERR members responded to the survey trough Google- CITIC-Xiangya. All biopsy samples were subjected whole genome amplifica-
Form. Data obtained were analysed in order to receive one answer from each tion(WGA). The validated NGS platform were used to detect the copy number
centre. Therefore, we collected answers from 42 centres. We asked our mem- and the mosaic were defined 30-70%. The complex mosaicism were defined as
bers how many fertility preservation cycles they have performed from 1st-Janu- three or more chromosomes with the copy number between 30% to 70%.
ary-2019 to 30th-April-2019 and how many in the same period of this year. The Participants/materials, setting, methods: Eighty-five patients opted to
remaining questions regarded the management of these cycles during the undergo warming, re-biopsy and re-vitrification in an effort to obtain a chance
pandemic. to transfer these embryos. Blastocysts resulting in a euploid diagnosis following
Main results and the role of chance: Albeit a general reduction of the the re-biopsy procedure were transferred with appropriate genetics counseling
number of procedures in many medical fields, our data show that fertility pres- in subsequent frozen embryo transfer (FET) cycles. The main outcome mea-
ervation was not hampered by lockdown. Comparing the number of procedures sures were euploid, implantation and live birth rates. The study was approved
during the same period in 2019 and 2020, results show a reduction of 19.6% in by the ethics committee of the Reproductive and Genetic Hospital of
2020. Interestingly, this reduction refers to the period previous Covid-19 emer- CITIC-XIANGYA.
gency. In fact, from 1st-January-2019 to 30th-April-2019 Italian centres performed Main results and the role of chance: Overall, 13451 blastocysts were
626 fertility preservation cycles. Conversely, during the same period in 2020, analyzed from 4359 PGT cycles, 253(1.9%) blastocysts from 243 cycles were
503 cycles were performed, of which 43% (74 female and 145 male preservation complex mosaic. Of these 243 cycles with complex mosaic embryos, 85 patients
procedures) during the lockdown. opted to re-biopsy these embryos. The warmed rate were 96.7% (88 /91) and
All patients have been screened with different methods: Triage (61%), RT-PCR all the re-biopsied embryos were assigned a genetic diagnosis. Fifty-five re-biop-
(33%) or both screening methods (6%). Eleven centres performed female pres- sied blastocysts from a total 53 individual patients were diagnosed as euploid
ervation cycles using open cryopreservation devices (91%) or closed system (55/88) and were suitable for subsequent FET. Of the 33 embryos that were
(9%). These samples have been stored in non-infected patients’ tanks (81%) or abnormal, the most common diagnosis was mosaic, other abnormalities included
in quarantine tanks (19%). 7 aneuploidy, 7 segmental aneuploidy, 2 complex mosaic. Sixteen patients have
In 5 centres some members of the staff resulted positive to the virus; not already undergone a subsequent FET of a re-biopsied single euploid blastocyst.
surprisingly 3 out the 5 interested centres are located in Lombardia, the most The survival rate for blastocysts undergoing second warming was 100% (16/16)
affected region by SarsCoV-2 in Italy. resulting in an ongoing pregnancy rate of 31.3% (5/16) and birth of two normal
Limitations, reasons for caution: Unfortunately, due to the abstract dead- healthy babies to date.
line, the survey remained opened only for six days. ART Societies recommen- Limitations, reasons for caution: But it should be noted that all the complex
dation helped to ensure safety in the execution of fertility preservation cycles. mosaic embryos re-biopsied in this study were with lower levels(30-50%) and
However, in the short-term we recommend to keep the guard up in order to the NGS platform for mosaic detection has been validated by known samples.
avoid the possibility of a return to phase1. At the time of writing the sample size is relatively modest.
Wider implications of the findings: Many reasons may have caused the Wider implications of the findings: The damage from laser, the quality of
decrease in the number of cases comparing 2019 to 2020: some not urgent the TE cells, the bias in the WGA and NGS may be the reason for the complex
oncological surgeries have been probably postponed and consequently their mosaic in first biopsy samples. Finally, we all endeavor to maximize the most
fertility preservation procedures. Moreover, because during the lockdown it was efficient route to a healthy birth for our patients in a single attempt.
strictly recommended to stay at home, many diagnoses might have been missed. Trial registration number: 81222007 and 31601183
O-282 Developmental potential and pregnancy outcomes of 210

SELECTED ORAL COMMUNICATIONS mosaic embryo transfers - a single center experience
SESSION 68: GENETIC DETERMINANTS OF EMBRYO QUALITY S. Madjunkova1, M. Madjunkov1, R. Antes1, R. Abramov1, S. Chen1,
08 July 2020 Parallel 4 12:00 - 13:00 H. Belakier1, V. Kuznyetsov1, C. Librach1
1
Create Fertility Centre, Reproductive Genetics, Toronto, Canada

Study question: What is the developmental potential and pregnancy outcomes Summary answer: CCs release microvesicles which are internalised from the
of mosaic embryos diagnosed by NGS based PGT-A and are there determinants oocytes and contain miRNAs possibly involved in the acquisition of oocytes’
that predict outcome? developmental competence.
Summary answer: Mosaic embryos have considerable implantation and devel- What is known already: During folliculogenesis oocytes grow and acquire
opmental potential. Type and level of chromosomal aberrations did not impact developmental competence inside the cumulus-oocyte-complex (COC) thanks
implantation and ongoing pregnancy rates for mosaic embryos. to a bidirectional communication with companion CCs. The nature of the
What is known already: Preimplantation genetic testing for aneuploidies(P- CC-determinants contributing to oocytes’ developmental competence remains
GT-A) using NGS in IVF has increased rates of implantation per transfer, but at poorly understood. We recently showed that when mouse CCs-free antral
the same time has increased mosaic embryo detection to ~20%. Current rec- oocytes are cultured to MII with a feeder layer (FL) of CCs isolated from devel-
ommendation is to consider mosaic embryos for transfer if there are no euploid opmentally competent surrounded-nucleolus (FL-SN-CCs) or incompetent not-sur-
embryos left. The evidence is limited on the developmental potential, implanta- rounded-nucleolus (FL-NSN-CCs) oocytes, the former acquire developmental
tion and birth outcomes of mosaic embryos. Some studies have suggested that competence to blastocyst, whereas the latter undergo developmental arrest.
level and type of chromosome aberrations may determine the implantation The hunt for FL-SN-CCs determinants points to the exocytosis of microvesicles
potential of mosaic embryos. This study aims to evaluate the implantation and containing functional molecules and to their fusion with or endocytosis by
pregnancy outcomes after mosaic embryo transfer detected at NGS resolution. oocytes.
Study design, size, duration: This is a single centre retrospective cohort Study design, size, duration: Between March-2019 and December-2019,
study where we analysed the clinical from 210 single mosaic embryo transfers SN-FL-CCs and NSN-FL-CCs were prepared from CD1 mouse SN-COCs and
from Jan. 2015-Dec. 2019. NSN-COCs, respectively, to investigate the release of microvesicles into the
Participants/materials, setting, methods: Clinical outcomes (implantation culture medium and their internalisation into CCs-free mouse oocytes. The
rate, ongoing pregnancy rate, miscarriage rate and birth outcomes) from 210 FL-CCs-derived microvesicles-specific miRNA content was screened from three
single mosaic embryo transfers from Jan. 2015-Dec. 2019 were available for cohorts of SN-FL-CCs and three of NSN-FL-CCs by RNA-sequencing.
analysis. NGS PGT-A analysis was performed using VeriSeq PGS (Illumina) kits. Sequencing was performed also on human FL-CCs produced between
CNV analysis was done using BlueFuse software. The sensitivity for mosaicism November-2018 and May-2019 from 6 IVF patients.
detection was established at 20%, and aberrations considered clinically relevant Participants/materials, setting, methods: Secreted-microvesicles were
were ≥10Mb in size and with ≥25% mosaicism. isolated by ultracentrifugation, sized and quantified with Nanosight. FL-SN-CCs
Main results and the role of chance: Overall implantation rate for mosaic were labelled with PKH67, co-cultured with CCs-free mouse antral oocytes and
embryos was 53.3%. Of the 210 transferred mosaic embryos 80.9% had mosaic microvescicles internalization was assessed by confocal-microscopy. Next
levels ≥25%-50% (Group 1) (58.6% with segmental mosaic (SM) gain or loss, Generation RNA-sequencing, followed by bioinformatics analysis, was conducted
41.4% with whole chromosome mosaic (WCM) gain/loss) and 19.1% had to screen FL-CCs-derived microvesicles-specific miRNAs in i) mouse FL-SN-CCs
mosaic levels of 50-70% (Group 2) (72.5% with SM and 27.5% with WCM). versus FL-NSN-CCs cohorts, and ii) human FL-CCs obtained from 6 maternal
There was no statistical difference between Group 1 and Group 2 embryos in age-matched cohorts of developmentally-competent (blastulation-rate per
implantation rates (IR) 55.3% vs 45%, ongoing pregnancy rates (OPR) 46.5% COC>30%, n=3) versus less-competent (blastulation-rate per COC<30%, n=3)
vs 30%, and miscarriage rates (MR) 11.8% vs 10% respectively. Two pregnancies oocytes.
2(1.2%) from Group 1 were ectopic. 70.9% of the ongoing pregnancies were Main results and the role of chance: After 15hr of maturation of mouse
from embryos diagnosed with SM (47.7% segmental losses, 28.4% segmental CCs-free antral oocytes upon FL-SN-CCs, a large number of microvesicles
gains, 23.9% complex segmental gains/losses), while the rest 29.1% had whole (9.65x1010/ml) was detected in the media, 77% were potentially exosomes
chromosome aberrations-WCA (24% trisomies, 48% monosomies and 28% (129.7nm mean diameter and positive to Alix-targeted labelling).
complex trisomy/monosomy). Similar distribution of SM and WCM were Confocal-microscope analysis on three separate experiments showed that a
observed among miscarried embryos. There was no predictive outcome based massive number of CCs-derived microvesicles passed through the zona pellucida,
on the chromosome involved in the SA mainly because of the small sample size. while only a few could penetrate the ooplasm.
Birth outcome is available for 72 ET and healthy babies were delivered in The comparison between the sequencing of miRNAs isolated from microve-
all cases. sicles released by mouse FL-SN-CCs versus FL-NSN-CCs highlighted 325 differ-
Limitations, reasons for caution: This is one of the largest studies presenting entially expressed miRNAs, 21 of which up-regulated (top-5: mmu-miR-5112,
pregnancy outcomes after mosaic embryo transfer, however the sample size is mmu-let-7a-1-3p, mmu-let-7c-2-3p, mmu-miR-28c, mmu-miR-28a-5p) and 23
still limiting the ability to correlate specific chromosomal aberrations with preg- down-regulated (top-5: mmu-miR-712-5p, mmu-miR-1195, mmu-miR-410-3p,
nancy outcomes. The sensitivity and specificity to detect mosaicism in our study mmu-miR-122-5p, mmu-miR-342-5p). Pathway analysis (conducted with the
applies to the assay used to diagnose mosaicism (NGS,VeriSeq-PGS-Kit). DIANA miRpath v3 software using the micro-T-CDS database of predicted
Wider implications of the findings: Our findings provide evidence that targets and the “genes union” merging algorithm) showed 27 and 42 pathways
mosaic embryos develop in healthy babies and support the hypothesis that low potentially controlled from the up-regulated and down-regulated miRNAs,
level mosaicism in early embryonic development may be a physiological phe- respectively.
nomena. Our results will aid to better genetic counseling on the risks of mosaic The comparison between miRNA sequencing data produced from microve-
embryo transfer, supporting their consideration for transfer. sicles secreted by human CCs of developmentally-competent or less-competent
Trial registration number: no trial registartion cohorts of oocytes highlighted 370 miRNAs, 25 of which differentially expressed,
6 up-regulated (hsa-miR-3675-3p, hsa-miR-4741, hsa-miR-640, hsa-miR-133a-5p,
hsa-miR-1909-3p, hsa-miR-6852-3p) and 19 down-regulated (top-5: hsa-miR-
O-283 Cumulus cells secrete microvesicles carrying miRNAs
6866-5p, hsa-miR-1303, hsa-miR-7110-5p, hsa-miR-2278 and hsa-miR-4639-5p).
that might contribute to the acquisition of oocyte developmental
The pathway analysis showed 12 and 8 pathways potentially controlled from the
competence
up-regulated and down-regulated miRNAs, respectively.
M. Zuccotti1, D. Cimadomo2, F. Cavalera1, L. Dusi3, S. Bertelle3, Limitations, reasons for caution: CCs-derived microvesicles internalization
B. Iussig3, F. Ubaldi, Maria2, L. Rienzi2, S. Garagna1 should be confirmed also in human oocytes. The sequencing data should be
1
University of Pavia, Dipartimento di Biologia e Biotecnologie ‘Lazzaro validated via more specific techniques, like qPCR and immunofluorescence, and
Spallanzani’, Pavia, Italy ; with a larger samples size of CCs. Possibly, CCs should be isolated from single
2
Clinica Valle Giulia, Genera centers for reproductive medicine, Rome, Italy ; oocytes, rather than cohorts.
3
Genera Veneto, Genera centers for reproductive medicine, Marostica, Italy Wider implications of the findings: The identification of CCs-secreted
microvesicles carrying miRNAs that are internalized by developing oocytes and
Study question: What do the cumulus cells (CCs) secrete during the germi- might govern the acquisition of developmental competence will have numerous
nal-vesicle to metaphase II (GV-to-MII) transition that might mediate the acqui- implications to improve: i) in-vitro-maturation, ii) treatment of patients suffering
sition of oocytes’ developmental competence? from reduced maturation rates (cancer, polycystic-ovarian-syndrome,

poor-ovarian-reserve and/or oocyte-maturation-arrest) iii) supplementation of was conducted in a single center. Finally, DNA fragmentation index assessment
culture media. is not included herein, albeit representing a valuable complimentary tool to
Trial registration number: not applicable evaluate sperm quality.
Wider implications of the findings: Different abnormal SA parameters may
O-284 Abnormal fertilization in ICSI and association with lead to different IVF cycle outcomes. Oligozoospermic semen samples featuring
abnormal semen parameters: A retrospective observational study at least one more abnormal parameter are associated with compromised blas-
on 1855 cases tocyst formation rates. Oligoasthenozoospermic and OAT patients present with
A. Pappas1, K. Pantos1, K. Sfakianoudis1, E. Maziotis2, A. Rapani2, lower clinical pregnancy rates. Depending on SA parameters a different IVF
E. Karantzali2, S. Grigoriadis2, P. Tsioulou2, P. Giannelou2, treatment strategy may be required.
T. Vaxevanoglou1, M. Chronopoulou1, M. Koutsilieris2, Trial registration number: Not applicable
M. Simopoulou2
1
Center for Human Reproduction, Genesis Athens Clinic, Athens, Greece ;
2
National and Kapodistrian University of Athens, Department of Physiology-
Medical School, Athens, Greece SELECTED ORAL COMMUNICATIONS
SESSION 69: BIOMARKERS OF DEVELOPMENTAL
Study question: Is abnormal fertilization following ICSI related to abnormal COMPETENCE
semen analysis (SA) parameters?
08 July 2020 Parallel 1 14:00 - 15:15
Summary answer: A SA entailing more than two abnormal parameters-par-
ticularly for oligozoospermia-may compromise normal fertilization, cleavage and
blastocyst formation rates, along with clinical pregnancy rates.
What is known already: Intracytoplasmic sperm injection (ICSI) efficiently
addresses male factor infertility. However, even when ICSI is employed, a positive O-285 An oocyte assessment tool using machine learning;
fertilization outcome may not be guaranteed. Undoubtedly, occurrence of abnor- Predicting blastocyst development based on a single image of an
mal fertilization patterns, mainly referring to abnormal pronuclei (PN) patterns oocyte
of extra or missing pronuclei, merits further investigation. This may be height- D. Nayot1, J. Meriano2, R. Casper1, K. Alex3
ened especially with regards to management of couples presenting with a trend 1
TRIO Fertility, Reproductive Endocrinology and Infertility, Toronto, Canada ;
for abnormal fertilization patterns. Identifying potential causative associations 2
TRIO Fertility, Embryology, Toronto, Canada ;
between specific abnormal SA parameters, their respective combination, and 3
FutureFertility, Embryology, Toronto, Canada
fertilization outcome following ICSI may be of added value for the IVF laboratory.
Study design, size, duration: This retrospective observational study included Study question: Can an Artificial Intelligence (AI) based image analysis tool
1855 ICSI cycles performed between 2014 and 2018. Medical records from predict the blastocyst development potential of oocytes better than the current
couples submitted to a first fresh autologous ICSI cycle between 2014 and 2019 standard?
were retrieved and included in the study. The principal inclusion criteria were Summary answer: Our AI image analysis tool outperformed all 17 embryol-
male age of 18-50 years old, female age of the partner 18-40 years old, and ogists by an average of 21.2% in predicting blastocyst development and, unlike
employment of the gonadotropin-releasing hormone (GnRH) long agonist stim- the embryologists, was 100% reproducible.
ulation protocol. What is known already: There are no morphological features of oocytes
Participants/materials, setting, methods: The participants were divided that have been validated to have prognostic value for further developmental
into groups, according to pathologies identified following SA, and the respective competence (Rienzi 2011). Currently there is no standardized or accepted visual
combinations of individual abnormal parameters, based on WHO criteria. Cases oocyte scoring system (Alpha 2011), and therefore limited feedback about
of normal SA served as the control group. Presence of two PN and extrusion oocyte quality is available to patients and clinicians.
of the second polar body indicated normal fertilization. Embryo transfer was Deep learning offers promise for the automation and standardization of
performed on day three or five, based on the number and quality of cleavage embryo quality assessment. There are several studies using machine learning in
stage embryos. R programming language was used for the statistical analysis. an attempt to automate embryo grading and improve embryo selection, but this
Main results and the role of chance: From the total of 1855 men, 162 is the first clinical application focusing on oocytes.
presented without any abnormal observations in the SA, serving as the control Study design, size, duration: The Violet (an AI image analysis tool) was
group. Two-hundred and ninety-five were diagnosed with asthenozoospermia, created with convoluted neural networks based on a retrospective dataset
229 with oligozoospermia, 83 with teratozoospermia, 232 with oligoastheno- (n=17,659 oocyte images). It’s able to predict fertilization and blastocyst devel-
zoospermia, 213 with oligoteratozoospermia, 284 with asthenoteratozoosper- opment with 91.2% and 63% accuracy respectively in an unbalanced dataset.
mia and 356 with oligoasthenoteratozoospermia (OAT). The results In a balanced test set of 300 oocytes, the Violet outperformed all 17 embry-
demonstrated that SA pathologies affect 2PN formation rate (p<0.0001), fer- ologists, from 8 IVF clinics, in accurately predicting fertilization (71.7% vs
tilization failure (p=0.0001), cleavage rate (p=0.0001) and blastocyst formation 58.9±4.3%; 21.8% increase) and blastocyst development (62.8% vs 52.2±3.7%;
rate (p<0.0001). No correlation was established between abnormalities and 20.2% increase).
1PN or 3PN formation rates. Regarding 2PN rate, the control group presented Participants/materials, setting, methods: N/A
with the highest (0.82±0.05) and the OAT group presented with the lowest Main results and the role of chance: In an unbalanced data set, the Violet
(0.60±0.08). The highest fertilization failure rates were reported in the oligoas- was able to predict fertilization and blastocyst development with 91.2% and 63%
thenoteratozoospermic (0.246±0.0749) and the lowest in the normal group accuracy respectively. It was especially effective at identifying negative cases with
(0.155±0.0511). The lowest cleavage rates were identified for oligoasthenozo- 99% accuracy when the confidence was > 70%
ospermic and oligoasthenoteratozoospermic patients (0.814±0.508 and Two validation studies were performed to compare the Violet to embryolo-
0.78±0.0999). The aforementioned groups along with oligoteratozoospermic gists. A balanced data set of 300 oocyte images was randomly selected from
patients presented with the lowest blastocyst formation rate (0.457±0.043, the test subset: 100 failed fertilization, 100 fertilized but did not reach blastocyst
0.453±0.068 and 0.459±0.035). Regarding clinical pregnancy data, only oligoas- stage, and 100 reached blastocyst stage.
thenozoospermic and oligoasthenoteratozoospermic patients were associated The Violet outperformed all 17 embryologists, from 8 IVF clinics, in accurately
with lower rates compared to the normal group (OR:0.538, 99.3225%CI:0.302- predicting fertilization (71.7% vs 58.9% ± 4.3%; 21.8% increase) and blastocyst
0.956; p-value=0.004 and OR:0.582, 99.3225%CI: 0.344-0.985; p-value=0.005, development (62.8% vs 52.2% ± 3.7%; 20.2% increase).
respectively). Following adjustment for number of blastocysts no statistically In a reproducibility study, 7 of these embryologists underwent the same task
significant difference was observed regarding the clinical pregnancy rate. 2-3 months later. The average accuracy remained close to chance, 53% ± 3.3%
Limitations, reasons for caution: The retrospective nature of this obser- with an intra-observer reproducibility of 81.4% for blastocyst formation, while
vational study may be correlated with bias, along with the fact that the study the Violet was 100% reproducible.

Limitations, reasons for caution: As with all AI image analysis tools, a larger Study question: Does the distribution of F-actin and MYO10 protein differ in
and more diverse data set is necessary to extrapolate findings. A prospective human preantral follicles from patients of different age and ovarian pathology?
multi-centre validation study is currently underway to validate the Violet Summary answer: Advanced age and endometriosis have an impact on the
technology distribution pattern of F-actin in granulosa cells and MYO10 protein in granulosa
Wider implications of the findings: The lack of a visual oocyte assessment cells and oocytes.
criteria is congruent with our findings that 17 senior embryologists were essen- What is known already: Direct markers of oocyte quality described so far
tially unable to predict blastocyst development (average 52.2%). Machine learning include spindle dysfunction and chromosomal misalignment during meiotic mat-
image analysis improves our ability to assess oocyte quality in an instantaneous, uration, whereas markers of the surrounding granulosa cells usually focus on
non-invasive, reproducible and more accurate method than the current standard. cell proliferation and death. Granulosa cells communicate with the oocyte via
Trial registration number: 16325-16:10:0722 actin-rich cytoplasmic extensions termed transzonal projections (TZPs) which
penetrate the zona pellucida and enable the oocyte to be supplied with essential
O-286 Fresh or vitrified oocytes from the same donor cohort: do nutrients and regulatory signals necessary for its growth. MYO10 protein likely
they differentially affect clinical outcomes? regulates filamentous (F-) actin assembly or function in TZPs. However, the
expression and distribution pattern of MYO10 protein and F-actin in women of
J. Maidana1, E. De Martino1, M. Papayannis1, P. Filardi1, C. Bisioli1,
different ages and clinical conditions remains unclear.
G. Terrado2, I. De Zúñiga2, M. Horton2, M. Bianchi2, H. Pettorossi2,
Study design, size, duration: A prospective study was conducted among
N. Passi2, M. Marcelli2, F. Sobral2, M. Gómez Peña1
1
21 women who underwent ovarian surgery and consented to fresh ovarian tissue
Pregna Reproductive Medicine, Embryology Laboratory, Buenos Aires, Argentina ;
2
donation in the period from 2017 to 2019. Ovarian follicles of primordial to
Pregna Reproductive Medicine, Reproductive Medicine, Buenos Aires, Argentina secondary stages were retrieved from 9 women aged 22 to 40 years. Four had
no ovarian pathology; the other five were diagnosed with endometriosis con-
Study question: Do clinical outcomes differ when using fresh or vitrified
firmed by histological analysis.
oocytes from the same donor cohort?
Participants/materials, setting, methods: Fresh tissue samples were
Summary answer: Fresh or vitrified oocytes lead to similar clinical outcomes
immediately delivered for follicle harvesting. Follicles with a morphologically
in an egg donation program.
normal oocyte and non-pyknotic granulosa cells were fixed and stained using
What is known already: The extended use of frozen oocytes in egg donation
phalloidin and anti-MYO10. Confocal microscopy was used to image equatorial
programs rest on the success of oocyte vitrification and the need to optimise
optical sections of each follicle. Parameters measured were the number of
synchronisation between donors and recipients. Oocyte vitrification has proven
MYO10 foci and intensity of F-actin in the granulosa cells and oocytes.
its efficacy, however the impact of thermal and osmotic stress on embryo devel-
Correlation between the oocyte diameter and the structures associated with
opment and clinical results still remains controversial.
TZP development (MYO10 foci and F-actin) was evaluated.
Study design, size, duration: We retrospectively compared embryo devel-
Main results and the role of chance: A total of 93 good-quality follicles
opment (fertilisation, embryo quality and blastocyst formation) and clinical out-
were included in the analysis. Mean oocyte diameter was 32.6 ± 6.8 µm. A
comes (implantation, pregnancy and miscarriage rates) of fresh and vitrified
correlation between oocyte diameter and MYO10 distribution (correlation esti-
oocytes cycles from the same cohort of 34 donors in 117 recipients between
mate (CE) 0.37; 95% confidence interval (95% CI) 0.08-0.61; P = 0.01) as well
January 2017 and December 2018.
as between oocyte diameter and F-actin intensity (CE 0.36; 95% CI 0.07-0.59;
Participants/materials, setting, methods: Recipient cycles were divided
P = 0.008) in granulosa cells was demonstrated in patients of 22-30 years of
into two groups: those who received fresh (n= 75) or vitrified oocytes (n= 42).
age. These correlations for both MYO10 distribution and F-actin intensity with
All oocytes (n= 679) were fertilised by ICSI and transfers were performed at
oocyte diameter were not observed in the group of 35-40 years (P = 0.15, P =
the blastocyst stage. Cycles with frozen embryo transfers or severe male factor
0.16, respectively). The intensity of F-actin in granulosa cells was also correlated
were excluded from this study. The mean number of embryos transferred was
with oocyte diameter in patients with normal ovaries (CE 0.31; 95%CI 0.02-0.55;
similar in both groups (1.01 vs 1.0). Chi-Square test was used as statistical data
P = 0.02). This correlation was not observed in patients with endometriosis
analysis.
(P = 0.83). Strikingly large MYO10 aggregates within the oocyte were significantly
Main results and the role of chance: Each recipient received 5.4 and 6.5
more common in cases with endometriosis compared to those with normal
MII oocytes on average in the synchronous (406 fresh) and the asynchronous
ovaries (14 (41.2%) versus 8 (17.8%); P = 0.02). The absence of correlation of
group (273 vitrified) respectively (p=NS). A total of 323 oocytes were warmed
MYO10 and F-actin distribution with oocyte diameter suggests a disorganization
and 273 (84.5%) survived. Although blastocysts formation rate (58.1 vs 48.4%,
of TZPs development in small growing follicles, which may contribute to poorer
p<0.05) and the rate of usable blastocysts (45.5 vs 35.6%, p<0.05) were higher
follicle quality in older women and patients with endometriosis.
in the fresh oocyte group, no differences were observed between fresh and
Limitations, reasons for caution: Only fresh ovarian samples were included
vitrified oocytes regarding fertilisation rate (80.5 vs 80.2%; p=NS), percentage
in the study. The effect of age and ovarian pathology on the structures associated
of good quality embryos (80.0 vs 86.0%; p=NS), clinical pregnancy (55.0 vs
with TZP development in frozen ovarian tissue is a matter for further studies.
45.0%; p=NS), miscarriage (2.0 vs 10.0%; p=NS) or implantation rates (55.0 vs
Wider implications of the findings: We suggest that MYO10 distribution
50.0%;p=NS).
and F-actin intensity in granulosa cells as well as the presence of MYO10 aggre-
Limitations, reasons for caution: Our study is limited by its sample size and
gates in oocytes can be used for quality assessment of ovarian follicles in women
retrospective design. Oocytes were donated by healthy, young women so these
of different ages and ovarian pathology.
results should not be generalised to other populations.
Wider implications of the findings: Although blastocyst formation was
significantly lower in the group of vitrified oocytes, we didn’t observe differences
O-288 Dynamic oxygen level (5%-2%) during human in vitro
in clinical outcomes. This could mean a possible stress effect or cell pre-condi-
embryo culture significantly improves usable blastulation and
tioning for better stress tolerance. Our study suggests that the reproductive
cumulative live birth rates in in vitro fertilization (IVF)
results should not be affected by donor oocyte vitrification.
Trial registration number: Not applicable S. Brouillet1, A. Andreeva1, A. Gala1, A. Ferrieres1, M. Anav1,
A. Fournier1, V. Loup2, N. Ranisavljevic2, S. Hamamah1
1
University of Montpellier - University hospital of Montpellier - INSERM U1203,
O-287 F-actin and MYO10 protein distribution: a novel tool in
preimplantation genetic testing- assisted reproductive techniques- CECOS,
assessing the quality of human growing follicles
Montpellier, France ;
A. Volodarsky-Perel1, S. Granados Aparici2, T. Tulandi1, 2
University hospital of Montpellier, preimplantation genetic testing- assisted
W. Buckett1, S. Krishnamurthy1, F. Williamson Mansour1, reproductive techniques- CECOS, Montpellier, France
J. Papillon Smith1, H. Clarke1
1
McGill University, Obstetrics and Gynecology, Montreal, Canada ; Study question: Does dynamic oxygen level (5-2%) during in vitro embryo
2
McGill University, Research Institute, Montreal, Canada culture improve IVF outcomes compared to continuous oxygen level (5%)?

Summary answer: Dynamic oxygen level significantly increases total and What is known already: THC, a cannabis plant derivative, is used for medic-
usable blastocyst formation rates as well as cumulative implantation, clinical inal and recreational purposes. It is the third most commonly used substance by
pregnancy and live birth rates in IVF. women of childbearing age, hence knowledge of the effect it has on reproduction
What is known already: In IVF, human embryos are usually cultured under is of utmost importance. THC exerts its effects via receptors of the endocan-
20% or 5% of oxygen. A dynamic oxygen exposure during in vitro culture may nabinoid system and can interfere with its regular functions which are required
represent the most physiologic system as in vivo oxygen tension is ≈5% for early for successful reproduction. Moreover, previous studies have shown THC alters
cleaved embryos in the oviduct and ≈2% for morulas/blastocysts in the uterus. methylation and histone modifications in sperm, brain, and blood cells. The levels
Moreover, ultra-low oxygen tension (2%) increases the expression of antioxidant of endocannabinoids have been measured in follicular fluid obtained during
enzymes in mouse embryos and enhances the proliferation rate of human tro- oocyte retrieval and are implicated in controlling folliculogenesis.
phoblast, protecting the blastocyst against an excess in oxidative stress during Study design, size, duration: Samples were previously biobanked from con-
its high metabolic activity and providing a favorable environment for blastocyst senting patients during IVF oocyte retrieval. Their follicular fluid was analyzed for
viability and implantation potential. patient exposure to phytocannabinoids, and to measure endoCBs. The correspond-
Study design, size, duration: A monocentric retrospective cohort study ing granulosa cells (hGC) were utilized to explore the dynamics of endoCB-Rs and
was performed from June 2014 to March 2019. A total of 120 couples were a methylating enzyme in the follicular niche. Cases were patients with at least one
enrolled. The study was approved by local institutional review board (2019_IRB- PC detected in their follicular fluid, and matched controls were negative for all three
MTP_05-12). All couples underwent one IVF cycle associated with continuous tested PCs (delta9-THC, 11-OH-delta9-THC, 11-COOH-delta9-THC).
oxygen exposure (5% from Days 0-6) and the subsequent IVF cycle associated Participants/materials, setting, methods: We performed liquid chroma-
with dynamic oxygen exposure (5% from Days 0-3 and 2% from Days 3-6) for tography-mass-spectrometry (LC-MS/MS) at the Analytical Facility for Bioactive
in vitro embryo culture. Molecules (SickKids Hospital, Toronto, CA) on FF obtained from dominant fol-
Participants/materials, setting, methods: The first objective was to licles between Jan 2018-July 2019. Cases were matched with controls by age,
evaluate total and usable blastocyst formation rates in both “continuous 5% BMI, AMH and trigger day E2 levels. Levels of endoCB-Rs (CB1R-APC, CB2R-
oxygen exposure” and “dynamic 5-2% oxygen exposure” groups. Blastocysts Alexa488; R&D Systems) and of the methylating enzyme DNMT3b (DNMT3b-
were scored using the Gardner grading system. The secondary objective was APC, Miltenyi Biotec), in corresponding hGC were assessed by flow cytometry
to evaluate cumulative implantation, clinical pregnancy and live birth rates (MACSQuant 10). Median fluorescence intensities were calculated (FlowJo10).
obtained after fresh and frozen-thawed morula/blastocyst transfers in both Main results and the role of chance: Out of 244 samples analyzed, 17 tested
“continuous 5% oxygen exposure” and “dynamic 5-2% oxygen expo- positive for PCs (6.5%) and these were matched with 15 samples that were negative
sure” groups. for all PCs tested (controls). The percentage of positive samples among our pop-
Main results and the role of chance: The maternal age (35.3 ± 4.4 vs. 33.5 ulation increased from 3.8% to 13.3% after legalization of cannabis in Canada in
± 4.6, p=0.003) and the total number of IVF cycles (2.7 ± 1.0 vs. 1.6 ± 0.9, October 2018 (chi²; p=0.02). The concentrations of PCs did not change following
p< 0.0001) were significantly higher in the “dynamic 5-2% oxygen exposure” legalization. There was no significant effect of PCs on maturation rate (mean case
group. All other clinical and biological parameters were similar in both groups. maturation rate 75.9% vs. control maturation rate 74.4%). In addition, the levels of
The blastocyst formation rate (54.8% vs. 44.4%, p=0.0007) and usable blastocyst endocannabinoids (including AEA, 2-AG, among others (10 in total)) did not differ
formation rate (32.8% vs. 21.8%, p<0.0001) were both significantly higher in the significantly between cases and controls. Next, the cell surface protein expression
“dynamic 5-2% oxygen exposure” group than in “continuous 5% oxygen expo- levels of endoCB-Rs in corresponding hGC was examined. Overall CB2R expression
sure” group. Implantation, clinical pregnancy and live birth rates seem increased was higher than CB1R, however, exposure to PCs did not significantly alter this
in the “dynamic 5-2% oxygen exposure” group compared to the “continuous expression. Taken together, this indicates that PC exposure might not alter endo-
5% oxygen exposure” group after fresh morula/blastocyst transfers, but the cannabinoid signaling significantly. DNMT3b, a DNA methylation enzyme, is involved
differences didn’t reach significance. The cumulative implantation (33.1% vs. in de-novo methylation and is crucial for epigenetic integrity. Interestingly, exposure
16.8%, p=0.016), clinical pregnancy (36.8% vs. 15.2%, p=0.006), and live birth of hGC to PC decreased DNMT3b positive events, indicating that cannabis expo-
(23.2% vs. 8.4%, p=0.004) rates were significantly higher in the “dynamic 5-2% sure appears to affect epigenetic machinery at the protein level.
oxygen exposure” group than in “continuous 5% oxygen exposure” group, Limitations, reasons for caution: Our study is limited by lack of details
respectively. regarding mode, frequency, and timing of PC consumption. A further limitation
Limitations, reasons for caution: This is a retrospective study. The selection is the small sample size, which warrants larger-scale studies to validate our find-
of embryos to be transferred and cryopreserved was based on embryo mor- ings. Finally, future studies should also focus on the effects of PC exposure on
phology criteria that might be different in other clinics. developing oocytes.
Wider implications of the findings: Improved IVF outcomes were obtained Wider implications of the findings: To our knowledge, this is the first study
in the same couples when dynamic oxygen exposure (5-2%) was used compared measuring cannabis derivatives in FF by LC-MS/MS. We show that consuming can-
to continuous oxygen exposure (5%), in spite of the negative impact of increased nabis does not appear to affect the endocannabinoid system in the developing follicle.
maternal age. Dynamic oxygen exposure (5-2%) may improve blastocyst forma- Lastly, we have implicated cannabis in disrupting epigenetic mechanisms in hGCs.
tion and implantation potential by preventing oxidative stress during IVF. Trial registration number: N/A
Trial registration number: 2019_IRB-MTP_05-12
O-289 Detection of Cannabis in Follicular Fluid and its impact on

Oocyte Quality, Endocannabinoid Signaling, and Epigenetics SELECTED ORAL COMMUNICATIONS
N. Fuchs Weizman1, B. Wyse2, P. Szaraz3, M. Defer2, S. Jahangiri4, SESSION 70: OVARIAN STIMULATION STRATEGIES IN IVF
C. Librach5 AND IUI
1
University of Toronto, Department of Obstetrics, Toronto, Canada ; 08 July 2020 Parallel 2 14:00 - 15:15
2
CReATe Fertility Centre, Research - Ovarian Biology Group, Toronto, Canada ;
3
CReATe Fertility Centre, Research - Cardiac Stem Cell Group, Toronto, Canada ;
4
CReATe Fertility Centre, CReATe BioBank, Toronto, Canada ;
5
O-290 Impact of controlled ovarian stimulation on vaginal and
CReATe Fertility Centre, Medical Director, Toronto, Canada endometrial microbiota in IVF cycles: a pilot study
Study question: How do phytocannabinoids (PC) in follicular fluid (FF) affect A.R. Carosso1, A. Revelli1, G. Gennarelli1, S. Canosa1, S. Cesarano1,
the dynamics of endocannabinoid receptors (endoCB-Rs) and of methyl-trans- N. Brunod1, C. Scarafia1, S. Leoncini1, L. Boatti2, E. Zanotto3, F.
ferase expression in the surrounding granulosa cells? Sidoti3, R. Cavallo3, C. Benedetto1
1
Summary answer: Marijuana exposure does appear not alter the dynamics Obstetrics and Gynecology 1U- Physiopathology of Reproduction and IVF Unit,
of the endocannabinoid system; however, it changes the epigenetic machinery in Department of Surgical Sciences- Sant’Anna Hospital- University of Torino, torino,
the follicular niche. Italy ;

2 1
University of Piemonte Orientale, SmartSeq s.r.l.-, Novara, Italy ; Monash University, Department of Obstetrics and Gynaecology, Clayton,
3
Division of Virology- University of Turin- Città della Salute e della Scienza di Australia ;
2
Torino, Public Health and Pediatrics Department, Turin, Italy Amsterdam UMC, Centre for Reproductive Medicine, Amsterdam, The
Netherlands ;
3
Augusta University, Department of Obstetrics and Gynecology, Augusta, U.S.A. ;
Study question: Does controlled ovarian stimulation (COS) modify the vaginal 4
Penn State College of Medicine, Department of Obstetrics and Gynecology,
and endometrial microbiota of women undergoing in vitro fertilization (IVF)?
Hershey, U.S.A. ;
Summary answer: COS modifies vaginal and endometrial microbiota inducing 5
UZ Leuven, Leuven University Fertility Center, Leuven, Belgium ;
a reduction of Lactobacillus abundance and an increased heterogeneity, leading 6
Gazi University Faculty of Medicine, Department of Obstetrics & Gynecology,
to the potential growth of pathogenic bacteria.
Ankara, Turkey ;
What is known already: COS is associated with raising estrogen levels and 7
Rijnstate Hospital, Department of Obstetrics and Gynecology, Arnhem, The
several studies report improved pregnancy rate when frozen embryos transfers
Netherlands ;
are performed, probably due to a better endometrial receptivity. Although 8
Hospices Civils de Lyon, Service de Biostatistique-Bioinformatique, Lyon, France
changes of the vaginal microbiota depending on hormone levels during a men-
strual cycle have been demonstrated, the impact of raising estrogens levels
associated with COS on the female microbiota is unknown. Furthermore, the Study question: Does ovarian stimulation with gonadotrophin or letrozole
effects of the presence of an active vaginal and endometrial microbiota on the increase live birth rate compared to clomiphene citrate (CC) in couples with
outcomes of IVF treatments are still controversial. unexplained or mild male factor infertility undergoing intrauterine
Study design, size, duration: In this pilot study, 15 women (age, 29-42 years) insemination (IUI)?
undergoing IVF or ICSI treatment attending the Reproductive Physiopathology Summary answer: Ovarian stimulation with gonadotropins results in higher
Center of the S. Anna Hospital in Turin (Italy) were included between July 2018 live birth rate compared to CC in couples undergoing IUI while evidence on
and October 2019. letrozole versus CC is insufficient.
Participants/materials, setting, methods: Patients underwent vaginal and What is known already: Ovarian stimulation in IUI aims to increase pregnancy
endometrial microbiota analysis in two moments: rates by increasing the number of dominant follicles. Ovarian stimulation can be
achieved with CC, letrozole and gonadotropins. Today IUI with ovarian stimu-
lation is the main source of multiple pregnancies. Multiple pregnancies carry the
- PRE-COS ANALYSIS: simulation of embryo transfer plus a vaginal swab risk of higher morbidity to mother and child. Individual participant data
were performed in the luteal phase of the cycle preceding COS. meta-analysis (IPD-MA) is considered as the gold standard for evidence synthesis
- POST-COS ANALYSIS: at the time of fresh embryo(s) transfer. which provides accurate assessments of outcomes from primary randomised
controlled trials (RCTs) and allows additional analyses for time-to-event
outcomes.
The distal extremity of the catheter tip and the vaginal swab were analyzed
Study design, size, duration: We performed an IPD-MA of relevant RCTs.
by 16S rRNA gene sequencing. To characterize the samples, the Shannon diver-
We searched PubMed, MEDLINE, EMBASE, CENTRAL and the Clinical Trial
sity index (SDI) was used.
Registration Database indexed up to 16 August 2018. No language restrictions
Main results and the role of chance: The genus most present in the PRE-
were applied.
COS vaginal samples was that of Lactobacillus, as in POST-COS analysis, although Participants/materials, setting, methods: We included RCTs that com-
with lower abundance (71.5 ± 40.6% and 61.1 ± 44.2 %, respectively). At the pared ovarian stimulation with CC, letrozole or gonadotrophins to each other
same time, an increase of pathogenic species, such as Prevotella (3.5 ± 8.9% Vs in IUI among couples with unexplained infertility or mild male factor infertility.
12 ± 19.4%) and Atopobium (5.7 ± 10.6% Vs 5.6 ± 9.4%), was observed. In the We contacted the authors of the identified trials to join and share their IPD. The
endometrium, the genus most represented was that of Lactobacillus, although primary effectiveness outcome was live birth and the primary safety outcome
the concentrations decreased between the two analyses (27.4 ± 34.5 % and was multiple pregnancy. The secondary outcomes included clinical pregnancy
25 ± 29.9 %, PRE-COS and POST-COS respectively), with a simultaneous and time to ongoing pregnancy leading to live birth.
increase of Prevotella and Atopobium (3.4 ± 9.5% Vs 4.7 ± 7.4 % and 0.7 ± 1.5
Main results and the role of chance: We identified 22 eligible RCTs (4624
Vs 5.8 ± 12 %). The Shannon indices evaluated at the PRE-COS and POST-COS couples). Authors from 6 RCTs provided IPD. After further excluding 411 couples
analysis were significantly different, indicating an effect of COS on vaginal and with infertility due to anovulation or other factors, we included 2,299 couples
endometrial microbiota diversity (p< 0.001 for both sites). Furthermore, the in this IPD-MA.
presence of a Lactobacillus dominated microbiota at POST-COS analysis in the Compared to IUI with CC, IUI with gonadotrophins increased live birth rate
endometrium was associated with a greater thickness at ultrasound monitoring (RR 1.28, 95%CI 1.06-1.55, 5 RCTs) and clinical pregnancy rate (RR 1.22, 95%CI
on the day of ovulation trigger (12 ± 0.8 mm Vs 8.8 ± 0.9 mm, p<0.005). 1.06-1.40, 6 RCTs), while it reduced time to ongoing pregnancy leading to live
However, an impact on pregnancy rates of vaginal and endometrial microbiota birth (HR 1.38, 95%CI 1.15-1.66, 5 RCTs). Data between the two groups on
was not observed. multiple pregnancy were inconclusive (RR 1.55. 95%CI 0.57-4.22, 5 RCTs). RCTs
Limitations, reasons for caution: Our research is a pilot study and it is with IPD and RCTs without IPD showed similar results in this comparison.
limited by the restricted number of patients included. Furthermore, the pres- One RCT compared letrozole versus CC in IUI. The evidence on IUI with letro-
ence of bacteria in the endometrium forming an active microbiota is still zole versus CC was inconclusive on live birth (RR 0.80, 95%CI 0.59-1.10), clinical
discussed. pregnancy (RR 0.79, 95%CI 0.60-1.04), multiple pregnancy (RR 1.13, 95%CI
Wider implications of the findings: Our results indicate that vaginal and 0.44-2.89) and time to ongoing pregnancy leading to live birth (HR 0.78, 95%CI
endometrial microbiota undergo relevant modification after COS. COS induces 0.55-1.11). RCTs with IPD and RCTs without IPD showed inconsistent results
higher instability at both levels that could justify the negative reproductive and on clinical pregnancy (group difference p=0.01) and none of the RCTs without
obstetric outcomes of a proportion of IVF treatments supporting the diffusion IPD reported live birth in this comparison.
of the “freeze all strategy”. Limitations, reasons for caution: The evidence between letrozole and
Trial registration number: not applicable CC are based on one RCT sharing the IPD. The difference between RCTs
with and without IPD may be due to the differences in study population or
O-291 Ovarian stimulation strategies in intrauterine insemination study quality.
for unexplained or mild male factor infertility – an individual Wider implications of the findings: Ovarian stimulation with gonado-
participant data meta-analysis trophins should be considered as the first-line treatment in couples with unex-
R. Wang1, N.A. Danhof2, M.P. Diamond3, R.S. Legro4, P. Karen5, M. plained or mild male infertility undergoing IUI, if intracycle monitor is available
Erdem6, T. Dankert7, E. Rene8, F. Van der Veen2, B.W. Mol1, M. Van and extra costs are acceptable. There is an urgent need for further RCTs to
Wely2, M.H. Mochtar2, *. On behalf of the IUI IPDMA compare letrozole and CC in IUI to confirm existing evidence.
Collaboration2 Trial registration number: CRD42017053966

O-292 In vitro maturation versus in vitro fertilization in women Summary answer: The egg collection performed after 36 hours of triptorelin
with high antral follicle count: a cost-effectiveness analysis acetate administration allows obtaining a higher number of metaphase II
alongside a randomised clinical trial (MII) oocytes
H. Vu Ngoc Anh1,2, N. Loc Minh Tai1,2, H. Tuong Manh1,2, D. Vinh What is known already: Both GnRH agonists (GnRHa) and hCG have proven
Quang1,2, W. Rui3, N. Robert J4, M. Ben3, V. Lan Ngoc1,2,5 to be effective in achieving ultimate oocyte maturation. Nevertheless, their
1 mechanism of action is different, and the hormonal dynamics of the ovulatory
IVFMD, My Duc Hospital, Ho Chi Minh City, Vietnam ;
2 peak and the molecular profiles differ between them. GnRHa faithfully repro-
HOPE Research Center, My Duc Hospital, Ho Chi Minh, Vietnam ;
3 duces the physiological characteristics of natural cycles. Therefore, the accurate
Monash University, Department of OB/GYN, Melbourne, Australia ;
4 time to achieve oocyte maturation could be different between the two drugs.
Robinson Research Institute, University of Adelaide, Adelaide, Australia ;
5 Consequently, the optimum moment to perform the follicular puncture after
University of Medicine and Pharmacy at HCM City, Department of OB/GYN, Ho
the GnRHa administration could be different from the 36 hours described for
Chi Minh, Vietnam
hCG. A study of the optimum time interval between ovulation triggering and
Study question: Is in vitro maturation (IVM) more cost-effective than in vitro follicular puncture is crucial
fertilization (IVF) in women with high antral follicle count (AFC)? Study design, size, duration: Randomised, controlled, single-blind clinical
Summary answer: IVM should be considered as cost-effective if society is trial carried out in a university-based IVF unit between September 2014 and
willing to accept €8,847 for foregoing an additional live birth. December 2017. Patients undergoing ovarian stimulation with FSH in a short
What is known already: Both IVF and IVM are treatment options in antagonist protocol were randomized to undergo egg collection after 30, 36
women with high AFC undergoing ART, mainly anovulatory women not respond- or 40 hours after ovulation trigger with 0.2 mg of triptorelin acetate. In patients
ing to ovulation induction. IVM has no risk of ovarian hyperstimulation syndrome undergoing fresh embryo transfer, luteal phase support was achieved by admin-
(OHSS) and is potentially more patient friendly. However, the cost-effectiveness istering 1500 IU of hCG and 400 mg of vaginal micronized progesterone
of IVM versus IVF has not been assessed. Participants/materials, setting, methods: Women aged 18-37 years with
Study design, size, duration: This cost-effectiveness analysis was based on a baseline FSH <10mIU/ml, AMH 5-45pmol/l, AFC 6-24 were included in the
data from a randomised clinical trial at IVFMD, My Duc Hospital, HCMC, Vietnam study. Patients with at least 5 follicles ≥16mm were randomized. Egg collections
(NCT03405701). Between December 2017 to April 2019, women were ran- were done separating follicles measuring more and less than 16mm. Embryo
domly assigned to one cycle of IVM or one cycle of IVF at IVFMD. The effec- transfer was performed according to usual clinical practice. The main outcome
tiveness measure for the CEA was the cumulative live birth rate after one variable was the number of MII oocytes retrieved and the number of MII oocytes
completed cycle including subsequent cryo-cycles within 12 months following retrieved per follicle measuring > 16mm the day of trigger
randomisation. Main results and the role of chance: A total of 121 patients were random-
Participants/materials, setting, methods: We collected data on ized to undergo ovulation triggering and egg collection after 30h (n=41), 36h
resource use related to treatment, medication, pregnancy from case report (n= 42) or 40 hours (n=38). The proportion of MII retrieved was significantly
forms. The primary economic analysis was performed from a health care lower after 30h of Decapeptyl administration in comparison with 36h and 40h
perspective. We measured costs relating to treatment (medication, compli- follicular aspiration times (30h: 6.63±4.14, 36h: 9.10±4.75 and 40h: 7.26±4.49;
cations, pregnancy, delivery). We additionally did the analysis from a societal p=0.03698). In addition to that, the ratio MII/follicles >16mm was higher 36h
perspective including non-medical costs. We calculated mean costs, effects, after trigger, compared to the 30h group (0.98±0.56 vs 0.73±0.39; p=0.05277).
averaged differences in costs and incremental cost-effectiveness ratios (ICER) MII rates were concordant with molecular analysis results: a significantly down-
using nonparametric bootstrap resampling to assess the effect of uncertainty regulation of genes related to oocyte maturation (AREG, BTC, CYP19A1,
in our estimates. EFNB2, EREG, PHLDA1, RGS2 and UGP2) were found in GCs of punctured
Main results and the role of chance: A total number of 546 women were follicles at 30h. No significant differences regarding pregnancy rates or live birth
randomized to the IVM group (n = 273) or the IVF group (n = 273). Cumulative live rates were found between groups
birth rates after one completed cycle were 112/273 (41.0%) in the IVM group vs. Limitations, reasons for caution: The external validity of this study could
160/273 (58.6%) in the IVF group (RR 0.70, 95%CI 0.59-0.83). OHSS did not be limited to patients undergoing the same type of ovarian stimulation. The
occur in the IVM group versus 2/273 (0.7%) in the IVF group. The mean total costs present study was not designed to detect differences in pregnancy or live birth
per couple were €5,684 for IVM and €7,240 for IVF (Absolute difference €-1,556, rates. Larger studies need to be performed in order to confirm the findings of
95%CI €-1,837 to €-1,274). The ICER for IVM compared with IVF per additional the present study
live birth was €8,847 (95%CI €8,634 - €9,083). Wider implications of the findings: The results of this study contribute to
Limitations, reasons for caution: We only compared one cycle of IVM improving IVF protocols in order to recover the maximum number of MII oocytes
versus one cycle of IVF. As data were collected from a single centre, unit costs and therefore, enhancing pregnancy rates. In addition, the use of GnRHa for
obtained might be less representative in other settings. ovulation induction would decrease the risk of OHSS associated with hCG
Wider implications of the findings: IVM is less effective, but less expensive stimulation protocols
than IVF, therefore, the use of IVM in women with high AFC depends on society’s Trial registration number: NCT02244151
willingness to accept foregoing an additional live birth.
O-294 Duration of infertility and IVF outcomes: analysis of 252
359 IVF cycles
O-293 Evaluation of Time Interval Between Ovulation Trigger Y. Beebeejaun1, T. Copeland2, I. Sarris1, M. Kamath3, S. Sunkara4
With Triptorelin Acetate and Oocyte Retrieval in IVF Cycles. A 1
King’s Fertility, Women’s Health, London, United Kingdom ;
Randomized Controlled Trial 2
University of California Los Angeles, Department of Health Policy &-
C. Díaz-García1, N. Garcia Camuñas2, A. Marzal3, M.J. Soriano4, J. Management-, Los Angeles, U.S.A. ;
Subira4, A. Garcia belda4, A. Pellicer5 3
Christian Medical College-, Department of Reproductive Medicine, Vellore, India ;
1
IVI London, Reproductive Medicine, London, United Kingdom ; 4
King’s College London, Women’s Health, London, United Kingdom
2
Hospital Universitario La Fe, IVF Unit, Valencia, Spain ;
3
IVI Valencia, Reproductive medicine, Valencia, Spain ; Study question: Does infertility duration affect clinical and perinatal outcomes
4
IIS La Fe, Reproductive Medicine Research Group, Valencia, Spain ; following IVF treatment?
5
IVI Roma, Reproductive Medicine, Rome, Italy Summary answer: The present study demonstrated an increased risk of low
birth weight when the duration of infertility exceeds 48 months. Further research
Study question: What is the optimal time interval between GnRH agonist validation is warranted.
trigger and follicular aspiration to maximise the number of mature oocytes What is known already: Infertility is associated with adverse reproductive
collected? outcomes and women with infertility have a higher risk of adverse perinatal

outcomes. Furthermore, risk of adverse perinatal outcomes is also higher following Study question: Is male factor infertility associated with an altered incidence
assisted reproductive treatments (ART) compared to spontaneous conceptions. of chromosomal abnormalities of potential relevance to subsequent preimplan-
Globally, when IVF is indicated for infertility, the duration of infertility varies when tation development?
women undergo treatment. There are several factors that influence when women Summary answer: Male factor infertility is associated with increased mosaic
have IVF. These factors could be other failed treatments before resorting to IVF, chromosome abnormalities, a potential consequence of the sperm’s role in
logistical or funding issues. With scant literature on whether infertility length could formation of the first mitotic spindle.
affect IVF outcomes, it is a matter of interest to explore this. What is known already: Infertility affects one in six couples worldwide with
Study design, size, duration: Anonymous data were obtained from the male factors contributing in ~50% of cases. Currently, the relative insensitivity
Human Fertilization and Embryology Authority (HFEA), the statutory regulator of methods used to assess the competence of male gametes represents a sig-
of ART in the UK. nificant limitation in evaluating the likelihood that a sperm sample will produce
Data from 1991 to 2016 comprising a total of 252 493 first fresh IVF ± ICSI viable embryos if used for oocyte fertilisation. The close relationship between
cycles were analysed based on the length of infertility (< 24 months vs 24 to 48 advancing female age and increasing oocyte aneuploidy rates has been well
months vs > 48 months). Multiple births were excluded for analysis of perinatal described. However, the extent to which the male gamete influences the genetics
outcomes. of the early embryo and its subsequent development is less clear.
Participants/materials, setting, methods: Perinatal outcomes of pre-term Study design, size, duration: 331 embryos produced by couples with male
birth (PTB), early PTB (ePTB) and low birth weight (LBW) and very LBW factor (MF) infertility (semen sample concentration <5million/ml with/without
(vLBW) as well as clinical outcomes of live birth rate, miscarriage and stillbirth testicular biopsy or epididymal aspiration) were evaluated, and their cytoge-
were analysed in singleton live births. Occurrence of a live birth at < 37 weeks netic status retrospectively compared to that of 1245 embryos derived from
gestation is defined as a PTB and at < 32 weeks gestation as ePTB. Birth weight infertile couples without MF evidence. Examination involved trophectoderm
< 2500 grams is defined as LBW and < 1500 grams as vLBW. (TE) biopsy followed by next generation sequencing (NGS) for preimplantation
Main results and the role of chance: Logistic regression analysis was per- genetic testing for aneuploidy (PGT-A). NGS enabled the accurate identifica-
formed adjusting for female age category, previous pregnancy, period of treat- tion of mosaic and non-mosaic aneuploidies in blastocysts generated by both
ment, cause of infertility (male factor, tubal disease, ovulatory disorder, patient groups.
endometriosis, unexplained) and number of oocytes retrieved. Our control Participants/materials, setting, methods: 517 couples having IVF with
group was those suffering from infertility for <24 months. PGT-A in 12 clinics participated. 69 patients (average female age 38.2 years)
Clinical outcomes: The live birth rate following single embryo transfer was underwent PGT-A due to MF infertility. The remaining 448 (average female age
25.8% (95% CI 25.6%- 26.2%) for those suffering from infertility for less than 24 40.2 years) underwent PGT-A due to other indications. Patients were divided
months, 24.6% (95% CI 24.3%-25.0%) for those suffering from infertility between into “younger” (MF- 32.8 years, others-33.7 years average female age) and
24 and 48 months and 21.9% (95% 21.6%-22.2%CI) when the duration of “older” (MF- 39.7 years, others- 40.6 years average female age) for data com-
infertility is over 48 months. The relative risk ratio (RRR) of miscarriage was parison. A highly validated targeted NGS approach was employed for chromo-
0.94 (p-value=0.09) and 1.03 (p-value=0.35) when the duration of infertility somal analysis.
was between 24-48 months and >48 months respectively The RRR of stillbirth Main results and the role of chance: More MF generated blastocysts
was 0.87 (p-value=0.44) and 0.88 (p-value=0.11) for the 24-48 months and > were euploid compared with those generated by patients undergoing PGT-A
48 months period respectively. for other indications (49% versus 32%, P<0.001). This is presumably a conse-
Perinatal outcomes: The relative risk of ePTB and vLBW was not seen to quence of lower meiotic error rates in the oocytes of MF couples, due to their
be associated with increased duration of infertility. However, the RRR of PTB lower average female age. Conversely, MF patients generated significantly
was 1.01 (p-value=0.84) and 1.11 (p-value=0.04) for the 24-48 months and >48 (P<0.001) more mosaic blastocysts (38%) compared with the remaining
months respectively. , For LBW, the RRR was 0.99 (p-value=0.78 in the 24-48 patients [17% 95%CI (2.906)], a difference that was detectable in both age
months and 1.13 (p-value= 0.01) in the >48 months cohort respectively. groups. This increased mosaicism incidence suggests that the MF generated
Limitations, reasons for caution: Limitations with observational data would embryos experience more mitotic errors post-fertilisation. No significant
apply to this study including residual confounding. This is the first study to address difference (P=0.118) in the incidence of segmental aneuploidies was observed
this study question. between the two patient groups. Of the 339 abnormalities scored in MF
Wider implications of the findings: The perinatal outcomes of offsprings generated blastocysts, 286 (84%) affected whole chromosomes and 53 (16%)
born following ART techniques is of clinical relevance for patients and clinicians. affected segments. A total of 1853 aneuploidies were identified in the blasto-
Trial registration number: Not applicable cysts generated by the patients undergoing PGT-A for other indications, and
1624 (88%) affected whole chromosomes, with the remaining 229 (12%) being
partial. This finding argues against sperm DNA fragmentation having a possibly
detrimental effect after fertilisation. MF infertility patients had a significantly
SELECTED ORAL COMMUNICATIONS lower no-transfer rate, compared to all remaining patients (16% vs. 39%
SESSION 71: ABOUT HOW SPERM QUALITY AND MALE respectively, P<0.001), presumably a consequence of the higher incidence of
INFERTILITY RELATE TO GENETICS euploid embryos in this group.
Limitations, reasons for caution: Classification was based on TE samples
08 July 2020 Parallel 3 14:00 - 15:15
biopsied from blastocysts during PGT-A. As only a fraction of the cells from
each embryo are assessed, some mosaic embryos may be incorrectly classified
as fully euploid or aneuploid. However, this misclassification is expected to have
O-295 The influence of poor semen parameters on embryonic little impact on the overall conclusions.
chromosome segregation Wider implications of the findings: Poor sperm parameter patients are
E. Fragouli1,2, D. Babariya1, G. Cutts1, V. Lozoya Garcia3, E. predisposed to generate mosaic embryos. This predisposition should be dis-
Fernandez Marcos1, L. Parnell1, A. Symon-Allen1, F. Bronet4, cussed during the counselling of couples considering PGT-A due to MF. Sperm
M. Florensa5, N. Prados6, A. Mercader7, N. Garrido3, D. Wells1,2 centrioles are responsible for the first mitotic spindle organisation. Impaired
1
IVI-RMA Global, Juno Genetics, Oxford, United Kingdom ; centriole function in suboptimal sperm might explain the increased post-fertili-
2
University of Oxford, Nuffield Department of Women’s and Reproductive Health, sation chromosome segregation error rates.
Oxford, United Kingdom ; Trial registration number: Not applicable
3
IVI-RMA Global, IVI Foundation, Valencia, Spain ;
4
IVI-RMA Global, IVI Madrid, Madrid, Spain ;
5
IVI-RMA Global, IVI Barcelona, Barcelona, Spain ; O-296 Single-cell genome-wide bisulfite sequencing for assessing
6
IVI-RMA Global, IVI Seville, Seville, Spain ; the role of DNA methylation in spermatozoa.
7
IVI-RMA Global, IVI Valencia, Valencia, Spain “Abstract withdrawn by the authors”

O-297 Genetic and Epigenetic Characterization of O-298 Average sperm head area: a novel risk factor for sperm
Globozoospermic Men to Tailor Assisted Reproductive Treatment aneuploidy in idiopathic male infertility
D. Tavares1, A. Parrella1, S. Cheung1, P. Xie1, Z. Rosenwaks1, G. M. Ruiz - Jorro1, M. Ferrer-Buitrago2, J.J. Bataller-Sánchez3, A.
Palermo1 Barberá-Alberola3, X. Vendrell-Montón4, C. Calatayud-Lliso5
1 1
Weill Cornell Medicine, The Ronald O. Perelman and Claudia Cohen Center for CREA Medicina de la Reproducción SL, Reproductive Andrology, Valencia, Spain ;
2
Reproductive Medicine, New York, U.S.A. CREA. Medicine de la Reproducción SL, Research and Development, Valencia,
Spain ;
Study question: Can we genomically characterize globozoospermia using 3
CREA Medicina de la Reproducción SL, Andrology Laboratory, Valencia, Spain ;
various biomarkers and nucleic acid sequencing to determine the best repro- 4
Sistemas Genómicos. Grupo Biomédico Ascires, Reproductive genetics, Paterna,
ductive treatment? Spain ;
Summary answer: Globozoospermia is associated with specific gene muta- 5
CREA Medicina de la Reproducción SL, Reproductive Medicine, Valencia, Spain
tions and imbalances. Bioassays can be used to assess this gamete’s capacity to
support embryonic development and tailor treatment. Study question: Is increased average of sperm head area (ASHA) a risk factor
What is known already: Round-headed spermatozoa lack an acrosome and of sperm aneuploidy in patients consulting for infertility?
have a defect in chromatin compaction, primarily diagnosed by transmission Summary answer: Increased ASHA values are associated with sperm aneuploidy
electron microscopy (TEM). The phenotype of globozoospermia is mostly in patients in whom altered meiosis during spermiogenesis is not initially suspected.
related to an autosomal recessive mutation and is frequently associated with What is known already: Sperm aneuploidy test is aimed at patients expe-
familial inheritance. While few globozoospermia genotypes have been identified, riencing recurrent implantation failure (RIF) or pregnancy loss (RPL). Likewise,
fortunately, this rare defect does not always have complete penetrance. We patients with high FSH levels due to a testicular failure are prone to accumulate
applied different bioassays and genomic studies to better characterize this gam- meiotic errors during spermiogenesis. Although sperm aneuploidy is detected
ete’s capacity to support embryonic development, as well as to tailor treatments in up to 15 % of infertile men, the risk of chromosomal aberrations increases
to achieve pregnancy. to 40% in patients with factors such as RIF, RPL or suspected testicular failure.
Study design, size, duration: Semen parameters, sperm ultrastructural Fluorescence in situ hybridation (FISH) is employed to analyze sperm aneu-
details, protamine content, sperm chromatin fragmentation (SCF), sperm cyto- ploidy. This costly test is usually requested after observing repeated reproduc-
solic factor, and sperm aneuploidy were assessed for 12 globozoospermic men. tive failures. Finding early indications to assess sperm aneuploidy remains a
The genome and transcriptome of 3 men were profiled and compared to spec- challenge.
imens from donors with proven fertility. ICSI was performed with or without Study design, size, duration: To determine the cut-off value for ASHA,
assisted gamete treatment (AGT) based on diagnostic results of the bioassays. we initially analyzed a total of 142 patients who showed normal FISH results
Clinical outcomes of conventional ICSI cycles and subsequent ICSI cycles with (control group). The positive predictive value of ASHA was further assessed
AGT were compared. in a first retrospective study, which included a total of 250 patients experiencing
Participants/materials, setting, methods: Semen analysis was performed RIF, RPL or testicular failure (Group 1). The validation of ASHA was assessed
on 12 patients. TEM surveyed acrosomal defects and centrosomal integrity. in a total of 22 patients who did not refer RIF, RPL or testicular failure
Aniline Blue assay quantified protamine content with ≤20% normal threshold. (Group 2).
SCF was measured by TUNEL with ≤15% normal threshold. Sperm cytosolic Participants/materials, setting, methods: Patients enrolled in CREA IVF
factor was determined by PLCζ assay with ≥30% normal threshold. Sperm aneu- program participated in the present study, after signing an informed consent
ploidy was assessed using FISH with <1.6% normal threshold. Genome and form. ASHA cut-off value was established according to the value at percentile
transcriptome were analyzed by NGS and RNA-seq, respectively. AGT was 95. Two semen samples were analyzed per patient according to WHO-V-manual.
performed by exposing both gametes to calcium ionophore. Sperm parameters, such as: sperm concentration, motility, morphology and
Main results and the role of chance: Globozoospermia was defined by the ASHA were evaluated using the CASMA commercial software ISAS. Samples
presence of round-headed spermatozoa exceeding 70%. Twelve globozoosper- were stained employing the Diff-quick kit. FISH analysis (5 chromosomes) was
mic men (35.8±5 years) had a concentration of 34.4±44x106/ml, 24±25% performed using Metafer-4 software.
motility, and normal morphology of 0.1±0.3%. Concurrent testing revealed Main results and the role of chance: ASHA cut-off value was established
abnormal protamine content of 53.8±24%, elevated SCF of 19.4±2%, insufficient as ≥14,8 µm2 in the control group. Group 1 revealed an incidence of altered
cytosolic factor of 6.0±4%, and a high aneuploidy rate of 4.4±5%. Complete FISH of 41,2%. Furthermore, ASHA showed lower variability amongst the
globozoospermia was confirmed in 3 men by TEM and the absence of PLCζ; studied samples (p<0,002) and higher positive predictive value to detect altered
genomic analysis identified 2 gene mutations: DPY19L2 and SPATA16. Epigenetic FISH (73,9%) than other sperm parameters such as concentration (56,5%),
analysis revealed 142 differentially expressed genes compared to the control. total sperm count (58,8%) and morphology (46,7%). To reduce the potential
We also found an overexpression of both mutant DPY19L2 and SPATA16 genes. bias associated with the retrospective data (patients with suspected risk of
To characterize their clinical outcomes, 7 couples (men, 35.8±5 years; women, sperm aneuploidy), the predictive value of ASHA was further validated in a
32.8±5 years) underwent 22 ICSI cycles yielding a fertilization rate of 31% prospective analysis. Sample size was adjusted to detect at least 30 % difference
(85/274), an implantation rate of 13.6% (6/44), and a clinical pregnancy rate from the basal 15 % incidence reported in a global population of infertile
(CPR) of 22.2% (4/18), with 1 pregnancy loss. For the remaining 3 couples with patients. All patients in Group 2 showed normal testicular function, with FSH
complete globozoospermia (men, 37.3±5 years; women, 36.6±6 years), histor- values lower than 10 mUI/ml (4,81 ± 2,83). Sperm parameters in group 2
ical ICSI cycles (n=3) yielded a 44% (8/18) fertilization rate, but no implantation. were: concentration (20,79 ± 26,47 million sperm/ ml), motility (44,95 ±
Once treated by AGT (n=4), there was a significant improvement, with a 50% 14,58) and morphology (1,55 ± 0,74%). The mean ASHA value in group 2 was
fertilization rate (12/24) (P<0.00001), a 22% implantation rate (2/9), and a 15,22 ± 0,64, with an incidence of sperm aneuploidy of 59,09%. This prospec-
33.3% CPR (1/3), resulting in 1 live birth. tive analysis reveals that patients with ASHA values > 14,8 µm2 show a higher
Limitations, reasons for caution: This is the first study that attempts to incidence of chromosomal aberrations in sperm than a global population of
utilize different bioassays and characterize the genome and epigenome of globo- infertile men (p<0,05).
zoospermic men in order to tailor reproductive treatment. While this preliminary Limitations, reasons for caution: The present study evaluates a limited
data is reassuring, the health of the resulting offspring should be investigated. number of patients. Further analysis in a broader population of patients with
Wider implications of the findings: The limitations of semen analysis are idiopathic infertility is needed to confirm the present findings.
more apparent in cases of globozoospermia. This study demonstrates the asso- Wider implications of the findings: ASHA is a sperm parameter which
ciation of this rare teratozoospermia to gene structure and function. indicates the risk of sperm aneuploidy from the first sperm analysis. Its assess-
Ultrastructural assays and genomic studies can be used to characterize this gam- ment contributes to reduce the time to advise PGT-A, and consequently the
ete’s capacity to support embryonic development and to tailor treatments max- time to conceive in some infertile couples. Our findings provide novel insights
imizing reproductive outcome. in the field of androgenetics and male infertility.
Trial registration number: not applicable Trial registration number: Not applicable

O-299 Attributing ICSI Fertilization Failure to the Responsible

Gamete
P. Chung1, S. Cheung1, P. Xie1, A. Parrella1, D. Keating1, Z. SELECTED ORAL COMMUNICATIONS
Rosenwaks1, G.D. Palermo1 SESSION 72: PREGNANCY LOSS: WHAT TO CONSIDER
1
Weill Cornell Medicine, The Ronald O. Perelman and Claudia Cohen Center for 08 July 2020 Parallel 4 14:00 - 15:15
Reproductive Medicine, New York, U.S.A.
Study question: Is there a way to identify and effectively treat the gamete
responsible for complete fertilization failure (CFF) with ICSI? O-300 Results of a highly significant, prematurely halted Dutch
Summary answer: Screening the male gamete for the presence of PLCζ may multicenter randomized double-blinded placebo-controlled
identify the specific gamete responsible for ICSI CFF and guide treatment in study of pretreatment with mifepristone to misoprostol in early
subsequent cycles. pregnancy failure (triple m)
What is known already: ICSI is capable of overcoming defects of the L. Hamel1,2, J. Van den Berg1, S. Coppus3, M. Snijders2, F.
zona pellucida and sperm acrosomal dysfunction, allowing fertilization. Vandenbussche4
However, although rare, CFF with ICSI can occur due to oocyte activation 1
Radboud University Medical Centre, Obstetrics and Gynaecology, Nijmegen, The
deficiencies, either oocyte-related or due to the absence of sperm-activating Netherlands ;
factors. Most treatment approaches of CFF with ICSI described in the lit- 2
Canisius-Wilhelmina Hospital, Obstetrics and Gynaecology, Nijmegen, The
erature involve different methods of oocyte activation (AOA), yielding Netherlands ;
inconsistent results. These approaches may overlook cases of CFF that occur 3
Maxima Medical Centre, Obstetrics and Gynaecology, Veldhoven, The Netherlands ;
despite apparently normal gamete characteristics and often due to ooplasmic 4
Kreiskrankenhaus Mechernich GmbH, Obstetrics and Gynaecology, Mechernich,
dysmaturity. Germany
Study design, size, duration: Over 20 years, couples with ≤10% ICSI fertil-
ization were identified. Seventy-six men were screened for the presence of PLCζ. Study question: Is, in early pregnancy failure (EPF), the sequential combination
Couples with oocyte-related OAD (n=52) underwent subsequent ICSI with of mifepristone with misoprostol superior to misoprostol alone in terms of
tailored stimulation. Those with sperm-related OAD (n=24) were further complete evacuation of the uterus?
screened by mouse oocyte activation test (MOAT) before undergoing subse- Summary answer: In early pregnancy failure, the sequential combination of
quent ICSI with assisted gamete treatment (AGT). Consenting men (n=4) under- mifepristone with misoprostol is highly superior to misoprostol alone in reaching
went a genomic evaluation by NGS. ICSI outcome was compared between complete evacuation of the uterus.
treatment and historical cycles of the same couple. What is known already: In case of EPF, women can choose from either
Participants/materials, setting, methods: A total of 114 couples, with expectant, medical or surgical management. Surgical intervention carries a risk
female partners ≤37 years of age and male partners with a sperm concentration of complications, however medical treatment appears to be a safe alternative.
of ≥1x106/ml, had fertilization rates of ≤10% despite injecting ≥3 oocytes (IRB When the Dutch common practice of at least one week of expectant manage-
0712009553). PLCζ assessment was performed, confirmed by MOAT and DNA/ ment is adhered to, current medical treatment with misoprostol alone has com-
RNA sequencing. In subsequent cycles, couples with oocyte-related OAD were plete evacuation rates just above 50%. Shortly after the start of this trial,
treated by modulating in-vivo/vitro maturation time, while sperm-related OAD Schreiber et al (NEJM 2018) reported in a non-blinded trial that pretreatment
cases were treated by exposing both gametes to calcium ionophore. Clinical with Mifepristone appears to be more successful when treating instantaneously
outcomes were compared. after EPF is diagnosed.
Main results and the role of chance: A total of 114 couples (maternal age, Study design, size, duration: This study is a multicenter, randomized, dou-
33.8±4 yrs; paternal age, 36.9±5 yrs) with a 9.1% ICSI fertilization rate were ble-blinded, placebo-controlled trial executed at 18 hospitals in the Netherlands.
identified. Consenting male partners were screened, and 52 couples were iden- Results of the pre-planned interim-analysis, performed after 50% (N=232) of
tified as having oocyte-related OAD. These couples underwent subsequent ICSI patients were enrolled, led to a premature halt of the study, due to highly sig-
cycles with tailored superovulation, yielding significantly higher fertilization (59.0% nificant outcomes. The study started on June 27th 2018, the 232nd inclusion was
vs. 2.1%; P<0.0001) and clinical pregnancy (28.6% vs. 0%; P<0.0001) rates. Thirty achieved in September 2019. When the decision was made to stop this trial
clinical pregnancies resulted in 25 deliveries and 5 pregnancy losses. A total of prematurely, in total 342 woman had been randomized.
32 babies were born, 15 boys and 17 girls, with no major or minor congenital Participants/materials, setting, methods: Women with a diagnosis of
malformations. EPF between 6-14 weeks gestation were included after at least one week of
Twenty-four couples (maternal age, 35.6±5 years; paternal age, 39.8±6 years) expectant management. They were randomized between pretreatment with
were confirmed as having sperm-related OAD following PLCζ and MOAT tests. mifepristone 600mg or placebo, both followed by two doses of misoprostol
In some men, DNAseq showed a PLCZ1 deletion, corroborating the initial 400μg orally on day three and four. Ultrasonography was performed 15-20
screening. Deletions of genes associated with spermiogenesis and embryo devel- days after treatment and, if required, repeated 4 weeks later to assess suc-
opment (PIWIL1, BSX, NLRP5), and the absence of subacrosomal perinuclear cessful treatment, defined as absence of gestational sac in utero and endome-
theca (PICK1, SPATA16, DPY19L), were also found. In these couples, AGT trial thickness <15 mm.
treatment provided higher fertilization (42.1% vs. 9.1%; P<0.05) and clinical Main results and the role of chance: After performing an interim-analysis
pregnancy (36% vs. 0%; P<0.05) rates compared to the historical cycles. Six of 50% of the pre-calculated required number of patients (n=232), as stated
patients have successfully delivered, with offspring displaying normal develop- in advance in the study protocol, complete evacuation of the uterus was found
ment at 3 years of age. to be 77,1% in the mifepristone group versus 52,9% in the placebo group
Limitations, reasons for caution: In this prospective cohort study, while (p=0.000). After 6 weeks of follow-up, the need for curettage was 10,2% in
we controlled for maternal age-related aneuploidy, confounding factors the mifepristone group compared to 31,9 % in the placebo group (p=0.000).
cannot be excluded with certainty. Although PLCζ and MOAT assays need Because these results met the predefined stopping rule, the Data Safety
further validation, they serve an important purpose in identifying gam- Monitoring Board instructed us to stop the study, since the difference between
ete-specific causes of CFF and in preventing the overuse of calcium iono- the two groups had already been proven beyond doubt, in favor of pretreat-
phore treatment. ment with mifepristone. There were no concerns regarding the safety of the
Wider implications of the findings: This is the first attempt to identify a patients enrolled in the study. At the ESHRE 2020 annual meeting, follow-up
gamete-specific OAD in couples with ICSI CFF. Those with an oocyte-related for all 342 randomized women will be completed, and a full analysis of the
OAD can be treated in a subsequent cycle with a modified superovulation pro- treatment effect can be reported.
tocol, preventing AGT overuse. For those with sperm-related OAD, genomic Limitations, reasons for caution: There is no consensus about the definition
analysis helped to identify the related genes involved. of ‘successful treatment’, or the optimal treatment regimen for EPF regarding
Trial registration number: not applicable dose or routes of administration of medical treatment for EPF.

Wider implications of the findings: In addition to adjustment of the Dutch O-302 First contact at implantation in humans: Endometrial
national guideline concerning EPF, these results contribute to the improvement epithelium induces trophectoderm differentiation to invasive
of medical management of EPF around the world. Especially in low income syncytiotrophoblast.
countries, this will lead to an extensive improvement of both the effectiveness P. Ruane1, L. Parsons1, T. Garner1, P. Babbington1, S. Kimber2,
and accessibility of proper medical treatment of EPF. A. Stevens1, M. Westwood1, D. Brison1, J. Aplin1
University of Manchester, Maternal and Fetal Health, Manchester, United
Kingdom ;
2
University of Manchester, Division of Cell Matrix Biology and Regenerative
O-301 Reproductive performance following application of Medicine, Manchester, United Kingdom
hyaluronic acid gel after dilatation and curettage for miscarriage
in women with at least one previous curettage
A. Hooker1, R. De Leeuw2, J. Twisk3, J. Huirne2 Study question: How does the human embryo breach the endometrial epi-
1
Zaans Medical Center ZMC, Department of Obstetrics and Gynaecology, thelium at implantation?
Zaandam, The Netherlands ; Summary answer: Embryo attachment to the endometrial epithelium induces
2
Amsterdam UMC- location VU University Medical Center- Amsterdam- the the formation of multinuclear syncytiotrophoblast from trophectoderm, which
Netherlands., Department of Obstetrics and Gynecology-, Amsterdam, The goes on to breach the epithelial layer.
Netherlands ; What is known already: A significant proportion of assisted reproduction
3
Amsterdam UMC- location VU University Medical Center- Amsterdam- the Netherlands., treatments fail due to unsuccessful implantation. The trophectoderm of the
Department of Epidemiology and Biostatistics, Amsterdam, The Netherlands blastocyst-stage embryo attaches to the endometrial epithelium before breaching
this barrier to implant into the endometrium. While historical histology suggested
Study question: Does intrauterine application of auto-crosslinked hyaluronic invasive syncytiotrophoblast after the embryo has breached the epithelium, in
acid (ACP) gel, following dilatation and curettage (D&C) improve reproductive vitro models are required to understand the processes of human embryo
outcome in women with at least one previous D&C. implantation.
Summary answer: Application of ACP gel after D&C for miscarriage in Study design, size, duration: Fresh and frozen human blastocyst-stage
women with at least one previous D&C.seems to have a favorable effect on embryos (n=46) and human trophoblast stem cells were co-cultured with an
subsequent reproductive outcome endometrial epithelial cell line to model implantation in vitro. Systems biology
What is known already: Application of auto-crosslinked hyaluronic acid approaches with published transcriptomic datasets were used to model implan-
(ACP) gel, in women who experienced at least one previous D&C undergoing tation in silico.
D&C for miscarriage, resulted in a significant lower rate of IUAs, 13.0% versus Participants/materials, setting, methods: Human embryos surplus to
30.6%, relative risk (RR) 0.43 (95% CI 0.22-0.83, P=.013, lower mean adhesion treatment requirements were consented for research. Day 6 blastocysts were
score and significant less moderate to severe IUAs. Although subsequent fertility co-cultured with Ishikawa endometrial epithelial cell line monolayers until day 8,
is a key outcome, data on reproductive performance following application of and trophoblast stem cell spheroids were co-cultured for 48h. Morphology was
ACP after D&C for miscarriage remain limited. assessed by immunofluorescence microscopy, and trophoblast differentiation by
Study design, size, duration: This was a follow-up study of a prospective RT-qPCR and ELISA. Human blastocyst, trophoblast stem cell and endometrial
randomized trial, conducted in one university and seven university-affiliated organoid transcriptomic datasets were used with hypernetwork analysis and
teaching hospitals in the Netherlands. Women with a miscarriage of < 14 weeks random forest machine learning to identify gene networks associated with
with at least one previous D&C, were randomized to D&C plus ACP gel (inter- implantation.
vention) or D&C alone (control). A hysteroscopy was performed after 8-12 Main results and the role of chance: The majority of embryos (37/46,
weeks to detect intrauterine adhesion (IUAs). In case IUAs were encountered, 80.4%) breached the Ishikawa cell layer, and syncytiotrophoblast was observed
adhesiolysis was executed. in all of these. Sites where a single point of breaching had occurred always
Participants/materials, setting, methods: To evaluate reproductive consisted of syncytiotrophoblast (7/7). Multiple (up to 7), independent syncy-
outcomes, participants of the PAPA-study were approached between tiotrophoblast regions were seen in 26/46 (56.5%) embryos. Human trophoblast
December 2017 and April 2018. Participants ( n=139) received questionnaires stem cell spheroids co-cultured with Ishikawa layers also exhibited synscytiotro-
by post or email at least 30 months after randomization. The questionnaires phoblast formation upon breaching the epithelium. RT-qPCR demonstrated
consisted of 45 items related to received interventions, complications, men- epithelium-induced upregulation of syncytiotrophoblast genes CGB (p=0.03),
strual pattern, contraceptive use, the desire to become pregnant, conception OVOL1 (p=0.04) and SDC1 (p=0.008), and ELISA revealed stimulation of hCG
and outcome of subsequent pregnancies. The primary outcome was ongoing secretion (p=0.03). Receptive endometrial organoid surface transcriptome was
pregnancy. used to identify trophectoderm surface binding partners and hypernetwork
Main results and the role of chance: The response rate in the intervention analysis established a group of 24 trophectoderm receptors that were highly
group was 93.1% and in the control group 95.5%. Baseline characteristics were connected within the transcriptome. Of these, 14 genes are differentially
comparable in both groups. The median duration of follow-up was slightly longer expressed in trophoblast stem cell-syncytiotrophoblast differentiation (p<0.047-
in the intervention group, 40.6 months (SD 2.7) versus 39.2 months (SD 3.6) in 8.7-9), and random forest machine learning assigned 20/24 genes to processes
the control group. Ongoing pregnancies were recorded in 74.6% in the inter- driving this differentiation pathway (p<0.01). In vitro embryo and stem cell mod-
vention group versus 67.2% in the control group, OR 1.33 (0.63-2.82). The els, and in silico models, all suggest that syncytiotrophoblast formation from
miscarriages rate was 20.9% versus 37.5% respectively, OR, 0.44 (95% CI 0.18- trophectoderm is induced by attachment to the endometrial epithelium and this
0.90, P=.05). The live birth, ectopic pregnancy and TOP rates were not signifi- cell type pioneers embryo invasion.
cantly different. In women wishing to conceive, ongoing pregnancies were Limitations, reasons for caution: In vitro and in silico models may not
recorded in 94.3% versus 71.7% , OR 4.84 (1.51-15.51) and more live births, recapitulate the dynamic interactions embryo-endometrial interactions that
86.8% versus 70.0% in the control group OR, 2.82 (95% CI 1.07-7.42; P=.04) occur in vivo. The influence of other cellular compartments in the endometrium,
The median time to conception leading to a live birth was 21.9 versus 36.1 including decidual stromal cells and leukocytes, was not recapitulated in
months, hazard ratio 1.46 (0.96-2.48). these models.
Limitations, reasons for caution: The main limitation is that the PAPA-study Wider implications of the findings: Understanding the mechanism of
was designed and powered for the presence of IUAs and not for long-term repro- human embryo breaching of the epithelium and the gene networks involved is
ductive outcomes. Therefor the outcomes should be interpreted with caution. crucial to improve implantation success rates after assisted reproduction.
Wider implications of the findings: The current study shows that applica- Moreover, early trophoblast lineages form the blueprint for the placenta and
tion of ACP seems to have a favorable effect on reproductive performance. thus underpin fetal growth trajectories, pregnancy health and even off-
Our results should be confirmed in a large prospective study of sufficient power. . spring health.
Trial registration number: NTR 3120 Trial registration number: NA

O-303 Comparison of DNA methylation patterns of parentally to 34 imprinted DMRs and we studied only the placenta tissue and no other
imprinted genes in placenta derived from IVF conceptions in two embryo derived tissues.
different culture media Wider implications of the findings: It has often been postulated that
C. Mulder1, T. Wattimury1, A. Jongejan2, C. De Winter-Korver1, imprinting mediates the effects of embryo culture conditions on pregnancy, birth
S. Van Daalen1, R. Struijk1, S. Borgman1, Y. Wurth3, D. Consten3, and child development. We did not detect any statistically significant effects of
J. Van Echten-Arends4, S. Mastenbroek1, J. Dumoulin5, S. Repping1, embryo culture conditions on methylation status of imprinted genes in the pla-
A. Van Pelt1, A. Van Montfoort5 centa, therefore other unexplored mechanisms may underlie these effects.
1 Trial registration number: Placental biopsies were obtained under
Amsterdam UMC - Location AMC, Center for Reproductive Medicine-
Netherlands Trial Registry number 1979 and 1298.
Amsterdam Reproduction and Development Research Institute ., Amsterdam,
The Netherlands ;
2 O-304 Risk factors for ectopic pregnancy after in vitro fertilization
Amsterdam UMC- location AMC, Bioinformatics Laboratory- Clinical
treatment: a case-control study
Epidemiology- Biostatistics and Bioinformatics- Amsterdam Public Health Research
Institute- Amsterdam UMC, Amsterdam, The Netherlands ; V. Trindade1, M.R. Hentschke1, V.C. Dornelles2, A.T.F. Kira1, T.
3
St. Elisabeth Hospital, Center for Reproductive Medicine, Tilburg, The Netherlands ; Colombo1, B. Cunegatto3, L. Okada3, A. Petracco4, J.D.R.
4
University Medical Center Groningen, Section of Reproductive Medicine, Michelon5, B.E.P. Da Costa2, M. Badalotti4
1
Groningen, The Netherlands ; Fertilitat - Center for Reproductive Medicine, Research, Porto Alegre, Brazil ;
5 2
Maastricht University Medical Center, Department of Obstetrics & Gynaecology- Pontifical Catholic University of Rio Grande do Sul, School of Medicine, Porto
GROW School for Oncology and Developmental Biology, Maastricht, The Alegre, Brazil ;
3
Netherlands Fertilitat - Center for Reproductive Medicine, Laboratory, Porto Alegre, Brazil ;
4
Fertilitat - Center for Reproductive Medicine, Director, Porto Alegre, Brazil ;
Study question: Is there a difference in DNA methylation status of imprinted 5
Fertilitat - Center for Reproductive Medicine, Medical Team, Porto Alegre, Brazil
genes in IVF derived placentas where embryo culture was performed in HTF
versus G5 medium? Study question: Is it possible to identify risk factors for ectopic pregnancy
Summary answer: We found no statistically significant differences in the mean after in vitro fertilization?
DNA methylation of imprinted genes in placentas derived from IVF conceptions Summary answer: Tubal factor, previous miscarriage, D3 embryo transfer,
cultured in HTF versus G5. two or more embryos transferred per cycle and oocyte recipient cycles were
What is known already: Animal studies indicate that the embryo culture associate with ectopic pregnancy after IVF.
environment affects the DNA methylation status of the embryo. In humans, What is known already: Ectopic pregnancy is an obstetrical disease that is
birthweight is known to be affected by the type of embryo culture medium used. potentially associated with maternal death at first trimester. It is one of the
The effect of embryo culture media on pregnancy, birth and child development well-known complications following IVF with embryo transfer (ET), as its inci-
may thus be mediated by differential methylation of parentally imprinted genes dence can reach 0.9-11% compared to 1–2% in spontaneous pregnancies. The
in the placenta. etiology of a higher EP rate in an IVF-ET cycle remains unclear and the strategies
Study design, size, duration: DNA methylation status of imprinted genes to decrease its rates are limited.Thus, it is important to identify accurate risk
was determined in human placenta derived from IVF conceptions exposed to factors for EP, as it represents not only a life-threatening event but also a lost
HTF or G5 embryo culture medium. Placenta samples (n=43 for HTF, n=54 for opportunity for couples who sought assisted reproduction treatments.
G5) were collected between 2010 and 2012 as part of a multi-centre randomized Study design, size, duration: Retrospective case-control study performed
controlled trial in the Netherland comparing these embryo culture media. at an assisted reproduction clinic in southern Brazil. To select the control group,
Placenta samples from 69 naturally conceived live births collected during 2008- a ratio of 1:4 was applied and collected from 4666 cycles that resulted in a clinical
2013 in the Netherlands were used as reference material. pregnancy after IVF-ET. In the end, 499 patients who underwent IVF cycle and
Participants/materials, setting, methods: To measure the DNA methyl- evolved to clical pregnancy were included. The data refers to a period from
ation status of differentially methylation regions (DMRs) associated with paren- 2001- 2019 and were collected from electronic records.
tally imprinted genes, we opted for an amplicon-based sequencing strategy on Participants/materials, setting, methods: The 499 patients were separated
an Illumina MiSeq sequencing platform. DNA was isolated and 34 differentially into groups: Group 1, who presented EP diagnose (n=90) and Group 2, intrauter-
methylated regions associated with well-defined parentally imprinted genes, were ine pregnancies (IUP) (n=409). Data were presented as mean ± standard deviation
amplified in a two-step PCR procedure before sequencing using MiSeq technol- or percentage. Student t test, Mann–Whitney U-test, and Chi-square test or
ogy. Sequencing data were analysed in a multivariate fashion to eliminate possible Fisher’s exact test were applied. Multiple logistic regression analysis was performed
confounding effects. to assess risk factors for EP. Statistical significance was defined as p < 0.05.
Main results and the role of chance: We found no statistically significant Main results and the role of chance: When compared Group 1 with Group
differences in the mean DNA methylation status in any of the imprinted DMRs 2 the following results were observed: Tubal factor infertility (35.6% vs. 21.1%,
in placentas derived from IVF conceptions cultured in HTF or G5 culture medium. p=0.005) (OR 2.0 [1.2-3.4], p=0.004); History of previous miscarriage (15.1%
We also did not observe any differences in the variance in methylation per vs.7.1%, p<0.05) (OR 2.0 [1.02–4.29], p=0.044); number of transfers in D3
amplicon between the two culture medium groups. A separate surrogate variable embryo (69.2% vs. 54.0 p=0.028)(OR 1.9 [1.08 - 3.33], p=0.025); two or more
analysis also demonstrated that the IVF culture medium was not associated with embryos transferred (OR 2.5 [1.12–5.70], p=0.025). Even though we excluded
the DNA methylation status of these DMRs. The mean methylation level and oocyte recipient patients (ORP) from the analyzes above, we found a difference
variance per CpG was equal between HTF and G5 placenta. Additional com- in ORP rate when comparing Group1 and group 2 (9.4% (10/106) vs. 3.0 %
parison of DNA methylation status of IVF placenta samples with naturally con- (13/434), p=0.007), (OR 3.3 [1.41 - 7.98] p = 0.005). No significant differences
ceived placenta samples revealed no statistically significant differences in mean were observed in the analysis of female and male ages, body mass index, number
amplicon and CpG methylation between G5, HTF and naturally conceived pla- of mature oocytes, sperm concentration, follicle-stimulating hormone,antral
centa. However, the number of placenta samples exhibiting outlier methylation follicle count, male factor, ovulation factor, previous EP, previous pelvic surgery,
levels was higher in IVF placenta compared to naturally conceived (p<0.00001). endometrial thickness, embryo classification, protocol with agonist or antagonist
Also, we were able to identify 37 CpG sites that uniquely displayed outlier or fresh versus frozen ET.
methylation in G5 placentas and 32 CpG sites that uniquely displayed outlier Limitations, reasons for caution: Retrospective study with limited number
methylation in HTF. In 8/37 (G5) and 4/32 (HTF) unique outliers CpGs, a of patients. In addition, even though results show that oocyte recipient patients
medium specific unique outlier could be directly correlated to outlier methylation seems more likely to have an ectopic pregnancy, this data should be carefully
of the entire amplicon. interpreted given sample size.
Limitations, reasons for caution: Due to practical reasons, not all placentas Wider implications of the findings: Findings demonstrated a higher risk
were collected during the trial and we collected the placentas from natural con- for EP in the group of patients who had tubal factor and previous miscarriage
ceptions from a different cohort, potentially creating bias. We limited ourselves history. D3 transfer and more than one embryo per transfer were also associated

with higher EP risk. In high risk patients, it is reasonable to advise blastocyst and adequately powered and the risk of bias reduced due to random allocation of
single embrio transfer. women to the intervention or control group.
Trial registration number: not applicable Limitations, reasons for caution: The long period for completion of the
trial is a limitation of the current study. Active endometriotic lesions were cau-
terized before any reproduction procedure. This treatment might hided any
beneficial effect of GnRH-a administration. Data on miscarriages and live birth
SELECTED ORAL COMMUNICATIONS rates are not available.
SESSION 73: ENDOMETRIOSIS AND ART Wider implications of the findings: This study is expected to guide
future management of infertile women with mild endometriosis undergoing
08 July 2020 Parallel 5 14:00 - 15:15
IVF-ET. According to our data, these women better proceed directly to
IVF-ET, after laparoscopy, since the administration of GnRH-a did not
improve their fertility.
O-305 Ultra-long administration of GnRH-a before in vitro Trial registration number: NCT1269125
fertilization does not improve the clinical pregnancy rate in
women with mild endometriosis. A prospective, randomized, O-306 Oocyte survival and clinical outcome is impaired in young
controlled trial endometriosis patients after fertility preservation (FP).
A. Kaponis1, E. Panagodimou1, G. Hadjopoulos2, M. Paschopoulos2, A. Cobo1, J. Giles2, S. Paolelli2, A. Coello1, B. Vallejo1, J. Serrano1,
I. Georgiou2, V. Paraskevaidis2, K. Zikopoulos2, K. Tsiveriotis1, F. J.A. García-Velasco3, J. Remohí2
Taniguchi3, G. Adonakis1, T. Harada3 1
IVIRMA-Valencia, IVF Laboratory-Cryopreservation Unit, Valencia, Spain ;
1 2
Patras University School of Medicine, Ostetrics & Gynecology, Patra, Greece ; IVIRMA-Valencia, Ob/Gyn, Valencia, Spain ;
2 3
Ioannina University School of Medicine, Obstetrics & Gynecology, Ioannina, IVIRMA-Madrid, Ob/Gyn, Madrid, Spain
Greece ;
3
Tottori University Faculty of Medicine, Obstetric and Gynecology, Yonago, Japan Study question: Is oocyte survival and clinical outcome impaired in endome-
triosis patients after FP when compared to elective freezers?
Study question: Is there any beneficial effect of GnRH-a administration, Summary answer: Oocyte survival, implantation, pregnancy and cumulative
post-laparoscopically, in the fertility of women with mild endometriosis who live birth rates are impaired in young (≤35y) patients with endometriosis.
underwent in vitro fertilization (IVF) procedures? What is known already: FP should be considered in women with endome-
Summary answer: The clinical pregnancy rate in mild endometriosis women triosis due to the progressive reduction in the ovarian reserve. In spite of the
who received GnRH-a, post-laparoscopy, did not improve compared with high incidence of the disease and the increasing number of endometriosis-women
women who did not receive this regime. who have oocytes vitrified for FP, very little is known about the efficacy of the
What is known already: GnRH-agonists have long been administered for strategy in these cases. Currently, there is evidence of successful outcomes
the treatment of endometriosis, however, their use postoperatively, in women achieved after oocyte vitrification for elective FP (EFP), although the age of the
with mild endometriosis has been linked with controversial results. Several patient is one of the most limiting factors. In this study, we compare the results
studies associated prolonged use of GnRH-a before IVF with a higher preg- achieved by endometriosis-patients after FP to those achieve by EFP patients
nancy rate, while others showed no statistically significant improvement. On according to age.
the other hand, long-term pituitary desensitization regimes may lower ovarian Study design, size, duration: Retrospective study including 485 (≤35y:
response to gonadotropins, affecting the success of assisted reproduction N=123 and >35y: N=518) women with endometriosis and 641 (≤35y: N=123
procedures. and >35y: N=518) EFP patients. All women included had their oocytes vitrified
Study design, size, duration: This is a prospective, randomized, controlled for FP and returned to use their oocytes to attempt pregnancy.
trial. Six hundred infertile women were recruited during a 15-year interval (May Participants/materials, setting, methods: The Cryotop method was used
2004- December 2018). Four hundred of the afore-mentioned women diag- for oocyte vitrification. Within the endometriosis group, 97.7% of patients were
nosed with mild endometriosis (ASRM stage I-II), while 200 from tubal factor diagnosed with stage III-IV endometriosis and 47.8% had ovarian surgery prior
infertility (group C). Patients with endometriosis were divided randomly in those to FP. Oocyte survival, implantation rate and clinical outcome were compared
who received GnRH-a, for three months before an IVF attempt (group A, between endometriosis and EFP groups according to age (≤35y vs <35y). Values
n=200) and those who did not (group B, n=200). are expressed as mean and 95% confidence intervals (CI). Chi-square tests were
Participants/materials, setting, methods: Women aged 29-38 years, were used to compare the categorical data. P<0.05 was considered as statistically
recruited from three tertiary University hospitals (Ioannina and Patras, Greece; significant.
Yonago, Japan). There were no demographic discrepancies between groups. Main results and the role of chance: Mean age of patients with endome-
Long protocol for ovulation induction was used. TNF-a, IL-1β, IL-6, IL-8, and triosis by the time of FP was 35.7 ± 3.7 and 37.6 ± 3.5 for EFP patients. Lower
IL-1ra levels were measured in the follicular fluid (FF) of all women. Fertilization outcomes were achieved in the group of women with endometriosis aged ≤35y
Rate (FR), Implantation Rate (IR), quality of embryos, and clinical pregnancy rate (P<0.05): oocyte survival was 85.1% (95%CI 83.8-86.5) vs. 91.4% (95%CI 89.6-
(PR) were evaluated and compared between the different groups. 93.2), implantation rate was 38.6% (95%CI 45.7-63.8) vs. 54.5% (95%CI 33.9-
Main results and the role of chance: Women with endometriosis who 43.3), clinical pregnancy rate was 49.2% (95%CI 42.9-55.5) vs.65.9% (95%CI
received the long-acting GnRH-a (group A) presented significantly decreased 57.9-73.9), ongoing pregnancy rate was 40.9% (95%CI 34.7-47.1) vs. 57.7%
concentrations of the FF’s cytokines compared with those who did not receive (95%CI 49.4-66), and CLBR was 61.9% (95%CI 56-67.8) vs. 68.8% (95%CI
similar treatment (group B). This applied to TNF-a, IL-1β, IL-6, and IL-8 59.4-78.2) respectively (P<0.05). In older patients (>35y) all the parameters
(p < 0.001). In women with endometriosis, FR and PRs were lower compared were comparable: 80.8% (95%CI 78.9-82.6) vs. 82.1% (95%CI 80.9-83.3) for
to women with tubal infertility. A statistically significant difference was observed oocyte survival, 28.3% (95%CI 23.1-33.5) vs. 33.8% (95%CI 28.1-39.4) for
only in the FR between women of group A and women of group B (group A: implantation, 41.4% (95%CI 34.0-48.8) vs. 36.5% (95%CI 32.5-40.5) for clinical
72.7, 95%CI: 70.50-74.90 vs. group B: 61.7, 95%CI: 59.20-64.20, p<0.05). pregnancy, 29.6% (95%CI 22.7-36.5) vs. 27.7% (95%CI 23.9-31.5) ongoing preg-
Embryo quality did not improve after administration of GnRH-a. The percentage nancy and 28.4% (95%CI 19.7-37.7) vs. 25.5% (95%CI 21.1-29.9) for CLBR in
of grade 1 embryos was comparable between the three groups (group A: 24.5, endometriosis vs EFP respectively (NS).
95%CI: 21.50-27.50; group B: 21.6, 95%CI: 18.80-24.40; and control group: 26, Limitations, reasons for caution: Retrospective nature of the study. The
95%CI: 22.70-29.30). IR did not differ significantly between women of the three lower number of patients diagnosed as stages I-II makes it difficult to compare
groups (group A: 18.4%, 95%CI: 14.6-22.2%; group B: 17%, 95%CI: 13-21%; by the stage of the disease.
group C: 19.3, 95%CI 15.5-23.1). Women of group A presented higher PR than Wider implications of the findings: The lower oocyte survival and the
women of group B however, this difference was not statistically significant (OR: impaired implantation and reproductive potential observed in young endome-
1.16, 95%CI: 0.73, 1.84, p=0.52). The study population is homogenous, triosis patients confirms the negative impact of the disease in the ovarian reserve

and in the oocyte quality. FP should be counseled to endometriosis patients at Study question: Is transvaginal endometriotic cyst aspiration better than lap-
young ages in order to increase their future pregnancy chance. aroscopic ovarian cystectomy for preservation of ovarian reserve?
Trial registration number: Not applicable Summary answer: Better ovarian reserve in transvaginal cyst aspiration group
achieve more number of oocytes and embryos resulting in significantly higher
O-307 ICSI treatment does not result in improved live birth rates cumulative pregnancy rate.
compared to conventional IVF in couples with endometriosis: An What is known already: Laparoscopic cystectomy causes decreased ovarian
analysis of 10,047 treatment cycles reserve due to loss of normal ovarian tissue and thermal damage leading to poor
reproductive outcome.
H. Kamali1, I. Gamaleldin2, A. Kaura3, P. Wilson4, V. Akande1
1
Study design, size, duration: In this Randomized, Control Trial over 2 years
Bristol Centre for Reproductive Medicine, Clinical team, Bristol, United Kingdom ;
2
( July 2017 to June 2019), 104 infertile patients of 20-36 years of age with
North Bristol NHS Trust, Women and Childrens Health, Bristol, United Kingdom ;
3
endometrioma were divided into- Group A and Group B comprising 52 patients
Imperial College NHS Healthcare Trust, Medical statistics, London, United Kingdom ;
4
in each group
Bristol Centre for Reproductive Medicine, Embryology, Bristol, United Kingdom Participants/materials, setting, methods: In this study Group A-52
patients had laparoscopic chocolate cystectomy followed by 4 weekly injection
Study question: Is ICSI associated with improved reproductive outcomes over GnRH-a depot(3.75 mg ) for 3months followed by IVF-ET treatment whereas
conventional IVF in couples with endometriosis related infertility (ERI) only? Group B-52 patients received 4 weekly Injection GnRH-a depot(3.75 mg ) for
Summary answer: Compared to conventional IVF, ICSI results in reduced 3months followed by transvaginal chocolate cyst aspiration and underwent
fertilisation failure but is not associated with improved implantation, clinical preg- IVF-ET treatment . Patients in both groups were evaluated for ovarian reserve,
nancy or live birth rates in ERI no. of oocyte retrieved, fertilization rate, No. of embryo, implantation rate,
What is known already: Endometriosis can affect up to 50% of women with pregnancy rate & outcome.
infertility. As well as anatomical distortion, the condition can also negatively affect Main results and the role of chance: In Group A & Group B, mean no of
oocyte quality, embryogenesis and implantation through altered folliculogenesis, baseline antral follicles and AMH were 2.1 ±0.9 Vs 4.8 ± 0.6 (p< 0.05) and 1.98
sperm phagocytosis, impaired fertilisation and embryotoxicity. ICSI, which is ± 0.8 Vs 3.1 ±1.2 (p< 0.01) respectively. Baseline FSH was significantly more in
typically used for male factor infertility and known to result in improved fertili- Gr A (9.1± 1.4 Vs 6.6±1.3, p= 0.04 ). The number of average oocyte retrieved
sation rates compared with conventional IVF, is increasingly being used in the was significantly more in Gr B (7.2±2.2 vs 4.8±2.8, p<0.05). Cumulative preg-
cohort of patients with ERI to try and overcome these effects. However, there nancy rate per ovum pick up cycle is significantly more in Gr B (60.4% vs 45.1%
are no large-scale studies directly assessing the benefit of ICSI over conventional , p<0.05).
IVF with regards to reproductive outcomes in women with ERI. Limitations, reasons for caution: As this study is of short duration with
Study design, size, duration: Anonymised data on all IVF and ICSI treatment small number of patients so widespread multicentric study comprising large
cycles performed in the UK from years 1991 – 2016 were retrospectively number of subjects is requirred to cone to a definitive conclusion.
obtained from the Human Fertility and Embryology Authority (HFEA). Women Wider implications of the findings: Treatment cost and patient’s morbidity
under 40 having their first treatment cycle using fresh embryos with a diagnosis is less in group B with better ovarian reserve leading to more number of oocytes
of ERI only were included. retrieved and number of good quality embryo available for transfer.
Participants/materials, setting, methods: Primary outcome was live birth Cumulative pregnancy rate following single ovum pick up cycle is significantly
rate. Secondary outcomes were fertilisation rate, implantation rate and clinical more in transvaginal cyst aspiration group.
pregnancy rates. Outcomes were further stratified by age group (<35, 35-37, Trial registration number: Not applicable
38-39) and time period to account for improved LBR over time. Data were
analysed using logistic regression and controlled for age, number of oocytes
collected, and number of embryos transferred. O-309 Endometrial scratching in women undergoing their first
Main results and the role of chance: A total of 10,047 women had ERI of In Vitro Fertilisation (IVF) cycle: results from the UK Multicentre
which 80.3% (8,075) had IVF compared with 19.7% (1,972) who underwent Endometrial Scratch Randomised Controlled Trial
ICSI. The proportion of couples with ERI having ICSI increased from 6.3% in
This abstract
M. Metwally 1
, R. has been
Chatters 2
chosen
, C. Pye1, M. for the 2press
Dimairo 2
, D. White2, S.
1991-2000 to 23.7% in 2011-2016. Rates of total fertilisation failure (TFF) were Walters , J. Cohen , T. Chater , K. Pemberton , T. Young2, R.
2 3 2
lower with ICSI compared with IVF (3.7% versus 6%, p-value <0.001). However, programme and will not be made public until the
Lomas1, E. Taylor1, S. Laird4, L. Mohiyiddeen5, Y. Cheong6
in women with ERI, ICSI did not result in an improved live birth rate (OR 1.05, abstract
1 is presented.
Sheffield teaching hospitals, Reproductive medicine, Sheffield, United Kingdom ;
CI 0.94-1.17, p-value 0.42), clinical pregnancy rate (OR 1.06, CI 0.95-1.17, 2
The University of Sheffield, Clinical Trials Unit, Sheffield, United Kingdom ;
p-value 0.33) or miscarriage rate (OR 1.07, CI 0.82-1.39, p-value 0.62) compared 3
Hull York Medical School, Hull Health Trials Unit, Hull, United Kingdom ;
with conventional IVF. The absence of improved reproductive outcomes with 4
Sheffield Hallam University, Biosciences and Chemistry, Sheffield, United Kingdom ;
ICSI compared to conventional IVF in women with ERI was consistent across all 5
Manchester University Hospitals, Reproductive Medicine, Manchester, United
age groups and different time periods. Kingdom ;
Limitations, reasons for caution: As a retrospective study, our analysis 6
The University of Southampton, Reproductive Medicine, Southampton, United
depends on previously recorded data, therefore certain key variables (endo- Kingdom
metriosis stage, presence/absence of endometriomas, clinic specific protocols
and duration of infertility) could not be accounted for. The reported cause of Study question: Does endometrial scratching (ES) in the mid-luteal phase
infertility is based on the treating clinician’s classification and may be inconsistent. prior to first time IVF/ICSI increase the chances of achieving a clinical pregnancy
Wider implications of the findings: In couples with ERI, ICSI results in and live birth?
reduced TFF compared with conventional IVF. However, this does not translate Summary answer: Endometrial Scratch performed prior to the first cycle of
to an improvement in post fertilisation reproductive outcomes, including live IVF/ICSI does not increase the chances of achieving a pregnancy and live birth.
birth rate. These findings do not support the use of ICSI over conventional IVF What is known already: Endometrial Scratch is currently being provided in
for treatment of couple with ERI. some fertility units where women are having IVF/ICSI for the first time. The
Trial registration number: NA most recent 2019 systematic review shows poor evidence to support this prac-
tice. Current studies are single centre, of relatively small size and/or involve
O-308 A prospective randomized comparative study of heterogeneous or unselected populations. Further evidence is therefore required
laparoscopic ovarian cystectomy and ablation versus transvaginal from a large multi-centre randomised controlled trial looking at a homogenous
cyst aspiration in infertile patients with Endometrioma population of only women undergoing first time treatment and expected to be
undergoing IVF – ET treatment good responders in order to minimise heterogeneity.
S. Saha1, H.H. Shirazee1, M. Murmu1 Study design, size, duration: A multicentre, pragmatic, open-label, individ-
1
North Kolkata Infertility & IVF Centre, Reproductive Medicine, Kolkata, India ually randomised parallel-group trial recruited women at 16 fertility centres

across the UK from July 2016 to Oct 2019. We planned to recruit 1044 women chromatin conformation and nucleoli), metaphase spindles and network of endo-
(522 per arm) to preserve a power of 90% and 5% two-sided type I error plasmic reticulum cisterns and mitochondria. It is believed that these morpho-
assuming a 30% live birth rate in treatment as usual (TAU), a 10% absolute dif- logical parameters can be useful for oocyte/embryo quality evaluation in IVF
ference between TAU and Endometrial Scratch (ES) arms (more likely to change protocols.
practice), and 5% dropout rate. Study design, size, duration: We tested the OCM on mouse oocytes and
Participants/materials, setting, methods: Women aged ≥18 and ≤37 embryos, focusing so far on immature (so-called germinal vesicle, GV) oocytes
undergoing their first IVF cycle, BMI <35 kg/m2 and expected to be good respond- and compacted morulas. We analyzed over 400 GV oocytes and over 400
ers were eligible. ES was undertaken in the mid-luteal phase prior to IVF/ICSI. morulas in order to verify whether OCM scanning affects their developmental
Women were randomised 1:1 to either ES or IVF/ICSI (TAU), using a web-based capabilities and whether morphological data obtained by OCM (chromatin con-
stratified block randomisation. The primary outcome was live birth (LBR); sec- formation in GVs and number of nuclei in morulas) correlate with their devel-
ondary outcomes included clinical pregnancy (CPR), implantation (IR), ectopic opmental potential.
pregnancy (ER), miscarriage (MR), preterm delivery (PDR), stillbirth (SBR) rates Participants/materials, setting, methods: GV oocytes were isolated from
and safety. ovaries whereas morulas were obtained by in vitro fertilization of ovulated
Main results and the role of chance: 1048 women were randomised TAU oocytes. OCM scans of GVs and morulas provided morphological information
(n=525) and ES (n=523). Mean age and BMI (standard deviation) was 32.5(3.4) that was correlated with their developmental potential tested as the ability to
years and 24.5 (3.3) kg/m2. Baseline characteristics were similar between arms. mature and react to fertilization (for GVs) or to achieve a blastocyst stage with
In the ES arm, 86.6% (453/523) received the ES procedure. IVF/ICSI was received well-differentiated embryonic cell lineages (for morulas). We also tested whether
by 94.1% (494/525) in the TAU and 95.0% (497/523) in the ES arm. For the scanned oocytes/embryos differ in their developmental capabilities from their
primary outcome, LBR was 37.1% (195/525) in the TAU and 38.4% (201/523) non-scanned counterparts.
in the ES (Unadjusted: difference, 1.3% [95%CI, -4.6% to 7.2%]; relative risk, 1.03 Main results and the role of chance: We showed that OCM allows for a
[95%CI, 0.89 to 1.21]; odds ratio, 1.06 [95%CI, 0.82 to 1.36]; P-value = 0.667). precise assessment of chromatin conformation in mouse GV oocytes (sur-
For the secondary outcomes, CPR was 40.6% (213/525) in TAU and 42.4% rounded by cumulus cells or denuded) and for the selection of oocytes with
(222/523) in ES arm (Unadjusted: difference, 1.9% [95%CI, -4.1% to 7.8%]; rel- so-called SN (surrounded nucleoli) chromatin conformation that have higher
ative risk, 1.05 [95%CI, 0.91 to 1.21]; odds ratio, 1.08 [95%CI, 0.84 to 1.38]; developmental potential. GVs scanned by OCM matured in vitro with the same
P-value = 0.538). All the other secondary outcomes (IR, EPR, MR, MBR, PDR, speed and efficiency as non-scanned control oocytes and developed similar Ca2+
and SBR) were all similar between arms. response to a fertilizing spermatozoon, indicating that OCM scanning did not
Adverse events and serious adverse events in women were similar between affect them negatively. We also showed that OCM provides data that allows for
arms. No deaths or neonatal deaths were reported. Only 1.2% (3/258) of born a precise nuclei (i.e. cell) count in compacted mouse embryos and that the
babies had severe congenital abnormalities reported in the TAU arm only. number of nuclei (cells) at the compacted morula stage correlates with the
Limitations, reasons for caution: The study did not include a sham proce- embryo’s ability to form high-quality blastocysts. Morulas scanned by OCM
dure in the Treatment As Usual arm but this is unlikely to have influenced the developed to the blastocyst stage with similar efficiency as their non-scanned
study outcomes Furthermore, the study is applicable to the demographic and counterparts, although the differentiation of the primitive endoderm (PE) cells
ethnic distribution of the population in the UK as no overseas centres were was slightly delayed. The number of PE cells in scanned embryos was lower at
included. day 4.0 than in non-scanned embryos, but the difference disappeared at day 5.0,
Wider implications of the findings: Endometrial scratch is currently per- suggesting that the developmental potential of the embryos was not significantly
formed for women undergoing first time IVF/ICSI in some centres. This study disturbed by the OCM.
provides conclusive evidence that Endometrial Scratch is not beneficial in this Limitations, reasons for caution: We tested OCM only on mouse embryos
population and suggests that it is time to stop this practice. and in the case of other species, additional tests are required. Moreover, further
Trial registration number: ISRCTN23800982 experiments testing the potential effect of OCM on the post-implantation
embryo development are necessary (and currently in progress).
Wider implications of the findings: Our data indicate that OCM is a non-in-
vasive imaging technique and that it provides morphological information about
SELECTED ORAL COMMUNICATIONS mammalian oocytes and embryos inaccessible for standard bright-field micros-
SESSION 74: OOCYTE AND EMBRYO EVALUATION copy. Therefore, it can be an interesting alternative for imaging techniques used
currently in assisted reproduction protocols.
08 July 2020 Parallel 6 14:00 - 15:15
Trial registration number: n/a
O-311 Development of an Advanced Quality Control Bioassay for

O-310 Optical coherence microscopy as a novel tool for quality
Assisted Reproductive Technologies
assessment of mammalian oocytes and embryos
K. Sakurai1, A. Tran1, V. Tovar1, R. Newman1, N. Ciray1
A. Ajduk1, M. Fluks1, A. Sobkowiak1, S. Tamborski2,
1
M. Szkulmowski2 Fujifilm Irvine Scientific, Research and Development, Santa Ana, U.S.A.
1
University of Warsaw, Faculty of Biology, Warsaw, Poland ;
2
Nicolaus Copernicus University, Institute of Physics, Torun, Poland Study question: Could embryos expressing GFP-tagged POU5F1, a transcrip-
tion factor critical for embryonic development during preimplantation develop-
Study question: We wished to examine whether optical coherence micros- ment, be used to detect adverse conditions resulting during ART practices?
copy (OCM), a novel technique of label-free live-cell imaging, can be used in the Summary answer: The Pou5f1-GFP embryos detected suboptimal conditions
oocyte/embryo quality assessment. as early as 48hours of exposure and subsequently after 96 hours.
Summary answer: OCM allows for non-invasive imaging of the intracellular What is known already: Growing concerns about safety of ART on human
architecture of oocytes and embryos. These morphological data can be predic- gametes and embryos have been raised by the observation of compromised
tive of oocyte/embryo developmental potential. embryo development and the outcome of the treatment during ART laboratory
What is known already: Fluorescence microscopy (including its confocal ver- procedures performed under suboptimal conditions. Incorporating evaluations
sion) allows for detailed structural and dynamic studies of single cells, but requires of gene expression essential for preimplantation development in an assay for
fluorescent markers to visualize cellular architecture and may cause short- and screening materials and conditions could benefit risk assessment. Zygotic Pou5f1
long-term photo-damage. Bright-field microscopy, although non-invasive, does gene expression, essential for mouse development, starts around the 4-cell stage
not provide detailed structural information. OCM is a promising alternative, as and later restricted to ICM. Mouse embryos expressing GFP under control of
it does not require sample pre-processing or labeling and is capable of providing Pou5f1 regulators formed a relevant functional assay for testing ART products
three-dimensional images of intracellular structures, such as nuclei (with prior to clinical implementation (Gilbert et al., 2016).

Study design, size, duration: Pou5f1-GFP transgenic mouse lines expressing Participants/materials, setting, methods: Study participants were similar
GFP-tagged POU5F1 were generated to utilize nuclear localization of POU5F1 with regard to age, BMI, parity, AMH levels, antral follicle count and number of
and to detect adverse culture conditions and epigenetic defect during preim- oocytes collected. FF cytokine protein levels were assayed using a commercially
plantation. Pou5f1-GFP expression were also used to visualize blastomere nuclei available multiplex array (Cytokine 30-Plex Human Panel, ThermoFisherScientific)
for cell counting in live cells. Pou5f1-GFP embryos were cultured for 96hrs under on the Luminex multiplex assay platform (Conway Institute, University College
optimal or suboptimal oil overlay to observe POU5F1-GFP expression at differ- Dublin). Data were analyzed using GraphPad Prism; cytokine levels were com-
ent stages of mouse embryo development (from 2PN to expanded/hatching pared using Kruskal–Wallis with Dunn’s multiple comparison test and Bonferroni
blastocyst). correction (p<0.05 considered significant).
Participants/materials, setting, methods: Pou5f1-GFP one-cell embryos Main results and the role of chance: FF levels of the inflammatory cytokines
(fresh or frozen) were cultured to blastocysts under uninterrupted conditions TNFa (p=0.037), IL-6 (p=0.031), IL-5 (p=0.021) and IL-10 (p=0.002) were
up to 96 hours in Continuous Single Culture Medium-Complete (CSCM-C, higher in follicles where the oocyte failed to develop to blastocyst (Group 1 and
FUJIFILM Irvine Scientific) with control or suboptimal oil overlay (5, 7.5, or 10% 2) than those where the oocyte developed to Day 5 blastocyst (Group 3).
adulterated oil) and observed daily. B6 one-cell embryos typically used in the Following Bonferroni correction to control for multiplex analysis (18 cytokines),
standard mouse embryo assay (MEA) were cultured in parallel. Embryo devel- only differences in IL-10 levels retained statistical significance (P<0.05). Area-
opment was evaluated at 48 hours (% ≥8-cell) and 96 hours (% Blastocyst) under-the-curve (AUC) of IL-10 as a predictor of oocyte development to a Day
quantitatively and qualitatively, respectively. 5 blastocyst in this patient cohort was found to be 0.73 (95% CI 0.618-0.85;
Main results and the role of chance: Transgenic mice expressing Pou5f1-GFP p<0.001). Nineteen FF samples had IL-10 levels <9pg/ml, 14 of which were
are viable and fertile, and successful germline transmission and temporally and from oocytes that successfully developed to blastocyst stage. IL-10 threshold
spatially regulated gene expression were confirmed. Zygotic Pou5f1-GFP gene levels <9pg/ml (sensitivity 48%, specificity 88%, prevalence rate 40% in this
expression started around the 4-cell stage and peaked after culturing for 72hrs, cohort) correspond to a positive predictive value (PPV) of 73.4% with 26.6%
consistent with earlier reports. The nuclear localization of POU5F1-GFP in chance of a false positive test result, a negative predictive value (NPV) of 72.1%
mouse embryos enabled a quantitative approach to the assay. The Pou5f1-GFP with 27.9% chance of a false negative result and an odds ratio (OR) of 7.093
embryos cultured with 5, 7.5, and 10 % suboptimal oil overlays showed a notice- (95% CI 1.57-4.32; p<0.001).
able delay in development (at 48hrs and 96hrs compare to the control and MEA Limitations, reasons for caution: The study cohort was limited in size, and
groups). Fresh and frozen Pou5f1-GFP embryos performed similarly in detecting clinical pregnancy rates and live birth outcomes are not yet available.
suboptimal conditions. Wider implications of the findings: A robust, non-invasive biomarker of
Limitations, reasons for caution: As observed with other mouse traits, oocyte competence would have a major impact on the prediction of ART suc-
embryo yields are correlated to harvesting efficiency. cess, including likely success of oocyte vitrification for an individual woman. This
Wider implications of the findings: Pou5f1-GFP embryos can be used in study shows that FF IL-10 may help predict the ability of an oocyte to fertilise
the one-cell mouse embryo assay to examine the effect of culture conditions and to reach blastocyst stage.
during preimplantation development. With the heightened sensitivity, the func- Trial registration number: not applicable
tional impact of materials and procedures on overall embryo development
including embryo health and defects can be adroitly evaluated in live cells.
Trial registration number: not applicable O-313 Evaluation of fragmented embryos – what is the best way
to predict its implantation potential?
S. Lahav-Baratz1, I. Blais1, M. Koifman1, M. Dirnfeld1,2
O-312 Follicular fluid levels of IL-10 are associated with oocyte 1
Division of Reproductive Endocrinology and IVF Lady Davis Carmel Medical
fertilization and early embryo development. Center- Haifa- Israel, Department of OB/GYN, Haifa, Israel ;
L. Hartigan1, L. Glover2, H. Groarke2, L. O’Shea3, M. Wingfield4 2
Technion – Israel Institute of Technology- Haifa- Israel, Ruth and Bruch Rappaport
1
Merrion Fertility Clinic and National Maternity Hospital- Dublin- Ireland, Merrion Faculty of Medicine-, Haifa, Israel
Fertility Clinic and National Maternity Hospital- Dublin- Ireland, Dublin, Ireland ;
2
Merrion Fertility Clinic, Merrion Fertility Clinic, Dublin, Ireland ; Study question: Can we predict the potential of fragmented embryos to
3
University College Dublin, School of Agriculture and Food Science, Dublin, Ireland ; implant, by using the general and our “in-house” model for embryo selection in
4
Merrion Fertility Clinic and National Maternity Hospital, Merrion Fertility Clinic a time-lapse incubator?
and National Maternity Hospital, Dublin, Ireland Summary answer: The algorithms developed for embryos evaluation by
morphokinetics, significantly increase the accuracy of a single embryo selection
Study question: Do follicular fluid cytokine profiles predict the developmental in our laboratory, including embryos with fragmentation.
potential of an oocyte in ART? What is known already: Fragmentation in IVF embryos has been a marker
Summary answer: Follicular fluid IL-10 levels were higher in follicles where for embryo deselection. Extensive fragmentation may be associated with reduced
oocytes failed to fertilize or develop to blastocyst than oocytes that developed blastocyst formation and with increased incidence of chromosomal abnormali-
to Day 5 blastocysts. ties. However, some of those embryos may yield pregnancy and delivery of a
What is known already: Embryo quality is greatly influenced by the oocyte healthy child. The cut-off of fragmentation rate which still enables achievement
and follicular microenvironment from which it originates. Follicular fluid contains of pregnancy is not defined clearly. Using models, based on time-lapse technol-
a range of factors that play important signaling roles, including steroid hormones, ogy, (general models, and lab-specific), may contribute to the selection of
metabolites, growth factors and cytokines. Inflammatory cytokines mediate func- embryos with fragments.
tional and structural changes associated with follicle growth and early oocyte Study design, size, duration: In this retrospective study, 4210 embryos
maturation. In previous work, our group identified follicular fluid AMH and pro- which were incubated to the blastocyst stage in the EmbryoScope between 2013
gesterone levels as predictors of oocyte and embryo quality and developmental and 2019 were analyzed. Three hundred seventy nine embryos, which had more
capacity. In this study, we adopted a similar approach to further investigate fol- than 5% fragmentation, were included in the study. Embryos were selected for
licular fluid cytokines and their association with oocyte development. transfer or freezing versus de-selection, based primarily on the general model
Study design, size, duration: This was a prospective study of women for day 5 provided by Vitrolife and then re-examined by our lab-specific
undergoing IVF/ICSI treatment in our fertility clinic.Women with diagnoses of algorithms.
PCOS or endometriosis were excluded (n=30).Follicular fluid (FF) was collected Participants/materials, setting, methods: Embryo fragmentations
from the first follicle aspirated during oocyte retrieval.The corresponding oocyte were measured using EmbryoScope tools by a senior embryologist.
was individually cultured to track its outcome.FF was assigned one of 3 groups: Percentage of fragmentation was documented twice for every embryo: at
Group 1: oocyte failed to fertilise (n=14); Group 2:oocyte fertilized but failed the first cell division and in their maximum volume. The patterns of frag-
to reach blastocyst (n=29);Group 3:oocyte developed into a Day 5 blasto- ments accumulation during embryos development were followed. Data was
cyst (n=29). analyzed using statistical methods for fragmentation with regards to patient’s

age, insemination method, blastocyst formation, embryos transfer or freez- What is known already: The mtDNA content of trophectoderm cells is
ing, known implantation data (KID) of the embryos, clinical pregnancy and related to the energy supply of the blastocyst, which could affect its implantation
live birth rates. ability. It has been proposed that the quantity of mtDNA in trophectoderm (TE)
Main results and the role of chance: Up to 32% fragmentation the specific biopsy cells can serve as a new biomarker of embryo viability. This information
model score and fragmentation percent was found to be an independent vari- could help to maximize the success rate of single embryo transfers (SET) in ART
ables. Embryos with up to 20 ± 12% fragmentation still had high score according by selecting embryos with the highest potential to achieve a live birth. However,
to the first division time and usually transferred or cryopreserved. Significant the application of a score based on mtDNA is controversial and highly discussed.
difference in the fragmentation rate was found between embryos which reached Study design, size, duration: A cohort of 1528 embryos from 349 couples
the blastocyst stage and embryos which failed: 15± 11% and 36± 20% respec- undergoing preimplantation genetic testing for aneuploidy between 2016 and
tively (p<0.0001). 2019 was retrospectively included. Mosaic embryos (between 20%-80%) were
Fragmentation appeared usually at the first division and worsening was excluded from this analysis. Blastocyst quality was established according to
observed during the 3th or 4th divisions. Gardner grading system and grouped in: high (3-6AA and 4-6AB), mid (any BB,
While no difference was found in fragmentation between embryos of standard 1-3AB and 1-2AA) and low (any AC, CA, BC, CB or CC) quality.
IVF or ICSI, age had a significant negative effect on fragmentation (p< 0.0001). Participants/materials, setting, methods: The embryo ploidy and the mtDNA
In this population of fragmented embryos, 33 of 379 embryos resulted in a content were determined using NGS and bioinformatics algorithms. The mtDNA
delivery of a healthy child, 104 were negative and for 242, information is not value was mathematically corrected according to embryo aneuploidy and gender.
available mainly due to freezing without thawing. βhCG > 15 IU was considered to be positive for biochemical pregnancy. Ongoing
In 92.8% of the patients more than 1 embryo was with fragments. pregnancy was defined as having a visible gestational sac with a fetal heartbeat at 8
In 64% of the patients fragmented embryos found in more than one cycle. All weeks after SET. For the statistical analysis T-test, Kruskal-Wallis and Pearson cor-
positive KID (known implantation data) had maximum of 30% fragmentation relation were applied. A p-value< 0.05 was considered statistically significant.
except one embryo with 43% fragmentation that was implanted and a healthy Main results and the role of chance: The average maternal age was 33.6
baby was born. (21-46 years). From the 1528 blastocysts assessed, 624 were euploid (40,8%),
Limitations, reasons for caution: Although fragmentation was analyzed 377 aneuploid (24,7%) and 527 mosaic embryos (34,5%). mtDNA was signifi-
meticulously by experienced embryologists, measurement may be less accurate cantly different between euploid and aneuploid embryos (p=0.0000038) and
during late divisions. Prospective randomized studies are required to confirm between embryos from younger (≤37 years, n=592) and older women (>38
these findings. years, n=409) (p=0.0002). A positive correlation of mtDNA with maternal age
Wider implications of the findings: We suggest that fragmented embryos was observed, but only in the group of older women the aneuploid blastocysts
should be examined primarily according to their division timing. A specific model contained higher mtDNA vs. euploid embryos (p=0.003). There was no signif-
can contribute to a better selection. Although rare, even embryos with as much icant difference in mtDNA between high, mid and low quality euploid embryos
as 40% fragmentations and appropriate division timing may develop into a blas- and between the different grades of TE (A, B and C). However, a difference was
tocyst which implant and results in a live birth. found between aneuploid and euploid embryos in the mid and low quality cat-
Trial registration number: 0010-19-CMC egories. Regarding the pregnancy outcome, 264 euploid embryos were trans-
ferred, resulting in 112 ongoing pregnancies (42%) and a 61% live birth rate
O-314 Assessment of the predictive value of mitochondrial DNA (n=69). There was no difference in mtDNA content between the positive βhCG
as a biomarker of embryo viability vs negative βhCG, the ongoing pregnancies vs the non-pregnant group and the
M. Galain2, M. Fabbro2, S. Menazzi2, R. Anria1, J. Ponte2, F. Nodar3, live birth vs abortions.
S. Papier1, C. Fernandez2 Limitations, reasons for caution: The study is limited by its retrospective
1 nature. A higher sample size or a prospective randomized design in future studies
Cegyr, Clinical, Buenos Aires, Argentina ;
2 would corroborate the current findings. Caution should be taken while compar-
Cegyr, Genetics, Buenos Aires, Argentina ;
3 ing these results to other reported studies since the same quantification meth-
Cegyr, Embryology, Buenos Aires, Argentina
odology is not always used.
Study question: Is the mitochondrial DNA (mtDNA) content related to female Wider implications of the findings: This study contributes evidence that
age, embryo morphology, ploidy, blastocyst implantation rate, pregnancy rate, mtDNA quantification in euploid embryos has no clinical impact on pregnancy
and live birth rate? outcome. mtDNA score adds no value when selecting which embryo to transfer.
Summary answer: There was a significant difference in mtDNA levels accord- It is unlikely that mtDNA quantification alone will be able to solve the complex
ing to embryo ploidy and maternal age, but no difference regarding implantation, scenery of embryo reproductive competence.
pregnancy and live birth outcomes. Trial registration number: not applicable

human
reproduction
ESHRE 2020 / Poster Viewing

Wider implications of the findings: The spermatozoa that were able to
POSTER VIEWING SESSION passthough COCs had higher sperm motility parameters, higher rates of normal
ANDROLOGY morphology, and lower DNA fragmentation. It may be advantageous to imple-
ment this process during ICSI.This study showed that implementing a sperm
selection method using an COCs model before ICSI may improve fertility
outcomes.
Trial registration number: HE 611250
P-001 Effect of sperm selection method by cumulus oophorus
complexes and conventional sperm preparation method on
sperm quality and DNA fragmentation for assisted reproduction P-002 Sperm retrieval surgery in male infertility cancer survivors
techonology can result in live births.
W. Naknam1, L. Salang1, J.S. Sothornwit-1, S. Amnatbuddee1, H. Hibi1, S. Miho1, S. Megumi2, F. Noritaka2, A. Yoshimasa2
K. Seejorn1, T. Pongsritasana1, S. Sookkasame1 1
Kyoritsu General Hospital, Urology, Nagoya, Japan ; 2Asada Ladies Clinic,
1
Department of obstetrics and gynecology Khon Khane university, obstetrics and Gyenecology, Nagoya, Japan
gynecology, Khon Khane, Thailand
Study question: To determine the viability of sperm retrieval surgery in
Study question: Are the effectiveness of sperm quality and DNA fragmenta- male infertility cancer survivors to restore the possibility of biological
tion different between sperm selection method by cumulus oophorus complexes fatherhood.
and conventional sperm preparation method ? Summary answer: Successful sperm retrieval was achieved in 40% cancer
Summary answer: Spermatozoa selected using COCs were likely to be effec- survivors, and 24% survivors became a biological fatherhood.
tive in terms of sperm quality and DNA fragmentation. What is known already: Sperm cryopreservation can be offered to men with
What is known already: Spermatozoa are commonly selected for intracy- cancer prior to undergoing gonadotoxic treatments such as chemotherapy.
toplasmic sperm injection (ICSI) visually under an optical magnification micro- However, cryopreservation of sperm prior to treatments is rarely done, whilst
scope based on motility and morphology. However, this method does not reveal after treatment, it is severely compromised due to azoospermia or ejaculatory
the genomic integrity of the spermatozoa.Cumulus oophorus complexs (COCs) dysfunction.
encircle the oocyte and contains cumulus cells and extracellular matrix (ECM). Study design, size, duration: To determine the viability of sperm retrieval
ECM is a main component in hyaluronic acid (HA), which is produced by COCs surgery in male infertility cancer survivors to restore the possibility of biological
after an LH surge.The head of mature spermatozoa have a hyaluronan specific fatherhood. Data from 1,525 male infertility patients who consulted in our clinic
receptor that bind with hyaluronan. During natural fertilization, only one healthy between 2009 and 2018 were analysed. From this database, a retrospective
spermatozoon can passthrough the COCs and zona pellucida and penetrate into analysis was done to identify patients with a history of past cancer treatment
the ooplasm for fertilization. and who underwent surgical sperm retrieval due to azoospermia or ejaculatory
Study design, size, duration: This was a prospective experimental study. dysfunction.
Thirty normal semen samples under the World Health Organization (WHO)’s Participants/materials, setting, methods: Twenty-five patients were diag-
2010 eligibility criteria were collected and processed using conventional sperm nosed as having azoospermia or ejaculatory dysfunction after cancer treatments.
preparation methods from June to October 2018. Type of cancers were testicular cancer (8 cases), hematologic cancer (6), col-
Participants/materials, setting, methods: The prepared sperm were orectal cancer (3), osteosarcoma (2), kidney cancer (2), extragonadal cancer
divided into two groups. Spermatozoa in the study group were selected based (2), gallbladder cancer (1), and rhabdomyosarcoma (1). Sperm retrieval surgery
on their ability to penetrate a layer of COCs via COCs selected model. In the was performed using vasal sperm aspiration in 4 cases, epididymal sperm aspi-
control group, spermatozoa were kept in culture medium under similar condi- ration (2), testicular microdissection (18), and onco-testicular sperm
tions. The selected-sperm were evaluated based on sperm quality and DNA extraction (1).
fragmentation. Retrieved sperm were cryopreserved for future ICSI treatment in our
Main results and the role of chance: center.
Thirty normal semen samples were recruited. Spermatozoa that were able Main results and the role of chance: Motile sperm were recovered in
to passthrough the COCs had significantly higher sperm motility param- 10 cases (10/25; 40%), whereas immotile sperm in 2. No sperm was obtained
eters than the control group (curvilinear velocity [VCL; 143.5 vs 122.2; in 13 (52%) cases. Six healthy deliveries were achieved with intracytoplasmic
P < 0.01], average path velocity [VAP; 83.6 vs 69.3; P < 0.01], straight-line sperm injection following use of motile sperm in each of six cases
velocity [VSL; 67.95 vs 60.45; P < 0.01]). The percentage of normal sper- (24%).
matozoa morphology in the COCs group was significantly higher than in Limitations, reasons for caution: The size of our sample was limited due
the control group (21.70 % vs 18.76%). In addition, there was significantly to the rarity of cancer cases in our clinic.
less DNA fragmentation in the COCs group than in the control group Wider implications of the findings: Successful sperm retrieval was
(18.83 vs 10.83). achieved in 40% cancer survivors, and 24% survivors became a biological
Limitations, reasons for caution: The limitation of this study, we did not fatherhood. On the other hand, some patients abandoned infertility
evaluate other sperm functions, such as sperm capacitation, acrosome reaction, treatment due to the costs associate with sperm retrieval surgery and fertility
and spermatozoa-zona binding. However, these processes are unnecessary in treatment.
ICSI because the ICSI usually bypass these processes. Trial registration number: Not applicable
BK-OUP-ESHRE2020_ABSTRACTS-200150-Poster.indd 144 22/06/20 8:44 PM

P-003 Quantitative analysis of the sHLA-G protein in seminal What is known already: Stress induces elevation in salivary cortisol and
plasma α-amylase. Prior studies have shown that women with high levels of
A. Schallmoser1, N. Sänger1 salivary α-amylase have reduced fertility compared to women with normal
1 values. A prior study of males suggested that higher levels of stress, as measured
University Hospital Bonn, Department of reproductive medicine, Bonn, Germany
by a four-item questionnaire, was associated with poorer sperm quality. The
Study question: To examine the paternal levels of the immunoregulatory impact of stress, as measured by salivary biomarkers, on male fertility is
sHLA-G protein in seminal plasma during IVF treatment and to investigate pos- not known.
sible correlations with age and pregnancy outcome of the female partner. Study design, size, duration: 94 infertile men with sperm concentration
Summary answer: High sHLA-G levels in seminal plasma of the male partner ≤15M/ml, motility ≤ 40%, or normal morphology ≤4% enrolled in the Males,
appear not to be required for pregnancy Antioxidants, and Infertility (MOXI) Trial, were included in this prospective
What is known already: Recent studies revealed that maternal and embryonic cohort study. Couples were followed for up to 6 months. Couples attempted
contributions impact on HLA-G protein expression and might contribute to to conceive naturally during the first 3 months and with clomiphene citrate with
pregnancy success or failure. intrauterine insemination in months 4 through 6 if not pregnant.
Also in seminal fluid, different levels of sHLA-G have been detected. Participants/materials, setting, methods: Men provided a first morning
Study design, size, duration: Retrospective study. 106 samples from male salivary sample and semen sample at baseline. Salivary samples were analyzed
donors were obtained between March and October 2018. for α-amylase and cortisol. Semen parameters and DNA fragmentation were
Participants/materials, setting, methods: Analysis of 106 paternal samples measured in the semen samples. The associations between salivary α-amylase
during IVF treatment. Semen was separated from seminal plasma using a density and cortisol and semen parameters or DNA fragmentation were assessed using
gradient consisting of a 90% lower layer and a 45% upper layer. Enzyme linked linear regression models adjusting for male age. Salivary levels were dichotomized
immunosorbent assay (ELISA), SDS-PAGE and Western blot were performed at the 80th percentile. Pregnancy and live birth rates in couples were compared
to analyze and confirm the sHLA-G values in paternal seminal plasma and TESE using chi square testing.
samples. Main results and the role of chance: Salivary levels of α-amylase were not
Main results and the role of chance: We observed a significant negative associated with semen parameters or DNA fragmentation. Salivary levels of
correlation of male age with total sHLA-G amount (P 0.023, R −0.221) in seminal cortisol were not associated with DNA fragmentation or normal sperm mor-
plasma. Testicular biopsy samples were analyzed and tested positively with phology. For every 1 unit increase in salivary cortisol the total sperm count
sHLA-G ELISA. No significant difference of sHLA-G levels in seminal plasma and increased by 18.1 million (95% CI: 4.5-31.73), and the total motile sperm count
pregnancy outcome of the female partner was detected. We observed a statis- increased by 12.6 million (95% CI: 4.5-20.6). Pregnancy rates did not differ for
tically significant difference between female age [P = 0.010, Mann-Whitney U partners of males in the highest quintile of α-amylase (26% and 27%, p= 0.92)
test] and pregnancy outcome. As expected, more ideal embryos and less non- or cortisol (44% and 24%, p=0.11) compared to partners of males with lower
ideal embryos were transferred in the pregnancy group. In univariate regression values. Live birth rates did not significantly differ for partners of males in the
only female age significantly predicted pregnancy outcome (OR 0.896, 95% CI highest quintile of α-amylase (21% and 20%, p=1.0) or cortisol (38% and 17%,
0.81-0.99, P = 0.026), while male age showed a trend to significance (P = 0.075). p=0.07) compared to partners of males with lower values.
Semen quality and sHLA-G were not significant. In multivariate logistic regression Limitations, reasons for caution: Men enrolled in the trial were infertile
including all parameters, a trend for an independent predictive value of female with abnormal semen parameters. These results are not generalizable to men
age was still detected (OR 0.911, 95% CI 0.81-1.03, P = 0.123). without infertility.
Limitations, reasons for caution: Study was retrospective and not random- Wider implications of the findings: Among infertile males, stress, as mea-
ized with a small sample size. sured by salivary α-amylase and cortisol, does not appear to negatively impact
Wider implications of the findings: We observed a significant correlation male fertility, and higher levels of cortisol may increase sperm count. Future
of male age with total sHLAG amount. Our analysis showed a wide spread of studies should investigate whether stress impacts semen parameters among men
sHLA-G protein levels in seminal plasma samples of male donors, supporting in the general population.
reports of previous studies. The determination of the origins of HLA-G in Trial registration number: Clinical Trials.gov NCT02421887
seminal fluid require further detailed studies.
Trial registration number: non applicable P-005 Alternations of sperm protein profiles to elucidate the
mechanism of impaired spermatogenesis by cancer chemotherapy
P-004 Biomarkers of stress and male fertility T. Takeshima1, K. Shinnosuke1, Y. Yasushi1
1
A. Steiner1, T. Spitzer2, F. Sun3, R.M. Coward4, K. Hansen5, Yokohama City University Medical enter, Department of Urology- Reproduction
S. Krawetz6, N. Santoro7, J. Trussell, C8 Center, Yokohama City- Kanagwa, Japan
1
Duke University, Obstetrics and Gynecology, Durham, U.S.A. ;
2
Naval Medical Center Portsmouth, Reproductive Endocrinology & Infertility, Study question: This study aims to analyze alterations in proteomic profiles
Portsmouth- VA, U.S.A. ; and validate selected protein biomarkers of spermatozoa in men with history
3
Yale School of Public Health, Collaborative Center for Statistics in Science, New of undergoing cancer chemotherapy.
Haven- CT, U.S.A. ; Summary answer: Cancer-associated protein was identified by liquid chro-
4 matography-mass spectrometer analysis and database searching. And the protein
UNC School of Medicine, Department of Urology, Chapel Hill- NC, U.S.A. ;
5 was validated by Western-blotting.
University of Oklahoma College of Medicine, Department of Obstetrics and
What is known already: As advanced cancer treatments have improved the
Gynecology, Oklahoma City- OK, U.S.A. ;
6 prognosis of cancer survivors, these treatments such as chemotherapy have
Wayne State University School of Medicine, Department of Obstetrics and
been known to cause harmful effect on fertile capacity. A few studies reported
Gynecology, Detroit- MI, U.S.A. ;
7 the alternation in proteins of spermatozoa between cancer patients and healthy
University of Colorado School of Medicine, Obstetrics and Gynecology, Aurora-
donor and some proteins with different expression levels were identified
Colorado, U.S.A. ;
8 between two groups.
Upstate University Hospital, Urology, Syracuse- NY, U.S.A.
Study design, size, duration: This research is a cross-sectional cell-line
Study question: Does stress, as measured by salivary α-amylase and cortisol, research of control versus treatment. A group of patients with a history of
negatively impact male fertility, as measured by semen parameters and couple anticancer drug administration in cancer diagnosis was assigned as “cancer
pregnancy and live birth rates? group”(n=3), and a fertile donor group was assigned as “control group”(n=3).
Summary answer: Biomarkers of stress in males are not negatively associated Written informed consent was obtained from all patients and this study design
with semen parameters or pregnancy and live-birth rates. Salivary cortisol is was approved by institutional review board of Yokohama City University
positively correlated with sperm count. Medical Center.

Participants/materials, setting, methods: The original diseases of cancer of 1.3 ( ±11.2) x 106. Furthermore, 24% of the participants achieved a TMSC
group were non-Hodgkin malignant lymphoma(n=2) and soft tissue tumor(n=1). greater than 5 million, giving them the possibility of intrauterine insemination,
Measuring the total sperm count by CASA, they were adjusted to 6 million in and 19% achieved a TMSC greater than 20 million.
all specimens, and protein concentration was adjusted by BCA assay. After trypsin Limitations, reasons for caution: The small number of participants and
digestion and desalting, the expressed proteins in spermatozoa were analyzed limited follow-up are the main limitations of this study
by LQ-MS/MS and database searching was performed in two groups. Validation Wider implications of the findings: Although advanced techniques of
was performed for the proteins with different expression levels by assisted reproduction are commonly offered as a first line therapy for men with
Western-blotting. severe oligospermia caused by varicocele, this study demonstrates that micro-
Main results and the role of chance: A total of 1,152 proteins and 5,268 surgical varicocelectomy can improve the SA parameters of such men, allowing
peptides were identified by global proteomics in both groups. Sorted by max these couples to choose less invasive treatments.
fold change of expressions (>5 folds) and ANOVA (p<0.01), 29 proteins were Trial registration number: not applicable
identified. Of these identified proteins, we focused on one protein, which is
cancer-associated protein highly expressed in digestive tract and urinary tract. P-007 Effect of a high fat diet on the spermatogenesis in an obese
In sperm of patients with cancer, this protein was overexpressed 54.8-folds more mouse model
than that of fertile donor. This protein co-works with other protein of T-cell L. Ramos1, D. Komninos2, G. Van der Heijden1,
proliferation factor. Regimens of cancer chemotherapy patients in cancer group T. Van Herwaarden3, A. Kiliaan4, I. Arnoldussen4
received were ABVD and IFM, ADM, and VCM therapy, respectively.By the 1
Radboudumc, Obst. & Gynaecology. div. Reproductive medicine intern 791,
Western-blotting, expression of this protein was validated.
NIJMEGEN, The Netherlands ;
Limitations, reasons for caution: Limitations of this research was inability 2
Radboud University, Faculty of Biology, Nijmegen, The Netherlands ;
to compare the sperm before and after the administration of cancer chemo- 3
Radboudumc, Medical Laboratory, Nijmegen, The Netherlands ; 4Radboudumc,
therapy, because spermatozoa were all cryopreserved for fertility preservation
Anatomy, Nijmegen, The Netherlands
before treatment.
Wider implications of the findings: It was speculated that T cell proliferation
Study question: Which changes in spermatogenesis are found in an obese/
was induced by interaction between these proteins after induction of cancer
overweight Ldlr-/-.Leiden mouse model by using a high fat diet (HFD) ?
chemotherapy. Functional analysis of these proteins would provide clue to the
Summary answer: This study demonstrates that diet induced obesity leads
mechanisms of impaired spermatogenesis after cancer chemotherapy.
to impaired testicular structure, probably due to the disturbance of the hypo-
thalamic-pituitary-gonadal (HPG) axis.
What is known already: Obesity can disturb one or more of these regulatory
P-006 Outcomes of Microsurgical Varicocelectomy in Men with factors, leading to impaired spermatogenesis and subsequently impaired fertility.
Severe Oligospermia To understand the effects of obesity on fertility at a cellular level, this study
F. Tenorio Lira Neto1, A.D.F. Cavalcanti Filho1, M.M.F. Santana1, analysed the testes of low-density lipoprotein receptor knock-out (Ldlr-/-.
T.D. Silva1, M.A.S. Freire1, E.D.P. Miranda2 Leiden) mice, which received a high fat diet and as such developed the typical
1
Instituto de Medicina Integral Prof. Fernando Figueira, Department of Urology, symptoms related to obesity.
Recife, Brazil ; Study design, size, duration: Knock-out mice with a normal diet (chow diet,
2
Universidade Federal do Ceará, Departmen of Urology, Fortaleza, Brazil n=15- control group) or a high fat diet (HFD, n=15) were used. Hormonal blood
levels, bodyweight of the animals at 36 weeks of age and analysis of adipose
Study question: Is microsurgical varicocelectomy useful in men with severe tissue (percentage of fat deposits) were measured. Differential diet started at 3
oligospermia? weeks of age until week 36 that the mice were sacrificed. Both testis (one fixed
Summary answer: Microsurgical varicocelectomy provides significant in Boiun and one snap frozen) were used for further analysis.
improvements in semen analysis parameters of men with severe oligospermia, Participants/materials, setting, methods: For histological evaluation,
allowing the couples to choose less invasive treatments. Sertoli cells, spermatocytes and spermatids were counted, and morphological
What is known already: According to the literature, men with severe oli- analyses of the testes were performed after haematoxylin-eosin stain. For anal-
gospermia (sperm concentration < 5 million sperm/mL) have a lower chance ysis of sperm cell quality, a TUNEL (terminal deoxynucleotidyl transferase dUTP
of semen analysis parameters (SA) improvement after microsurgical varicoce- nick end labelling) staining was performed on testes slices to quantify the apop-
lectomy (MV) when compared to men with mild or moderate oligospermia. totic cells. In addition, a chemical staining with chromomycin A3 (CMA3) was
However, recently, some authors have shown that MV can improve the SA performed on spermatozoa obtained from the epididymis to examine the chro-
parameters in some severe oligospermic men, potentially allowing for less inva- matin compaction of the sperm cell.
sive and costly assisted reproductive techniques. Main results and the role of chance: At 36-weeks of age, the control group
Study design, size, duration: This is a retrospective cohort study in 38 mean weight was 37.82 ± 1.23 grams while the HFD fed mice was 49.56 ± 0.90
consecutive men with severe oligospermia and clinically palpable varicocele who grams. The HFD fed mice had significantly more mesenteric and inguinal fat
underwent MV by two male fertility specialists between June 2016 and compared to the mice on a control diet ((p<0.001). Plasma measurements of
September 2019. cholesterol, triglycerides, insulin, leptin, and testosteron were al significantly
Participants/materials, setting, methods: We included those men and higher in HFD fed mice. No significant differences in the relative and absolute
who had at least one postoperative SA, and excluded those who were azoosper- number of seminiferous tubules in the testis slides between both groups was
mic. Baseline demographic and clinical characteristics, intraoperative findings, observed. But, the HFD fed mice had a significantly increased number of loosely
pre- and postoperative SA parameters, and SA improvement were evaluated. arranged cells in the seminiferous tubules, which a lower spermatids:spermato-
Semen analysis Improvement was defined as an increase of more than 10% in cytes ratio. The morphology of the seminiferous tubules in the testes was aber-
total progressive motile sperm count (TMSC). rant and the spermatogenic cycle of the seminiferous epithelium was disturbed
Main results and the role of chance: Thirty-eight participants were included in the testes of the HFD mice.
in the study. The mean age of the participants was 34 (±9) years, and bilateral No significant increase of TUNEL-positive cells were observed between
varicoceles were present in 70% of the men. Regarding preoperative SA, the groups. CMA3 measurements (chromatin condensation) did not differ between
median sperm concentration was 1.4 (±3.1) x 106/mL, the median progressivel groups (HFD mice= 3.5% ± 2.0; control=(2.4% ± 0.89) but it was increased
motility was 10 (±25) %, and the median TMSC was 0.2 (± 1.3) x 106. compared to the wild type C57BL/6 mice (aprox.1% CMA3-positive cells).
Intraoperative findings were, mean number of ligated veins of 11.9 on the left Limitations, reasons for caution: In this study only mice with a LDL-receptor
side and 9.3 on the right side; mean diameter of the largest vein of 3.5 mm on mutation were used. Our findings can be partly related to this mutation effect
the left side and 2.6 mm on the right side. After a median follow-up of 120 (±60) in the cells next to dietary feeding. The addition of a wild type mice with a normal
days, 79% of the participants improved their TMSC, with a median improvement of HFD diet will help to evaluate the effect of fat consumption.

Wider implications of the findings: These results are a substantial addition P-009 Semen exosomes upregulated trophocytes migration
to the emerging evidence of the negative impact of obesity on male fertility, and invasion and its effects on trophocytes angiogenesis via the
which is of great societal impact and will help with counselling and treatment of Notch1/NF-κB signaling pathway
infertile couples. Z. Xiao1, P. Jiali1
Trial registration number: Not applicable 1
Wuhan University, Renmin Hospital, Wuhan, China
P-008 Evaluation of a fully automated semen quality analyzer Study question: Does semen exosomes (Exos) affect trophocytes (Tros)
(LensHooke™ X1) for home-based monitoring biological behaviors and angiogenesis? And how does it work?
A. Agarwal1, M.K. Panner Selvam1, C.T. Hsu2 Summary answer: Semen Exos could promote Tros migration, invasion and
1
Cleveland Clinic, American Center for Reproductive Medicine, Cleveland, U.S.A. ; angiogenesis via the Notch1/NF-κB signaling pathway, have no significantly effect
2
The Taiwanese Association of Andrology, Male Infertility, Taipei City, Taiwan on cell cycle and apoptosis.
R.O.C. What is known already: At present, researches on Tros mainly focus on the
maternal, including hormone level, inflammatory factors, growth factors, etc.
Study question: Can LensHooke™ X1 serve as a home-based semen quality The study on the regulation of Tros biological behavior by semen may provide
monitoring (SQM) device and offer accurate test results? a new direction for elucidating maternal-fetal interface modeling.
Summary answer: LensHooke™ X1 semen quality analyzer offers not only Studies have shown that intrauterine infusion of SP can improve the implantation
a user-friendly experience for at home use, but it also provides accurate and rate of IVF-ET. Semen Exos mainly exists in SP, it’s considered to be an important
reproducible results. medium for micro-environmental regulation of reproductive function. RNA-seq
What is known already: At-home semen analysis is a choice for men who studies have shown semen Exos can regulate the expression of genes in endo-
are reluctant to visit the diagnostic laboratory for semen testing. Although home- metrium. However, the effect of semen Exos on Tros has not been studied.
based semen quality analyzers (HBSQA) are available in the market, the infor- Study design, size, duration: 40 cases of normal quality semen (according
mation regarding post-surgical SQM using HBSQA is not available in the literature. to the WHO 5th edition standards) were collected to extract semen Exos at
Most HBSQA tests for only one or a few parameters at a time and their results Renmin Hospital of Wuhan University. To investigate whether semen has an
cannot replace laboratory analysis of semen specimen. LensHooke™ X1 semen effect on Notch-1 expression in villus, a total of 12 villus (6 cases of pregnancy
quality analyzer, a fully automated portable device, measures more than 3 semen by IVF-ET as test group and 6 cases of pregnancy in natural way as the control)
parameters and provides real-time results for user, making it a good candidate were collected for immunohistochemical staining.
for semen quality monitoring. Participants/materials, setting, methods: Exos were extracted and iden-
Study design, size, duration: tified from 40 volunteers and Notch-1 were measured in villus from 6 patients
(1) Accuracy evaluation: 20 participants performed sperm concentration and conceived through IVF-ET and 6 patients conceived naturally. The human tro-
motility analysis using X1. phoblast cell line (HTR-8/SVneo cells) were co-cultured with 10ug/ml semen
(2) Reproducibility, precision and usability evaluation: To simulate at home SQM, Exos for 48h. Then CCK-8 assay, transwell assay, flow cytometry, and angiogen-
45 participants (28 Male, 17 Female) measured the concentration of latex esis assay were performed to determine cell proliferation, migration, invasion,
beads using X1 at home device. Tests were performed once a week for a apoptosis, cell cycle and tube formation. Q-PCR and western-blot analysis were
period of 5 months (20 readings). Following the completion of the trial, performed to determine RNA and proteins.
45 questionnaires were filled out to evaluate user experience. Main results and the role of chance: Exos were successfully extracted from
Participants/materials, setting, methods: semen and identified by transmission electron microscopy, NTA analysis and
(1) Semen analysis was conducted on 3 seminal aliquots from each leftover western-blot analysis. After co-cultured with semen Exos, HTR-8/SVneo cells
specimens. Sperm concentration and total motility were initially assessed by migration, invasion and tube formation were increased significantly (P<0.05,
medical technologist using Makler™ counting chamber. Participants with no n=3). And the expression of HTR-8/SVneo cells biological function-related
experience in lab work (n=10) and trained-lab professionals (n=10) subse- mRNA and protein, like Notch-1, NF-κB-VEGF, VEGFR-1, VEGFR-2 were sig-
quently performed semen analysis on these samples using X1 analyzer. nificantly increased (P<0.05, N=3). While there were no significant changes in
(2) To evaluate user-experience of X1 device, we recruited 45 participants of cell cycle and apoptosis (P>0.05, N=3). Mechanistic studies showed Notch-1
different age and educational background. signaling pathway played an important role. The expression of Notch-1 in villus
Main results and the role of chance: from patients conceived by IVF-ET was significantly lower than the control group.
(1) For the evaluation of accuracy in semen analysis, the results of semen testing Furthermore, the semen Exos promoting effects could be significantly attenuated
by either the participants with no experience in lab work (n=10) or by Notch-1 inhibitor.
trained-professionals (n=10) presented high degree of accuracy in concen- Limitations, reasons for caution: It remains unclear that which substance
tration (90 % vs 93 %) and total motility (90 % vs 90 %) compared to refer- in semen Exos is the upstream signal of notch-1 signaling pathway.
ence value. There were no significant differences between concentration and Wider implications of the findings: The study of roles of semen Exos of
motility results between the two groups (p = 0.84; p = 0.94). This suggests maternal-fetal interface modifying is of great significance not only in exploring
that users can operate X1 and obtain accurate results without hands-on the origin of reproduction, but also in preventing and controlling pathological
experience or additional training. pregnancy, improving IVF-ET pregnancy outcome, developing new contracep-
(2) Results from 45 participants showed a very high reproducibility (>98.6 %) tives, and improving the reproductive rate of rare animals.
for the measurement of latex beads concentration. Moreover, the question- Trial registration number: not applicable
naire shows 96% (±5.73%, 95% CL) positive feedback on the overall impres-
sion of X1 and up to 98% (±4.09%, 95% CL) satisfaction rate with an average P-010 Easy, efficient and safe sperm separation for IUI and IVF
score of 3.32 out of 4 points. The current data indicates that LensHooke™ A. Arav1, Y. Barak2, A. Komsky-Elbaz3, D. Reches4, D. Kalo4,
X1 is an accurate, reproducible and user-friendly device. Z. Roth4, Y. Natan5, P. Patrizio6
Limitations, reasons for caution: Use of latex beads instead of semen 1
Fertilesafe Ltd., R&D, Nes-Ziona, Israel ;
samples to evaluate the reproducibility and usability of LensHooke™ X1 may 2
Dr. Yona Barak Laboratory, Laboratory, Rosh Haayin Il, Israel ;
not mimic the results of a semen sample. 3
Department of Animal Science- Faculty of Agricultural- Food and Environmental Quality
Wider implications of the findings: LensHooke™ X1 is an easy-to-use Sciences- the Hebrew University of Jerusalem-, Animal Science, Rechovot, Israel ;
SQM device which appears to be well suited for long term monitoring of semen 4
Department of Animal Science- Faculty of Agricultural- Food and Environmental
quality after reproductive surgery as well as for short term SQM. Future studies Quality Sciences- the Hebrew University of Jerusalem-, Animal Science, Rechovot, Israel ;
to evaluate the performance of LensHooke™ X1 for post-medical or surgical 5
Fertilesafe, Animal Science, , Israel ;
intervention specimens will be useful. 6
Obstetrics- Gynecology & Reproductive Sciences- Yale Fertility Center & Fertility
Trial registration number: CS17027 Preservation, In Vitro Fertilization, New Haven, U.S.A.

Study question: Is it possible to do easy and simplified sperm separation Participants/materials, setting, methods: Semen samples were taken from
procedure using a new device named E. Sep. 50 patients who were diagnosed with normospermia. The sperms were prepared
Summary answer: E.Sep is an efficient and easy to use device allowing sperm with the Swim-Up Method, and spread slides were prepared, and the İmmune
separation from semen for IUI/IVF directly into a syringe by thermotaxis through Fluorescent Staining Method was used for CatSper protein. The Student’s t-test
micropores. and Pearson Correlation Test were used for statistical analyses.
What is known already: Conventional separation of sperm includes various Main results and the role of chance: The CatSper Protein was detected
technologies such as two-step washing technique, density gradient centrifugation with the IHC Method in Principal Peace Region with the middle part. The relation
(DGC) and swim up. These techniques expose sperm to several noxious con- of CatSper-positive sperms with sperm morphology and concentration was
ditions such as: reactive oxygen species (ROS), contaminants present in the found to be statistically significant (p<0.05). No statistical significance was
media or freezing solutions, air, pipetting, centrifugation steps, temperature shifts. detected with sperm motility (p>0.05). The present study examined the effects
Study design, size, duration: Experimental research carried out in two of CatSper Channels on reproduction, and showed that it will open new ways
different centers using animal and human sperm samples. to treat male infertility and for contraceptive purposes.
Participants/materials, setting, methods: Frozen/thawed bull semen Limitations, reasons for caution: The inclusion of only 50 normospermic
(n=5) were mixed and processed (3 replicates) with ‘swim-up’ using three dif- patients, and working with Swim Up samples in the study caused limitations in
ferent techniques: regular swim-up (RSU); E.Sep in horizontal and E. Sep in terms of the sampling.
vertical position. Spermatozoon mitochondrial membrane potential (ΔΨm), ROS Wider implications of the findings: The results of our study have the quality
levels and acrosome membrane integrity were evaluated by flow cytometry, of pioneering future studies to investigate the relations of CatSper-expressing
using specific kits (IMV Technologies). In humans, 3 fresh ejaculates were pooled cells with pregnancy. It is aimed with a new study project that IVF results are
and processed as 2 separated samples. Sperm concentration, motility and mor- investigated in terms of male infertility and unexplained infertility.
phology (WHO 2010) was assessed before and after separation. Trial registration number: None
Main results and the role of chance: The animal experiments showed
significantly higher ΔΨm, reflected by the ratio of polarized/depolarized P-012 Intra-Uterine Insemination (IUI): Is there an upper cut-off
mitochondria, in E. Sep sperm groups compared to RSU (0.7 ± 0.30 vs. level of the Number of Motile Spermatozoa Inseminated (NMSI)?
0.3 ±0.01; P < 0.02). The ΔΨm tended to be higher in vertical E. Sep group An analysis of 2.642 IUI cycles.
(2.1± 0.45; P <0.1), compared to horizontal E. Sep. The oxidation level, reflected L. Delaroche1, H. Caillou2, F. Lamazou1, E. Genauzeau1, P. Meicler1,
by the percentage of viable spermatozoa exhibiting ROS, was significantly lower P. Oger1, C. Dupont3, P. Humaidan4
in horizontal E. Sep (23.5% ± 5.83) and vertical E. Sep (4.8% ±1.24) compared 1
Ramsay Santé, Hôpital Privé de Parly 2- IVF Center, Le Chesnay, France ;
to RSU (53.0% ± 2.00 ; P <0.008 and P<0.0007 , respectively). Acrosome 2
Capionis, Capionis, Bordeaux, France ;
integrity was significantly higher in horizontal E. Sep (13.1 ± 3.06%) and vertical 3
Sorbonne Université, Saint Antoine Research center- INSERM équipe
E. Sep (29.4± 6.41) compared to RSU (3.2 ± 1.61%; P<0.02 and P<0.05, respec-
Lipodystrophies génétiques et acquises. Service de biologie de la reproduction-
tively).In the first human sperm pilot study (pool of 3 samples) we performed
CECOS- AP-HP- Hôpital Tenon, Paris, France ;
horizontal E. Sep. After 18 min at 37°C the number of motile sperm recovered 4
Aarhus University, The Fertility Clinic- Skive Regional Hospital Faculty of Health,
in the syringe (volume 0.2mL) was 6x106 with 12% normal morphology as com-
Skive, Denmark
pared to 3% normal morphology and 9.6 x106 motile sperm in the equivalent
Study question: Does an upper cut-off level of the NMSI exist, above which
0.2 mL unprocessed semen sample. In the second trial we recovered 4 x106 total
the Live Birth Rate (LBR) is negatively affected?
motile sperm compared to 6 x106 in the unprocessed semen sample.
Summary answer: No upper cut-off level for NMSI was found in IUI cycles.
Limitations, reasons for caution: Further studies with higher number of
However, the LBR correlated positively with the NMSI until 30 million.
patients and various sperm quality parameters, are currently planned.
What is known already: Several authors tried to determine a lower threshold
Wider implications of the findings: This easy and safe method can be use
of post-washed motile sperm in IUI. However, to date no data have been pub-
for IUI, IVF and ICSI.
lished regarding a possible upper cut-off level of motile sperms, above which
Trial registration number: not clinical trial
the LBR is negatively affected. Importantly, IVF laboratories are often used to
diluting sperm preparations when the NMSI exceeds 10 million. The objective
P-011 Examination of Spermatozoa CatSper membrane channel of this study was to determine the predictive value of the post-washing NMSI
proteins by immunohistochemical method and investigation of on the LBR.
their relationship with sperm parameters in normozoospermia Study design, size, duration: A total of 2.642 IUI cycles carried out from
S. Yazıcı1, T. Irez2 January 2010 to July 2018 with an NMSI >1 million were retrospectively analyzed.
1
T.C Biruni University, Histology and Embryology, İstanbul, Turkey ; Patients under the age of 43 years were stimulated, using either clomifene citrate,
2
Yeniyüzyıl University, Histology and Embryology, İstanbul, Turkey FSH or combinations. Sperm was prepared by swim-up selection. IUI cycles using
donor sperm or frozen-thawed sperm were excluded. The institutional review
Study question: Is the expression of Sperm CatSper protein related to sperm board approved the study and all patients gave written informed consent to the
parameters? use of their data.
Summary answer: It was determined that the concentration of the sperma- Participants/materials, setting, methods: A multivariate logistic regression
tozoa is related to the CatSper positive sperm ratio.No significant relations were model was used to assess the influence of NMSI of >10-20, >20-30 and >30
detected with the Sperm motility. million on LBR compared to a NMSI of >1-10 million. In addition to NMSI, this
What is known already: CatSper is a sperm-specific, voltage-dependent, model was adjusted for all clinically relevant factors, i.e. having demonstrated
Ca+2-selective, pH-sensitive and positive-load channel, and also provides the their influence on LBR as part of a preliminary selection step, using univariate
passage of calcium ions. CatSper Cation Channels contribute directly to the logistic regression. The odds ratio (OR) and their 95% bilateral confidence inter-
mobility of the sperm, regulate acrosome reaction, and balance the intracellular val from the multivariate model were calculated.
pH. It is considered that the mutations and disorders in any subunit of the Main results and the role of chance: A total of 2.642 IUI cycles in 1.036
CatSper structure cause infertility. In a trial that was conducted in mice, the loss couples were included. Mean female age was 33.5 ± 4.4 years at the time of the
of α-subunits caused infertility. For this reason, it is an important cation channel first IUI. The mean NMSI per cycle was 9.0 million [1-97]. The LBR increased
for sperm hyperactivation and male fertility. with the NMSI until 30 million without any maximum threshold (AUC = 0.5438).
Study design, size, duration: In this prospective and controlled study, the Using NMSI categories, the LBR per IUI cycle was 14.4%, 17.8%, 22.9% and 6.8%
18-35 year-old normozoospermic male patients who applied to the Cebeci for NMSI between >1-10, >10-20, >20-30 and >30 million, respectively
Hospital UYTE Center of Ankara University, Cebeci Hospital, between (p=0.003). Univariate analysis showed that the NMSI, female age, the number
December 2018 and August 2019 were included. The Semen analysis was carried of mature follicles and the estradiol level on day of ovulation trigger, the cycle
out according to WHO 2010 Criteria. rank and the etiology individually influenced the LBR. Multivariate analysis,

adjusted for clinically relevant factors, showed that the LBR was 1.49 and cycle. For each 5 cm increase in men’s waist circumference, the odds (95% CI)
1.76 times higher when IUI was performed with a NMSI of >10-20 million of implantation, clinical pregnancy and live birth per initiated cycle decreased by
(OR [95%CI] = 1.49 [1.10; 2.01]) and of >20-30 million (OR [95%CI] = 1.76 14% (2-24%), 12% (2-21%) and 9% (1-17%), respectively, after accounting for
[1.07; 2.91]), respectively compared to IUI with an NMSI of >1-10 million anthropometric and demographic characteristics of both partners. Results were
(p=0.0119). comparable when waist circumference was modeled in categories using tertiles
Limitations, reasons for caution: Although more than two thousand cycles of the observed distribution, when using WHO suggested cutoffs for abdominal
were included and the main outcome was LBR, the design was retrospective obesity, when restricting analyses to men with a normal BMI, and when restricting
and all cycles were performed in a single center. the analyses to fresh embryo transfer cycles.
Wider implications of the findings: The LBR after IUI can be optimized by Limitations, reasons for caution: As is the case of all observational studies,
inseminating a maximum of motile sperms. IUI preparations should not be diluted residual confounding cannot be ruled out. In addition, it is not known whether
when more than 10 million motile sperms are obtained. It remains to be deter- these findings are generalizable to couples attempting conception without med-
mined whether a dilution is necessary beyond 30 million. These preliminary ical assistance.
results call for a prospective RCT. Wider implications of the findings: Men’s abdominal adiposity, as measured
Trial registration number: COS-RGDS-2019-09-004-DELAROCHE-L by waist circumference, may adversely impact ART outcomes, even in the
absence of obesity as defined by BMI cutoff values. These results suggest that
P-013 Men’s waist circumference in relation to infertility central obesity may be an independent risk factor for male factor infertility.
treatment outcomes among couples undergoing assisted Trial registration number: not applicable
reproductive technologies
H. Bian1,2, L. Mínguez-Alarcón3, I. Souter4, J. Chavarro, E.1,5,6 P-014 Testicular sperm extraction (TESE) and genetic analysis
1 feedback in a non-obstructive azoospermic man presenting a
Harvard T.H. Chan School of Public Health, Department of Nutrition, Boston,
46,XY/46,XX constitutional chimerism.
U.S.A. ;
2
Peking University School of Public Health, Institute of Reproductive and Child X. Ferraretto1, J.F. Hermieu2, M.A. Llabador1, E. Amar3,
Health and Department of Epidemiology and Biostatistics, Beijing, China ; A. Hercent4, J.P. Siffroi4
3 1
Harvard T.H. Chan School of Public Health, Department of Environmental AP-HP - Hôpital Bichat-Claude Bernard, Service de Biologie de la Reproduction,
Health, Boston, U.S.A. ; Paris, France ;
4 2
Massachusetts General Hospital Fertility Center and Harvard Medical School, AP-HP - Hôpital Bichat-Claude Bernard, Service d’Urologie, Paris, France ;
3
Vincent Department of Obstetrics and Gynecology, Boston, U.S.A. ; Cabinet d’Andrologie Victor Hugo, Andrologie Urologie, Paris, France ;
5 4
Harvard T.H. Chan School of Public Health, Department of Epidemiology, AP-HP – Hôpital d’Enfants Armand Trousseau, Département de Génétique
Boston, U.S.A. ; Médicale, Paris, France
6
Channing Division of Network Medicine, Brigham and Women’s Hospital and
Harvard Medical School, Boston, U.S.A. Study question: What is the sex ratio of spermatozoa incoming from sper-
matogenesis of a man presenting a 46,XY[70%]/46,XX[30%] constitutional
Study question: Is there an association between men’s waist circumference chimerism?
and ART outcomes? Summary answer: Sex ratio of testicular spermatozoa analysed by fluorescent
Summary answer: Men’s waist circumference was inversely related to preg- in situ hybridization (FISH) was 1:1, confirming that spermatogenesis is only
nancy and live birth rates among couples undergoing ART independently of conducted by 46,XY spermatogonia.
men’s and women’s body mass index (BMI). What is known already: Constitutional chimerism results from in utero fusion
What is known already: Female adiposity, overall and central, has been of two different zygotes in one single embryo. However, its prevalence is not
related to worse ART outcomes. Likewise, men’s obesity has been related to well known, and only a few cases of 46,XY/46,XX chimerism have been
poor semen quality and may also have a negative effect on ART outcomes. Waist reported to date. Moreover, no clinical case has been published on the explo-
circumference as a measure of central obesity is also negatively associated with ration of spermatogenesis in 46,XY/46,XX chimerism by FISH analysis per-
semen quality parameters in men of subfertile couples. Whether men’s waist formed on spermatozoa after TESE.
circumference impacts a couple’s fertility independently of their and their part- Study design, size, duration: We studied a 35 years old patient with
ner’s BMI is unknown. non-obstructive azoospermia. The patient was not issued from a twin preg-
Study design, size, duration: We utilized data from the Environment and nancy and had no history of heterologous stem cell transplantation. He presented
Reproductive Health (EARTH) Study, a prospective cohort study, which recruited a normal male phenotype but with a clinical bilateral testicular hypotrophy
subfertile couples seeking evaluation and treatment at the Massachusetts General (left=4mL and right=8mL). Hormonal exploration results were: FSH=16.0UI/L,
Hospital Fertility Center. Measurement of waist circumference was introduced LH=10.5UI/L, Testosterone=3.68ng/mL and Inhibin B=18pg/mL.
to the study in October 2009. This analysis includes 179 couples undergoing Genetic analyses performed on peripheral blood cells showed a
ART utilizing their own gametes, with complete anthropometric information, 46,XY[70%]/46,XX[30%] chimeric karyotype, and no chromosome Y azo-
recruited through January 2019. ospermic factor (AZF) deletion.
Participants/materials, setting, methods: Men’s and women’s height, Participants/materials, setting, methods: A bilateral TESE was performed
weight and waist circumference were measured on-site at baseline by trained by a senior urological surgeon. Sperm extraction was performed using swim-up
personnel. Clinical treatment outcomes were extracted from the medical and centrifugation. A few normal motile spermatozoa were retrieved on both
records. We analyzed the association of men’s waist circumference and clinical sides and frozen. Remaining fresh testicular tissue was used for sperm analysis
outcome measures using cluster-weighted generalized estimating equation mod- by FISH using direct hybridization probes coding for X (DXZ1) and Y (DYZ3)
els to account for repeated treatment cycles of the same couple and adjust for chromosomes and for chromosome 18 (D18Z1) as control of haploidy. FISH
men’s age, BMI, height, race, education level, smoking status, and women’s age, was performed separately on spermatozoa retrieved from right and left testic-
BMI, waist circumference and height. ular tissue
Main results and the role of chance: Men’s median (interquartile range, Main results and the role of chance: On the right side, 96 nuclei were
IQR) age, waist circumference, and BMI were 36.6 years (32.9-40.1), 95 cm observed and results showed nearly identical rates of X and Y bearing sperma-
(89-103) and 26.5 kg/m2 (24.2-29.1), respectively. The correlation coefficient tozoa: nuc ish(DXZ1x1,D18Z1x1)[45]/(DYZ3x1,D18Z1x1)[51]. Similar results
of men’s waist circumference and BMI was 0.58 (P <0.0001). The correlation were found on the left side after the analysis of 100 sperm nuclei: nuc ish(DX-
coefficients between men’s and women’s BMI and waist circumference were Z1x1,D18Z1x1)[49]/(DYZ3x1,D18Z1x1)[51].
0.29 and 0.40 (P <0.0001), respectively. Men’s waist circumference was unrelated These results highlight a 1:1 sex ratio on spermatozoa analysed from both
to fertilization rate overall and when IVF and ICSI cycles were separately exam- testicular tissues, thus strongly suggesting that spermatogenesis was initiated
ined. However, men’s waist circumference was inversely related to clinical out- from 46,XY spermatogonia. Performing FISH separately on both sides limited
comes, including implantation, clinical pregnancy and live birth rates per initiated the risk of error and the role of chance.

Frozen testicular sperm was used in intracytoplasmic sperm injection (ICSI) technologies to overcome current clinical limitations, to improve patient care
and resulted in the birth of a healthy boy following the first embryo transfer. and ultimately treatment outcomes.
Limitations, reasons for caution: Given that phenotypes are highly Trial registration number: Not applicable
variable in this type of patient and are also related to the proportion of the
two cellular contingents, our results in terms of presence of testicular sper- P-016 Improvement of semen parameters by pre-conception care
matozoa and genetic analysis cannot be applied to all patients with for male partners of infertility couples
46,XX/46,XY chimerism. A. Komiya1,2, N. Uchida3, A. Kuga3, M. Ibayashi3, K. Hiraoka3,
Wider implications of the findings: This clinical case shows that it is possible K. Kawai3, T. Ichikawa1
to find usable sperm with a 1:1 sex ratio after TESE. These data can be used to 1
Chiba University Hospital, Urology, Chiba, Japan ;
inform future patients with this genetic abnormality about the safety and potential 2
Kameda IVF clinic Makuhari, Urology, Chiba, Japan ;
success of this procedure, without transmitting this genetic abnormality to their 3
Kameda IVF clinic Makuhari, Reproductive Medicine, Chiba, Japan
offspring.
Study question: This study was conducted to analyze whether lifestyle mod-
ification as a part of preconception care could improve semen parameters among
P-015 A systematic review of at-home semen analysis the male partners of couples who visited a fertility clinic.
technologies: A potential supplement to the fertility clinic? Summary answer: Lifestyle modification as a part of pre-conception
F. Willendrup1, E. Andersen1, C. Panayotidis2 care for male partners improved semen parameters without urologic
1
ExSeed Health ApS, Denmark, Copenhagen, Denmark ; intervention.
2
Gynaecology, Gynaecology, Isle of Wight, United Kingdom What is known already: Pre-conception care is the provision of biomedical,
behavioral and social health interventions to women and couples before con-
Study question: The aim was to determine the use and validity of current ception occurs. It aims at improving their health status, and reducing behaviors
at-home semen tests (HST), and their screening potential of male subfertility. and individual and environmental factors that contribute to poor maternal and
Summary answer: Most HST’s are user friendly, accurate and effective diag- child health outcomes. Such care is also required for male partners of infertility
nostic tools for primary and secondary care services. couples. Recent Japan’s National survey for male infertility revealed that 42.1%
What is known already: Semen analysis remain the main tool for diagnosis of male factor infertility was idiopathic, suggesting that at least some of these
of male fertility status. Conventional spermograms use either automated or were caused by inappropriate lifestyle factors.
manual evaluation under a microscope. A basic predictive parameter is the Total Study design, size, duration: This is a single-arm, single-center, retrospective
Motile Sperm Count, which correlates well with pregnancy outcomes. HST observation study for a duration of one year after June in 2018 with 203 con-
provide an immediate result on basic semen parameters increasing the flexibility secutive subjects.
and easiness for the men. Although HST technologies are recently commercial- Participants/materials, setting, methods: Non-azoospermic male part-
ised the general gynaecologists, practitioners and fertility specialists are not aware ners of infertility couples were enrolled and asked about their lifestyles. Median
about their existence. The use of HST in primary and secondary care could age of male patients was 35 years old. These patients were promoted to modify
improve the early correction of subfertility conditions and potentially improve their inappropriate lifestyles as pre-conception care before urologic evaluation.
pregnancy rates. The lifestyles include factors such as smoking, chronic alcohol use, inappropriate
Study design, size, duration: The research protocol followed the published underwear use, excessive body weight, length of abstinence and so on. Semen
methodology for Preferred Reporting Items for Systematic Review and Meta- parameters was compared between before and after the promotion by student
Analysis Protocols (PRISMA-P), including their extension for Diagnostic Test T or chi-square tests.
Accuracy (PRISMA-DTA). The literature search included online databases with Main results and the role of chance: Forty-seven men (23.2%) were current
a predefined search strategy. Additional search was performed across online smokers. 94 (46.3%) were chronic alcohol users. The lifestyles elevating scrotal
markets to find available tests not yet included in scientific reporting. All published temperature including fitted underwear and/or excessive time bathtub/sauna
material up to and including 12th of January 2020 was included. use were found in 150 (73.9%). Body mass index (BMI)=or>30 was found in
Participants/materials, setting, methods: All available HST’s were 16 (7.9%). Blood tests revealed hypogonadism in 27 (13.3%), zinc deficiency
detected via research online, including any relevant applications in App Store in 30 (14.8%), abnormal lipid metabolism in 58 (28.6%), abnormal liver function
(iOS) and Google Play (Android). All articles published on testing methods or in 42 (20.7%). Palpable varicoceles were found in 41 (20.2%). Erectile dysfunction
devices used for analysing semen samples at-home or outside the conventional was found in 80 (39.6%). Most of patients (200, 98.5%) had at least one factor.
laboratory were included. Each technology and test were reviewed based on Then, changes in semen parameters were compared before and after the pro-
adapted predefined checklist from PRISMA-DTA. motion of lifestyle modification. Median intervals of semen analyses were
Main results and the role of chance: 12 home sperm tests were identified 28 days. Abstinence was decreased from 3.7 to 2.8 days in average (p=0.0135).
and included in this review. Across the technologies, seven test methods were Oligozoospermia was decreased from 99/203 cases (48.8%) to 77/197 (39.1%,
identified for the semen analysis, including manual microscope, automated micro- p=0.0511). Asthenozoospermia was decreased from 160/203 (78.8%) to
scope, microfluidics antibody reaction, paper-based antibody reaction, paper- 91/196 (53.6%, p<0.0001). Oligoasthenozoospermia was decreased from
based antibody reaction, microfluidic centrifugation, and mail-in tests. Some of 81/203 (39.9%) to 47/196 (23.7%, p<0.0001). Total motile sperm count was
them were smartphone enabled while others were paper-based. The accuracy improved from 18.7±27.3 million to 33.1±44.6 million (mean±standard devia-
of these tests was between 95-98 % and the “time until results” ranged between tion, p<0.001). However, such improvement was not significant in those with
a few seconds to several days. palpable varicocele (4.7±25.0 million increase, p=0.2488), hypogonadism
The majority of HST’s give qualitative information on either the concentration (22.3±58.4 million increase, p=0.0685), BMI=or>30 (33.1±69.2 million increase,
or motile concentration. Most of the HST provide only the user with interpre- p=0.0753), and zinc deficiency (8.3±34.2 million increase, p=0.1947).
tation of the single sperm parameter result. Limitations, reasons for caution: This study was a single-arm, single-center,
New high tech HST’s include more in-depth analysis of several parameters. retrospective study with relatively small sample size. Results could be different
It is suggested by the authors that HST’s can now aid further in improving patient if the study is conducted by double-arm, multi-center and prospective fashion
experience and fertility awareness. in larger sample size.
Limitations, reasons for caution: Material for some of the tests is unreliable, Wider implications of the findings: Most of male partners of infertility cou-
incomplete and subject to commercial imperatives. The study did not assess ples had lifestyle and/or medical factors to decrease semen quality. Promotion of
user experience and reproductive outcomes. lifestyle modification should be attempted before and during urological/medical
Wider implications of the findings: Our review shows that some HST are intervention. However, a short-term modification was not enough in those with
now measuring multiple sperm parameters, as well as reaching a high accuracy, varicocele, hypogonadism, zinc deficiency and excessive BMI.
making them valid tools for semen analysis. We propose a way for these Trial registration number: not applicable

P-017 A new small-molecule inhibitor of BRDT shows reversible Summary answer: Sperm selected by microfluidic sorting are associated with
male contraception effect in mice model significant increase in live birth rate, clinical pregnancy rate and reduced miscar-
X. Li1, S. Wu1, J. Hou1, M. Yin2, W. Xu3 riage rate.
1 What is known already: DNA damage is unrecognisable in living sperm prior
Sichuan university, West China Second University Hospital, West China School of
to insemination and an increased sperm DNA fragmentation index has been
Medicine ;
2 associated with lower fertilization rates, impaired embryo development and
Central South University, Department of Dermatology- Huan Engineering
reduced pregnancy rates. Standard semen processing techniques are associated
Research Center of Skin Health and Disease- Hunan Key Laboratory of Skin
with centrifugation, which may induce reactive oxygen species and DNA damage.
Cancer and Psoriasis- Xiangya Hospital, Changsha, China ;
3 Microfluidic systems are capable of working with small volume samples and have
Sichuan university, Joint Laboratory of Reproductive Medicine- SCU-CUHK- Key
high sensitivity and low response time. This technique helps to improve the
Laboratory of Birth Defects and Related Diseases of Women and Children-
efficiency of sample preparation, enable consistency in embryo culturing and
Ministry of Education- West China Second University Hospital, West China School
reduce human error. It has been demonstrated that microfluidic technique could
of Medicine
provide sperm with significantly reduced DNA damage.
Study question: To date, a reversible, reliable and oral medication to male Study design, size, duration: A prospective randomised control study was
contraceptive has not been successfully applied in the clinic. conducted from 1st August 2017 to 31st December 2018. One hundred and
Summary answer: We have discovered a dose-dependent small molecule nighty eight patients were randomised by computer generated list and divided
Inhibitor of BRDT that can reversibly achieve male contraception through oral into 2 groups. Group A (n=100) , in which sperm were processed by microfluidic
medication in animal experiments. sperm sorter while in group B (n=98), sperm were processed by density gradient
What is known already: Effective male contraceptives include condoms and technique and morphologically normal motile sperm were injected by
vasectomy, but these methods are not ideal for all men. The bromodomain Intracytoplasmic sperm injection (ICSI) technique in all mature oocytes.
(BRD) and extra-C terminal domain (BET) protein family consists of four mem- Participants/materials, setting, methods: The study period included all
bers (BRD2, BRD3, BRD4 and BRDT). These bind to acetyl lysine (KAc) residues normozoospermic patients with high DNA fragmentation index (>25% ) while
on the tails of histones H3 and H4, and regulate chromatin structure and gene oligospermic, asthenozoospermic samples, patients with poor ovarian reserve
expression. The testis-specific BET member, BRDT, is essential for the normal and advanced age were excluded from the study. All A grade embryos were
progression of spermatogenesis and mutations in the Brdt gene result in com- vitrified and transferred in frozen embryo transfer cycle. Both groups were
plete male sterility in mice model. Therefore, promising method acts by blocking compared on the basis of fertilisation rate, day 3 grade A embryo development
Brdt involved in the process of spermatogenesis for male contraception. rate, clinical pregnancy rate, miscarriage rate and live birth rate.
Study design, size, duration: We used 60 male mice divided into six groups Main results and the role of chance: Cycle characteristics (female age,
(0mg/kg, 1mg/kg, 2mg/kg, 3mg/kg, 4mg/kg, 5mg/kg) treated daily from 6 to length of stimulation, gonadotrophin dose, number of oocytes and number of
14 weeks with NHWD-870, a potent BRDT inhibitor. After drug or vehicle transferred embryos) were similar in both groups.
treatment, mice were either sacrificed or mated to females while continuing to Between the two groups, there was a significant increase observed in group
receive NHWD-870. A over group B in day 3 grade A embryo development rate (60% vs. 38%,
Participants/materials, setting, methods: C57BL/6 mice model, gene p-0.003), clinical pregnancy rate (62% vs. 41%, p-0.004) and live birth rate
cloning, mouse contraceptive studies, hematoxylin-eosin staining, sperm counts (46% vs. 29%, p-0.011), while a significant decrease in miscarriage rate ( 12% vs.
and motility, sex hormone radioimmunoassay, immunohistochemistry, western 25%, p-0.028). On the other hand there was no statistical difference observed
blot, Immunofluorescence. in fertilisation rate (82% vs. 78%, p-0.80).
Main results and the role of chance: NHWD-870 was the most potent Limitations, reasons for caution: Larger randomised control studies are
inhibitor of BRDT with a biochemical IC50 of 2.5 nM, 13.1nM, 2.3nM, 13.93nM needed to strengthen these results.
on GC1, GC2, TM3 and TM4 cell lines proliferation. We tested spermatogenic Wider implications of the findings: We have demonstrated that sperm
effects of NHWD-870 administered to male mice. High dose groups (4mg/kg sorted by microfluidic not only helps in selection of sperm with better DNA
and 5mg/kg) had a significant reduction to 74.4% in testis weight compared integrity but also increased live birth rate. Using it in routine practice can help
control. Epididymal sperm number was reduced to 5% of control and sperm in reducing the negative effect of reactive oxygen species and thus improve
motility was decreased to 3% of control. Moreover, high dose group showed pregnancy rate and live birth rate.
sterility mated with normal females. Biochemical study showed that the BrdT Trial registration number: MCDH/2017/35
expression is dose-dependantly decreased in these mice model. Low dose (1mg/
kg, 2mg/kg and 3mg/kg) groups had no difference in testis weight and fertility P-019 Male dietary fat intake and fecundability: a preconception
but sperm number and motility was decreased compared with control. After cohort study
withdrawal the drug about 25 days, the high dose group males had recovery of L.A. Wise1, A. Wesselink1, S. Willis1, E. Mikkelsen2, K. Tucker3,
the fertility, testis weight and sperm number. A.S. Dam Laursen2, K. Rothman1, E. Hatch1
Limitations, reasons for caution: Mice treated with NHWD-870 showed 1
Boston University School of Public Health, Department of Epidemiology, Boston,
decreased level of testosterone in serum. High dose drug also lead to decreased U.S.A. ;
body weight after 6 weeks treatment. 2
Aarhus University Hospital, Department of Clinic Epidemiology, Aarhus, Denmark ;
Wider implications of the findings: By optimizing the structure of the 3
University of Massachusetts at Lowell, Biomedical & Nutritional Sciences, Lowell,
NHWD-870, it could be possibly used in human as an oral medication for U.S.A.
reversible male contraception.
Trial registration number: Not applicable Study question: To what extent does male dietary fat intake influence
fecundability?
Summary answer: We observed little overall association between intakes of
P-018 Live birth rate of patients where sperm selected using total fat, and most fat subtypes, and fecundability.
microfluidic technique in high DNA fragmentation index sperm What is known already: Higher intakes of saturated fat and trans fatty acids
samples have been associated with poor semen quality and low testosterone levels,
F. Khan1, K.D. Nayar1, G. Kant1, M. Singh1, S. Mishra1, S. Gupta1, whereas higher intakes of omega-3 fatty acids have been associated with
R. Bhattacharya1, K.D. Nayar1 improved semen quality in some studies. There have been few preconception
1
Akanksha IVF Centre- Mata Chanan Devi Hospital, Reproductive Medicine, New cohort studies of male diet and fecundability.
Delhi, India Study design, size, duration: Pregnancy Online Study (PRESTO) is an ongo-
ing North American prospective preconception cohort study. Analyses were
Study question: Does microfluidic sorting technique help in increasing live restricted to 446 couples attempting conception for ≤6 cycles at enrollment
birth rate in patient with high DNA fragmentation index (DFI) sperm samples? during 2013-2020.

Participants/materials, setting, methods: Men aged ≥21 years completed nmol per ejaculate] than that in non-normozoospermic group [347.58(447.61)
an online baseline questionnaire on demographic, medical, and anthropometric nmol per ejaculate] (P<0.001). Mean value of DFI in normozoospermic group
factors. Ten days after enrollment, they completed a validated food frequency was6.97% (6.07%), which was lower than DFI in abnormal group [14.51%
questionnaire (DHQ II). Their female partners completed bimonthly follow-up (13.41%)] (P<0.001) Content of L-carnitine in semen was positively corelated
questionnaires for up to 12 months or until pregnancy. We used proportional with total sperm count, percentage of progressively motile sperm and percentage
probabilities regression to estimate fecundability ratios (FR) and 95% CIs for of morphologically normal spermatozoa, whose coefficient of correlation was
associations of % energy from total fat and fat subtypes with fecundability, adjust- 0.580 (P<0.001), 0.150 (P<0.001) and 0.131 (P<0.001) respectively. Moreover,
ing for energy and other covariates. L-carnitine in seminal plasma was negatively corelated with DFI, whose coefficient
Main results and the role of chance: Intakes of total fat, saturated fat, of correlation was 0.065 (P<0.001).
mono-or poly-unsaturated fat, or trans fat intake were not appreciably associated Limitations, reasons for caution:
with fecundability. FRs (95% CIs) comparing the top vs. bottom quartiles of It is a single-center study and the above results were from retrospective analysis
dietary fat intake were 1.00 (0.73-1.36) for trans fatty acids, 1.15 (0.84-1.57) for Wider implications of the findings:
omega-3 fatty acids, 0.78 (0.56-1.08) for omega-6 fatty acids, and 1.23 The above results can be used as references in oral L-carnitine of treatment in
(0.93-1.64) for the ratio of omega-3 to omega-6 fatty acids. improving semen paraments and the choice of antioxidants in reducing sperm
Limitations, reasons for caution: We observed little overall association DNA fragmentation.
between male intake of total fat, and most fat subtypes, and fecundability. Weak Trial registration number: Not Applicable
associations were seen for intakes of omega-6 fatty acids and the ratio of
omega-3 to omega-6 fatty acids, though associations were imprecise.
P-021 The relationship between seminal leukocytes and oxidative
Wider implications of the findings: These data contribute to the limited
stress markers in semen
existing literature on the association between male diet, specifically fat intake,
and fecundability. Although numbers were small, these findings indicate that male S. Kuroda1, T. Takeshima1, Y. Yumura1
1
dietary intake of fat is not a strong determinant of fecundity. Yokohama City University Medical Center, Department of Urology- Reproduction
Trial registration number: NICHD grants R21HD072326, R01 HD086742, center, Yokohama, Japan
and R03 HD090315
Study question: Does leukocyte concentration in semen and semen param-
P-020 L-carnitine in seminal plasma correlated with semen eters correlate with oxidative stress markers?
parameters and sperm DNA fragmentation Summary answer: Seminal leukocytes concentration was strongly correlated
L. Ye1, W. Huang1, M. Song1, J. Han1, Y. Peng1, T. Wu2, Y. Zeng3,4 with ROS level in semen, and ORP was more negatively correlated semen param-
1
Shenzhen Zhongshan Urology Hospital, Department of Andrology and Urology, eters than ROS.
Shenzhen, China ; What is known already: Reactive oxygen species (ROS) in semen has been
2
Shenzhen Zhongshan Urology Hospital, Department of Reproductive Immunology, reported to have negative effect to male fertile capacity, and recent studies
Shenzhen, China ; reported the efficacy of oxidation-reduction potential (ORP) which reflects the
3
Shenzhen Zhongshan Urology Hospital, Fertility Center, Shenzhen, China ; balance of oxidants and antioxidants in semen. The source of ROS in semen is
4
Shenzhen Zhongshan Urology Hospital, Shenzhen Key Laboratory for considered as immature spermatozoa and seminal leukocytes though the detail
Reproductive Immunology of Preimplantation, Shenzhen, China is still unknown. Pyospermia, globally defined as the presence of more than one
million leukocytes in 1 mL of semen, is considered as an indicator for genital
Study question: Is there any relationship between L-carnitine in seminal plasma infection and inflammation.
and sperm count, motility, morphology and DNA integrity? Study design, size, duration: Between November 2018 and July 2019, 50
Summary answer: L-carnitine in seminal plasma was positively corelated with infertile males who visited our hospital were enrolled. All patients underwent
sperm count, progressively motile sperm, morphologically normal sperm as well semen analysis and measurement of ROS and ORP levels and the concentration
as sperm DNA fragmentation. of leukocytes in semen. The correlation between these values were analyzed
What is known already: L-carnitine, presenting in testis and epididymis, is retrospectively.
involved in sperm maturation and it is used as antioxidants in therapy of idiopathic Participants/materials, setting, methods: ROS level in semen was mea-
male infertility. Evidence showed that oral L-carnitine can improve sperm param- sured using Monolight 3010TM Luminometer. Before and after adding 40 mL of
eters including total sperm count, concentration, progressive motility, vitality, 100 mmol/L luminol to 500 mL of semen sample, the subtraction of the inte-
and morphology in infertile men. Moreover, recently publications indicated that grated chemiluminescence was measured between 0 and 200 seconds. ORP
L-carnitine preserves the sperm membrane and DNA integrity. But limited level per sperm concentration (sORP) was measured using MiOXSYS SystemTM.
researches investigated the relationship of L-carnitine in semen and sperm The concentration of leukocytes were evaluated using myeloperoxidase staining
parameters as well as sperm DNA fragmentation. (Endtz test). The relationships between these values and semen parameters
Study design, size, duration: Data of semen analysis from January 2017 to were evaluated using non-parametric correlation analysis.
December 2019 were involved and divided into normozoospermic group and Main results and the role of chance: The average patient’s age was 37.1
non-normozoospermic group according to WHO guideline. Total content of (20-53). ROS level was positively correlated with leukocyte concentration
L-carnitine in seminal plasma, sperm DNA fragmentation index (DFI) and semen (ρ=0.646, p<0.001) although sORP didn’t show significant correlation. Both
parameters including sperm numbers, motility and morpology were retrospec- ROS level and sORP showed negative correlation with sperm concentration and
tively analyzed. For all the included study subjects, the leukocytes count in semen motility significantly, although the Spearman correlation coefficient was higher
less than 1 × 106/ml. in sORP than in ROS (ρ= -0.873 vs -0.425, -0.547 vs -0.228, respectively). sORP
Participants/materials, setting, methods: Sperm count and motility were was also negatively correlated with straight velocity and mean amplitude of
analyzed by computer assisted sperm analysis (CASA) system, while sperm lateral head displacement (mean ALH). When the positive cut-off value of ROS
morphology was examined after Diff-Quik rapid staining procedure. L-carnitine level was set 4332 RLU as previously reported, ROS was positive more than
in seminal plasma was determined by high-performance liquid chromatography. 1.1×105/ml of leukocytes in semen according to ROC curve (AUROC= 0.862) .
DFI was calculated from sperm chromatin structure analysis (SCSA). Content Limitations, reasons for caution: This study is retrospective and single
of L-carnitine and DFI of normozoospermic group abnormal group and was center analysis with small number.
compared by Mann-whitney U test. Bivariate correlation analysis was used to Wider implications of the findings: Seminal leukocyte was considered as
explore the relationship between L-carnitine and sperm paraments and DFI. a source of ROS in semen. ROS was positive even under the global definition
Main results and the role of chance: A total of 4437 subjects were enrolled of pyospermia, thus fewer leukocytes than definition may have adverse effects
in the study and normozoospermic group contained 2591 cases while non-nor- to sperm quality through ROS generation. ORP have potential as keener bio-
mozoospermic group contained 1846 cases. Median value of L-carnitine in sem- marker to reflect the sperm quality than ROS.
inal plasma was significantly higher in normozoospermic group [458.78(439.92) Trial registration number: not applicable

P-022 Does Testicular Sperm Aspiration have a place in the 3

Imperial College Healthcare NHS Trust, Urology, London, United Kingdom ;
Management of Non-Obstructive Azoospermia Patients? 4
Guy’s and St Thomas’ NHS Foundation Trust, Urology, London, United Kingdom ;
5
A. Majzoub1, K. Khalafalla1, H. Elbardisi1, S. Alsaid1, M. Arafa1 Imperial College Healthcare NHS Trust, Andrology, London, United Kingdom
1
Hamad Medical Corporation, Urology, Doha, Qatar
Study question: What constitutes the semen microbiome (SM) and what is
the impact of the semen microbiome on male fertility and semen parameters?
Study question: Are there any clinical predictors that could help in selecting
Summary answer: Specific bacterial species are associated with worsening
non-obstructive azoospermia (NOA) candidates for testicular sperm aspira-
sperm quality, whilst others are protective, but the association with infertility is
tion (TESA)?
uncertain
Summary answer: TESA may be offered for patients with a serum FSH level
What is known already: Male factor is attributable in up to 50% of cases of
<7.5IU/L and average testicular size >7.75ml
infertility. It is widely known that bacterial infection can impair sperm function.
What is known already: Microsurgical testicular sperm extraction (m-TESE)
In-vitro studies demonstrate that bacteria can reduce sperm function through
is the gold standard surgical sperm retrieval technique allowing magnified eval-
uation and biopsy of a large area of testicular tissue thereby increasing the overall mechanisms such as motile sperm agglutination or induction of apoptosis. The
sperm retrieval rate (SRR). However, while a small quantity of testicular tissue use of next-generation sequencing(NGS) techniques has provided a better
is presumably extracted during m-TESE, the procedure can have detrimental understanding of the microbiome and has led to the emergence of new thera-
effects on testicular function evidenced by the resulting temporary or permanent peutic approaches, and recent studies have highlighted the importance of the
state of hypogonadism postoperatively. Furthermore, microTESE is expensive, changes in the microbiome(dysbiosis) to female reproductive health. Several
time-consuming and requires a skilled microsurgeon who may not be available emerging studies have investigated the impact of the semen microbiome on
in certain places. These facts advocate that TESA may still be of value in the sperm function and fertility.
management of NOA patients. Study design, size, duration: A systematic search was conducted in accor-
Study design, size, duration: This was a retrospective study of 394 patients dance with the Preferred Reporting Items for Reviews and Meta-analysis
with NOA who underwent their first trial of surgical sperm retrieval over a (PRISMA) statement. The systematic review was registered with PROSPERO
period of 5 years. Patients with NOA based on the presence of one or more (ID CRD42019124483). The databases MEDLINE, OVID and PubMed were
of the following: 1) histopathology denoting defective spermatogenesis; 2) high searched to identify English language studies related to the identification of bac-
gonadotropins; 3) scanty number of sperm retrieval; 4) genetic abnormality teria in the semen of infertile and fertile men, between 1st January 1999 and 15th
March 2019. Two independent investigators screened abstracts and selected
were included. We excluded patients with proven obstructive azoospermia or
studies and extracted data for meta-analysis
who performed surgery due to causes other than azoospermia.
Participants/materials, setting, methods: 54 studies were included, with
Participants/materials, setting, methods: Data including age, testicular
50811 subjects. Observational studies were included that identified the seminal
size, serum hormones, genetic tests and histopathologic results were collected.
All patients underwent a staged retrieval procedure starting with TESA and microbiome in male subjects, and it’s association with fertility and semen param-
eters. We included studies that analysed semen according to the 1999 or 2010
proceeding to m-TESE. Multivariate logistic regression analysis was used to assess
WHO manual, and where bacteria were identified using NGS, culture or poly-
the impact of different factors on the outcome. ROC curve was performed to
merase chain reaction (PCR). We excluded animal studies. The National Institute
identify the cut-off value for significant variables. The likelihood for positive
of Health quality assessment tool was used to grade the studies.
TESA-SRR was calculated for each significant variable and for all variables com-
Main results and the role of chance: NGS enabled the detection of greater
bined using regression analysis.
variety of bacterial genera than culture/PCR. The SM was reported to be rich
Main results and the role of chance: TESA was the sole procedure in 31%
and diverse in both fertile and infertile men. It is uncertain if the microbiome
of patients while two-thirds of patients underwent m-TESE. Overall, SRR with
differs between these groups as only 1 NGS paper studied this (n=77 partici-
TESA was 26.7% while with m-TESE was 42.3%. Patients with positive TESA had
pants), and found no difference.
significantly larger testicular size (10.39 ± 0.6 vs. 7.15 ± 0.3, p<0.001) and sig-
Three of the four NGS studies reported clustering of the seminal microbiome
nificantly lower FSH (6.44 ± 0.7 vs. 14.1 ± 0.8) and LH levels (4.03 ± 0.3 vs.
with a predominant species. All three studies reported that Lactobacillus and
7.15 ± 0.4 (p<0.001 for both) than those with a negative TESA. The highest
Prevotella were dominant in respective clusters. Lactobacillus was associated
SRR was observed in men with hypospermatogenesis (52.6%) followed by
with improvements in semen parameters, and had a protective effect on quality.
Maturation arrest (34.2%), Sertoli cell only (13.2%) and tubular hyalinization
Conversely Prevotella appeared to exert a negative effect on sperm quality.
(0%) (p<0.001). Multivariate analysis revealed that only testicular size (OR 1.084,
Bacteriospermia negatively impacted sperm concentration (difference in
95%CI 1.001-1.174, p=0.046) and serum FSH (OR 0.924, 95%CI 0.863-0.989,
means(MD) -15.6,95%CI -22.65to-8.65) and progressive motility(PM)(MD
p=0.024) had a significant impact on SRR with TESA. A testicular size cut-off of
-8.105,95%CI -13.568to-2.642). There was an increased prevalence of
7.75ml (sensitivity 62.9%, specificity 63.4%, AUC=0.674) and FSH cut-off of
Ureaplasma urealyticum (UU) in infertile men(OR 2.25, 95% CI 1.47-3.46). UU
7.5IU/L (sensitivity 64.6%, specificity 64%, AUC=0.718) were the best predic-
negatively impacted concentration (MD -13.85,95%CI -20.93to-6.67) and mor-
tors of TESA SRR. TESA was positive in 39.5% of patients with FSH < 7.5 IU/L
phology (MD -2.82,95%CI -3.546to-2.1). There was no difference in the prev-
(OR 3.247, p< 0.001), 34.6% of patients with testicular size > 7.75 ml (OR
alence of chlamydia trachomatis (CT) between fertile and infertile men(OR 1.11,
2.916, p<0.001) and 42.5% of patients with both FSH < 7.5 IU/L and testes size
95%CI 0.75-1.65), and CT had no significant impact on routine semen parame-
> 7.75 ml (OR 3.419, p<0.001)
ters(concentration:MD -8.723,95%CI -19.255to1.809,PM:MD -1.727,95%CI
Limitations, reasons for caution: The main limitations are the retrospective
-5.567to2.112, morphology:MD 0.449,95%CI -1.601to2.5). Enterococcus fae-
design of the study.
calis(EF) negatively impacted total motility(MD -11.03,95%CI -17.845to-4.22),
Wider implications of the findings: Investigating the predictors for positive
and Mycoplasma hominis(MH) negatively impacted concentration, PM and
SRR with TESA may provide important prognostic evidence that can be used
morphology(p<0.05).
during patient counselling to aid the couple in making sound treatment decisions.
Limitations, reasons for caution: The quality of the evidence ranged from
fair to low quality, and although 50,811 total subjects were studied, the numbers
in the NGS studies were small (n=385). There was a high degree of heteroge-
P-023 The semen microbiome and the impact on sperm function neity in some of the individual meta-analyses and therefore caution must be
and male fertility: a systematic review and meta-analysis observed when interpreting the results.
L. Farahani1,2, T. Tharakan2,3, T. Yap4, J. Ramsay3,5, C. Jayasena2,5, Wider implications of the findings: NGS is a superior method of charac-
S. Minhas3 terizing the SM. UU, EF, MH and Prevotella negatively impact sperm, whereas
1
Imperial College Healthcare NHS Trust, Obstetrics & Gynaecology, London, Lactobacillus appears to protect sperm, paving the way for novel therapies
United Kingdom ; (e.g.probiotics). The evidence regarding the impact of microbiome on fertility
2
Imperial College, Metabolism- Digestion and Reproduction, London, United is inconclusive. Couples should be informed that further studies are needed.
Kingdom ; Trial registration number: not applicable

P-024 Is there an endocrine contribution to the sexual dysfunction dysfunction and anejaculation during coïtus. Semen parameters are often
seen in end stage renal disease patients ? impaired in SCI patients mostly spermatozoa motility, vitality and typical forms
S. Al Said1, H. Elbardisi1, K. Khalafalla1, M. Arafa1, A. Majzoub1 associated with an elevated concentration of leukocytes. SCI patients are at high
1 risk of urinary tract infections and positive spermoculture generating an inflam-
matory syndrome (IS) and exposing spermatozoa to oxidative stress (OS).
Study question: Does hormonal disturbance in end stage renal disease patients Fertility is a major concern in this population and many of them fear that the
affect their sexual function ? delay post-SCI can negatively impact sperm quality.
Summary answer: Endstage renal disease is commonly associated with sexual Study design, size, duration: 35 SCI patients aged from 18 to 60 years old
dysfunction that is more likely to be attributed to organic causes rather than able to ejaculate by masturbation or in response to penile vibratory stimulation
solely to endocrine disturbances (PVS) have been enrolled. A prospective longitudinal study was conducted over
What is known already: Male sexual dysfunction is commonly prevalent in 18 months with 4 visits to collect medical information focusing on uro-genital
patients with end stage renal disease (ESRD) and has been partly attributed to tract and concomitant treatments. At each visit a semen analysis was performed
the concurrent state of hyperprolactinemia and hypogonadism, often observed with the evaluation of seminal IS and OS. Thirty-five SCI patients have been
in this patient population. The aim of this study was to evaluate the hormone included.
profile and sexual function of ESRD patient, using validated questionnaires, in Participants/materials, setting, methods: Semen analysis was conducted
attempt to explore this association. according to WHO recommendations. IS was quantified through granulocyte
Study design, size, duration: This was a prospective study which included concentration evaluated by leucoscreen® test and considered positive when
98 patients with end stage renal disease (ESRD) who followed in the outpatient >1million/ml and seminal plasma elastase measured by ELISA with a 500 ng/ml
department of a tertiary medical centre over a period of 1 year. Patients receiving positive threshold. OS was assessed using the Tunel assay methodology and
treatment for hyperprolactinemia or those known to have an endocrine disorder analyzed by flow cytometry to measure spermatozoa DNA fragmentation with
were excluded in addition to patients receiving medical or surgical treatment for a 40% positive threshold. Data were compared using Wilcoxon signed rank and
erectile dysfunction (ED) or premature ejaculation (PE). the Mann-Whitney-Wilcoxon tests.
Participants/materials, setting, methods: Full history details with com- Main results and the role of chance: SCI patients mean age of was
plete physical general and genital exam was conducted on the included patients. 29.4 ± 6.4 years with mean age at SCI 21.8 ± 8.3 years. Semen concentration
After filling the international index for erectile function-5 and the Arabic index was normal (155.1±231.2 million/ml) associated to a decrease in progressive
for PE questionnaire, morning serum samples were taken from patients to mea- motility (13.6±12.4%) and vitality (22.7±19.3%) and an increase in morphological
sure testosterone and prolactin levels. Descriptive statistics was used to report abnormalities (typical forms 11.5±11.2%). Mean round cell semen concentration
frequency or means of variables. Chi-square test was used to examine associa- was increased (38.3±37.6 million/ml) as well as granulocyte concentration
tions between categorical variables. P<0.05 was considered statistically (46.4±121.1 million/ml). IS and OS were highly positive with a median elastase
significant. quantified at 477.4 ng/ml and the DNA fragmentation at 50%. Semen parameters
Main results and the role of chance: Out of the 98 ESRD patients, 72 were compared between patients i) positive and negative for IS and OS and
(73.6%) were treated with hemodialysis, 13 (13.2%) with peritoneal dialysis and ii) in absence or presence of a urogenital i.e. urinary and/or seminal tract (UGI)
13 (13.2%) with medical treatment only. Diabetes mellitus was observed in all infection. iii) taking into account the lesion level, the completeness of the lesion,
patients (type 1, 52%; and type 2, 48%), while hypertension, coronary heart the time elapsed since SCI and age at injury. We could not find any difference in
disease and dyslipidemia were detected in 97.1%, 34.3% and 25.5%, respectively. semen parameters in the presence of IS and OS nor when UGI was present.
The mean age, serum testosterone and prolactin levels were 52.4 ± 12.1 years, Moreover, neither completeness nor level of the lesion nor time post SCI nor
12.95 ± 6.5 nmol/L and 514.2 ± 592.8mIU/L. Results of the PE index ques- age at injury did seem to influence semen quality. Semen parameters at the first
tionnaire revealed that 86 (87.7) patients had PE, 9 (9.1%) probable PE and 3 and the last visit were compared in 17 patients and showed no change over time.
(3.1%) no PE. With IIEF-5, ED was detected in 96 patients; it was severe in 23 Limitations, reasons for caution: The study was conducted in a small sam-
(23.5%), moderate- severe ED in 29 (29.4%), mild-moderate ED in 30 (30.4%) ple. Only 28 semen samples were collected at first visit (V1) versus 11 at V4
and mild in 14 (14.7%). Patients were divided into 2 groups, one according to indicating a high number of patients lost of follow-up. In some cases planed
prolactin levels (normal and high) and other according to testosterone level (low semen analysis has been hindered because of the decrease in semen volume
and normal). 55 patients had high prolactin while 33 had low testosterone levels. characteristic of SCI.
No significant differences were observed in IIEF or PE index levels between Wider implications of the findings: Within the limits of the study, there was
patients with low/normal testosterone and normal/high prolactin. no significant decline in semen quality overtime. Even preliminary, these findings are
Limitations, reasons for caution: This is a single center study. essential to reassure SCI men about their future fertility. The present study is not
Wider implications of the findings: Since hormonal disturbance doesn’t in favor of routine sperm freezing for fertility preservation in SCI patients.
affect sexual dysfunction, definitive treatment for sexual dysfunction should be Trial registration number: NCT02144558
started including medical or surgical options.
Trial registration number: NA P-026 The Efficacy of Repeat Micro TESE after Failed First
Attempt in Men with Nonobstructive Azoospermia
P-025 Prospective follow-up of semen parameters in spinal cord- E. Bakircioglu1, O. Ozman2, S. Tosun3, B. Numan3, C. Sami3,
injured patients (FertiSCI): a pilot study O. Cenk3
C. Chalas1, L. Jilet2, V. Drouineaud1, C. Patrat1, J.P. Wolf1, 1
Istanbul Florence Nightingale Hospital, Urology, Istanbul, Turkey ;
F. Giuliano3 2
Health Sciences University- Gaziosmanpaşa Taksim Education and Research
1
Hôpital Cochin-Port Royal, Biology of Reproduction, Paris Cedex 14, France ; Hospital, Urology, Istanbul, Turkey ;
3
2
URC/CIC Paris Descartes Necker Cochin, Biostatitics, Paris, France ; Istanbul Florence Nightingale Hospital, In Vitro Fertilization Center, Istanbul,
3
Hopital R. Poincare APHP, Neuro-urology, Garches, France Turkey
Study question: Are men with spinal cord injury (SCI) at risk for semen param- Study question: What is the efficacy of repeat micro-TESE in men with
eters impairment overtime according to the lesion characteristics and associated non-obstructive azoospermia who failed the first micro-TESE?
genital inflammatory state? Summary answer: Repeat micro-TESE was enabled retrieved sperm in 18.4%
Summary answer: Neither genital inflammatory state, completenesslevel of of men with NOA who failed the first micro-TESE.
the lesion,age nor time post SCI, seemed to exert any influence on semen quality What is known already: Testicular sperm extraction combined with ICSI is
over time. the only way for the patients with non-obstructive azoospermia to have their
What is known already: Spinal cord injury (SCI) primarily affects young men own genetic offspring. Micro-TESE promises high rates of surgical sperm retrieval
who have not experienced fatherhood yet. SCI often results in erectile compared to methods such as testicular sperm aspiration and conventional TESE.

Salvage micro-TESE operation showed successful sperm recovery after failed Wider implications of the findings: Our study shows that TEERT is highly
conventional TESE in many studies in the literature. prevalent in infertile men with OA and those with TEERT are much likely having
Study design, size, duration: This was a retrospective cohort study of 125 their sperm retrieved by a sperm retrieval technique. This finding suggests the
men who failed the previous micro-TESE in other institutions that underwent predictive value of TEERT in the prediction of OA and sperm retrieval.
repeat micro-TESE between November 2014 and July 2018. Clinical parameters Trial registration number: not applicable
of the patients were compared for the success of sperm recovery between the
patients who were successful for sperm recovery and who have not.
Participants/materials, setting, methods: All of the patients underwent P-028 Bicarbonate-dependent alkalinization of acrosomal pH
repeat micro-TESE operation. The age of the patients, duration of infertility, occurs during human sperm capacitation
serum FSH and total testosterone levels and testicular volumes were compared "Abstract withdrawn by the authors"
between the patients who have successful sperm recovery and who have not.
Main results and the role of chance: Sperm was successfully recovered in P-029 Efficacy of Magnetic Activated Cell Sorting (MACS) in
23 of 125 (18,4%) men with repeat micro-TESE. The age of patients Sperm Selection for Intracytoplasmic Sperm Injection (ICSI)
(33.9±4.7,33.7±5.5; P=0.86), the duration of infertility (7.1±4.3, 6.4±3.5;
T.T.H. Tran1, T.T.C. Bach1, V.T. Phung1, T.T. Tran1, T.L. Dinh1
P=0.47) serum FSH (22.0±13.8, 15.4±13; P=0.29) and total testosterone level
1
(3.4±1.1, 3.1±1.6; P=0.37) did not show a statistical difference between men Hospital of Post and Telecoms, Assisted reproductive technology center, Hanoi,
who have successful sperm recovery and who have not. However, testicular Vietnam
volume was significantly lower in men who have successful sperm recovery
(8.2±5.4 ml) compared to men who have not (11.3±5.3 ml, p=0.01). Seven of Study question: This study investigated if the application of MACS would
14 (50%) men who have diagnosed Klinefelter syndrome (KS) sperm recovery have a critical effect on ICSI outcome, especially with patients having multiple
was successful with repeat micro-TESE. unsuccessful IVF cycles.
Limitations, reasons for caution: It is a retrospective cohort study. Official Summary answer: The results of this study suggested favorable influence on
records of the micro-TESE operations that the patients had previously were patients with multiple failed IVF cycles and highly promising for others.
insufficient. What is known already: MACS is reported as a possible procedure to help
Wider implications of the findings: Our cohort is the largest repeat micro- increase ICSI outcome by using annexin V to separate apoptosis sperms. The
TESE series in patients who have non-obstructive azoospermia with a failed first separation of apoptosis sperms is believed to result in a positive change in sperm
attempt. The chance of sperm recovery may enable valuable information for quality.
the patients before the decision to repeat micro-TESE. Study design, size, duration: 114 patients were enrolled in this
Trial registration number: Not applicable study between February 2018 to January 2020. We analysed the outcome of
52 multiple returning cycle couples who has undergone one MACS/ICSI cycle
P-027 Tubular ectasia of epididymis and rete testis among (group 1) and their most recent non-MACS/ICSI cycle (group 2). We also
azoospermia patients accessed the outcome of 62 couples who has only has one cycle of IVF done
(group 3).
H. Nguyen1,2, H.H.T. Nguyen1, C.K. Trinh1, T.A. Nguyen1,
Participants/materials, setting, methods: Group 1 and 2 comprised of
V.H. Dinh1
52 multiple returning cycle couples who has undergone one MACS/ICSI cycle
1
Andrology and Fertility Hospital of Hanoi- Vietnam, Andrology department, Hanoi, and one non-MACS/ICSI cycle. 62 participants in our group 3 has only one IVF
Vietnam ; cycle and has specifically requested for MACS/ICSI to be performed. In all
2
Vinmec International General Hospital, Andrology department, Hanoi, Vietnam MACS/ICSI cycles, ejaculated semen was undergone density gradient centrifu-
gation in combination with MACS. Collected sperms were used for routine ICSI.
Study question: To identify the rate of tubular ectasia of epididymis and rete T-Test and Chi-square test were applied.
testis (TEERT) in infertile men with obstructive and non-obstructive azoospermia Main results and the role of chance: Group 1 has higher fertilization rate
(OA and NOA). (67%) than group 2 (59%). Group 2 with non-MACS procedures saw 16 embryo
Summary answer: Rates of TEERT in patients with OA and NOA were 87.8% tranfers for the first time and 04 for the second, performed on X couples with
and 22.5%, respectively. 8 couple having no embryos; cumulatively, positive beta-HCG rate was 13%.
What is known already: TEERT is a condition mainly relating to the obstruc- Whereas in group 1, all but 1 couples had embryos; 19 couples have had embryo
tion of seminal tract. With high resolution ultrasound and the routine use of transferred, resulted in 45 % positive beta-HCG rate; In group 3, the FR and
scrotal ultrasonography, this condition is increasingly diagnosed. However, it is positive beta-HCG rate respectively was 78% and 51%. In conclusion, there
often unclear about the clinical significance of this diagnosis. were significant differences in the result of group 1 and 2. For group 3 where
Study design, size, duration: This monocentric, retrospective study was patients requested to perform MACS , the result was also very promising.
conducted on 203 infertile men with azoospermia who visited Andrology and Limitations, reasons for caution: The sample size of this study participants
Fertility Hospital of Hanoi (AFHanoi) from November 2017 to October 2019 is small. Further studies with more control groups with sperm and eggs quality
for seeking treatment on infertility. analytics is needed to confirm our methodology.
Participants/materials, setting, methods: Patients were chosen to take Wider implications of the findings: According to this study, the application
part in the study if they had information of clinical examination, serum folli- of MAC/ICSI is a novel and easy to perform clicician technique with promising
cle-stimulating hormone (FSH), testosterone, scrotal ultrasonography performed results.
at AFHanoi. Participants had also to undergoing any of these sperm retrieval Trial registration number: Not applicable
procedures i.e. PESA, TESE and micro-TESE in the same hospital.
Main results and the role of chance: There were 123 (60.6%) patients with
P-030 Severe male factor affects the blastocysts ploidy status
OA and 80 (39.4%) patients with NOA. Rates of tubular ectasia of epididymis
and rete testis in the whole sample was 62.0% (126/203 men). In men with OA, E. Zhylkova1, O. Feskov2, O. Somova3, O. Zozulina4, A. Feskova5
1
rate of TEERT was 87.8% (108/123 men), whereas in men with NOA this rate Centre of Human Reproduction Sana-Med, Genetic laboratory, Kharkiv, Ukraine ;
2
was 22.5 (18/80 men). Interestingly, among those with tubular ectasia almost Center of Human Reproduction “Clinic of Professor Feskov A.M.”, IVF
all cases had the condition observed in epididymis (106/108 cases, 98.0%). department, Kharkov, Ukraine ;
3
However, only 2 cases (2%) had the condition observed in rete testis. As Center of Human Reproduction “Clinic of Professor Feskov A.M.”, IVF
expected, sperm retrieval rate in those with TEERT was as high as 96% Department, Kharkiv, Ukraine ;
4
(121/126 men). Center of Human Reproduction “Clinic of Professor Feskov A.M.”, IVF
Limitations, reasons for caution: This study is a retrospective cohort study Department, Kiev, Ukraine ;
5
with all associated inherent biases. The findings of this study should be confirmed Kharkiv National Medical University, Department of Obstetrics- Gynecology and
with further prospective study. Pediatric Gynecology, Kharkiv, Ukraine

Study question: Does the severe male factor affect the blastocysts ploidy Main results and the role of chance: Treatment of infertile men with high-
status and IVF outcome? dose vitamin D did not affect sex steroid levels. There was no difference in serum
Summary answer: Severe male factor of infertility affects the blastocysts’ levels of testosterone, estradiol, LH or FSH between vitamin D and placebo
aneuploidy. Sperm parameters affect blastocyst morphology. The IVF outcome treated men. However, a predefined subgroup analysis of men with vitamin D
does not depends on severe pathospermia. insufficiency and high PTH at baseline had significant higher free testosterone
What is known already: Male factor of infertility is responsible for up to 60% (∆24%; p=0.027) and free testosterone/LH ratio (∆34%; p=0.048) after 150
of fertility problems worldwide. The development of intracytoplasmic sperm days vitamin D and calcium treatment compared with the placebo-treated men.
injection (ICSI) has allowed patients with severe oligozoospermia to have chil- Limitations, reasons for caution: Our biggest limitations are the small num-
dren. The question about the effect of severe male infertility on embryo aneu- ber of men in the subgroupanalyses. Our findings should be verified in large
ploidy and IVF results is still discussable. Almost 60% of the blastocysts in IVF clinical trials preferably in men with persistent vitamin D insufficiency character-
are aneuploid. Existing data regarding the relationship between embryo mor- ized by low serum 25-OHD and high PTH.
phology and its ploidy are conflicting. Wider implications of the findings: Our findings suggest an increased
Study design, size, duration: The effect of severe male factor of infertility awareness on vitamin D status in infertile men which may be important to avoid
on the early embryo development was studied. The correlation of sperm param- deterioration of their Leydig Cell function
eters with the morphology of blastocysts and PGD and IVF outcomes was Trial registration number: NCT01304927
examined. The study’s protocol was approved by the Center’s IRB.
Participants/materials, setting, methods: Totally the DNA samples of P-032 Study on the specific piRNAs in human seminal plasma as
406 preimplantation blastocysts from 117 men diagnosed with severe male factor a non-invasive diagnostic indicator before microsurgical testicular
of infertility and normal karyotype 46,XY were examined using the method of sperm extraction
next generation sequencing (NGS). ICSI/IMSI procedure was done. The rates
X. Huang1, S. Xie1
of blastocysts euploidy and IVF outcome were compared with 299 biopsied
1
blastocysts from 67 patients with normal sperm parameters. A chi-squared test Center for Reproductive Medicine- Tongji Medical College- Huazhong University
and Spearman coefficient were used to perform statistics analysis. of Science and Technology- China, Department of Andrology, wuhan, China
Main results and the role of chance: The rate of blastocysts aneuploidies
was significantly higher in group of patients with severe male factor of infertility Study question: Clinical studies have not found an effective non-invasive test
comparing with the control group (51.1% vs. 33.8% respectively, χ2 = 6.125, to accurately determine the presence of sperm in the testicles of non-obstructive
χ2critic. = 3.841, P = 0.014). There was a significant positive correlation between azoospermia (NOA) patients
the blastocysts euploidy rates and sperm concentration (rs = 0.16, P < 0.05). A Summary answer: Except AZFa and AZFb microdeletion for a small group
significant positive correlation between sperm motility and blastocysts morphol- of NOA, no other reliable diagnostic technique has been widely accepted
ogy was proved (rs = 0.17, P < 0.05). The transfer of one euploid blastocyst nowadays.
was performed in each case for patients in both studied groups. There was no What is known already: piRNAs are a series of specific small molecule nucleic
significant difference in IVF outcomes in both groups. The clinical pregnancy rates acid produced in the testis of different mammals. Previously, we compared
were 53.8% and 63.2% in group of patients with severe male factor of infertility piRNAs expression profiles in the testicles of NOA patients with successfully
and in control group respectively (P>0.05). identified sperm and those without sperm by using second-generation sequenc-
Limitations, reasons for caution: Blastocyst biopsy can be carried out for ing. We identified and screened a variety of piRNAs with significant differences
the embryos with the high quality of morphology. between the two groups.
Wider implications of the findings: The formation of the blastocyst and Study design, size, duration: We randomly selected 120 NOA patients who
embryo implantation may be blocked at various stages of development. were interested in micro-TESE and collected their ejaculated semen prior to
Performing the PGD test using the high resolution techniques for patients with microsurgery for piRNAs detection.
severe pathospermia results in high IVF outcomes. The necessity of PGD for Participants/materials, setting, methods: According to our previous
patients with severe male infertility is proved. sequencing results, we screen out eight specific piRNAs. Before Micro-TESE
Trial registration number: No number performed for a total of 120 NOA cases, these piRNAs in human seminal plasm
were detected by Fluorescent quantitative PCR. Each specimen was tested three
P-031 Vitamin D supplementation improves the Leydig Cell times. Patients were classified into two groups (successful sperm retrieval [SSR]
function in infertile men with persistent vitamin D insufficiency and unsuccessful sperm retrieval [USR]) after the operation of micro-TESE.
Statistics were conducted according to the results of micro-TESE and seminal
R. Holt1
plasma piRNAs detection.
Main results and the role of chance: Three piRNAs DQ578561, DQ576926
Study question: Can vitamin D supplementation improve the Leydig Cell
and DQ577970 were found to have significantly higher expressions in the SSR
function in infertile men?
group (51 cases) than the USR group (69 cases).
Summary answer: Yes, high-dose vitamin D supplementation had a beneficial
Limitations, reasons for caution: A relative small sample without patho-
effect on Leydig Cell function in infertile men with low vitamin D and high PTH.
logical classification of testicular tissue in the patients with NOA.
What is known already: Vitamin D is important for calcium homeostasis and
Wider implications of the findings: piRNA DQ578561, DQ576926 and
bone health, but the spectrum of vitamin D related effects has expanded in
DQ577970 are expected to be new diagnostic indicators for non-invasive pre-
recent years. All actions of vitamin D are mediated by the interaction between
diction of testicular residual sperm in NOA patients, which need further clinical
activated vitamin D (calcitriol) and the vitamin D receptor (VDR) which is
verification with a larger sample size in multiple centers.
expressed in human Leydig cells and thus makes a direct effect of vitamin D on
Trial registration number: No. 31671551
steroidogenesis plausible. Analyses of VDR knock-out (KO) mice revealed
impaired reproductive and endocrine function compared with wildtype mice.
Furthermore, a few cross-sectional studies reported a positive association P-033 A new container with higher success rate for the
between vitamin D and testosterone. cryopreservation of testicular tissues and motile spermatozoa
Study design, size, duration: A single-center, randomized clinical trial with from mice and testicular cancer patients
307 infertile men randomized to receive placebo or a single dose of cholecal- K. Nakata1, S. Wakayama2, M. Iijima3, T. Wakayama2, H. Nakata3,
ciferol 300,000 IU followed by 1400 IU + 500 mg of calcium daily for 150 days. A. Mizogami3, N. Yamashita4
Participants/materials, setting, methods: Men included in the randomized 1
Yamashita shonan yume clinic, Reproductive Medicine Reserch Center, Kanagawa,
clinical trial were initially referred to our andrological outpatient clinic due to infer- Japan ;
tility and subsequently invited to participate in the study. The primary endpoint was 2
University of Yamanashi, Faculty of life and environmental sciences, Yamanashi,
change in semen quality, while serum testosterone was a secondary endpoint. Japan ;

3
Kanazawa University, Integrative Cancer Therapy and Urology- Division of Cancer Study question: Can Tobacco smoke alter the expression level of H2BFWT,
Medicine- Graduate School of medical Science, Kanazawa, Japan ; TNP1, TNP2, PRM1, and PRM2 genes?
4
Yamashita Shonan Yume Clinic, Doctor division, Kanagawa, Japan Summary answer: Can Tobacco smoke alter the expression level of H2BFWT,
TNP1, TNP2, PRM1 and PRM2 genes in spermatozoa?
Study question: Is the new container effective for the freezing and thawing What is known already: Various studies were focusing on the mechanisms
of the testicular tissues (TT) and motile spermatozoa? by which the environmental and lifestyle factors especially smoking influence on
Summary answer: The new container is effective for the cryopreservation the sperm genome and epigenome.
of TT in order to preserve motile spermatozoa from mice and testicular cancer Testis-specific histones like H2BFWT, transition proteins TP1 and TP2 and
patients. protamines P1 and P2 are the main nuclear proteins that have a crucial role
What is known already: Most men diagnosed with testicular tumors are during the protamination of sperm genome during spermiogenesis. Protamination
between ages 15-40. Although the European Association of Urology recom- leads to the elimination of histones carrying epigenetic signals. Thus, protami-
mends all testicular cancer patients are referred for semen analysis and cryo- nation take part in the epigenetic regulation of the spermatozoa and any factor
preservation prior to treatment, it was reported that 6-24% of testicular cancer changing protamination may be considered as an epigenetic signal like DNA
patients were azoospermia, 50% were Oligozoospermia. Because testicular methylation and histone modification influencing the transcription regulation
cancer patients have a low sperm count and may not be able to collect enough after fertilization.
sperm for ART, TT banking is recommended. Commonly, straws or vials have Study design, size, duration: A prospective controlled trial carried between
been used for cryopreservation of TT. There are no studies referring to the August 2016 and April 2018 at the Department of Obstetrics and Gynecology,
comparison between different methods. But after thawing, motility of sperm is University of Saarland, Germany. 167 semen samples were included in this study
very low. and divided into 54 non-smokers (G1) and 113 heavy-smokers (G2).
Study design, size, duration: Mouse TT was divided into nine groups to Participants/materials, setting, methods: After semen purification, total
examine the effect of the combination of freezing speed (slow vs. rapid), freezing RNA was isolated using Isolate II DNA/RNA/Protein kit then concentration
medium (Cellbanker1 vs. Fertiup), and freezing vessel (New container vs. and purity were checked with Nanodrop spectrophotometer ND-2000c.
Cryotube), and control (Fresh), repeatedly. That is, SCN, SFN, RCN, RFN, SCC, RT-qPCR technique was used for the quantification of the expression level of
SFC, RCC, RFC and Control. TT from Testicular cancer patients was divided the five studied genes using miScript reverse transcription and QuantiTect SYBR
into two groups using slow freezing, with (Cellbanker1 vs, Sperm freezing) and Green PCR Kits. Protamine deficiency (CMA3+) was assessed by Chromomycine
the new container, that is SCN and SSN. CMA3 staining and sperm DNA fragmentation (sDF) by TUNEL assay.
Participants/materials, setting, methods: Mouse and testicular cancer Main results and the role of chance: In G1, the sperm count (88.09±
patient’s TT were moved into the physiological saline solution and cut into 2-3 63.42 mill/ml), progressive motility (27.31±21.78%), normal morphology
mm pieces with the scissors. Mice TT was frozen in 9 groups, and testicular (10.87±12.11%), were significantly higher (p<0.01) than G2 (62.17±
cancer patient’s TT was frozen in 2 groups. After thawing, we observed sperm 51.68 mill/ml; 14.86±10.95%; 4.01±2.88%, respectively).
motility, sperm membrane damage and Caspase-3 assay in mice and testicular Whereas, CMA3+ (23.50±14.70%) and sDF (17.41±14.59%) in G1 were
cancer patients. Also In vitro and in vivo development after embryo transfer were significantly lower (p<0.01) than G2 (33.58±21.34%; 27.55±20.01%,
assessed in mice. respectively).
Main results and the role of chance: The average number of mouse motile Protamine mRNA ratio was significantly higher in G1 in comparison to G2
spermatozoa after thawing was examined. The motility rate was 25.7±13.6 % (0.11 ± 0.84 vs. 0.60 ± 1.08; p=0.001). Unlike G1, the protamine mRNA ratio
in SCN. PI positive rate was 20.0±10.0 in SCN. SCN was significantly higher significantly correlates with CMA3+ (r=0.413, p=0.0001) and sDF (r=0.302,
than the other groups in motility and PI positive rate (P<0.05). The highest rate p=0.003) in G2.
was obtained when TT was frozen using slow freezing, Cellbanker1 and New Moreover, the relative amounts of each studied gene mRNA (mean delta ct)
container[若山1] . After ICSI, the blastocyst rates were 54.9±13.6 % in SCN, were differentially expressed between G1 and G2 and this difference was highly
48.4±25.7 % in SFN, 40.8±16.2 % in SCC, 20.6±25.4 % in SFC and 64.2±10.8 significant (p<0.01). Besides, H2BFWT, TNP1, TNP2, PRM1, and PRM2 genes
% in Control, respectively. SCN was significantly higher than SFC (P<0.05). were down-regulated in spermatozoa of G2 compared to G1 (Fold change <0.5).
Additionally, we obtained fetuses 22.4% in SCN, 22.8% in SFN, 15.4% in SCC, Limitations, reasons for caution: The size number of the sample.
14.3% in SFC and 42.0% in Control. Five testicular cancer patient’s tissues were Wider implications of the findings: Studied genes are expressed in a
frozen by SCN or SSN. After thawing, sperm motility rates of 5 patients were well-organized chronological manner and any alterations from an internal or
18.9±13.2% (477/2681) in SCN and 29.6±13.2% (1415/4832) in SSN. SSN external factor like smoking my altered this mechanism and thus altered sper-
was significantly higher than SCN (P<0.05). PI positive sperm rates of 3 patients miogenesis and sperm function.
were 53.3±20.1% (293/460) in SCN and 29.6±12.5% (159/469) in SSN. SSN Trial registration number: not applicable
was significantly higher than SCN (P<0.02). In Caspase-3 assay of 3 patients,
negative cell number was 127 in SCN, 255.5 in SSN and 1046.5 in control,
respectively. P-035 Sperm DNA integrity and human papillomavirus (HPV)
Limitations, reasons for caution: This is a basic study on a relatively small infections: a controversy that could be resolved by a new
sample size with limited conditions. The confirmation using larger samples under molecular approach
various conditions may be required. Furthermore, although healthy mouse off- R. Schillaci1, T. Notari2, G. Capra3, M.A. Ragusa4, M.C. Roccheri4,
spring was obtained, we need to check the safety of using TT frozen-thawed A. Perino3, A. Ferrigno4, L. Bosco4
sperm in ART. 1
Centro Interaziendale di Procreazione Medicalmente Assistita- Ospedale
Wider implications of the findings: The findings of this study indicate
“V. Cervello”- Università di Palermo- Italy, Department of Sciences for Health
that the new container allows checking inside the seminiferous tubule using
Promotion and Mother-Child Care ‘G. D’Alessandro’ PROSAMI, Palermo, Italy ;
microscope. Because it’s made of polydimethylsiloxane and the bottom is trans- 2
Check Up Research Unit - Polydiagnostic and Day Surgery Centre - Salerno, Check
parent and very thin. It is also useful for a small number of spermatozoa
Up Research Unit - Polydiagnostic and Day Surgery Centre, Salerno, Italy ;
(K. Nakata, et al. 2019) and testicular tissues cryopreservation. 3
University of Palermo, Department of Sciences for Health Promotion and
Trial registration number: No
Mother-Child Care ‘G. D’Alessandro’ PROSAMI, Palermo, Italy ;
4
University of Palermo, Department of Biological- Chemical- and Pharmaceutical
Sciences and Technologies STEBICEF, Palermo, Italy
P-034 Impact of tobacco smoking on the expression level of
H2BFWT, TNP1, TNP2, PRM1 and PRM2 genes in spermatozoa
Study question: The aim was to determine if HPVs affect spermatozoa DNA
H. Amor1, M.E. Hammadeh1 integrity. To resolve the discrepancy regarding the association between HPV
1
University of Saarland, Obstetrics and Gynecology, Homburg/Saar, Germany infections and sperm DNA damage.

Summary answer: To elucidate if HPV impairs DNA integrity, we suggest to spermatozoa travels through the epididymis. And hence it is widely understood
investigate HPV DNA positivity in spermatozoa by a differential lysis procedure and accepted that testicular spermatozoa have a relatively lesser degree of DNA
before TUNEL assay. fragmentation. Surgical sperm retrieval techniques have become more refined
What is known already: HPV can influence human fertility, in fact it has been over the years and now yield good quality testicular spermatozoa
demonstrated that infected couples undergoing assisted reproduction techniques Study design, size, duration: Data was collected retrospectively from 2015,
showed an increased risk of pregnancy loss compared with non-infected till 2018. SCSA (Sperm Chromatin Structure Assay) was done to assess the DNA
counterparts. fragmentation Index (DFI) of male partner of all couples who had one failed IVF
Recent studies revealed that DNA Fragmentation Index (DFI) was not different cycle. Those with DFI>30 were identified. Couple with female partners age>35
in HPV-infected and in non-infected semen samples. Other evidences reported years were excluded. The Fertilisation, Blastocyst, Implantation, Clinical
statistically significant differences. Therefore, it remains controversial whether Pregnancy, Miscarriage, Live Birth rates, along with Perinatal and neonatal out-
HPV infection in semen is associated to DNA damage. comes of the subjects were then compared.
Moreover, it is known that HPV DNA can be found in the different semen Participants/materials, setting, methods: 605 subjects were included in
components, then it is necessary to investigate if patients defined as HPV-positive the study based on the pre-requisites Those, who discontinued treatment, or
have viral DNA contained in the spermatozoa. used Donor sperms for ICSI, were excluded. Couple who chose ICSI (Intra
Study design, size, duration: From April 2015 to October 2019, a total of Cytoplasmic Sperm Injection) with ejaculate (n=14) were included in the control
105 patients: 56 HPV positive and 49 HPV negative, male partners of women group, and the study group included all who chose ICSI with TESA obtained
with High Grade Squamous Intraepithelial Lesions (HSILs) positive for high risk spermatozoa (n=37). TESA was done after taking all aseptic precautions and
(HR) HPVs were enrolled in an observational study. following the defined procedural Standard Operating Protocol.
Participants/materials, setting, methods: Patients who met the following Main results and the role of chance: The embryo parameters, Fertilisation
inclusion criteria were enrolled: age between 18 and 50 years; partner positive Rate (TESA = 96.59%; Non-TESA = 89.88%) & Blastocyst Rate (TESA = 33.65%;
for HR-HPV types; sperm concentration after swim up procedure that exceeded Non-TESA = 40.95%) in both groups were comparable. The implantation rates
1 million/ml. Exclusion criteria were: varicocele; cryptorchidism; other genital were higher for the study group (TESA= 60.0%; Non-TESA = 23.1%) in the
infections; chemo/radio therapy. Seminal parameters were evaluated according control group. Pregnancy induced hypertension and IUGR were not observed
to 2010 World Health Organization guidelines and sperms were then separated in both the groups. The Clinical pregnancy (TESA = 63.2%; Non-TESA = 28.6%)
by swim-up technique. HPV-DNA was detected and genotyped by reverse and Live birth rates (TESA = 42.1%; Non-TESA = 28.6%) were better in the
hybridization. DFI was evaluated by TUNEL assay. study group. 4 subjects in the TESA group have ongoing pregnancy There were
Main results and the role of chance: As we have previously demonstrated, no adverse perinatal and neonatal outcomes in the study group. Though
no statistically significant effect of HPV infection on sperm parameters was Blastocyst rates seemed better in the Non-TESA group, reproductive outcomes
observed when we compared HPV-positive total semen samples with respect were better in the TESA group. And also, the perinatal and neonatal outcomes
to HPV-negative counterparts. However samples showing higher viral DNA were confirming the safety of using TESA obtained spermatozoa for ICSI in
ratio had lower percentage of progressively motile sperms. couples with raised DFI.
In our experience, DFI values are not different in HPV positive and in negative Hence, based on the data obtained from the study, it is safe to assume that
samples (17% vs 20% respectively). the use of testicular spermatozoa for raised DFI seems to be a safe and beneficial
This result could be explained by the following considerations. Our previous intervention.
data showed that HPV DNA can be identified in every fraction of semen, by Limitations, reasons for caution: The major limitations of this study is that
differential lysis procedure: spermatozoa, somatic cells and seminal plasma. this is a retrospective study with a small sample size, along with the fact that
Different samples can contain HPV DNA in different fractions and a sample could TESA is an invasive intervention
result HPV-positive also in absence of viral DNA into spermatozoa. Therefore, Wider implications of the findings: Male partners with raised DFI could
it is necessary to determine which semen fraction contains viral DNA in order be offered TESA as a treatment option, benefiting couples with optimal repro-
to classify samples as positive or negative with respect to spermatozoa infection ductive outcomes.
before to perform TUNEL assay, since DFI is evaluated exclusively into Trial registration number: Not Applicable
spermatozoa.
Limitations, reasons for caution: Low number of samples because of the P-037 Body mass index and age correlate with antioxidant
difficulty to find male partners agreeable to the enrollment in the study. supplementation effects on sperm quality: Post-hoc analyses from
Wider implications of the findings: Semen parameters and DFI may have a double-blind placebo-controlled trial
no significant differences with respect to HPV positivity detected by conventional G.M. Busetto1, F. Del Giudice1, A. Virmani2, E. De Berardinis1,
procedures. Therefore, it is advisable to perform the differential lysis procedure A. Agarwal3
in HPV-positive patients to verify in which fraction the virus is located and as a 1
Sapienza University Policlinico Umberto I, Urology, Roma, Italy ;
consequence its potential effect on male fertility. 2
Alfasigma HealthScience, Nutraceuticals, Utrecht, The Netherlands ;
American Center for Reproductive Medicine, Andrology Center, Cleveland, U.S.A.
P-036 Will the use of testicular spermatozoa optimise Study question: Is there any correlation between age and body mass index
reproductive outcomes? (BMI) with the efficiency of medical therapy for infertile men?
R. Mani_Raghupathy1, K.C. Mantravadi2, D. Rao Gedela3 Summary answer: Medical therapy with compounds is more effective in
1
Oasis Centre of Reproductive Medicine, Clinical Embryology, Hyderabad, India ; subjects with varicocele younger than 35 years and with BMI <25.
2
Oasis Centre for Reproductive Medicine, Clinical Embryology, Hyderabad, India ; What is known already: The decline of male fertility is an emerging problem
3
Oasis Centre for Reproductive Medicine, Reproductive Medicine, Hyderabad, and causes for these changes are lifestyle factors and global changes in our
India eating habits with increasing evidence of obesity. Spermatozoa are vulnerable
to lack of energy and oxidative stress as a result of elevated levels of reactive
Study question: Will TESA (Testicular Sperm Aspiration) be a beneficial pro- oxygen species. Therefore, it is essential that appropriate nutrients are available
cedure for couples with raised DFI in the male partner. during maturation. At present, no studies investigated the correlation of age
Summary answer: Testicular spermatozoa obtained surgically from individuals and body mass index (BMI) with the efficiency of a medical therapy in infer-
with raised DFI, will benefit the couple in achieving optimal reproductive tile men.
outcomes Study design, size, duration: The present analysis stems from the extensive
What is known already: Raised DFI (DNA Fragmentation Index) is one of database that has been created to determine the effects of antioxidant supple-
the major causes of male infertility and is known to affect fertilisation, embryo mentation on semen quality. This database includes 104 infertile patients with
quality, implantation, miscarriage & pregnancy rates after assisted reproduction. oligo- and/or astheno- and/or teratozoospermia with an average age of 32.5
ROS is known to attack and damage the spermatozoal DNA happens as years, enrolled in a randomized, double-blind, placebo-controlled trial. Included

are 52 patients with grade I-III varicoceles and 52 patients without varicocele Seminal antioxidant profile (reduced Glutathione (GSHr); Oxidized Glutathione
that were divided into supplementation or placebo groups. (GSSG); Glutathione-S-transferase (GST)), and total protein sulfhydryl (P-SH)
Participants/materials, setting, methods: In accordance with the random- concentrations were measured spectrophotometrically.
ization schedule, subjects received 2 packets of either supplement or placebo Main results and the role of chance: Patient with isolated teratozoosper-
daily for 6 months. Semen parameters were evaluated in a standard semen mia, when compared to fertile donors, showed significantly increased level of
analysis at the beginning of the treatment (V1) and after completing 6 months single sperm DNA breaks, and higher proportions of spermatozoa with phos-
of therapy (V2). Pregnancy rate was included as a secondary outcome. The phatidylserine externalization and mitochondrial depolarisation. Among the
present post-hoc analyses were carried out on the samples as categorized by different studied oxidative stress seminal parameters, the rates of seminal GSHr,
age/BMI and presence/absence of varicocele. GST and P-SH were significantly decreased in the patient group. However, the
Main results and the role of chance: One of the primary aims of this study seminal levels of GSSG and GST have decreased, but only GST didn’t showed
was to correlate the results of the semen analysis with BMI and age. In particular, a significant difference. Interestingly, significant relationships were found between
we wanted to see if aging and obesity status would decrease efficacy of the the studied apoptotic markers and the rate of atypical sperm forms with the
supplementary antioxidant treatment on main sperm parameters (see Tables incidences of head abnormalities. Furthermore, positive inter-correlations were
1-5). For BMI, a significant difference was observed in the BMI <25 group with found between sperm DNA defects, impaired seminal antioxidant profile and
varicocele for total sperm count (p=0.0272) and progressive motility (p=0.0159). the sperm apoptotic markers.
No statistical significance was observed in the combined classes. The results Limitations, reasons for caution: Further combined analysis of oxidative
were partially confirmed by carrying out the Chi-Square test on the data arranged stress, apoptotic markers and nuclear defects should provide complementary
as “Responder/Non Responder”. As for the total sperm count, in both the BMI measurements for the evaluation of sperm quality and could contribute to pro-
<25 and the combined varicocele group (i.e. BMI <25 and age <35) a statistical vide adequate reproductive and genetic counselling for hypofertile patients with
difference was observed (p=0.0066 and p=0.0078 respectively). These post-hoc isolated polymorphic teratozoospermia.
analyses suggest that the nutritional supplement seems to be more effective in Wider implications of the findings: Sperm DNA defects as well as apop-
subjects younger than 35 years with a BMI below 25. tosis and seminal oxidative stress are interlinked in the context of teratozoosper-
Looking at other parameters, patients treated with compounds obtained a mia, and constitute a unified pathogenic molecular mechanism
statistically significant improve of sperm parameters for the following items: total Trial registration number: not applicable
count, progressive and total motility, morphology.
As a secondary outcome, 12 pregnancies occurred during the follow-up time:
P-039 In spermatozoa collected after pellet swim up, when total
10 in the supplementation group and 2 in the placebo group.
dna fragmentation is higher than 15%, the normal morphologically
Limitations, reasons for caution: Even as a double-blind placebo-controlled
spermatozoa population shows an increased dna damage.
study with very strict inclusion and exclusion criteria, we did not include sperm
DNA fragmentation. Also, an oxidative stress measure such as ORP was not G. Ruvolo1, A. Ferrigno2, M.C. Roccheri2, E. Cittadini1, A. Pane1, C.
included. There also may be other factors besides aging and obesity involved, Simonaro1, B. Ermini3, I. Camera3, A.M. Bruccoleri1, M. Manno1,
including lifestyle, associated disease and fat distribution. A. Lo bue1, L. Bosco2
1
Wider implications of the findings: In addition to earlier findings regarding Centro di Biologia della Riproduzione, Centro di Biologia della Riproduzione,
improved sperm parameters in supplemented patients, these post-hoc analyses Palermo, Italy ;
2
suggest that antioxidant supplementation seems to be more effective on improv- University of Palermo- Palermo- Italy, Department of Biological- Chemical- and
ing sperm parameters in subjects aged less than 35 years old and with BMI Pharmaceutical Sciences and Technologies STEBICEF-, Palermo, Italy ;
3
below 25. CIPA- Centro Italiano di Procreazione Assistita, CIPA- Centro Italiano di
Trial registration number: NCT04177667 Procreazione Assistita, Roma, Italy
P-038 New insights into the physiopathology of teratozoospermia Study question: We investigated the DNA Fragmentation Index (DFI) in motile
and its association with sperm DNA defects, apoptotic alterations normal morphologically spermatozoa comparing samples with total DFI < 15%
and oxidative stress Vs ≥ 15% collected after pellet swim up
O. Ammar1, M. Mehdi2, A. Sallem2, M. Muratori3 Summary answer: In the case of DFI ≥15% the percentage of normal mor-
1
Faculty of Medicine University of Monastir-, Laboratory of Histology Embryology phologically spermatozoa with fragmented DNA is significantly higher than the
and Cytogenetic LR 18-ES 40-, Monastir, Tunisia ; population with DFI < 15%
2
Faculty of Medicine- University of Monastir, Laboratory of Histology Embryology What is known already: Intracytoplasmic sperm injection (ICSI) is widely
and Cytogenetics LR 40 ES 18, Monastir, Tunisia ; used in the treatment of male infertility. Only morphologically normal sperma-
3
Center of Excellence DeNothe, Department of Experimental and Clinical tozoa are mainly used by embryologists to fertilize an oocyte. Different papers
Biomedical Sciences “Mario Serio”- Unit of Sexual Medicine and Andrology, have reported that spermatozoa with apparently normal morphology may have
Florence, Italy DNA fragmentation. These evaluations suggest that it is possible that nor-
mal-shaped spermatozoa but with DNA fragmentation could be easily selected
Study question: This study set out to determine the level of sperm nuclear to fertilize oocytes during ICSI. It is known that the presence of an increased
DNA damage in patients with isolated poymorphic teratozoospermia and exam- proportion of normal spermatozoa with damaged DNA is negatively associated
ining its relationship with oxidative stress and apoptosis. with embryo quality affecting both pregnancy and implantation outcomes
Summary answer: Decreased seminal antioxidant profile may be an important after ICSI.
factor involved in the mechanism of sperm cell death-mediated DNA breaks in Study design, size, duration: We designed an observational study on 70
teratozoospermic semen. male patients. We speculated that the examination of DNA integrity in motile
What is known already: Sperm morphological defects is associated with and morphologically normal sperm, collected after pellet swim up, could provide
apoptosis. useful information concerning sperm competence, rather than the DFI evaluation
Study design, size, duration: A total of 89 patients was divided into two in the raw seminal sample. We analyzed data from January 2019 to December
groups, men with isolated teratozoospermia (n = 69) and men with normal 2019. The aim is to demonstrate that DFI in normal morphologically spermato-
semen parameters (n = 20) as controls. zoa, could be indicated as predictive parameter of ICSI success.
Participants/materials, setting, methods: Sperm DNA breaks were eval- Participants/materials, setting, methods: DFI and traditional semen
uated by using acridine orange staining. The proportion of viable spermatozoa parameters (WHO, 2010), were evaluated in all patients. DFI was calculated
with mitochondrial transmembrane depolarization was detected by fluorescence using in situ TUNEL assay in at least 250 spermatozoa. By means of NIS-Elements
microscopy through the use of MitoPT-JC-1 staining method. Bivariate Annexin BR 3.10 image analyzer software (Nikon) using images of the same field (light,
V/ 6-CFDA analysis was then carried out to measure the percentage of both fluorescence and “merged”) it was possible to evaluate sperm morphology
viable and dead spermatozoa with phosphatidylserine (PS) externalization. associated with DNA fragmentation. Data were analyzed using the Kruskal-Wallis

test, a non-parametric ANOVA, confirmed by restrictive Bonferroni correction Main results and the role of chance: Fertilization rate was comparable
using the Dunn’s test. (p>0.05) among fresh and frozen samples for both group A (fresh: n=24,
Main results and the role of chance: In this observational study we included 67%fertilization rate / frozen: n=33, 62% fertilization rate) and group
70 oligoasthenospermic patients undergoing ICSI. The patients were classified B(fresh: n=23, 47%fertilization rate / frozen: n=38, 43% fertilization rate),
in 2 groups according to the sperm DFI: Group A (n=35) included those who with group A spermatozoa (n=27, 64%fertilization rate) performing signifi-
had a DFI < 15% in the population of sperm collected after swim up. In group cantly better compared to group B spermatozoa (n=91, 51%fertilization
B (n= 35) patients with a DFI ≥ 15%. We did not find any statistical difference rate), p<0.05.
between the two groups in the traditional sperm parameters like density, motility Group A spermatozoa produced significantly more good day 3 embryos com-
and morphology. pared to Group B (p<0.05), in particular 1,56 additional good quality embryos.
We observed that, in Group A, the average value of the total of sperm DFI Interestingly, fresh Group B spermatozoa performed better than frozen Group
was 9.32% while in Group B was 24.71 % (p< 0.0001). When the analysis was B spermatozoa, resulting in significantly more good quality embryos on day 3
restricted only to spermatozoa with normal morphology, it was observed that (p<0.05).
among patients of Group B the DFI value was 13.6%, while in A Group the Furthermore, there was a significant decrease in good quality day 3 embryos,
average DFI value was 2.2%, with a strong statistical difference (p<0.0001). DFI if both morphology and motility were graded as low/immotile (0.75 and 0.45
calculated on motile, normal morphologically spermatozoa can provide an less good quality embryos respectively, p<0.05).
important information on the probability and risk of injecting, during ICSI pro- Although there was a trend for higher cycle cancellation rate in group B com-
cedure, a sperm with normal morphology but with fragmented DNA. This risk paring to Group A, either fresh or frozen, the difference was not statistically
is higher if the sperm population collected after pellet swim up has a DFI higher significant.
than 15%. Overall, although LBR was not affected by any parameter examined, the num-
Limitations, reasons for caution: This type of analysis only provides a ber of good quality embryos available for transfer was affected by both the
prediction to select a sperm with fragmented DNA, but does not allow the quality of testicular spermatozoa used for ICSI and cryopreservation in low
selection of single spermatozoa with intact DNA to be used for ICSI. Further quality samples.
studies are needed to correlate these data with the clinical outcome. Limitations, reasons for caution: The evaluation of “presence” and “mor-
Wider implications of the findings: Our results suggest that the evaluation phology” as good/average/bad could have a subjective complexion. However,
of DFI in morphologically motile normal sperm selected after pellet swim up this variation is eliminated through the grouping of samples.
appears to be a more accurate strategy to evaluate the sperm competence, with Accumulative LBR was not calculated, as LBR was based only on the first transfer.
the aim to improve the ICSI outcomes, than the traditional evaluation of sperm Wider implications of the findings: Since there is cryopreserved testicular
DFI in the whole seminal sample. tissue of good quality, there is no added value in proceeding to another surgery.
Trial registration number: not applicable However, in low quality samples, the use of fresh testicular spermatozoa could
alter the final outcome, since cryopreservation affects the number of available
good quality embryos.
P-040 The impact of motility, morphology and presence of Trial registration number: Not applicable
testicular spermatozoa on fertilization, embryo development and
live birth rates, in fresh and frozen testicular samples
P-041 The effect of prolonged incubation of sperm at testis
C. Oraiopoulou1, A. Vorniotaki1, E. Taki1, A. Papatheodorou1, M. temperature (35°C) versus room temperature (26°C) on semen
Moissidou1, N. Christoforidis1, A. Chatziparasidou1 parameters
1
Embryolab IVF Clinic, Embryolab, Thessaloniki, Greece M. Mehrafza1, C. Ostadian2, A. Hosseini3, A. Eftekhari3,
M. Asgharnia1, S. Aghajani3, H. Vahabzadeh3, M. Gholami3,
Study question: Does cryopreservation or the quality parameters of testicular
A. Raoufi4, S. Samadnia4, E. Hosseinzadeh4
spermatozoa in fresh or frozen samples have an impact on fertilization rate, 1
embryo development and live birth rate? Mehr Fertility Research Center- Guilan University of Medical Sciences- Rasht- Iran,
Summary answer: Although live birth rate (LBR) is not directly associated to Gynaecology, Rasht, Iran ;
2
any parameters examined, morphology and motility of testicular spermatozoa Mehr Fertility Research Center- Guilan University of Medical Sciences- Rasht- Iran,
influence the number of available embryos. Andrology, Rasht, Iran ;
3
What is known already: Almost 5% of couples undergoing IVF treatments Mehr Fertility Research Center- Guilan University of Medical Sciences- Rasht- Iran,
are confronted with azoospermia and are counseled towards TESE-ICSI cycles. Embryology, Rasht, Iran ;
4
At the same time, it has been reported that there is no influence of the use of Mehr Fertility Research Center- Guilan University of Medical Sciences- Rasht- Iran,
cryopreserved testicular sperm in fertilization rate and live birth rate and in the Research, Rasht, Iran
presence of motile spermatozoa, high embryo quality and pregnancy rates are
expected. Motility of the spermatozoa during the ICSI procedure has been Study question: Does prolonged incubation of sperm at 35° C versus room
associated to live birth, while other studies claim that motility of either fresh or temperature (26 ° C) affect semen parameters and DNA fragmentation
frozen/warmed testicular spermatozoa is the only parameter associated to index (DFI)?
ongoing pregnancy. Summary answer: The concentration and motility of spermatozoa were
Study design, size, duration: A retrospective cohort study between 01/2014 significantly higher in room temperature than 35°C. However, Temperature had
and 12/2017 was performed in Embryolab IVF Clinic, Greece, including 108 no effect on DFI after 24 h.
TESE-ICSI treatment cycles. Logistic regression analysis was used to explore the What is known already: Currently, cryopreservation is used routinely for
influence of variables (fresh-frozen/warmed testicular tissue, presence/motility/ prolonged storage of sperm even for one day, which, despite its high cost, can
morphology of testicular spermatozoa) in fertilization rate, embryonic develop- affect the quality of sperm samples. If long-term incubation of sperm in the
ment and LBR. Women above 38 years old, frozen oocyte cycles, PGT cycles laboratory environment is possible without affecting its quality, it will be possible
and couples with abnormal karyotypes were excluded from the study. to manage the patient’s treatment with higher quality and with greater choice.
Participants/materials, setting, methods: Morphology, presence and Study design, size, duration: In the present experimental study, sperm sam-
motility were graded as: good/motile(1 grade), average/twitcher(2 grades), ples were collected from 40 participants referred to Mehr Medical Institute,
low/immotile(3 grades) and the sum of grades represented the total quality Rasht, Iran, from September 2019 to December 2019.
score for the testicular spermatozoa used for ICSI. Group A included cases Participants/materials, setting, methods: Each semen sample was
with up to total grade 4, while Group B included cases with total grade 5 or divided into two equal parts and was subjected to swim-up procedures. One
higher. Embryo quality was evaluated up to day 3 (good quality: more than group was incubated at 35°C and the other at room temperature, in the
5 blastomeres, less than 20% fragmentation). LBR was calculated per first darkness. Both groups were evaluated for number, motility (Grade A and B)
transfer. and morphology at 45 min, 24 h and 48 h intervals. Statistical analysis was

performed using repeated measure analysis of variance (ANOVA) and stu- Wider implications of the findings: Long-term obstruction is not related
dent´s t-test. to a sperm retrieval by conventional TESE and clinical pregnancy, however,
Main results and the role of chance: Sperm concentration (P = 0.007) and embryonic development were not similar as the other OAs. In CBAVD couples,
motility (P <0.001) was significantly higher in room temperature than 35°C, at the selection of motile sperm from testicular tissue could be a critical key to
three intervals of 45 minutes, 24 hours and 48 hours. However, the mean values succeed and rationale for good embryonic development.
of normal morphology spermatozoa was not significantly different between the Trial registration number: not applicable
two groups at three intervals of 45 minutes, 24 hours and 48 hours (P = 0.08).
Also, there was no significant difference in DFI at 26 and 35°C after 24 h (P = 0.2). P-043 Sperm count affects cumulative live birth rate of assisted
Limitations, reasons for caution: At the time of the abstract preparation, reproduction cycles in relation to ovarian response
20 samples had not been read for DFI. C. Zaca’1, G. Coticchio1, N. Tarozzi1, M. Nadalini1, C. Lagalla1,
Wider implications of the findings: Sperm morphology was significantly A. Garolla2, A. Borini1
reduced following prolonged incubation. 1
9.Baby, Family and Fertility Center, Bologna, Italy ; 2University of
Trial registration number: not applicable Padova, Department of medicine - Unit of andrology and reproductive
medicine, Padova, Italy
P-042 Clinical outcomes in patients with congenital bilateral
absence of the vas deferens (CBAVD) undergoing testicular sperm Study question: Does sperm quality, as assessed by prewash total sperm count
extraction-intracytoplasmic sperm injection (TESE-ICSI) (TSC), affect cumulative success rates in assisted reproduction cycles?
T. Ishikawa1, S. Mizuta1, K. Yamaguchi1, H. Matsubayashi1, Summary answer: Sperm quality impacts on the efficacy of IVF treatments
T. Takeuchi2, K. Kitaya1 primarily in cases of reduced ovarian response, but not in normal or high
1
Reproduction Clinic Osaka, Reproductive Medicine, Osaka, Japan ; responders.
2
Reproduction Clinic Tokyo, Reproductive Medicine, Tokyo, Japan What is known already: The clinical outcome of treatments is influenced by
numerous intrinsic and extrinsic factors. The female gamete is recognised as the
Study question: What is the fertilization rate, embryonic development, and single most important factor affecting the ability of the preimplantation embryo
clinical outcome for TESE-ICSI using testicular spermatozoa among obstructive to implant and develop. For this reason, oocyte legacy has overshadowed the
azoospermia (OA) including CBAVD couples? sperm role in embryogenesis. Regardless, TSC has been described as being highly
Summary answer: Clinical pregnancy rate (CPR) per embryo transfer (ET) predictive of male health in general and reproduction in particular. Its relative
by testicular sperm did not differ significantly between CBAVD and other OA importance in determining the efficacy of IVF treatments remains uncertain and
patients. therefore demands thorough assessment, especially in the light of possible inter-
What is known already: OA is caused by congenital including bilateral absence actions between the male and female gametes
of the vas deferens (obstructed in the long term) or acquired (vasectomy, inguinal Study design, size, duration: Reported data concern a retrospective cohort
hernioplasty in childhood, etc.). It is known that long-term obstruction is related study carried out between January 2009 to December 2013 involving 765 cou-
to a defect of spermatogenesis, however, clinical outcomes after ICSI with tes- ples undergoing complete ICSI cycles, i.e. whose all embryos were transferred
ticular sperm in the etiology OA, classified as congenital or acquired causes have or disposed. Couples were characterised by male infertility, while female age
hardly been investigated. was less than 36 years to minimise the well-documented maternal age effect on
Study design, size, duration: We performed a retrospective study based treatment outcome. Couples with a combination of female and male infertility
on two reproduction centers in Japan and evaluated 206 NOA patinets including factors were excluded.
50 cases with CBAVD performed in our clinic between September 2013 and Participants/materials, setting, methods: The cohort was grouped
December 2019. In addition, a total of 108 TESE-ICSI cycle with 47 couples for according to male partner’s TSC into five groups, according to values dis-
CBAVD and 293 TESE-ICSI cycles with 138 couples for other obstruction were cussed in the World Health Organization guidelines: A)< 0.1x106 (189/765
performed. Sperm retrieval rate was 100% and TESE-ICSI was performed with cycles,24.7%); B)0.1x106 to 1x106(144/756 cycles,18.8%); C)1x106 to
only motile spermatozoa. 5x106(150/756 cycles,19.6%); D)5x106 up to 10x106 (103/756
Participants/materials, setting, methods: The diagnosis of CBAVD is cycles,13,5%); E)10x106 to 39x106 TSC(179/756 cycles,23,4%).Groups
based on normal-size testes (>16mL), normal FSH and nonpalpable vas deferens. were also analysed according to the number of oocytes retrieved. Mean
The diagnosis of OA also required confirmation of normal spermatogenesis by age and number of oocyte was comparable among groups.Preimplantation
testicular biopsy. TESE-ICSI cycles were evaluated in embryonic development genetic testing treatments and cycles in which spermatozoa were surgically
rates and CPR. Additionally, paternal and maternal age, numbers of oocytes recovered were excluded.
retrieved, baseline FSH, LH and E2 levels of women, and FSH, LH and Main results and the role of chance: Mean age and number of retrieved
Testosterone of men were compared between the two groups. oocyte was comparable among groups. The mean number of embryos across
Main results and the role of chance: The wives age at ICSI was 33.9±4.4 the different TSC groups was also similar (3.0±1.6, 2.8±1.7, 3.4±2.2, 3.1±1.8
years for CBAVD and 35.9±5.1 years for other obstruction. The sperm retrieval and 3.2±1.9, respectively; P=0.066). Cumulative live birth rates (CLBR) were
rate with TESE was 100%, in which motile spermatozoa were retrieved and used progressively higher with increasing TSC, reaching a plateau in groups with higher
for ICSI for all patients. Two pronuclei (2PN), blastocysts development, and TSC (22.2%, 27.1%, 34.7%, 36.9%, 36.9%, respectively; P=0.01). On the con-
good-quality blastocysts rates were 57.2%, 44.8%, and 19.6% in CBAVD and trary, miscarriage rates were comparable ranging from 14.7% to 25% (P=0.5).
64.0%, 53.3%, and 23.0% in other obstruction, respectively. 2PN and blastocysts Furthermore, cumulative outcome rates were comparatively assessed in indi-
development rate in CBAVD was significantly lower than in other obstruction vidual TSC groups in relation to different oocyte yield (1 to 5; 6 to 10, >10). As
(P<0.001). However, no significant difference was found in good-quality blasto- expected, in the most severe TSC condition (≤0.1x106) CLBR were progressively
cysts rates. Embryo transfer was performed 44 couples with CBAVD patients higher as the number of retrieved oocytes increased (5.3%, 21.5% and 30.2%,
undergoing TESE-ICSI which were divided into clinical pregnancy positive (n=37) respectively; P=0.008). Similar trends were observed in the outcome of groups
and negative (n=7) groups. There was no difference for paternal and maternal with TSC values of >0.1x106 to 1x106 and >1x106 to 5x106. On the contrary,
age, numbers of oocytes retrieved, baseline FSH, LH and E2 levels of women, in groups characterized by TSC of >5x106 to 10x106 and >10x106 to 39x106,
and FSH, LH and Testosterone of men. CPR per ET cycle and per couple were CPR and LBR did not increase as a function of the number of retrieved oocytes.
similar in CBAVD (40.2% and 84.1%, respectively) and in other obstructive Limitations, reasons for caution: The study design is retrospective and
(41.3% and 79.4%, respectively). requires further refinement to control for factors that may impact clinical
Limitations, reasons for caution: There was a lack of accurate term of outcome.
obstruction with respect to not CBAVD OA patients. Some other obstructive Wider implications of the findings: This study highlights the general
patients could be congenital but not acquired. Additionally, the safety and screen- importance of sperm TSC for the efficacy of assisted reproduction treatments.
ing for congenital malformations among these children has not been fully The consequences of sperm quality on clinical outcome emerge decisively in
investigated. case of reduced ovarian response, suggesting a relationship between ovarian

response and oocyte ability to compensate for paternal-derived Summary answer: The use of microfluidic sperm sorting chip allows the
deficiencies. selection of highly motile sperm with very low levels of DNA fragmentation.
Trial registration number: Not applicable What is known already: DNA fragmentation represents the last event of
cell apoptosis. The use of sperm cells with DNA fragmentation for oocyte
P-044 Effect of paternal age on outcomes in ART cycles: A insemination is correlated with a negative paternal effect on embryo develop-
systematic review and meta-analysis ment and an increased miscarriage rates.
G. Morris1, D. Mavrelos1,2, E. Theodorou1, M. Campbell-Forde1, Both microfluidic sorting (MS) of unprocessed semen and Magnetic activated
D. Cansfield1, E. Yasmin1,2, P. Sangster1,2, W. Saab1, P. Serhal1, cell sorting (MACS) performed after density gradient centrifugation were
S. Seshadri1 reported to improve the sperm quality by selecting non apoptotic cells. However,
1
Centre for Reproductive and Genetic Health, Medical, London, United Kingdom ; it is uncertain which procedure leads to the lowest DNA fragmentation
2
University College London Hospitals NHS Foundation Trust, Reproductive index (DFI).
Medicine Unit, London, United Kingdom Study design, size, duration: The study investigates whether microfluidic
sorting of unprocessed semen improves sperm selection as compared to stan-
Study question: Could paternal age have an independent effect on the clinical dard preparation procedures (sperm washing and density gradient centrifugation)
outcomes of ART cycles? or MACS in split samples after routine diagnostic semen analysis. The primary
Summary answer: Autologous oocytes studies suggest increased male age outcome was sperm DNA fragmentation index and the secondary outcome was
reduces live birth and increases miscarriage. Donor oocytes studies found no sperm motility.
impact of male age on ART outcome. Participants/materials, setting, methods: Sperm samples from 6 different
What is known already: There is no effect of paternal age on clinical out- patients were used. Each sample was split and subjected to 4 different sperm
comes in donor oocyte studies. However, studies where autologous oocytes selection techniques: 1) sperm washing (SW), 2) density gradient centrifugation
were used, have demonstrated conflicting results with some suggesting that (DGC), 3) DGC followed by MACS (Miltenyi Biotec, Germany) and 4) MS
increased male age has a deleterious effect. (Fertile plus, Koek Biotechnology).
Study design, size, duration: A systematic review and meta-analysis of ten DNA fragmentation was analyzed using the Sperm-Chromatin-Dispersion Assay
autologous oocyte cohort studies (including 11004 cycles) and 11 donor oocyte (GoldCyto sperm kit, Goldcyto Biotech corp.). The DFI was calculated on at
cohort studies (including 10338 cycles). least 500 cells by the Sperm Class Analyzer CASA-system (Microptic, Spain).
Participants/materials, setting, methods: The Cochrane Central Citation Main results and the role of chance: The median DFI for the 4 processing
Index Register of Controlled Trials (CENTRAL), MEDLINE, EMBASE, NHS techniques were: 18.3% (range 13.9-87.6) for SW; 14.6% (range 6.1-71.7) for
evidence and conference abstracts were searched for randomized controlled DGC; 11.8% (range 3.9-46.9) for MACS, and MS: 0.7% (range 0-34.2) for MS.
trials (RCT) and observational studies (OBS) that addressed the impact of pater- The samples processed by microfluidics showed a significantly lower DNA frag-
nal age on ART published up to December 2018. We searched reference lists mentation rate compared to the other techniques (Friedman test, P<0.05).
of relevant articles and hand-searched relevant conference proceedings/ The median progressive motility for the 4 processing techniques were: 47.5%
abstracts. Data for women aged 39 years or under were extracted and analysed. (range 29-54) for SW; 58% (range 27-61) for DGC; 46% (range 3-78) for MACS
Main results and the role of chance: Clinical pregnancy rates were found and 92.5% (range 82-96) for MS.
to be statistically higher when the paternal age was under 40 years in autologous The samples processed by microfluidics showed a significantly higher progres-
oocyte studies (OR 1.68, [1.24 – 2.28]; p = 0.0009 ) however there was no sive motility (Friedman test, P<0.05).
difference in clinical pregnancy in the same age category when donor oocyte Limitations, reasons for caution: The major limitation of the study is the
studies were analysed (OR 0.95, [0.84 – 1.07]; p = 0.41). Livebirth rate (LBR) low sample size although the advantage of MS is obvious. Consequently, there
was reported in three autologous oocyte studies (2926 cycles) and five donor is a lack of variation between the samples regarding sperm quality. As this obser-
oocyte studies (7648 cycles). LBR rate was found to be significantly increased vational study was conducted on semen samples for diagnostic analysis, data on
when male age was under 40 years in autologous oocyte studies (OR 2.37, [1.15 clinical outcome are not available.
– 4.85]; p = 0.02) but no difference in LBR was found in donor oocyte studies Wider implications of the findings: Microfluidic sorting of sperm selects
(OR1.03, [0.80 – 1.32]; p = 0.84). Miscarriage rate was reported in two autol- a population with significantly lower DNA fragmentation index and higher pro-
ogous oocyte studies (970 cycles) and four donor oocyte studies (3741 cycles). portion of progressive motility compared to standard selection methods or
Miscarriage was found to be more likely with male age over 40 years in autolo- MACS. Moreover, MS offers the advantage of using unprocessed semen, as such
gous studies (OR 0.57, [0.42 – 0.77]; p = 0.0003). In donor oocyte studies there reducing the negative impact of centrifugation as compared to standard sperm
was a trend towards increased miscarriage when male age was above 50 years selection methods.
however this effect did not reach statistical significance (OR 0.71, [0.48 – 1.04]; Trial registration number: not apllicable
p value = 0.08).
Limitations, reasons for caution: The autologous oocyte studies are rela-
tively heterogeneous in populations with varied female ovarian reserve and P-046 Enzymatic digestion by Collagenase-IV improves quality
differing male age group divisions. The analysis was limited to cycles including of testicular sperm retrieved in Non-Obstructive Azoospermic
women under 39 years of age to mitigate this heterogeneity. (NOA) patients
Wider implications of the findings: Men over the age of 40 may have a E. Piatti1,2, E.M. Colpi3, G. Caminiti2, F. Castiglioni2,4,
reduced chance of a successful outcome following ART when using autologous E. Bernasconi5, P. Testa5, V. Monti5, M. Bellavia4, G. Momi6,
oocytes. This effect does not appear to be present when donor oocytes are G.M. Colpi2,4
studied although men over the age of 50 years may still have an increased chance 1
ProCrea, Andrology, Lugano, Switzerland ;
of miscarriage. 2
Clinica San Carlo, Andrology, Paderno Dugnano, Italy ;
Trial registration number: n/a 3
Gynecentro Ticino, Gynaecology, Lugano, Switzerland ;
4
ProCrea, Reproductive Unit, Lugano, Switzerland ;
P-045 Microfluidics is highly effective in selecting a sperm 5
Clinica San Carlo, Reproductive Unit, Paderno Dugnano, Italy ;
subpopulation with low DNA fragmentation index. 6
ProCrea, IVF Laboratory, Lugano, Switzerland
K. Wouters1, L. Vandenberghe1, A. Racca1, D. Jankovic1, G.
Verheyen1, I. Mateizel1 Study question: Does Enzymatic Digestion alone have positive effects on
1
UZ Brussel, Centre for reproductive medicine, Brussel, Belgium isolation of human sperm from surgical testicular samples, compared to standard
mechanical treatment?
Study question: Is microfluidic sorting of unprocessed semen better in select- Summary answer: Enzymatic Digestion alone by Collagenase-IV seems a valid
ing non-DNA fragmented sperm compared with other semen preparation method to increase the total surgically retrieved viable sperm count suitable
techniques? for ICSI.

What is known already: When available, surgically-retrieved testicular-sperm What is known already: According to the Russian Association of human
(Testi-Sperm) used with ICSI offer an opportunity to father children even to reproduction data after the transfer of embryos obtained by ICSI pregnancy
NOA patients. The treatment of testicular fragments is crucial, both for a positive occurs in 36.2 % of cases and perinatal miscarriage reaches 78.1 %. The use of
retrieval and for the total Testi-Sperm number obtained and storable for ICSI. hyaluronate for the study of male germ cells in the ICSI procedure allows select-
Enzymatic treatment following mechanical mincing of testis fragments was ing of sperm without DNA damage and abnormalities in the chromosomal set,
reported to increase retrieval success rate and final Testi-Sperm count. thereby increasing the chances of successful fertilization and meanwhile according
Collagenase-IV, a Sertolian protease, was shown to provide higher yelds of to a large systematic review devoted to comparing the effectiveness of ICSI and
Testi-Sperm cells. However, a critical factor for ICSI success is sperm motility, PICSI in male infertility factor, which included 2918 studies, there are no statis-
as a proof of sperm viability, especially after cryopreservation. tically significant differences between these two methods.
Study design, size, duration: A prospective blind observational study was Study design, size, duration: Retrospective multiparametric comparative
performed on 24 NOA patients. In each, the testicular microfragments from analysis of 284 ICSI procedures and 200 PICSI procedures performed from 2015
microdissection-TESE were divided into two same-weight parts: the first under- to 2019 in the ART Department of “IDK Medical company”, (Samara, Russia)
went mechanical mincing, the second was treated by Collagenase-IV solution was performed.
alone (never previously reported). Viability was assessed by Eosin-Y test (WHO Participants/materials, setting, methods: The following performance
2010), applied to small cell suspension aliquots; and concentration by a counting parameters were evaluated: fertilization rate, blastocyst formation rate, embryo
chamber. All samples were prepared by one biologist and then blind-evaluated freezing rate, embryo utilization rate, implantation frequency rate, pregnancy
by another. rate, live-birth rate, cumulative pregnancy rate, cumulative live-birth rate.
Participants/materials, setting, methods: The surgical samples were Main results and the role of chance: The fertilization rate in ICSI was 86%,
placed in Petri dishes containing HAM’s F10 medium: the first half, mechanically in PICSI - 82%. The embryo freezing rate and blastocyst formation rate in ICSI
minced to obtain a homogeneous cell suspension, and the second half, immersed were 28% and 47% respectively, in PICSI - 33% and 51%. The differences between
in 1 ml Collagenase IV solution. Both were incubated at 37°C for 120’; then, two methods are statistically insignificant (p>0,05). The embryo utilization rate
both treatments were stopped by adding 5 ml HSA 0.5% medium, the tubes in ICSI was 48%, in PICSI - 58%. The differences in these indicators are statistically
centrifuged for 10’ at 500g for washing, the pellets re-suspended with fresh significant (p=0,043). While assessing the implantation frequency rate it was
medium and analyzed. found that in ICSI it has become 44.6%, in PICSI it has become 45%. The preg-
Main results and the role of chance: For each treatment, we measured nancy rate and the live-birth rate after ICSI were 47% and 33% respectively, and
the Total Count of the Testi-Sperm retrieved, the percentage of Viable Testi- the pregnancy rate and the live-birth after PICSI were 48.5% and 35% respec-
Sperm (by Eosin-Y test on over 100 cells), and consequently the Total Count of tively. The differences in these indicators are statistically insignificant (p>0,05).
Viable Testi-Sperm actually available for cryopreservation or ICSI. Statistical While analyzing the cumulative pregnancy rate and the cumulative live-birth rate
analysis was performed by Wilcoxon-paired rank test. the following data was provided: in ICSI these indicators were 53% and 34%,
Viable Testi-Sperm were [X±SD; (range)]: 31.9±9.7% (19-63%) and respectively, in PICSI - 81.5% and 56.5% respectively. The differences in these
67.2±8.1% (52-80%) for mechanically and Collagenase-IV treated samples, indicators are statistically significant (p=0,038).
respectively (p<0.01). Limitations, reasons for caution: Absent.
Total Testi-Sperm retrieved were [M (range)] 0.49∙106 (0.06-2.78∙106) and Wider implications of the findings: Multiparametric analysis makes it pos-
0.77∙106 (0.14-2.3∙106)(p<0.01), and Total Viable Testi-Sperm retrieved were sible to evaluate the effectiveness of ART programs with great confidence. There
0.16∙106 (0.01-1.0∙106) and 0.54∙106(0.08-1.59∙106) (p<0.01) for mechanically are no significant differences between ICSI and PICSI for most of the analyzed
and Collagenase-IV treated samples, respectively. indicators, which is consistent with the literature data. But cumulative perfor-
Therefore, Collagenase-IV treatment alone gives a Total Count of Viable mance indicators demonstrate the advantage of PICSI over conventional ICSI.
Testi-Sperm useful for cryopreservation or ICSI significantly higher than that of Trial registration number: not applicable
the standard mechanical treatment.
Limitations, reasons for caution: Our results on the higher Viable Total
Testi-Sperm Count retrieved by Collagenase-IV alone versus mechanical treat- P-048 Which is better for sperm incubation before IVF: lower or
ment invite future studies on microdissection-TESE samples, especially to confirm higher oxygen?
the maintained functional competence of Testi-Sperm in ICSI. E. Yalcinkaya1, B. Alyuruk2, F. Martinez Pastor3
Wider implications of the findings: The enzymatic-only method in pro- 1
Adatip Hospital, Assisted Reproduction Unit, İstanbul, Turkey ;
cessing Testi-Sperm during microdissection-TESE can offer higher positive Testi- 2
Kocaeli University, IVF Unit, Kocaeli, Turkey ;
Sperm retrieval rates and higher number of viable cells available for 3
INDEGSAL, University of Leon, Leon, Spain
cryopreservation or ICSI in NOA patients, especially in those with the poorer
prognosis. Study question: Do lower levels of oxygen result in less oxidative stress on
Trial registration number: None sperm cells before fertilization in bull?
Summary answer: This pilot study reported a higher oxidative stress due to
increased mitochondrial activity under high oxygen during incubation.
P-047 Multiparametric comparative analysis of the effectiveness What is known already: Fertilization of bovine oocytes is still carried out
of conventional ICSI and physiological ICSI (PICSI) among couples under high oxygen levels in most of the laboratories although embryo culture is
with male infertility factor performed under lower oxygen tension due to the solid evidence supporting
D. Rusakov1, O. Shurygina2, A. Petrova3, T. Bykova3, A. Baizarova3 higher blastocyst development rates. However, incubation of sperm cells under
1
Samara state medical University- IDK medical company, Department of histology lower oxygen concentration might be crucial in terms of sperm function consid-
and embryology- ART Department, Samara, Russia C.I.S. ; ering that these cells are susceptible to oxidative stress due to their inadequate
2
Samara state medical University- IDK medical company, Department of histology cell repair system against excessive production of reactive oxygen species (ROS).
and embryology - Department of reproductive medicine clinical embryology and Study design, size, duration: This was a pilot study for comparing the effects of
genetics - ART Department, Samara, Russia C.I.S. ; two different levels of O2 on sperm cells during capacitation. The sample of the study
3
IDK medical company, ART Department, Samara, Russia C.I.S. included 10 cryopreserved bull semen samples from different breeders. Treatment
groups were named as LO (low oxygen; 5% O2) and HO (high oxygen; 20% O2).
Study question: Multiparametric comparative analysis of the effectiveness of Participants/materials, setting, methods: Two straws were thawed per
conventional ICSI and physiological ICSI (PICSI) in couples with male infertility each bull and sperm cells were left to swim up for 1 hour. Then, swam sperm
factor performed. cells were divided equally into two; and aliquots were assigned to both treatment
Summary answer: PICSI compared to ICSI reduces the number of embryo goups to incubate for 5-6 hours. Variables of motility, progressive motility, mito-
losses at the laboratory stage and increases cumulative pregnancy rate and cumu- chondrial activity, capacitation status and intracellular ROS were measured by
lative live-birth rate. flow cytometry at 0 h and 5-6 h.

Main results and the role of chance: Compared to starting levels, both modifications, semen samples treated with myo-inositol or untreated were ana-
groups showed significantly decreased total and progessive motility, nonsignifi- lysed by transmission electron microscopy. Despite the treatment seems to not
cant increase in capacitation status, significant decrease in the percentage of induce appreciable ultrastructural changes, it may be highlighted that sperm
active mitochondria and statistically significant decreases in intracellular ROS. surface as well as the observed environment are clearly free from any amorphous
The decrease in total and progressive motility was found to be significantly higher fibrous material, which instead appears as background in untreated sperm sam-
in LO compared to HO (22.6% vs 32.8% and 24.4% vs 18.6%; p<0.05, respec- ples. This finding may confirm the ability of myo-inositol to disrupt the mucoid
tively). Moreover, the percentage of capacitated cells were found to be higher masses of the seminal fluid, usually able to entrap sperm and any other seminal
in HO compared to LO; but the difference was not statistically significant fluid components, thus reducing sperm motility and capacitation efficiency.
(p>0.05). At the end of incubation, mitochondrial activity was observed to be Limitations, reasons for caution: Large-scale studies should be required to
slightly higher in HO than LO; but the difference was not again statistically sig- assess whether an addition of myo-inositol improves sperm motility, also in
nificant (p>0.05). Interestingly, intracellular ROS was found to be decreased oligoasthenozoospermic men. Moreover, the mechanism by which myo-inositol
following incubation; and it was found to be statistically lower in LO group ameliorates mitochondrial activity should be further evaluated.
compared to HO (p<0.05). Wider implications of the findings: These data, if confirmed in future larger
Limitations, reasons for caution: The main limitation of our study was bull studies, will disclose that myo-inositol ameliorates key feature of human sperm
variability and the limited size of the study sample. (motility, mitochondrial functions, DNA integrity).
Wider implications of the findings: Considering that ROS may lead to Trial registration number: None
problems in reproductive processes when present in excess amounts, use of
lower oxygen also during capacitation was found to keep intracellular ROS levels P-050 Impact of sperm quality to achieve pregnancies in oocyte
significantly lower as desired. Any further effects on the quality of spermatozoa donation cycles
need to be evaluated by fertilization and IVF outcomes. M. Cozzolino1, I. Hervás1, R. Rivera-Egea1, A. Pellicer1, N. Garrido1
Fundacion IVI, Valencia, Valencia, Spain
P-049 In vitro myo-inositol treatment significantly ameliorates Study question: When should be considered sperm replacement in case of
motility, mitochondrial respiratory efficiency and DNA integrity of the first failure of oocyte donation?
human sperm Summary answer: Replacement patients by donors sperm significantly
P. Piomboni1, R. Ponchia1, B. Semplici1, L. Governini1, P. Artini2, E. increase live birth rate and cumulative live birth rate (CLBR) either in oligo-as-
Casarosa2, I. Marzi2, A. Luddi1 thenoteratozoospermia or in suboptimal/normal sperm.
1
Siena University, Molecular and Developmental Medicine, Siena, Italy ; What is known already: Oocyte donation is an extremely successful treat-
2
University of Pisa, Experimental and Clinical Medicine, Pisa, Italy ment in assisted reproductive technology (ART), unfortunately, any couples
remain still barren despite normal sperm and regular uterus morphology. The
Study question: Does the in vitro treatment with myo-inositol affects the use of ICSI in clinical practice has modified the indications of sperm donation
mitochondrial function and DNA integrity of human sperm in both basal and use, currently, the ESHRE suggests the donation of sperm after at least three
capacitated condition? failed cycles of ICSI with the sperm of the couple. A repetitive clinical question
Summary answer: The in vitro treatment with myo-inositol of normozoosper- that remains unsolved so far, concerns to when a couple should be recommended
mic samples, before and after swim-up capacitation, leads to a significant ame- to change patient’s by donor sperm after oocyte donation failed treatments
lioration of sperm motility and DNA integrity Study design, size, duration: A retrospective study in university-affiliated
What is known already: Mammalian fertilization is a multifaceted and com- ART centers was conducted between January 2000 and April 2019. 6065 cycles
plex process because the site of semen deposition is far from the site of fertil- of oocyte donation were included in the study, with a total of 7757 embryo
ization and sperm need to have an adequate motility to reach the oocyte. Sperm transfers and 14119 embryos transferred.
motility, as well as capacitation, hyperactivation and acrosome reaction, requires Participants/materials, setting, methods: Couples in which male part-
energy and, to this regard, mitochondria play a pivotal role in the aerobic pro- ners have oligoasthenoteratospermia or suboptimal/normal sperm were
duction of energy in comparison with the glycolytic pathway. At same time, scheduled for oocyte donation cycles after the failed first attempt, using
DNA integrity must be preserved during sperm capacitation and related pro- patients or donors sperm.Oligoasthenoteratospermia was defined as the pres-
cesses. These features may be affected by in vitro procedure for sperm selection ence of less than 5 million total progressive motile spermatozoa in the ejacu-
and capacitation in assisted reproductive techniques. late. Obstructive azoospermia was excluded. Live birth rates per embryos
Study design, size, duration: Human ejaculated sperm were analyzed transferred and embryo transfers, and cumulative live birth rates (CLBR)
according to WHO guidelines between September 2019 - January 2020; a total considering oocyte consumed in the previous donation cycles using patient or
of 40 normozoospermic samples were selected in order to evaluate the effect donor sperm.
of an in vitro treatment with myo-inositol, before and after swim up selection. Main results and the role of chance: After the first failure of the oocyte
Isolated sperm from all specimens were used to measure the respiratory control donation cycle our results confirm that in patients with oligoasthenoteratosper-
ratio (RCR) and the DNA oxidation; Sperm ultrastructure was analysed. All mia or suboptimal/normal sperm, replacement patient’s sperm by donor sperm
patients had given their informed consent. as soon as increased live birth per embryo transfers or the number of embryos
Participants/materials, setting, methods: Normozoospermic human semen transferred. The live birth rate in couples with oligoasthenoteratospermia
samples (n=40) collected at the Siena University Hospital were processed for the showed that replacement patient by donor sperm OR 2.18 CI95% (1.7-2.8)
oxygraphic analysis, before and after swim up selection, with or without treatment p<0.001 when considered for embryo transfers and OR 2.46, CI95% (1.9-3.2)
myo-inositol (2 or 20 µg/µl). To this end, sperm cells were demembranated by p<0.001 considering for embryos transferred. In couples with suboptimal/nor-
hypotonic swelling and the RCR was calculated. DNA sperm oxidation measure- mal sperm replacement of patient sperm with donor showed an increase live
ment by 8-OHdG and ultrastructural evaluation by transmission electron micros- birth for embryo transfers OR 2.21, 95%CI (1.7-2.8), p<0.001 and for embryos
copy were performed in myo-inositol treated and untreated sperm samples. transferred OR 2.43, CI 95%(1.9-3.1), p<0.001. The same results were reached
Main results and the role of chance: Our data highlight that myo-inositol in the third cycles of donation with improving live birth rate for embryo transfers
treatment significantly increased sperm progressive motility, in a dose dependent OR 1.51, CI95% (1.0-2.2), p=0.033 and embryos transferred OR 1.77, CI 95%
manner (p<0.05), both in basal and swim-up selected samples. Moreover, the (1.2-2.6), p=0.004 in oligoasthenoteratospermia, in case of suboptimal/normal
consumption of O2 measured by oxigraphic analysis was significantly raised by sperm was OR 1.40, CI95%(1.0-2.0), p=0.039 for embryo transfer and OR 1.53,
increasing doses of myo-inositol in vitro treatment.The concentration of CI 95%(1.0-2.4), p=0.013 for embryo transferred.The survival curves of CLBR
8-OHdG, an early marker of DNA oxidation, in untreated sperm was significantly with the patient and donor sperm considering oocyte by oocyte showed a
higher than those of sperm supplemented with myo-inositol, confirming its pro- statistically significant difference (p<0.001), replacing sperm patients after fifteen
tective role against DNA oxidative damage during the in vitro manipulation/ in oligoasthenoteratospermia and twenty oocytes donated, in suboptimal/nor-
incubation of human sperm. To evaluate any possible ultrastructural mal sperm.

Limitations, reasons for caution: The principal limitation of our study was Wider implications of the findings: We would recommend to verify patho-
the lack of andrological studies at the molecular level, such as the sperm DNA logical results using those markers in order to increase precision diagnosis among
fragmentation test or sperm FISH test, which could have identified a clearer NOA patient in whom sperm was not retrieved. These results may be used for
indication for the gamete change for each patient and the retrospective nature considering re-TESE or re-microTESE.
of the study. Trial registration number: not required
Wider implications of the findings: Prospective studies taking into account
an andrological study should confirm the possibility of changing the patient’s
semen after the first failed donation. P-052 Outcome of IVF/ICSI procedures in male cancer
Trial registration number: not applicable patients: retrospective analysis of procedures performed between
2004 – 2018
P-051 Distinct expression levels of spermatogenic markers and T. Papler1, M. Štimpfel2, E. Vrtacnik Bokal3, S. Drobnič2
growth factors in testicular tissue of Klinefelter patients with 1
University Medical Center Ljubljana, Department of Human Reproduction,
negative sperm retrieval
Ljubljana, Slovenia ;
E. Lunenfeld1, M. Azab2, S. Kleiman3, R.R. Hauser4, M. Huleihel.2 2
University Medical Centre Ljubljana, Department of Human Reproduction,
1
The Center of Advanced Research and Education in Reproduction CARER-Faculty Ljubljana, Slovenia ;
of Health Sciences Ben-Gurion University of the Negev- Beer-Sheva- Israel. and 3
Univeristy Medical Centre Ljubljana, Department of Human Reproduction,
Soroka Medical Center, Department of Obstetrics and Gynecology, Beer - Sheva, Ljubljana, Slovenia
Israel ;
2
The Center of Advanced Research and Education in Reproduction CARER- Faculty
of Health Sciences- Ben Gurion University of the Negev, The Shraga Segal Study question: Study question: Is the clinical outcome of IVF/ICSI cycles
Departement. of Microbiology- Immunology and Genetics-, Beer Sheva, Israel ; comparable when fresh or cryopreserved semen from male cancer patients is
3
Male Fertility Clinic and Sperm Bank- Lis Maternity Hospital- Tel Aviv Sourasky used for fertilization?
Medical Center School of Medicine Tel Aviv University, Obstetrics and Gynecology, Summary answer: There are no statistically significant differences in
Tel Aviv, Israel ; clinical outcomes of IVF/ICSI procedures when fresh or frozen semen is
4
Male Fertility Clinic and Sperm Bank- Lis Maternity Hospital- Tel Aviv Sourasky being used
Medical Center- School of Medicine Tel Aviv University, Obstetrics and Gynecology, What is known already: Due to improved survival of men after oncological
Tel Aviv, Israel treatment it has become important to enable them a quality life after the
treatment is finished. Fertility preservation and a chance to have biological
Study question: Can we use spermatogenic markers and testicular growth children represent an important aspect of quality of life. In men we can
factors to predict spermatogenesis in testicular biopsies of Klinefelter patients preserve fertility by cryopreserving semen prior to the start of oncological
without sperm? treatment. Outcomes of IVF procedures are comparable when we use fresh
Summary answer: The expression of oct4 and CSF-1 were high and in a or frozen semen in healthy infertile couples. Data on the use of fresh or frozen
positive correlation with the expression of crem-1 and protamine, in the exam- semen from oncological patients and the outcomes of IVF procedures how-
ined biopsies. ever, are scarce.
What is known already: Klinefelter syndrome (KS) is the most prevalent Study design, size, duration: We retrospectively analysed the outcome of
genetic disorder, occurring at about 0.2% of males and 11% of the azoospermic IVF/ICSI cycles where cryopreserved or fresh semen from male oncological
patients. At birth, KS patients have normal amount of testicular germ cells. Until patients was used for oocyte fertilization. Analysis included 226 IVF/ICSI cycles
age 10, spermatogonial cells (SCs) are present in the testes, but can be found in performed between 2004 and 2018, 138 with cryopreserved semen and 88 with
40-50% of the adult patients. The presence of SCs (OCT4 and MAGE-A4) was fresh semen, respectively. We had no exclusion criteria with regard to female
reported. It is suggested that spermatogenic arrest occurs at the spermatogo- age (aged 23-43 years) or female causes of infertility.
nium or primary spermatocyte level in KS boys, which undergo apoptosis instead Participants/materials, setting, methods: Groups (cryopreserved versus
of differentiation. The expression of growth factors in testicular biopsies of fresh semen) were compared in terms of fertilization rate, number of embryos,
Klinefelter patients is not yet clear. blastocysts and cryopreserved embryos. Pregnancy, live birth and miscarriage
Study design, size, duration: The expression levels of the pre-meiotic (oct4, rates were determined to determine which type of semen used yields better
cd9, gfr-a1, a-6-integrin, sall4, c-kit), meiotic (crem-1) and post-meiotic (prota- clinical outcomes. To determine the differences between the two groups,
mine) markers as well as the colony stimulating factor-1 (CSF-1) (growth factor) Pearson’s chi-square test and two-tailed t-test were used. Statistical significance
were examined in testicular biopsies of 17 Klinefelter patients (28-40 years old), was set at p < 0.05.
with no sperm in their testicular tissue. Main results and the role of chance: The cryopreserved semen and fresh
Participants/materials, setting, methods: 17 Klinefelter patients semen group of patients were comparable for mean female age (31.7 ± 4.7 vs.
underwent conventional TESE and no sperm was retrieved. qPCR analysis 32.5 ± 4.3; p=0.65), mean number of retrieved oocytes per aspiration (10.2 ±
was used to quantify the expression levels of the extracted RNA from the 5.6 vs. 10.1 ± 6.6; p=0.71), fertilization rate (56.9% vs. 59.1%; p=0.35) and
17 examined biopsies in whom sperm was not retrieved. The levels of the number of embryos (629 vs. 423; p=0.34). There was however, statistically
markers were expressed as fold of increase compared to GAPDH in the significant higher number of blastocysts (211 vs 116; p=0.018) and cryopre-
same tissue sample. served embryos (124 vs 51; p=0.001) in the group where cryopreserved semen
Main results and the role of chance: Our results show that in 14 biopsies, was used. In terms of clinical IVF outcomes, there were no differences in preg-
we examined low expression levels of oct4 - a pre-miotic marker (fold of nancy (34.2% vs. 28.6%; p=0.43), live birth (24.8% vs. 24.3%; p=0.92) and mis-
increase was less than 5), CREM - a miotic marker (fold of increase was less carriage (27.5% vs. 10%; p=0.18) rates between cryopreserved and fresh
than 6) and CSF-1 (fold of increase were less than 18) but no expression of semen group.
protamine. However, in three biopsies that highly expressed oct4 (29, 39 and Limitations, reasons for caution: In the fresh semen group, timing from
61 fold) and CSF-1 (162, 86 and 88 fold, respectively), the expression levels of finished oncological treatment to IVF procedure should be taken into account
crem-1 were high (50, 21 and 28 fold, respectively) and also protamine (1364, to get a clearer perspective about semen quality. Also, the type of cancer and
20 and 466, respectively). No such correlation was found between the expres- oncological treatment should be considered to get a clearer picture.
sion levels of the pre-meiotic markers cd9, gfr-a1, a-6-integrin, sall4, c-kit and Wider implications of the findings: Semen cryopreservation prior to onco-
the expression levels of the meiotic marker crem-1 and the post-meiotic marker logical treatment represents an efficient way for fertility preservation in men with
protamine. cancer. Furthermore, it seems that the use of cryopreserved semen is better in
Limitations, reasons for caution: The number of the patients need to be terms of number of blastocysts obtained and number of embryos cryopreserved
increased and the expression levels of the markers need to be confirmed by after IVF procedure.
immunostaining. Trial registration number: Retrospective analysis.

P-053 Impact of slow freezing on sperm nuclear quality and S. D’Andrea1, C. Castellini1, E. Minaldi1, M. Totaro1, G. Cordeschi1,
telomere length S. Francavilla1, F. Francavilla1, A. Barbonetti1
C. Gouhier1, H. Pons-Rejraji1,2, S. Carlet-Dollet2, B. Pereira3, 1
University of L’Aquila, Pubblic Health- Clinical- Life and Envirenomnental Science,
A. Tchirkov4, F. Brugnon1,2 L’Aquila, Italy
1
CHU Clermont-Ferrand- Hôpital Estaing-, Laboratoire de Biologie du
Développement et de la Reproduction- AMP- CECOS, Clermont-Ferrand, France ; Study question: What is the relationship between the degree of sperm-au-
2
Faculté de Médecine, Université Clermont Auvergne- Inserm 1240- IMoST, toimmunization, assessed by IgG-mixed antiglobulin reaction (MAR) test, with
Clermont-Ferrand, France ; natural and intra-uterine insemination (IUI)-assisted live births?
3
CHU Clermont-Ferrand, DRCI- Délégation Recherche Clinique et Innovation, Summary answer: Infertile couples with 100%-positive MAR-test, where
Clermont-Ferrand, France ; natural live births were much less than at lower degree of positivity (50%-99%),
4
CHU Clermont-Ferrand- Hôpital Estaing-, Service de Cytogénétique Médicale, could be successfully treated with IUI.
Clermont-Ferrand, France What is known already: Although the World Health Organization (WHO)
has recommended the IgG-MAR test as an integral part of semen analysis, con-
Study question: Does slow freezing affect human sperm telomere length? sidering 50% antibody-coated motile spermatozoa as the clinically-relevant
Summary answer: Slow freezing of human sperm does not alter telomere threshold, the predictive value of the degree of positivity of the MAR test above
length despite of increasing DNA oxidation and fragmentation. such a cut-off on the occurrence of natural pregnancies still remains largely
What is known already: Sperm telomere length (STL) is a potential interesting undetermined. Furthermore, the effectiveness of IUI in cases of strong sperm-au-
biological marker to assess male fertility. While its relationship with standard toimmunization is not yet well-established.
sperm parameters is still controversial, recent studies have shown correlations Study design, size, duration: This was a historical cohort study on 108
between STL and DNA alterations, notably DNA fragmentation and oxidation. infertile couples with ≥50% positive MAR test in men, who had attended a
Likewise, STL is positively related with pregnancy rate. As STL is transmitted to university/hospital’s andrology/infertility clinic for the management of couple
offspring, its alteration could lead to pathologies that may worsen from gener- infertility from March 1994 to September 2017.
ation to generation. Human sperm freezing is an indispensable tool for male Participants/materials, setting, methods: The IgG-MAR was carried out
fertility preservation despite of inducing nuclear spermatic alterations. as an integral part of semen analysis. The patients were divided into two groups:
Nevertheless, nothing is known about its impact on STL. 100%– and 50%–99%-positive MAR test. Post-coital test (PCT) was performed
Study design, size, duration: This prospective study was conducted in all the couples and IUI was offered as the first-line treatment. Laboratory and
between January and December 2018 and approved by the “Germethèque” other clinical data were retrieved from the computerized database. Data on
Scientific Committee and the Ethical Committee under the French IRB CPP subsequent pregnancies were obtained by contacting patients over the
Sud-Est6. Samples were surplus semen obtained from 63 men undergoing telephone.
routine semen analysis at the Center for Reproductive Medicine. STL mea- Main results and the role of chance: Eighty-four men (77.8%) were suc-
surement was carried out on all the 63 fresh samples, and for 30 of those also cessfully contacted by telephone and agreed to participate. Forty-four men
after cryopreservation. In addition, nuclear quality was measured before and belonged to the group of 100%-positive MAR test, while 40 showed lower
after freezing-thawing cycle. MAR test positivity. The couples with a 100%-positive MAR test showed a
Participants/materials, setting, methods: The 63 subjects (35 normozo- natural live birth rate per couple (LBRcouple), that was considerably lower than
ospermic) had a mean age of 34±0.6 years and a BMI of 27.4±0.8. Spermatozoa that observed at a lower degree of positivity (4.5% vs. 30.0%; p = 0.00001).
were slowly frozen in high security straws using freezing medium and program- Among the clinical variables, a significant difference was observed only for the
mable freezer. STL was measured for all fresh and frozen-thawed samples by PCT outcome, which was poorer in the 100%-positive MAR test group. Better
qPCR. For 10 samples, STL before and after freezing-thawing was additionally PCT outcomes (categorized as negative, subnormal and good) were positively
assessed by qFISH. For another 10, sperm DNA oxidation (8-OHdG residues correlated to the occurrence of natural live births (6.3%, 21.7%, and 46.2%,
immuno-detection) and fragmentation (TUNEL assay) were measured by flow respectively; p = 0.0005 for trend), for which the sole independent negative
cytometry. predictor was the degree of sperm-autoimmunization. IUI was performed as
Main results and the role of chance: We did not measure any significant the first line treatment in 38 out of 44 couples with 100%-positive MAR test,
impact of freezing on the mean telomere length of sperm samples, whether it yielding 14 live births (LBRcouple = 36.8%). In couples with lower MAR test
was analyzed by qPCR (3.25 ± 0.15 arbitrary unit (a.u.) vs. 3.46 ± 0.2 a.u., n = positivity, the LBRcouple after IUI was similar to the natural LBRcouple (26.9%).
30) or by qFISH (0.84 ± 0.1 a.u vs. 0.84 ± 0.04 a.u, n=10). qFISH method Limitations, reasons for caution: We cannot exclude uncontrolled variables
highlighted STL heterogeneity within each sperm sample. The cryopreservation that may have affected natural pregnancies during the follow-up, or a selection
tended to modify STL distribution (segmented by intervals of 0.25 a.u.) even if bias in the comparison of outcomes, due to the lack of randomization even
it was not statistically significant. though the clinical variables were not different in couples that had or had not
In addition, slow freezing-thawing cycle decreased sperm motility and vitality undergone IUI.
(p <0.05 for both, n=10), and increased DNA oxidation (average oxidation Wider implications of the findings: While 100%-positive IgG-MAR test
intensity 839 ± 210 a.u. vs. 1445.2 ± 202 a.u., p < 0.05, n=10) and fragmentation can represent the sole cause of a couple’s infertility, which could be successfully
(23.1 ± 5.4 % vs. 39 ± 6 %, p <0.005, n=10). Finally, we did not measure any treated with IUI, a lower degree of positivity may only represent a contributing
significant correlations between STL and the analyzed spermatic nuclear markers. factor to be considered in addition to conventional prognostic factors, including
Limitations, reasons for caution: Standard parameters of analyzed samples the PCT outcome, in decision making “treat or wait”.
were normal or slightly altered because the different measurements required a high Trial registration number: not applicable
number of spermatozoa. Nevertheless, fertility preservation is proposed to men
with potentially altered sperm parameters, therefore it would be essential to measure
P-055 Cell-free seminal mRNA as a marker of complete
cryopreservation impact on semen of oligo-astheno-teratozoospermic patients.
obstruction in azoospermic men
Wider implications of the findings: Although slow freezing increased DNA
oxidation and fragmentation, it did not alter STL. To our knowledge, this was M. Jeseta1, P. Ventruba1, W. Kranc2, K. Jopek3, J. Žáková1, I. Crha1,4,
the first study to assess the impact of cryopreservation on STL. Compared to B. Kempisty2,3
1
qPCR that only gives a mean STL, qFISH provides the distribution of STL within Faculty of Medicine- Masaryk University Brno and University Hospital Brno,
a semen sample. Department of Gynecology and Obstetrics, Brno, Czech Republic ;
2
Trial registration number: not applicable Poznan University of Medical Sciences, Department of Anatomy, Poznan, Poland ;
3
Poznan University of Medical Sciences, Department of Histology and Embryology,
P-054 Relationship between the degree of sperm- Poznan, Poland ;
4
autoimmunization with natural and intra-uterine insemination- Faculty of Medicine- Masaryk University Brno and University Hospital Brno,
assisted live births Department of Nursing and Midwifery, Brno, Czech Republic

Study question: Is it possible to use cell-free mRNA (cfs-mRNA) in seminal occurring only in a minority of individuals. No factors predicting recovery were
plasma as a diagnostic biomarker of obstruction in azoospermic patients? elucidated.
Summary answer: Our results showed that the seminal plasma samples from What is known already: Escalated BEACOPP (bleomycin, etoposide, doxo-
non-obstructive azoospermic donors differed significantly in contents of cell-free rubicin, cyclophosphamide, vincristine, procarbazine, prednisone), a multiagent
mRNA when compared to obstructive azoospermic men. chemotherapy regimen, was introduced in the 1990s for the treatment of
What is known already: Extracellular cell-free mRNA has been detected in advanced-stage Hodgkin’s lymphoma. It demonstrates superior prognostic out-
various body fluids including seminal plasma. This mRNA is actively released comes in progression-free survival, overall survival and complete remission when
from living cells or can be released from dying cells. Cell-free seminal mRNA compared to the preceding first-line protocol, ABVD (doxorubicin, bleomycin,
(cfs-mRNA) can be used as a biomarker in many non-invasive diagnostic appli- vinblastine, dacarbazine). These benefits over ABVD appear to come at the cost
cations as it contains tissue-specific and stable mRNA. In azoospermic males, it of increased germinal toxicity and impaired spermatogenesis in male patients.
is very important to know if there is an obstructive problem or not. However, The degree of impairment and the recovery of this fertility post-treatment
to the best of our knowledge there is no existing model that can accurately remains in question, and considering the youth of many Hodgkin’s lymphoma
predict successful sperm retrieval by MESA or TESE. Analysis of cfs-mRNA is a survivors, is clinically significant.
promising tool for such prediction. Study design, size, duration: A retrospective longitudinal study examining
Study design, size, duration: In this study we evaluated cfs-mRNA compo- semen and hormonal analysis of 28 male patients with advanced-stage Hodgkin’s
sition in samples of seminal plasmas from men with non-obstructive azoospermia lymphoma (Ann Arbor stage IIB, III or IV) treated with escalated BEACOPP.
(NOA) and obstructive azoospermia (OA). A total of 8 patients (4-NOA and Pre- (n=28) and post-treatment data (n=19) was evaluated. 19 patients engaged
4-OA) aged 28 to 40 years were included in this prospective study from spring in post-escalated BEACOPP analysis which occurred at no uniform time,
to autumn 2019. The seminal plasma samples were collected after centrifugation between one and six years post-treatment. The number of post-treatment anal-
of ejaculate. The cfs-mRNA content was determined by the microarray analyses. yses undertaken by each patient varied from one to four with a median of two.
Participants/materials, setting, methods: The non-obstructive azoosper- Participants/materials, setting, methods: Included patients were those
mia group included men who were cryptozoospermic in previous years and with histologically-confirmed advanced-stage Hodgkin’s lymphoma who pre-
azoospermic in this analysis, the obstructive azoospermia group included men sented to Concord Repatriation General Hospital’s (CRGH) Andrology
after vasectomy. RNA was isolated from a total of 8 samples. Two samples (with Department between January 2009 and June 2017 for semen cryopreservation
the highest concentration and quality) were selected for each group. Further and analysis prior to commencing escalated-BEACOPP therapy, without age
analyses were performed using GeneChip ™ Human Gene 2.1 ST Array Strip limits. As a retrospective study, data was obtained from medical records (stage,
microarrays. The statistical analysis was performed in the R programming lan- treatment) and semen cryopreservation data of the CRGH Andrology depart-
guage with the Bioconductor package. ment. Each analysis involved seminal fluid analysis, hormonal analysis (testoster-
Main results and the role of chance: Statistical analysis consisted of com- one, LH, FSH, SHBG) and testicular volume measurement.
paring cfs-mRNA content from men with non-obstructive azoospermia (NOA) Main results and the role of chance: In pre-treatment analysis, gonadal
and men with obstructive azoospermia (OA). In the next step, a total of 762 dysfunction was present in one in three patients with 68% of patients having
upregulated genes in NOA (fold above 2) were selected. Most of the upregulated normospermia and 32% having azoospermia or other dyspermia. In the identi-
genes belong to processes such as: cellular component organization or biogenesis fication of factors which may affect baseline sperm quality, only the presence of
(GO:0071840), cellular metabolic process (GO:0044237), cellular component B symptoms (weight loss ≥ 10%, night sweats or fevers ≥ 38˚C) was found to
organization (GO:0016043), organelle organization (GO:0006996). These pro- have significant effect on pre-treatment semen quality (P<0.05).
cesses belong to the GO BP (Gene Ontology Biological Process). The genes All 19 patients undergoing post-treatment follow-up were azoospermic at
that showed the largest change in expression (largest fold) are these: VTRNA1-1 first analysis after escalated BEACOPP. Serum LH (P<0.01) and FSH (P<0.001)
(Vault RNA1-1), MT-TS1 (Mitochondrially Encoded tRNA Serine 1), MT-TV were significantly increased, while testosterone and SHBG concentrations were
(Mitochondrially Encoded tRNA Valine), RNU6-441P (RNA, U6 Small Nuclear 441, not significantly changed.
Pseudogene), MT-TC (Transfer RNA Mitochondrial Cysteine), RNU1-83P (RNA, U1A Three patients (15.8%) recovered spermatogenesis with a mean recovery
Small Nuclear), PCGEM1(Prostate-specific gene), SEMG2 (Semenogelin 2), ND1 time of 44 months. Spermatogenesis recovery in these three patients
(NADH dehydrogenase, subunit 1), MT-TF (mitochondrially encoded tRNA phenylal- occurred at 36.5 months (Patient A), 44.6 months (Patient B) and 50.8 months
anine) and PIP (prolactin induced protein) which was previously considered as a (Patient C) post-treatment.
specific protein marker of non-obstructive azoospermia. To ascertain whether potential predictors of spermatogenesis recovery exist,
Limitations, reasons for caution: A limitation is the number of samples a binary logistic regression was performed on patient age (P=0.414), the pres-
included and analysed in this study which slightly reduced the power of statistical ence of B symptoms (P=0.715), pre-treatment total sperm count (P=0.126)
analysis. The obtained result will be validated on more samples using the quan- and the number of escalated-BEACOPP cycles received (P=0.261). None of
titative RT-qPCR method. these factors were found to contribute to the likelihood of spermatogenesis
Wider implications of the findings: These results suggest that the analysis recovery.
of cell-free seminal mRNA is a very promising tool and that the cfs-mRNA could Limitations, reasons for caution: Primary limitations are 1) the small sam-
be used as a non-invasive biomarker for identifying the complete obstruction in ple-size, which limited statistical power and subgroup analysis, and 2) the retro-
case of azoospermia. spective nature, culminating in significant loss to follow-up and no schedule of
Trial registration number: Supported by MH CZ – DRO (FNBr, 65269705) analysis. Records also contained limited data on other possible risk factors for
and Ministry of Health, Czech Republic projects no. NV18-08-00291 and impaired spermatogenesis including radiotherapy, pre-treatment ESR, and bulky/
NV-18-08-00412. mediastinal disease.
Wider implications of the findings: These results suggest that, while
impaired spermatogenesis is initially universal after escalated-BEACOPP, sper-
P-056 Spermatogenesis in advanced-stage Hodgkin’s lymphoma matogenesis recovery is evidenced in a proportion of patients in the years fol-
patients treated with escalated BEACOPP: a retrospective study lowing. This research will allow appropriate evidence-based risk-counselling of
of impaired fertility and recovery advanced-stage Hodgkin’s lymphoma patients considering escalated-BEACOPP
E. Williams1, V. Jayadev1 and facilitate more informed patient decision-making.
1
Concord Repatriation General Hospital, Andrology, Sydney, Australia Trial registration number: Not applicable
Study question: To determine the effect of escalated-BEACOPP on sper-

matogenesis in Hodgkin’s lymphoma patients and to investigate both the poten- P-057 Immunohistochemical Investigation of CatSper Ca2 +
tial for spermatogenesis recovery and factors predicting recovery. Channel Proteins in Normozoospermia and OAT patient groups in
Summary answer: Escalated-BEACOPP causes significant spermatogenesis Progesterone Applied Culture Media.
impairment in Hodgkin’s lymphoma patients, with spermatogenesis recovery N. Köprülü1, T. Irez2

1
Biruni University, Institute of Health Sciences- Department of Clinical Embryology, results were compared. A subset of 62 patients who underwent an IVF cycle at
Istanbul, Turkey ; T0 with unexplained <70% fertilization rate (sperm count >1x106/ml, woman
2
Istanbul Yeni Yuzyıl University, Medical Faculty- Clinical Embryology, age <38 years, inseminated oocytes >6) took the antioxidant supplements.
İstanbul, Turkey Outcomes of the paired T0 and T90 IVF cycles were compared.
Participants/materials, setting, methods: Patients’ diet was supplemented
Study question: To investigate effect of progesterone, which is a strong che- with myo-inositol (1000 mg), alpha-lipoic acid (800 mg), coenzyme Q10
moattractant candidate, on sperm tail motions that evaluate hyperactivation and (200 mg), selenium (83 mcg), zinc (15 mg), vitamin B2 (2.8 mg), B6 (2.8 mg),
CatSper expression in fertile and infertile cases. B12 (5 mcg). Sperm analysis was assessed according to WHO manual. Semen
Summary answer: Progesterone-treated sperms effected Catsper Channels OS was measured by MioxSIS® (Aytu BioScience). The IVF cycles were per-
and caused hyperactivation in infertile and fertile cases. formed according to conventional procedures using fresh gametes of each cou-
What is known already: Sperm chemotaxis causes intracellular calcium ple. Wilcoxon and chi-square tests compared data before and after the
increase and hyperactivation motility as a result of Ca2 + flow into the cell with antioxidant treatment.
the activation of odor receptors. CatSper channels are cation channels necessary Main results and the role of chance: Statistical analyses showed a significant
for fertility in rodents. Chemoattractants provide CatSper cation channel protein increase of sperm concentration (P=0.0307) and motility (P=0.0003) after
expression in the sperm principal peace region. Rabbit studies have shown that 90 days of treatment. After therapy with antioxidants it was observed a decreas-
progesterone produces CatSper expression. ing trend of OS levels in semen (normalized oxidation-reduction potential
Study design, size, duration: Sperm chemotaxis causes intracellular calcium (mV/106 ml sperm). T0: 8.9+8.6; T90: 6.1+7.7). As regard as the outcomes of
increase and hyperactivation motility as a result of Ca2 + flow into the cell with the IVF cycles, the fertilization rate was significantly higher in the T90 cycles with
the activation of odor receptors. CatSper channels are cation channels necessary respect to T0 ones (P<0.0001). Moreover, results demonstrated a significant
for fertility in rodents. Chemoattractants provide CatSper cation channel protein improvement in the embryo quality in the cycles performed at T90, in terms of
expression in the sperm principal peace region. Rabbit studies have shown that higher embryo top quality rate (P=0.0001) and blastulation rate (P<0.0001).
progesterone produces CatSper expression. Any T0 IVF cycle did not result in a successful pregnancy. Conversely, in the T90
Participants/materials, setting, methods: After standard semen analysis, cycles we obtained 11/62 clinical pregnancies (30%) with an implantation rate
sperm prepared in two groups by the swim-up method was kept at 37 ° C for of 19%.
45 minutes in culture medium containing progesterone. Motility and CatSper Limitations, reasons for caution: These data will be confirmed in a larger
IHC were evaluated before and after incubation. sample size to correct primary endpoints for putative confounders. We also
Main results and the role of chance: Progesterone was found to have foresee to deeply investigate the effect of the antioxidant treatment on semen
important chemoattraction ability for hyperactivation effect on the sperm tail OS status and ultimately on sperm DNA integrity. Lastly, a prospective study
movement on normospermic and oligoasthenoteratospermic sperm that integ- will determine true clinical value and limitations.
rity in luteal phase doses. Infertile group showed less progressive motility, less Wider implications of the findings: The outcomes of this pilot study suggest
hyperactivation and less CatSper expression than fertile Normospermia group. that treatment of sub-fertile men with combined myo-inositol and alpha-lipoic
(p<0,05). Progesterone administration caused higher rate of hyperactivation acid may have beneficial effect on sperm quality and pregnancy rates. Therefore,
and CatSper positivity in normospermic cases than OAT group (p <0.05). such a “nutraceutical” diet supplementation may become an additional tool to
Limitations, reasons for caution: We conducted our study in 56 cases. It reduce the semen OS, thus improving the success of IVF.
is beneficial to conduct the study in larger groups. Trial registration number: not applicable
Wider implications of the findings: The results of our study may be the
pioneer of studies investigating the relationship between CatSper expression P-059 Improvement in mitochondrial integrity and fertility
and sperm hyperactivation. A new research project is planned to examine IVF potential in men with Rheumatoid arthritis by yoga based lifestyle
results in terms of infertility and unexplained infertility in men. intervention
Trial registration number: 0 S. Gautam1, R. Kumar2, U. Kumar3, R. Dada1
1
All India Institute of Medical Sciences, Anatomy, New Delhi, India ;
P-058 Combined treatment with myo-inositol and alpha-lipoic 2
All India Institute of Medical Sciences, Urology, New Delhi, India ;
acid of sub-fertile men significantly improves semen competence 3
All India Institute of Medical Sciences, Rheumatology, New Delhi, India
in IVF cycles
F. Sozzi1, P. Canepa1, E. Maccarini1, C. Massarotti1, C. Rizzo1, D.L. Study question: Can a yoga based lifestyle intervention bring about alterations
Alessandro2, M. Gallo2, E. Licata2, P. Anserini1, R. Rago2, P. Scaruffi1 in mitochondrial integrity and fertility potential in men with Rheumatoid
1
IRCCS Ospedale Policlinico San Martino, UOS Fisiopatologia della Riproduzione arthritis?
Umana, Genova, Italy ; Summary answer: Yoga not only reduces disease severity, minimizes usage
2
Ospedale Sandro Pertini - ASL Roma 2, UOSD Fisiopatologia della Riproduzione e of drugs but also increases mitochondrial copy number and improves mitochon-
Andrologia, Roma, Italy drial integrity.
What is known already: The complex mechanism of Rheumatoid arthritis
Study question: Does combined treatment with myo-inositol and alpha-lipoic with infertility in men involves interactions between endocrine, immune,
acid of sub-fertile patients improve semen competence in IVF cycles? and reproductive systems. Association of autoimmunity with dysregulated
Summary answer: Sub-fertile men’s diet oral supplementation with combined androgen (hypogonadism) levels may cause transient infertility in men.
myo-inositol and alpha-lipoic acid improved sperm parameters and semen repro- Furthermore, usage of disease-modifying antirheumatic drugs (DMARDs)
ductive competence in IVF cycles. like cyclophosphamide, methotrexate, sulphasalazine etc. may result into
What is known already: Human infertility affects 15% of couples in child- decreased quantity and quality of sperm, mitochondrial dysfunctions,
bearing age, of which 40% is due to male factors. Alterations of sperm number, reduced fertility potential and ultimately permanent infertility. These drugs
motility and morphology are mostly associated with male infertility. Some 30% can cross blood-testis-barrier and induce changes in sperm impairing sper-
to 80% of sub-fertile men may be associated to oxidative stress (OS) that damage matogenesis. Complementary and alternative medicine like yoga reduces
spermatozoa. Furthermore, negative relations between OS and low success of seminal oxidative stress and its consequences like DNA fragmentation in
IVF techniques has been observed. Antioxidant oral supplements are widely sperm nuclear/mitochondrial genome.
used in sub-fertile patients. We previously reported that a combination of nutra- Study design, size, duration: Sixty males with RA were enrolled in this
ceuticals (myo-inositol, alpha-lipoic acid, folic acid, betaine, and vitamins) signifi- 12-week prospective, open-label, single-arm exploratory study, designed to
cantly improves sperm parameters in sub-fertile men. explore the impact of yoga based lifestyle intervention (YBLI) on mitochondrial
Study design, size, duration: This pilot, multi-centric study was performed integrity and fertility potential in men with Rheumatoid arthritis.
in 2018-2019 including 124 idiopathic, sub-fertile, 26-53 years aged men. Semen Participants/materials, setting, methods: The participants were evaluated
analysis was performed before (T0) and after 90 days (T90) of treatment and for pre (day 0) and post (12th week) levels of inflammatory markers: IL-6, IL-17A

and soluble HLA-G. Mitochondrial health was assessed by calculation of mito- Study question: The aim of the present study was to assess the impact of
chondrial copy number variation and transcripts associated with mitochondrial high risk HPV infection on semen parameters and sperm DNA fragmentation
integrity: AMPK, PRC-1, TFAM, SIRT-1. Sperm parameters as per WHO 2010 index in infertile men.
guidelines and reactive oxygen species (ROS), DNA fragmentation index (DFI), Summary answer: High risk HPV seminal infection is probably responsible
8-hydroxy-2’-deoxy guanosine (8-OHdG) were estimated. of lower sperm motility and higher DNA fragmentation index (DFI).
Main results and the role of chance: The mean levels of ROS (p<0.05***), What is known already: HPV infection is a common sexually transmitted
DFI (p<0.05**) and 8-OH2dG (p<0.05**) levels were significantly reduced after disease, related to genital warts and cancer. In males, human papilloma virus
12 weeks of yoga intervention. We observed reduction in mean levels of CRP DNA has been found in the seminal fluids. HPV is commonly present in sperm
(p<0.05***), IL-6 (p<0.05**), IL-17A (p<0.05**) and soluble HLA-G (p<0.05**) samples. However, whether the prevalence of high risk HPV in semen is asso-
at 12 weeks compared to baseline level (day 0). There was a significant increase ciated with modified semen parameters and sperm DNA fragmentation index
in mitochondrial copy number and increased expression of transcripts that main- has yet to be elucidated.
tain mitochondrial integrity after 12 weeks with respect to the baseline levels Study design, size, duration: From January to December 2018, about 505
(day 0). infertile couples were analysed at the Human Reproduction Unit of Ioannina
Limitations, reasons for caution: Compliance of patients for Yoga based University Hospital. A total of 335 clinical and laboratory data from male partners
lifestyle intervention was poor, hence we enrolled large number of patients to were included in the study. Semen parameters and sperm DNA fragmentation
achieve the desirable sample size. index were compared among those with or without HPV in spermatozoa.
Wider implications of the findings: Adoption of yoga based lifestyle inter- Participants/materials, setting, methods: A real time polymerase chain
vention as an integral part of our lifestyle may hold the key to increase mito- reaction assay was performed to detect the presence of HPV. All patients under-
chondrial copy number, increase the expression levels of transcripts that maintain went a semen analysis. The amount of DNA fragmented sperm, expressed in
mitochondrial integrity and its associated consequences on physical, mental and DFI, was valuated using the sperm chromatin dispersion test. Semen parameters
reproductive health. and sperm DNA fragmentation index were compared among high risk HPV
Trial registration number: N/A positive and HPV negative men.
Main results and the role of chance: 49 (14,6%) of the total semen samples
P-060 TDG involves in the key maintenance pathways of neonatal were HPV positive. Overall, 38/335 (11,3%) and 11/335 (3,3%) patients had
spermatogonial cells high risk HPV and low risk HPV, respectively. HPV31 was the prevalent type.
The amount of DNA fragmented sperm between high risk HPV positive and
T.L. Lee1
1
negative males was different. Sperm motility was lower (P=0.007) and SDF values
The Chinese University of Hong Kong, School of Biomedical Sciences, Sha Tin, were higher (P=0.003) in infected men compared to those without HPV.
Hong Kong Limitations, reasons for caution: Main limitation is the relatively small sam-
ple size.
Study question: The role of TDG in spermatogonial cell development during Wider implications of the findings: The results suggest that high risk HPV
neonatal stage infection probably impairs sperm DNA quality. These observations point out
Summary answer: TDG involves in key pathways associated with sper- the importance of testing seminal HPV presence in everyday clinical practice.
matogonial cell maintenance at neonatal stage. Trial registration number: None
What is known already: Active DNA demethylation through 5-formylcy-
tosine (5fC) and 5-carboxylcytosine (5caC) by TDG is essential for mouse
embryonic and stem cell development, but its role remains elusive in SSC biology. P-062 Permanent testicular metabolomic and lipidomic
Therefore, we examined the functional role of TDG in male germ cell develop- signatures induced by the adoption of a high-fat diet in childhood
ment using our recent established germline-specific TDG knockout animal. and sperm quality in adulthood
Study design, size, duration: 6-day old balb/c mouse with TDG KO (KO)
L. Crisostomo1, L. Rato2, I. Jarak1, R. Videira1, K. Starcevic3,
were compared against the wild type and hemizygous controls.
T. Masek3, J. Raposo4, O. Pedro5, M.G. Alves1
Participants/materials, setting, methods: 6-day old balb/c mouse strain 1
was used. Isolation of neonatal spermatogonial population was performed by Institute of Biomedical Sciences Abel Salazar- University of Porto, Laboratory of
OCT4 GFP signal. Cell Biology- Department of Microscopy, Porto, Portugal ;
2
Main results and the role of chance: Using KO TDG animal model, we University of Beira Interior, Faculty of Health Sciences, Covilhã, Portugal ;
3
found that TDG knockout males expressed a progressive loss of fertility. University of Zagreb, Veterinary Medicine, Zagreb, Croatia ;
4
Transcriptome analysis by RNA-seq revealed altered gene expression of a subset Nova University Lisbon, Public Health Department, Lisbon, Portugal ;
5
of retinoic acid target genes, including Stra8 and Cyp26a1. Furthermore, we QOPNA & LAQV- University of Aveiro, Chemistry Department, Aveiro, Portugal
found that the demethylation of Stra8 during the KIT transition was impaired
Study question: Can a diet intervention in young mice fed high fat diet improve
after depletion of TDG in Oct4-GFP+ undifferentiated spermatogonia. These
sperm quality in adulthood?
results indicate that TDGmediated active demethylation regulates the differen-
Summary answer: The adoption of high-fat diets from childhood to early
tiation of undifferentiated spermatogonia through the coupling of the retinoic
adulthood causes irreversible metabolic and lipidomic changes in testis, reflected
acid signaling pathway.
in poorer sperm quality.
Limitations, reasons for caution: The role of TDG in adult SSC stage was
What is known already: The prevalence of overweight and obesity has
not studied.
soared worldwide in recent decades, fueled by poor dietary habits as the adop-
Wider implications of the findings: A key question in SSC biology is
tion of high-fat diets (HFD). Besides, the adoption of HFD often occurs in
whether and how epigenetic programs contribute to the cellular identity of SSC
childhood, triggering the onset for these conditions and associated comorbidities,
and specific gene expression programs. The study provides details about the
such as diabetes, at increasingly younger age. Thus, men can spend most of their
molecular regulation of TDG in neonatal spermatogonial cell development and
sexual development suffering from a metabolic disease, with unknown conse-
will allow the development of future male infertility treatment.
quences for future sperm quality. A change in diet habits may prevent the pro-
gression of the disease, but it is not known whether it can rescue potential
negative effects of HFD to male fertility.
P-061 The presence of high risk human papilloma viruses in Study design, size, duration: Hereby, we used a rodent model to study diet
semen affects seminal parameters and sperm DNA quality in intervention during early adulthood, after childhood obesity, to correct HFD
infertile men and how it affects the metabolic and reproductive health later in life. For that,
E. Mastora1, A. Zikopoulos1, E. Hatzi1, I. Mpoumpa1, I. Georgiou1, after weaning (day 21 after birth), 3 groups of 12 mice each were fed with dif-
K. Zikopoulos1 ferent diet regimen during 200 days: CTRL – standard mucedola; HFD – high-fat
1
University of Ioannina, Department of Medicine, Ioannina, Greece diet; HFDt – high-fat diet for 60 days, then replaced by standard mucedola.

Participants/materials, setting, methods: Animals were then sacrificed, miR-20a-5p might represent a new source of non-invasive diagnostic biomarkers
and tissues collected and weighed. Epidydimal sperm parameters (sperm counts, for idiopathic male infertility.
motility, viability and morphology) were evaluated. Animals biometric, metabolic Study design, size, duration: From January 2018 to December 2019, 24
(fasting glucose, ipGTT, ipITT, HOMA2) and endocrine (serum insulin, testos- infertile couples who referred to our ARTs Centre were prospectively enrolled.
terone, estrogen, gonadotrophins) parameters were studied. Testicular metab- All the patients were included into two groups: Group 1 comprised infertile men
olites were extracted and characterized by 1H-NMR and LC-MS. Testicular with NOA, Group 2 involved healthy normozoospermic men belonging to cou-
mitochondrial (citrate synthase, complex I, II and IV) and antioxidant activity ples with female infertility tubal factor who achieved pregnancy using IVF or ICSI.
(CAT, SOD, GSR, GPx) were also evaluated by colorimetric and fluorometric Participants/materials, setting, methods: The expression of circulating
methods. miR-20a-5p was assessed by RT qPCR in plasma samples. A relative quantification
Main results and the role of chance: Our results showed an induction of strategy was adopted using the 2-ΔCqmethod to calculate the target miR-20a-5p
a pre-diabetic state due to consumption of a HFD. Based on HOMA2, diet expression with respect to miR-16-5p as an endogenous control. Total cell-free
intervention in early adulthood prevents the development of metabolic syn- RNA extracted from 0.5 ml plasma using the mirVana PARIS kit (ThermoFisher
drome in adulthood, although insulin resistance is maintained. No changes were Scientific, USA) was submitted to RT-qPCR using TaqMan Advanced miRNA
observed in endocrine function. Importantly, sperm parameters were not recov- cDNA Synthesis Kit and TaqMan® Advanced miRNA Assays (ThermoFisher
ered after diet intervention. Spermatozoa motility and viability were still affected Scientific) according to the manufacturer’s instructions.
in mice fed HFD during childhood, even after a dietary intervention, particularly Main results and the role of chance: Group 1 included 14 patients, Group
head defects. HFD, even if transient, promoted changes in testicular metabolome 2 comprised 10 men. No genetic abnormalities and significant comorbidities
and lipidome. Notably, it affected the glutamate, pyruvate, ethanol and ammonia were detected in all the patients enrolled. Mean male age was 35.6 ± 4.2 years.
metabolism, and promoted the storage of unsaturated fats. Interestingly, no Considering the Group 1, mean FSH value was 19.4 ± 7.8 IU/l, mean LH value
differences were found in mitochondrial and antioxidant enzyme activities. The was 8.5 ± 3.4 IU/l, mean TT was 12.5 ± 3.9 nmol/l, mean TSH was 2.0 ± 1.1
progression of metabolic syndrome due to HFD was found to be correlated mIU/l and mean PRL was 10.5 ± 3.2 ng/ml. Mean right testicular volume was
with the changes in testicular metabolism. In turn, testicular metabolic remodeling 8.9 ± 5.2 ml, mean left testicular volume was 8.2 ± 4.5 ml. All NOA patients
was strongly correlated with sperm parameters. Therefore, a causal relation underwent testicular sperm extraction (TESE). Successful sperm retrieval (SR)
between the adoption of HFD in childhood and poorer sperm quality in adult- with cryopreservation was found in 8/14 patients (overall SR rate: 57.1%). Mean
hood is not just a stochastic event. sperm concentration was 0.001 ± 0.0001 x 106/ ml. Mean non progressive
Limitations, reasons for caution: Despite using this mouse model, we are motility was 0.2 ± 0.6 %. Mean number of biosystem straws collected was 3.2
unable to state whether normal sperm quality could be achieved even later in ± 2.0. All healthy normozoospermic men showed serum hormonal levels and
life, or if the adoption of HFD in adulthood triggers the same deleterious effects testicular volume in the normal range. Mean miR-20a-5p value was 0.25 ± 0.20
in sperm parameters. and 0.06 ± 0.02 in the Group 1 and Group 2, respectively. Thus, the relative
Wider implications of the findings: Diet intervention reverses some of the expression of miR-20a-5p was significantly higher in patients affected by NOA
detected changes but cannot overcome the long-term consequences of HFD than in healthy normozoospermic control subjects (p=0,026).
to sperm quality. Our conclusions must raise awareness about tackling childhood Limitations, reasons for caution: Further larger studies are needed to
obesity, to avoid irreversible damage for the reproductive health of the future evaluate the correlation between this marker and testicular histopathological
fathers, with unpredicted effects to their progeny. findings in order to increase its usefulness as a predictor in the clinical practice.
Trial registration number: The authors declare no conflict of interest. this Wider implications of the findings: Blood plasma miR-20a-5p could rep-
work was supported by the Portuguese Foundation for Science and Technology: resent a potential non-invasive diagnostic biomarker in infertile patients with
L. Crisóstomo (SFRH/BD/128584/2017), M.G. Alves (IFCT2015 and PTDC/ non-obstructive azoospermia who undergo TESE. A possible correlation of this
MEC-AND/28691/2017), QOPNA (UID/QUI/00062/2019) and UMIB (UID/ marker with testicular histopathological findings could allow the clinician to cor-
Multi/00215/2019) co-funded by FEDER funds (POCI/COMPETE 2020); by rectly counsel the azoospermic patients in performing surgery for fertility
the Portuguese Society of Diabetology: L. Crisóstomo and M.G. Alves (“Nuno purpose.
Castel-Branco” research grant); and by the Amadeu Dias Foundation: M.G. Alves Trial registration number: not applicable
and P.F. Oliveira.
P-064 A new modified method: Aniline Blue (AB) stain in live

P-063 Blood plasma miR-20a-5p expression as a potential non- spermatozoa reveals that all motile sperms not contain histones.
invasive diagnostic biomarker in patients with non-obstructive (Preliminary study. EcoFoodFertility Project).
azoospermia “Abstract withdrawn by the authors”
G. Cito1, P. Pinzani2, R. Fucci3, R. Picone3, F. Salvianti2, C. Giachini3,
P. Falcone3, E. Micelli3, A. Cocci1, P. Verrienti1, A. Minervini1, P-065 Novel PMFBP1, TSGA10 and SUN5 variants in acephalic
M. Carini1, A. Natali1, M.E. Coccia3 spermatozoa syndrome and the overcoming this male infertility
1
Careggi Hospital- University of Florence, Dept. of Urology, Florence, Italy ; L. Weina1, W. Ni2, Z. Huan3, Y. Silu1, L. Ge3, L. Guangxiu3, L. Gang1
2
Careggi Hospital- University of Florence, Molecular and Clinical Biochemistry Unit, 1
Reproductive and Genetic Hospital of CITIC-Xiangya, Andrology Laboratory,
Florence, Italy ; Changsha, China ;
3
Careggi Hospital- University of Florence, ARTs Center, Florence, Italy 2
Central South University, Institute of Reproduction and Stem Cell Engineering,
Changsha, China ;
Study question: To evaluate the blood plasma miR-20a-5p expression in infer- 3
Reproductive and Genetic Hospital of CITIC-Xiangya, Reproductive center,
tile patients with non-obstructive azoospermia (NOA) compared to healthy Changsha, China
normozoospermic men.
Summary answer: The overexpression of blood plasma miR-20a-5p seems Study question: What are the genetic diagnosis and effective therapeutic
to be directly related to damaged spermatogenesis, including conditions of NOA. strategies to overcome infertility in acephalic spermatozoa syndrome?
What is known already: Recently, alterations in the expression of specific Summary answer: We identified four novel mutations in PMFBP1, TSGA10,
microRNAs (miRNAs) in semen have been linked to altered spermatogenesis and SUN5 in three out of eleven patients. These male infertility could be over-
suggesting their expression could be used as potential infertility biomarkers and come by ICSI.
explain the molecular mechanisms underlying the altered spermatogenesis. In What is known already: Acephalic spermatozoa are a rare teratozoospermia
previous animal studies dysregulation of miR-20a-5p was found to be under-ex- which leads to male infertility. The vast majority of acephalic spermatozoa cases
pressed in the low motile sperm and potentially target genes associated with cell are idiopathic, and 52.63% (51.22%, 21/41 SUN51-5; 50%, 7/14 PMFBP16 7;
apoptosis and spermatogenesis alteration. Considering previous findings with 100%, 1/1 TSGA108; 100%, 1/1 BRDT9) have been associated with definitive
miRNAs in human plasma, serum and urine, we believe that the expression of genetic etiology. Considering that negative/negative results are rarely published,

the previous study population mainly covers Hefei and Xiamen, and the contri- Study design, size, duration: Founder (F0) male Sprague Dawley rats
bution of these genes to acephalic spermatozoa might not be as high as described (12 per group, 48 total) were weaned (Day21) onto control (C) or HFD (H),
previously. or micronutrient supplemented versions of these (CS; HS). At 19 weeks of age,
Study design, size, duration: This genetic study used whole-exome and they were mated with CD females. 48 F1 offspring (one from each F0 pairing)
Sanger sequencing to discover and identify related-genes mutations, and was were weaned onto chow diet, generating four F1 groups. At cull (30 weeks for
followed by a series of studies to certificate potentially damaging. F0, 23 weeks for F1), right testis was harvested and stored at -80º.
Participants/materials, setting, methods: 11 patients with acephalic sper- Participants/materials, setting, methods: Testis was assayed for the level
matozoa were recruited from our hospital in China in the first half of year 2019. of lipid peroxidation (MDA), glutathione and the activity of superoxide dismutase
Genomic DNA samples from the individuals were extracted from peripheral (SOD), catalase (CAT), glutathione peroxidase (GPx) in F0 founders and MDA,
blood. All samples were subjected to whole-exome sequencing and variant fil- SOD, CAT in F1 offspring according to kit protocols. Susceptibility index of
tering, using Sanger sequencing to identify mutations. Identified mutations were prooxidative damage was calculated in F0 testis from the ratio SOD:CAT+GPx.
further investigated using bioinformatics, papanicolaou staining, CASA, trans- Statistical analyses were performed using IBM SPSS v23.0 software. Data were
mission electron microscopy(TEM), scanning electron microscopy (SEM) and presented as mean±SEM and analysed by two-way ANOVA followed by post
RT-PCR.We evaluated their available clinical data and pregnancy outcomes. hoc LSD.
Main results and the role of chance: We identified one compound het- Main results and the role of chance: In founders, HFD significantly increased
erozygous variant (c.361C>T[p.Gln121Ter],c.2089-1G>T) in PMFBP1, one body weight (H: 765.1±16.5 vs C: 632.5±15.1 g; p= 0.0001) which was nor-
homozygous mutation (c.1739A>C[p.Gln580Pro]) in TSGA10, and one homo- malized by supplementation (HS: 601.9±11.8 g vs H; p=0.0001). Supplemented
zygous variant (c.772C>T[p.Arg258Cys]) in SUN5 in three unrelated infertile HFD fed fathers had reduced testicular lipid peroxidation (HS: 7.5±2.1 vs H:
men.The remaining were new gene results that should be further validated by 12.8±1.7 nmol/g testis; p=0.01). No significant effect of HFD was observed in
animal models. RT-PCR in blood revealed that mutation causes loss of the wild- testicular lipid peroxidation. HFD fed fathers had reduced testicular enzymatic
type splice site, which leads to aberrant splicing of PMFBP1 mRNA and consec- antioxidant activity (SOD & GPx: H vs C; p=0.002) while supplementation
utive formation of the PMFBP1 alternative transcript (p.Ile697Leufs*257). All of restored these activities (HS vs H; p=0.002). Moreover, supplementation
these mutations cause protein truncations or lead to point mutations and have increased a major cellular antioxidant, glutathione (HS: 4179.7±197.1 vs H:
a recessive inheritance pattern. All patients with acephalic spermatozoa had 3421.2±194.8 nmol/g testis; p=0.004). No significant effect of HFD was
different phenotypes and proportions caused by their different ultrastructure. observed on testicular glutathione levels. Susceptibility index of prooxidative
Patients with p.Arg258Cys in SUN5 showed 70% acephalic spermatozoa and damage was significantly increased by chronic HFD intake (H: 1.7±0.2 vs C:
25% microcephalic spermatozoa but no so-called round-headed like spermato- 1.1±0.1; p=0.002) and normalized by supplementation (HS: 1.2±0.1 vs H;
zoon,which suggest that a variant in SUN5 does not result in microcephalic p=0.004). No effect of HFD and supplementation was observed in F0 testicular
spermatozoa in patient 3. There are other pathogenic genes for his multiple catalase activity. In F1 testis, offspring from HFD fed fathers had increased tes-
malformations in spermatozoa, which need to be further explored. PMFBP1, ticular SOD activity (H: 67.2±6.5 vs C: 49.6±6.1 U/mg protein; p=0.04)
TSGA10 and SUN5 may associate with the centrosome and basal body and whereas, offspring from HS fathers had decreased SOD activity (HS vs H;
participate in completing the coupling apparatus, ultimately resulting in acephalic p=0.02). No effects of paternal diet and supplementation were observed in
spermatozoa but not delayed or stagnated embryonic development. PMFBP1, offspring testicular lipid peroxidation level and catalase activity.
TSGA10 and SUN5 mutation-associated male infertility could be successfully Limitations, reasons for caution: F0 testis was harvested at 30 weeks rather
overcome by ICSI in humans. than at mating (19 weeks). As we have no measurement of sperm DNA damage
Limitations, reasons for caution: Our pilot study was intended as proof in F0 and F1 males, it is difficult to comment on underlying mechanisms of
of principle, and the number of patients is low. Although these patient defects transgenerational effects of paternal obesity at this stage.
could be resolved by microinjection of the tailless sperm head into the oocyte, Wider implications of the findings: HFD induced paternal obesity may
further studies are needed to validate the safety of offsprings. affect offspring testicular antioxidant capacity which might relate to metabolic
Wider implications of the findings: We summarized known data on the and reproductive complications in future generations. Our micronutrient sup-
genetic contribution of genes to acephalic spermatozoa, expanded the mutation plementation might be a novel intervention to restore antioxidant capacity in
spectrum, and analyzed the variety of phenotypes for each type of mutation. obese father and their offspring. Supplementation of non-obese individual may
This work lays the foundation for the genetic diagnosis, genetic counseling and have deleterious effects in next generation.
treatment of acephalic spermatozoa syndrome in China,for reproductive Trial registration number: Not applicable
options.
Trial registration number: Not applicable. P-067 Seminal oxidative stress and sperm DNA fragmentation
in men from couples with idiopathic recurrent pregnancy loss or
P-066 Effect of high fat diet induced paternal obesity and infertility
micronutrient intervention on offspring testicular antioxidant J.M. Kold1, M.R. Dalgaard1, F. Dardmeh2, H. Alipour2,
capacity O.B. Christiansen1
M.M. Billah1, S. Khatiwada1, V. Lecomte1, M.J. Morris1, 1
Aalborg University Hospital, Department of Obstetrics and Gynaecology, Aalborg,
C.A. Maloney1 Denmark ;
2
1
UNSW Sydney, School of Medical Sciences, Sydney, Australia Aalborg University, Department of Health Science and Technology, Aalborg,
Denmark
Study question: Does paternal obesity perturb testicular antioxidant capacity
in father and offspring? If so, can we mitigate this transgenerational effect by Study question: Are seminal oxidative stress (OS) and sperm DNA fragmen-
micronutrient intervention in father? tation (SDF) correlated, and can they explain idiopathic infertility or recurrent
Summary answer: Obesity-related reductions in testicular SOD and gluta- pregnancy loss (RPL)?
thione peroxidase activity in fathers and increased SOD activity in offspring were Summary answer: OS and SDF levels are not different between case groups
ameliorated by micronutrient intervention in obese fathers. and fertile controls and OS correlates to SDF in infertile but not in RPL cases.
What is known already: Obesity has tripled in males of reproductive age What is known already: Around 15% of all couples experience infertility
since 1970. As well as affecting individuals, obese people can produce offspring while 1-2% experience RPL. Approximately 25% of infertility and 40% of RPL
that are predisposed to metabolic and reproductive complications. One possible cases are considered idiopathic. Studies have reported that seminal OS and SDF
mechanism is oxidative stress which increases sperm DNA damage, alters sperm appear frequently in men from couples with infertility or RPL. Furthermore, the
epigenetic profile thus affecting future generations. It is known that obesity is use of sperm with a high level of SDF has been associated with poor artificial
associated with higher levels of oxidative stress. We sought to reduce this effect reproductive technology outcome. However, there is no consensus on the
by providing a micronutrient supplement (consisting of metabolites of one- impact of and whether to test for seminal OS or SDF in cases of idiopathic RPL
carbon metabolism) targeting oxidative stress to prospective rat fathers. or infertility.

Study design, size, duration: This clinical case-control study aimed to include formula 2 (low concentration of ATM/ATR-enhancer, n = 25). Semen param-
30 men in each of the two case groups and 30 fertile controls. The data collection eters and sperm DNA fragmentation were determined before and after 13
and assessments were according to the protocol planned between June 2019 weeks of treatments.
and July 2020 at a tertiary university centre for infertility and RPL treatment. Participants/materials, setting, methods: All patients included presented
Participants/materials, setting, methods: Semen samples from male part- high values of DSB. Patients receiving medication or other fertility treatments
ners of couples with idiopathic infertility (n=23), idiopathic RPL without con- before the start of the study were excluded. DSB and SSB were analysed through
comitant infertility (n=20), and fertile men (n=29) were assessed for SDF using the Neutral and Alkaline CometFertility assay (CIMAB, Spain), respectively.
sperm chromatin dispersion test, concentration, motility and morphology by the Semen parameters including concentration, total sperm count and progressive
Sperm Class Analyzer (Microptic S.L., Spain) computer aided sperm analysis motility were determined according to the last World Health Organization
(CASA) system. Seminal OS was measured as static oxidation-reduction potential guideline.
(sORP) using Male Infertility Oxidative System (MiOXSYS, Aytu BioScience Inc, Main results and the role of chance: DSB values were significantly reduced
USA). sORP were normalised to semen concentration. by all three treatments (p=0.042). The greatest reduction was observed in
Main results and the role of chance: The infertile and RPL groups, showed Formula 1, which reduced 27.1%. Formula 2 and the Control group reduced
no significant difference in the levels of OS (p=0.59 and p=0.67, respectively) 19.9% and 15.8%, respectively. SSB was reduced by all three treatments. Formula
or SDF (p=0.85 and p=0.11 respectively) when compared to fertile controls. 1 significantly reduced SSB 19.5% (p=0.040). Even not statistically significant,
There was a significant correlation between OS and SDF in the infertile group Formula 2 and the Control group reduced SSB 6.8% and 8.6%, respectively.
(R=0.404, p=0.028) but not in RPL group (R=0.225, p=0.18). Additionally, in Regarding seminal parameters, Formulas 1 and 2 increased sperm concentration
the infertile group 84.6 % of men with a high level of OS (normed sORP 2.11% (p=0.046) and 53.55% (p=0.200) respectively. The Control treatment
>1.38mV/106 sperm mL) had a high level of SDF (> 15 %), whereas 60 % with reduced 20.14% this parameter (p=0.160). Total sperm count was improved by
low OS had a low level of SDF. The infertile men were from couples diagnosed experimental treatments with an increase of 17.41% (p=0.135) and 113.13%
with idiopathic infertility based on traditional (manual) semen analysis, but nev- (p=0.012) for Formulas 1 and 2, respectively. The Control group reduced 23.64%
ertheless analysed by CASA 78.3% had a motility below the World Health this parameter (p=0.001). Formulas 1 and 2 showed a moderate improvement
Organisation (WHO) reference values. in progressive sperm motility (9.10% and 1.08%, respectively) despite not being
Limitations, reasons for caution: Several of the men diagnosed with idio- statistically significant (p=0.072 and 0.067, respectively). Not significantly, the
pathic infertility had a motility below the WHO reference values. Thus, this study Control treatment reduced 14.14% the progressive motility (p=0.200).
had not only examined men from couples with idiopathic infertility as intended None of the experimental ATM/ATR-enhancement-based treatments showed
but also men who may have occult male factor infertility. any adverse effects on sperm genome integrity or other semen parameters.
Wider implications of the findings: OS determined as normed sORP may Formula 1 proved to be the most effective treatment to reduce sperm DNA
be useful to evaluate the risk of SDF in men with infertility. Several men diagnosed fragmentation. On the other hand, Formula 2 proved to be the best treatment
with idiopathic infertility had low motility when samples were assessed using improving seminal parameters.
CASA, indicating that these men had been misdiagnosed with idiopathic infertility Limitations, reasons for caution: Patients compliance with treatment cannot
instead of male factor infertility. be guaranteed. For this reason, patients were excluded from the study when
Trial registration number: Ethical committee approval no. N-20190023 there was a suspicion that the intake of the treatment did not follow the protocol
instructions.
P-068 A new oral treatment based on the ATM/ATR DNA repair Wider implications of the findings: DSB are a major cause of chromosomal
pathways enhancement reduce sperm DNA damage and improve abnormalities in a zygote, cause a delay in embryo development and impair
semen parameters embryo implantation. Prevention of sperm DNA fragmentation through effective
J. Franco1, S. Lara-Cerrillo2, Á. Vives-Suñé3, M. Ballester-Ferrer3, oral treatments focused on improving DNA damage repair and/or activating
T. Lacruz-Ruiz2, C. Rosado-Iglesias2, M. Velasco-Álvarez4, apoptosis systems would be of great interest to patients affected by DSB.
E. Rodríguez-Ferradas4, A. Jiménez-Macedo5, J. Carrera-Rotllan5, Trial registration number: Does not apply.
A. García-Peiró2
1
Hospital Ruber Internacional, Human Reproduction, Madrid, Spain ; P-069 Association of serum metabolomic profile by nuclear
2 magnetic resonance spectroscopy with semen parameters.
CIMAB- Barcelona Male Infertility Centre, Research & Development, Sant Quirze
del Vallés - Barcelona, Spain ; A. Taylor1, K. Al Rashid2, M.A. Lumsden2, N. Goulding1, D. Lawlor1,
3
Hostpital Universitari Dexeus, Reproducció Assistida, Barcelona, Spain ; S. Nelson2
4 1
Instituto Vasco de Fertilidad - Grupo IVF Life, Reproducción asistida, San UNiversity of Bristol, Bristol Medical School, Bristol, United Kingdom ;
2
Sebastián, Spain ; Universiy of Glasgow, School of MEdicine, Glasgow, United Kingdom
5
Unitat d’Endocrinologia Ginecològica, Reproducció assistida, Vic, Spain
Study question: To assess the association of 155 circulating metabolic mea-
Study question: Can double-strand sperm DNA breaks and semen parameters sures relevant to lifestyle and metabolic health with sperm parameters (as mea-
be improved by oral administration of antioxidants and ATM/ATR-enhancers? sured by concentration, motility and total motile sperm count (TMSC)).
Summary answer: A new treatment based on ATM/ATR-repair-pathways Summary answer: An extensive range of metabolites were not associated
enhancement significantly reduced double-strand sperm DNA breaks and with semen parameters, highlighting the potential limited contribution of lifestyle
improved semen parameters. modification to improve sperm count or motility.
What is known already: Double-strand breaks (DSB) take place during male What is known already: Several observational studies and small randomised
meiosis to enable DNA recombination and packaging. These breaks must be controlled trials (RCTs) have investigated the association between metabolic
repaired later to ensure sperm genome integrity. However, defective chromatin health, nutritional supplements and lifestyle modification on sperm parameters
condensation, incomplete apoptosis and oxidative stress can increase both DSB with some, but not all studies, suggesting that improved metabolic health may
and single-strand breaks (SSB). Several studies demonstrated that DSB are a be associated with improved semen parameters. However, these studies have
major source of chromosome abnormalities causing miscarriage. The ATM/ been limited to a range of metabolic measures with limited adjustment for con-
ATR kinases play an essential role in genomic maintenance through the repair founders, and recent large well-conducted RCTs of supplementation have not
of DSB and the activation of cell-cycle checkpoints. Moreover, seminal param- observed an improvement in sperm parameters.
eters are improved by oral treatments including antioxidants. The enhancement Study design, size, duration: Cross-sectional study of 325 men prospectively
of ATM/ATR-repair-pathways could be a new strategy to improve sperm quality. recruited between 1 April 2017 and 31 March 2019.
Study design, size, duration: This prospective and multicentric study Participants/materials, setting, methods: Men intending to undergo
included 71 infertile patients that were randomly divided into three groups, assisted conception at a University Hospital, had a detailed demographic, life-
receiving a different treatment: Control group (Androferti, n = 21); Experimental style, fertility and medical history and semen analysis. Non-fasting serum lipids,
formula 1 (high concentration of ATM/ATR-enhancer, n = 25); Experimental lipoprotein subclasses, and low-molecular weight metabolites (including amino

acids, glycolysis and inflammatory markers) were quantified by NMR spectros- Limitations, reasons for caution: Sample size is still low even after collecting
copy. Multivariable linear and logistic regression was used to examine the data for 2 years and more patients are required to better address this issue.
associations of serum metabolic profiles (exposures), with functional sperm Wider implications of the findings: Sperm DNA fragmentation may be a
concentration, motility and TMSC (outcomes) with adjustment for potential predictor of reproductive outcomes of IUI.
confounders. Trial registration number: Not applicable
Main results and the role of chance: Participants were mean 37.2 (SD 5.7)
years and had a median sperm concentration of 35 million/ml (IQR 15, 69
million/ml) and median motility of 53% (IQR 42,67). 76% of men had a TMSC P-071 The impact of physical activity on semen analysis
>15 Million, 10% 5-15 Million and 14% <5 Million. In both univariate and adjusted parameters in healthy males undergoing fertility investigation
analyses an extensive range of lipids and lipoproteins, acetate, beta-hydroxybu- N. Vlahos1, A. Philippou2, E. Maziotis1, E. Alexopoulou1,
tyrate, creatinine, albumin, glucose and the amino acids; alanine, glutamine, P. Giannelou2, A. Rapani1, S. Grigoriadis1, A. Gryparis1, P. Bakas1,
glycine, histidine, isoleucine, leucine, valine, phenylalanine and tyrosine did not C. Siristatidis1, D. Tzanakaki1, M. Koutsilieris2, M. Simopoulou1
show strong statistical evidence of associated with sperm concentration, motility, 1
Aretaieio Hospital- National and Kapodistrian University of Athens, Second
or the odds of having a reduced or low TMSC (all _PBonferroni > 0.0029). Department of Obstetrics and Gynecology, Athens, Greece ;
Limitations, reasons for caution: Residual confounding may have resulted 2
National and Kapodistrian University of Athens- Medical School, Physiology,
from crude questionnaire measurements, though we cannot think of masking Athens, Greece
confounders. Sperm parameters were measured on a single sample, in accor-
dance with recent WHO guidance, and by non-automated techniques which Study question: Do levels of physical activity (PA) and metabolic equivalent
may have introduced random measurement error that could have attenuated (METs) correlate to parameters of semen analysis in healthy males?
to the null. Summary answer: Men reporting with moderate levels of PA present with
Wider implications of the findings: We did not find any robust associations higher sperm concentration, progressive and total motility, compared to men
between a range of established and novel metabolic measures with semen presenting with low levels of PA.
parameters. Replication of our observed null results is critical given current What is known already: The potential association between semen analysis
interest in improving semen parameters through lifestyle modification. parameters and environmental as well as lifestyle factors-including physical activ-
Trial registration number: Not applicable ity-has been extensively investigated in literature. Recent studies demonstrate
that moderate physical activity may improve semen analysis parameters.
P-070 The effect of sperm DNA fragmentation index on the However, discordant conclusions are reported on this topic. Furthermore, exist-
outcome of intrauterine insemination ing evidence is of poor quality as evident in recent systematic reviews and
meta-analyses. This underlines the requirement for conducting large well-de-
D. Gunasheela1, R. Nayak1, B.S. Indrani1, L. Sasikala1, P. Ajitha1,
signed studies. The aim of this study was to investigate the possible effect of
W.Y. Son2
1
physical activity on semen parameters, evaluating the metabolic equivalent, in
Gunasheela Surgical & Maternity Hospital, Obst & Gyne, Bangalore, India ; healthy males undergoing fertility investigation.
2
McGill University Health Centre MUHC- Royal Victoria Hospital- Canada, Study design, size, duration: A total of 223 men were recruited for this
Embryology, Montreal, Canada present prospective observational single-center study from November 2015 to
October 2017. Participants were 18-40 years old, with a Body Mass Index (BMI)
Study question: Can sperm DNA fragmentation affect reproductive outcomes of 18.5-29.9 kg/m2. Men subjected to medication administration as part of a
of intrauterine insemination? therapeutic regime, with current or previous diagnosis of cancer, endocrinolog-
Summary answer: High sperm DNA fragmentation (DFI: > 30%) is associated ical or genetic disorders, with a history of diseases that may affect the repro-
with poor reproductive outcomes in couples undergoing IUI. ductive system and azoospermic men were excluded from the study.
What is known already: The pregnancy outcome after IUI procedures was Participants/materials, setting, methods: The participants were inter-
unpredictable because several possible factors were involved in the process. A viewed according to the Global Physical Activity Questionnaire (GPAQ). They
number of studies concerning the association between DFI and pregnancy out- were divided into 3 groups -namely, low, moderate and high PA- according to
come after IUI have been performed. Some studies showed that high DFI was their physical activity levels as suggested by the World Health Organization
positively correlated with low pregnancy rate after IUI. However, several other (WHO). Semen analysis was performed according to WHO 2010 guidelines.
reports have found that DFI is not correlated with pregnancy outcome in IUI. Main results and the role of chance: Forty men were categorized in the
The predictive value of DFI in reproductive outcome of IUI remains Low PA group, 117 in the moderate PA group and 66 in the high PA group. No
controversial. statistically significant difference between the three groups was observed regard-
Study design, size, duration: This prospective observational study included ing the age, BMI, smoking status and the number of days of abstinence. No
413 women who underwent IUI cycle from October 2017 to October 2019. correlation was observed between the levels of METs and sperm concentration,
The male partners of the infertile couple underwent sperm-DFI by TUNEL or total motility or the percentage of normal morphologically spermatozoa. The
SCD assay. Depending on DFI, they were divided into three groups; Group 1 level of METs were negatively correlated with progressive motility (p=0.01).
(DFI-negative:<15%), Group 2 (DFI-fair positive:15-30%) and Group 3 (DFI-high The Kruskal-Wallis test revealed differences between the groups regarding pro-
positive: > 30%). The clinical & ongoing/live birth rates were compared among gressive (p=0.001) and total motility (p=0.04). In the post-hoc analysis, the
the three groups. P < 0.05 was considered statistically significant moderate PA group presented with higher total motility (43.08±18.34 vs
Participants/materials, setting, methods: This study was conducted in 36.43±16.01; p=0.01) and progressive motility (31.86±17.69 vs 25.75±16.50;
infertile couples attending our ART center, who planned for IUI and tested DFI p=0.04) when compared to the low PA group. The three groups-low, moderate
((n= 413), with also signed the consent form. The exclusion criteria were men and high PA-did not differ statistically significantly regarding neither sperm con-
not consenting to undergo the DFI test, those with oligozoospermia (< 15 centration (26.04±21.04 vs 55.53±37.16 vs 47.86±32.83; p=0.25), nor normal
million/ml) and cases of donor semen. Ovulation induction in IUI cycles was morphologically spermatozoa (12.35±6.69 vs 10.64±7.65 vs 10.82±7.13;
performed by clomiphene citrate 50–100 mg or Letrozole 5 mg with or without p=0.80). The moderate PA group presented less risk for abnormal semen anal-
human menopausal gonadotropin (hMG), or hMG alone. ysis compared to the low PA group (RR:0.65;95%CI:0.52-0.81;p=0.01). The
Main results and the role of chance: Out of 413 men who had tested DFI, high PA group presented with similar risk for abnormal semen analysis compared
80 (19.3 %), 247 (59.8%) and 86 (20.8%) men were Group 1, 2 and 3, respec- to either the low (RR:0.82;95%CI:0.66-1.03;p=0.10) or the moderate PA group
tively. The clinical pregnancy and ongoing/live rates in Group 3 (11.6%, 10/86; (RR:1.26;95%CI:1.00-1.60;p=0.06), albeit marginally.
8.1%, 7/86) was significantly lower than those of Group 1 (17. 5% 14/80; 15.0%, Limitations, reasons for caution: The present study corresponds to a single
12/80) and Group 2 (16.6%, 41/247; 14.2%, 35/247) (P < 0.05). However, time-frame regarding semen analysis, as a second evaluation was not performed.
there was no statistically significantly difference in the reproductive potential Furthermore, the small sample size and the single-center nature of this study
between Group 1 and Group 2. present as limitations.

Wider implications of the findings: The moderate PA group presented Study question: Does unilateral microsurgical subinguinal varicocelectomy
with a higher total and progressive motility, as well as with a lesser risk regarding have a favorable effect on the fertility potential of patients with severe
abnormal semen analysis compared with the low PA group. To conclude, results oligospermia?
presented herein demonstrate that moderate PA may improve semen quality, Summary answer: Microsurgical sub inguinal varicocelectomy improves
especially spermatozoa motility, buttressing existing published data. semen parameters of patients with severe oligospermia improving their fertility
Trial registration number: not applicable outcome.
What is known already: Severe oligospermia or a sperm concentration below
P-072 Zona pellucida selects spermatozoa with good DNA 5 million/ml, is an extreme derangement of male fertility potential that is often
fragmentation managed with assisted reproductive therapy. Intracytoplasmic sperm injection
R. Ganeva1, D. Parvanov1, D. Vaneva2, M. Vasileva3, G. Stamenov4 (ICSI) is usually preferred in such type of patients in order to maintain a higher
1 success rate during the invitro fertilization (IVF) cycle. The effect of
Nadezhda Woman’s Health Hospital, Research and development, Sofia, Bulgaria ;
2 Varicocelectomy on patients with oligoasthenoteratospermia has been exten-
Nadezhda Woman’s Health Hospital, Andrology, Sofia, Bulgaria ;
3 sively investigated with proven improvement, however its impact on patients
Nadezhda Woman’s Health Hospital, Embryology, Sofia, Bulgaria ;
4 with severe oligospermia remains to be elucidated.
Nadezhda Woman’s Health Hospital, Obstetrics and gynecology, Sofia, Bulgaria
Study design, size, duration: This was a retrospective study of 114
Study question: To evaluate and compare the sperm DNA fragmentation in patients diagnosed with severe oligospermia between January 2016 till January
normozoospermic patients after swim up preparation and zona pellucida binding 2018, who underwent unilateral microsurgical sub inguinal varicocelectomy.
sperm isolation. Patients with genetic abnormalities, history of varicocelectomy, or those who
Summary answer: Zona pellucida selected spermatozoa from normozoosper- received any infertility related therapy before surgery were excluded from
mic patients have significantly lower DNA fragmentation than native semen and the study.
those prepared only by swim up. Participants/materials, setting, methods: Patients were evaluated
What is known already: Sperm DNA fragmentation is associated with fer- pre-operatively and 6 months following surgery. Data regarding age, testicular
tilisation and implantation success. It was suggested that routinely applied meth- size, serum hormones (Testosterone, LH, FSH) and semen analysis results were
ods, such as swim up, lead to selection of spermatozoa with low DNA collected. Categorical data was presented as numbers (percentages) while
fragmentation. Another technique for sperm selection is based on sperm zona numerical data was presented as mean ± SEM. Data was compared using
pellucida binding. The sperm zona adhesion abilities were suggested as a predic- Wilcoxon Signed Ranked Test. A p value <0.05 was considered significant.
tor for the fertilisation ability of the semen. Recent study has shown that using Main results and the role of chance: The patients’ mean age was 36.8 ±
sperm bound to zona pellucida proteins for ICSI leads to higher pregnancy rates 5.2 years. A statistically significant improvement in sperm concentration
and fewer miscarriages than conventional sperm selection. However, data about (p<0.001), total motility (p<0.001) and progressive motility (p=0.003) was
sperm DNA fragmentation after zona pellucida selection is still absent. observed after surgery. The pre-surgical total motile sperm count (TMSC) was
Study design, size, duration: This observational study involved 29 normozo- 1.5±0.34 million sperm and was significantly increased following surgery to
ospermic patients of MHAT “Nadezhda”, Sofia, Bulgaria, between December 2019 10.5±2.0 million sperm (p< 0.001). No significant differences were noted in
and January 2020. Semen samples were analysis performed according to WHO hormonal levels following surgery.
2010. Native semen was liquefied and was subjected to swim up. The isolated Six months following varicocelectomy, the sperm concentration improved in
motile spermatozoa were allowed to adhere on acid solubilized zona pellucida 79 patients (69.3%); 16/114 (14%) patients showed normal sperm concentration
coated petri dishes. Sperm DNA fragmentation (SDF) was analysed in each patient’s while 26/116 (22.8%) patients improved to the oligospermia category. The
native semen, swim up prepared samples and zona bound spermatozoa. TMSC increased to above 5 million in 28 patients (24.5%).
Participants/materials, setting, methods: SDF of fresh semen, swim up Limitations, reasons for caution: The main limitation is the retrospective
prepared samples and zona bound spermatozoa was done using Halosperm G2 design of the study.
test kit (Halotech DNA SL, Madrid, Spain). Data was checked for normal distri- Wider implications of the findings: Documenting an improvement in the
bution using Kolmogorov-Smirnov test. Spearman correlation analysis was used fertility potential of patients with severe oligospermia is important as it would
for comparison of the SDF and semen analysis parameters. Differences between change their management plans. A good percentage of patients may become
the groups were analyzed by Wilcoxon paired test. Statistical analysis was per- candidates for intrauterine insemination or even natural conception instead of
formed using IBM SPSS ver.21. P<0.05 was considered significant. intracytoplasmic sperm injection.
Main results and the role of chance: The percentage of SDF of fresh semen, Trial registration number: NA
swim up prepared samples and zona bound spermatozoa did not correlate to
the conventional semen analysis parameters (sperm concentration, motility and
morphology) of the studied normozoospermic patients (p>0.05). However, the
P-074 Effect of Abstinence Period on Seminal Oxidative Stress in
SDF values of the fresh semen were found to be 32±5%, which is slightly above
Infertile men
the previously estimated cut off for good DNA fragmentation (30%).
Following sperm preparation by swim up and zona binding the SDF values were M. Arafa1, K. Khalafalla1, A. Majzoub1, A. Agarwal2, S. Alsaid1,
reduced significantly (p=0.018, p=0.017 respectively). Moreover, the median SDF H. Elbardisi1
1
percentage in the swim up prepared samples and zona bound spermatozoa were Hamad Medical Corporation, Urology, Doha, Qatar ;
2
found to differ significantly (Median: 15% vs. 10%; p=0.01). In addition, zona bound Cleveland clinic, American Center for Reproductive Medicine, Cleveland-, U.S.A.
spermatozoa from all patients had DNA damage 3-4 times lower than the esti-
mated cut off value for good SDF.Limitations, reasons for caution: Results Study question: Does the duration of abstinence period affect seminal oxi-
from this study need to be confirmed by larger group of samples. dative stress in infertile men?
Wider implications of the findings: Sperm zona selection provides sper- Summary answer: Shorter abstinence period significantly decreases seminal
matozoa with good DNA fragmentation. Zona selection technique may be used oxidative stress with subsequent improvement of motility, progressive motility
to improve the impact of abnormal SDF on the reproductive outcomes even in and normal morphology.
normozoospermic patients. What is known already: The duration of abstinence may affect the semen
Trial registration number: not applicable quality. While longer abstinence is associated with more semen volume and
sperm concentration, it negatively affects total and progressive motility. WHO
P-073 Effect of Unilateral Microsurgical Varicocelectomy on guideline stated that abstinence should be 2-7 days. Different studies have sought
fertility outcome and treatment plans of patients with severe to determine the optimal time frame for ejaculatory abstinence, however the
oligospermia results are often found to be contradictory. Also, there are no studies discussing
H. Elbardisi1, K. Khalafalla1, A. Majzoub1, S. Alsaid1, M. Arafa1 the effect of abstinence on seminal oxidative stress measured by oxidation reduc-
1
Hamad Medical Corporation, Urology, Doha, Qatar tion potential.

Study design, size, duration: This retrospective study included 255 patients varicocele were significantly older than patients with No-varicocele (p=0.006).
presenting with male factor infertility to a tertiary medical center over a period No significant differences between the 2 groups with regards to testicular size
of 2 months. The inclusion criteria were patients who did semen analysis with (right, left), reproductive hormones (Testosterone, LH, FSH, Estradiol) and BMI
an abstinence of less than 2 days or more than 5 days. were observed.
Participants/materials, setting, methods: Patients were grouped into
group A (76 patients) with abstinence < 2 days and group B (177 patients) with
abstinence > 5 days. Semen analysis was done according to the 5th edition WHO
guidelines. ORP was determined using the MiOXSYS system (Aytu BioScience, Non Varicocele Varicocele
Parameters P Value
Englewood, CO). SDF was measured by sperm dispersion method (Halosperm). group group
The results were compared by Wilcoxon rank sum test and paired T test. p value
< 0.05 was considered significant. Age (years) 35.3±0.74 38.2±0.623 0.006
Main results and the role of chance: The patients’ mean age was 35.9±7.6 BMI (Kg/m²) 31.5±1.6 30.4±0.8 0.12
years. ORP was significantly lower in group A than in group B (2.5±3.0 vs
Testosterone (nmol/L) 14.8±1.06 15.8±0.62 0.38
5.3±10.5mV/106 sperm). Sperm concentration was lower in group A than group
B but the difference was non-significant (32.9±25.7 vs 37.7±28.6millions/ml). LH (IU/L) 7.23±0.62 7.03±0.43 0.79
Total and progressive motility were also better in group A than group B FSH (IU/L) 13.85±1.26 13.53±0.78 0.83
(53.6±18.1% vs 49.2±18.8%, 12.5±10.9% vs 11.6±11.5% respectively) but again
E2 (pg/ml) 110.15±5.8 122.5±7.4 0.296
the differences were insignificant. Abnormal form was slightly and insignificantly
less in group A than group B (93.7±9.8% vs 93.8±9.0%). Sperm DNA fragmen- Left Testes size (cm³) 7.02±0.54 7.06±0.3 0.94
tation was less in group A than group B (28.8±21.2% vs 31±15.9%) but the Right Testes size (cm³) 7.9±0.6 7.0±0.3 0.12
difference was not significant.
Limitations, reasons for caution: The main limitation is the retrospective
design of the study.
Wider implications of the findings: The significant improvement in seminal Limitations, reasons for caution: The main limitation is the retrospective
oxidative stress with the short abstinence may introduce a new treatment tech- design of the study.
nique in infertile men were couples can be counseled to engage in frequent sexual Wider implications of the findings: Patients with NOA who have a clinical
intercourses hoping to benefit from the improvement in semen parameters. varicocele should be thoroughly counselled before treatment decisions are made.
Trial registration number: NA The expected clinical benefit in face of the invasiveness and the extra cost of
the varicocelectomy should be discussed with the patients before undergoing
testicular sperm retrieval.
P-075 Threshold-dependent effects of energy restriction as a
novel potential therapy to increase sperm concentration in men
with obesity: A Randomised Controlled Study
“Abstract withdrawn by the authors” P-077 Impact of the assessment of sperm DNA damage on IVF
and ICSI outcome: role of intra-individual variation
P. Christensen1, E. Parner2, A. Birck3
P-076 Does the presence of varicocele Influence microsurgical 1
SPZ Lab A/S, Clinical Research, Copenhagen, Denmark ;
testicular sperm retrieval in men with non-obstructive 2
Aarhus University, Department of Public Health- Section for Biostatistics, Aarhus
azoospermia (NOA)?
C, Denmark ;
K. Khalafalla1, M. Arafa1, H. Elbardisi1, S. Alsaid1, A. Majzoub1 3
SPZ Lab, Clinical Research, Copenhagen OE, Denmark
1
Study question: How does intra-individual variation in the assessment of
Study question: Can the outcome of sperm retrieval in patients with non-ob- sperm DNA damage affect the outcome of IVF and ICSI treatments?
structive azoospermia be affected by an untreated clinical varicocele? Summary answer: Intra-individual variation in assessment of sperm DNA
Summary answer: The presence of clinical varicocele did not influence the damage plays a very important role for the outcome of IVF and ICSI treatments.
sperm retrieval rate in this population of NOA patients. What is known already: The impact of sperm DNA damage on the outcome
What is known already: Varicocele is prevalent in approximately 5% of men of assisted reproduction remains controversial. The lack of agreement in the
with NOA, the pathophysiology of azoospermia and its relation to varicocele is literature is partially due to a diversity of test methods as well as poorly stan-
still under investigation. It is still debatable whether varicocele contributes to dardized protocols. Methods based on flow cytometry have the potential of
spermatogenesis disruption and affects sperm retrieval rates in NOA. Recent being very robust and precise. However, large differences in intra-individual
studies have reported a significant influence for varicocele on patients with variation have been reported for different laboratories assessing sperm DNA
non-obstructive azoospermia (NOA) suggesting varicocelectomy prior to damage based on the SCSA protocol. Some of this variation may have a biological
testicular sperm extraction (TESE) procedures. background, but can to a large extend be due to differences in laboratory per-
Study design, size, duration: The charts of 448 patients who presented formance and quality control procedures.
to the Male infertility clinic from 2011 – 2016 with Non-obstructive azo- Study design, size, duration: Simulated power calculations using 10,000
ospermia and underwent microsurgical TESE were included and retrospec- replications were based on our previous retrospective analysis of the impact
tively studied. Exclusion criteria for this study were prior varicocelectomy, of sperm DNA damage on 406 clinical cases for IVF and ICSI (Christensen
TESE, chemotherapy or radiation therapy, patients with abnormal genetics et al., Hum Reprod. 2013;28:i128-P-026). Intra-individual variation (CV
or chromosomal deletions. within male: CVw) for assessment of sperm DNA damage in our laboratory
Participants/materials, setting, methods: Patients demographics, clinical was calculated to 16.5% using data from a previous randomized and dou-
and laboratory data (age, BMI, estradiol, FSH, LH, Prolactin, Testosterone) as ble-blind controlled trial (Blomberg-Jensen et al., J Clin Endocrinol Metab
well as the sperm retrieval outcome were compared between non-obstructive 2018;103:870-81).
azoospermic patients with and without varicocele. Data was compared using Participants/materials, setting, methods: The median percentage of
Wilcoxon Signed Ranked Test. A p value <0.05 was considered significant. sperm with damaged DNA (DFI) was 15.9% with a total CV of 60.2%.
Main results and the role of chance: Varicocele data was available in 222 Calculations of true pregnancy rates for IVF and ICSI were estimated using a
charts, 157 patients had a clinical varicocele and 70 had no varicocele. Sperm CVw of 0.0%. The true pregnancy rates were then used in calculations of statis-
were retrieved from 57% of patient with Varicocele (n=89) compared with tical power. We considered scenarios of CVw of 16.5% plus increases of CVw
45.3% of patients (n=38) with No-Varicocele. The difference in sperm retrieval with 50% and 100%. A 2-sided chi2-test was used for comparison of pregnancy
between both groups was not statistically significant (p=0.09). Patients with rates (significance level 0.05, power 80%).

Main results and the role of chance: For IVF treatments, true pregnancy Main results and the role of chance: Euploidy rates of PGT-A blastocyst
rates (CVw = 0) were calculated to 47.7% (DFI≤15) and 22.2% (DFI 15 to 25). with AOA and without AOA after ICSI were compared in each group. Maternal
For ICSI treatments, true pregnancy rates were calculated to 49.5% (DFI<25) and paternal age had no significant differences between two groups, with AOA
and 24.7% (DFI≥25). Using our laboratory’s CVw of 16.5%, the sample size and without AOA. Euploidy rates of each group are below. (Groups with AOA-
needed for comparison of pregnancy rates was estimated to n=169 for an IVF Group 1: 34.88%(43/15), Group 2: 33.33%(78/26), Group 3: 32.46%(228/74),
study. A study with ICSI-treatments would require a sample size of n=336. When Group 4: 38.71%(31/12), Groups without AOA- Group 1: 30.77%(13/4), Group
CVw was increased with 50%, the required sample sizes increased to n=207 (IVF) 2: 33.33%(48/16), Group 3: 34.36%(2363/812), Group 4: 30.77%(26/8)). In
and n=437 (ICSI). A 100% increase in CVw gave corresponding sample sizes of severe male factor group, Group 1 and Group 4, cases with AOA had higher
n=260 (IVF) and n=569 (ICSI). A small intra-individual variation in the assessment euploidy rate than cases without AOA. However, there was no significant dif-
of sperm DNA damage is essential for correct diagnosis and treatment. In conference stastistically in Group 1(34.88% vs. 30.77%, p>0.05) and Group
trast, a high variation may lead to misclassification of patients and potentially 4(38.71% vs. 30.77%, p>0.05). In oligozoospermia group, Group 2, euploidy
suboptimal treatment. When the impact of sperm DNA damage on assisted rate of cases with AOA was no different from cases without AOA(33.33% vs.
reproduction outcomes is to be studied, a high degree of intra-individual variation 33.33%, p>0.05). Also, there was no significant difference in normal sperm group,
increases the required sample size markedly. Without a sufficient sample size, Group 3(32.46% vs. 34.36%, p>0.05). This study showed no significant differ-
the true biological relationship may remain concealed. To produce reliable and ences in euploidy rate between cases with and without AOA.
valuable results having low intra-individual variation, laboratories performing Limitations, reasons for caution: Even though there were numerous cases
tests of sperm DNA damage should ensure good quality control to minimize in our center, we had limits in analyzing significant differences among cases. It is
potential laboratory errors. because there were a few PGT-A cases with AOA in male factor infertility
Limitations, reasons for caution: None. patients. Furthermore, we only concerned cases treated with calcium ionophore
Wider implications of the findings: Laboratories performing test of sperm among various AOA in this study.
DNA damage should declare the level of uncertainty of the test used. Differences Wider implications of the findings: Although our study needs to be sup-
in quality control procedures between laboratories are likely to play a significant ported by further study, this study revealed that AOA contributes to increasing
role in interpretation of results even when the same method is used and may euploidy rates in severe male factor infertility patients. Consequently, it is
impact assisted reproduction treatment outcome. expected that AOA could increase clinical pregnancy rates by increasing euploidy
Trial registration number: Not applicable rates in severe male factor infertility patients.
P-078 Is there an effect of artificial oocyte activation on euploidy
of embryo in patients with male factor infertility? P-079 Very low concentration of aged cerium dioxide
H. Jeong1, K.Y. Kang1, H.S. Choi1, E.H. Moon1, Y.R. Park1, nanoparticles (CeO2NPs) induce DNA damage in human and rat
S.H. Lee1, E.A. Park1, J.Y. Lee1, H.S. Kim1, K.A. Lee2, J.J. Ko2, gametes after in vitro exposure
I.S. Kang1, M.K. Koong1, Y.S. Kim1, M.J. Kim1 M. Cotena1, M. Auffan2, V. Tassistro1, C. Uboldi1, J. Rose2, J. Perrin3
1
1
CHA Fertility Center Seoul Station, Fertility laboratory, Seoul, Korea- South ; Faculty of Medecine- Aix Marseille Université- Equipe Biomarqueurs-
2
CHA University, Department of Biomedical Science- College of Life Science, Environnement- Santé - Institut Méditerranéen de Biodiversité et d’Ecologie UMR
Pocheon, Korea- South 7263- IRD 237, Faculty of Medicine, Marseille, France ;
2
European Centre Research And Teaching In Geosciences of the Environment
Study question: The purpose of this study is to identify euploidy rate in CEREGE - CNRs- IRD- Collège de France- INRA- Aix-en-Provence- France,
severe male factor patients treated with artificial oocyte activation(AOA) ED-Environnement Durable, Marseille, France ;
after ICSI. 3
Aix Marseille University- AP-HM La Conception- Centre Clinico-Biologique
Summary answer: There was an insignificant difference of euploidy in severe d’Assistance Médicale à la Procréation - CECOS- 13385 Marseille Cedex 5-
male factor groups treated AOA after ICSI with preimplantation genetic France, Faculty of Medicine, Marseille, France
testing-aneuploidy(PGT-A).
What is known already: In many studies, it is known that sperm abnor- Study question: Do aged CeO2NPs induce DNA damage in human and rat
malities result in abnormal sperm decondensation, aberrant pronuclear gametes after in vitro exposure?
development, migration, apposition and first mitosis. It has been reported Summary answer: In vitro gametes exposure to the lowest concentration of
that treatment of AOA in patients with male factors improves these prob- CeO2NPs tested induces a significant increase in DNA damage compared to the
lems. Also, AOA leads to a cascade of events including extrusion of the highest doses.
second polar body, decondensation of a haploid set of chromosomes and What is known already: Cerium dioxide nanoparticles (CeO2NPs) are an
initiation of embryonic development. Thus, sperm abnormalities and oocyte active catalyst and are widely used as diesel additive to increase fuel economy.
activation are important in fertilization. Generally, AOA is used for low quality They are released in the atmosphere after engine combustion. The Organization
oocytes and sperm, and the low fertilization rate and frequent fertilization for Economic Cooperation and Development included CeO2NPs in the priority
failure. list of nanomaterials requiring urgent evaluation. CeO2NPs can transfer to the
Study design, size, duration: 25,427 IVF-ICSI cycles were performed testicles and epididymis after inhalation in rats. In vitro exposure of human and
at our center from January 2016 to December 2019. From those studies, mouse sperm and cumulus oocyte complexes (COC) to low concentrations of
only 2,285 cycles were performed with preimplantation genentic testing pristine CeO2NPs (10 µg.l-1) induce significant DNA damage. Nevertheless,
and 750 cycles were performed with PGT-A. Among them, the cycles with pristine CeO2NPs are altered by combustion and the potential hazard of aged
artificial activation after ICSI were only 103 cases. The embryo euploidy CeO2NPs exposure remains unexplored.
rate was evaluated in cases of blastocyst PGT-A. Statistical analysis was Study design, size, duration: Pristine CeO2NPs were extracted from
performed using the chi-squared test. P values p<0.05 were considered EnviroxTM diesel additive, combusted at 850°C, (average combustion tempera-
significant. ture in a diesel engine) and physically characterized. Chemical (un)stability in
Participants/materials, setting, methods: IVF-ICSI cycles that performed FerticultTM embryo culture medium was assessed by Inductively coupled plasma
blastocyst biopsies were sorted from cycles performed with PGT-A and these mass spectrometry (ICP-MS). Rat gametes were collected in epididymis and
were divided into four groups depending on the concentration of the sperm; oviducts after cervical dislocation euthanasia of 60 days old males and 26 days
Group 1: severe oligozoospermia(<1xml), Group 2: oligozoosper- old females (after ovarian stimulation). Human frozen sperm from fertile donors
mia(1≤~<15xml), Group 3: normal(≥15xml), Group 4: TESE sperm(sperm were purchased from Germetheque biobank (France).
retrieved from azoospermia patients). Each group was additionally divided into Participants/materials, setting, methods: Human and rat gametes were
two groups; with or without activation. We used 10µM/L calcium iono- exposed in vitro to CeO2NPs [1 to 1.103 µg.l-1] during 1 hour at 37°C, 5%CO2.
phore(A23187; Sigma-Aldrich) in culture medium for the AOA for 5min at 30min DNA damage was analysed by alkaline comet assay (ACA) and quantified by
after ICSI. Olive Tail Moment (OTM) in COC and by %Tail DNA in sperm. CeO2NPs-cells

interaction was assessed in human sperm by Transmission Electron Microscopy Participants/materials, setting, methods: All patients had a history of
(TEM). Oxidative stress was evaluated using 2,7-dichlorodihydrofluorescein reproductive implantation failures and/or miscarriages. When classified accord-
(H2DCF) probe detected by Flow Cytometry and expressed as Mean ing to WHO criteria, 29 samples were normozoospermic, 82 were moderate
Fluorescence Intensity (MFI). oligoasthenoteratozoospermic (OAT) (sperm count ≥5 million and < 15 million
Main results and the role of chance: In human sperm, exposure to 1 µg.l-1 spermatozoa/ml) and 28 were severe OAT (sperm count <5 million sperma-
CeO2NPs induced a significant increase of DNA damage (mean %Tail DNA±SEM tozoa/ml). The FISH analysis was performed for nine chromosomes using probes
= 32.06 ± 0.54) compared to the unexposed control (11.96 ± 0.30) (p < specific for the chromosomes X, Y, 13, 15, 16, 17, 18, 21 and 22.
0.0001). TEM analysis showed big CeO2NPs aggregates at 100 µg.l-1 but close Main results and the role of chance: TZI was significantly higher in severe
proximity between cells and NPs at 1 µg.l-1. Oxidative production was increased OAT samples (2.1±0.2, range 1.5-2.4) when compare with normal (1.7±0.1,
in human sperm exposed to 1 µg.l-1 CeO2NPs (MFI±SEM = 16.9% ± 1.5), range 1.4-1.9) (P= 1.11E-8) and moderate OAT samples (1.9±0.2 range 1.5-2.5)
compared to the negative control (2.7% ± 1.4). (P= 2.20E-5). TZI was also significantly higher in moderate OAT compared with
In rat oocytes and follicle cells, exposure to 1 µg.l-1 CeO2NPs induced signifi- normal samples (P= 1.19E-6).
cantly higher DNA damage (mean±SEM OTM = 10.26±0.36 and 4.91±0.29, The frequency of total sperm aneuploidy was in inverse correlation with sperm
respectively) compared to the unexposed controls (2.21±0.18 and 0.72±0.05, concentration (R= -0.38, P= 5.36E-6), progressive motility (R= -0.41, P= 4.57E-
respectively) and to higher CeO2NPs concentrations (p < 0.0001). In rat sperm, 7), total motility (R= -0.25, P= 0.0026) and morphology (R= -0.36, P= 1.04E-5.
exposure to 1 µg.l-1 CeO2NPs induced significantly higher DNA damage (mean When considering the sperm typology, the frequency of total aneuploidy was
%Tail DNA±SEM = 21.23±0.37) compared to unexposed control (11.06±0.24) significantly higher in severe OAT (5.01±1.82%, range 2.62-9.04%) when com-
and to higher CeO2NPs concentrations (p <0.0001). pared with normal (2.24±0.79%, range 0.63-4.22%) (P= 9.31E-9) and moderate
DNA damage was inversely proportional to the CeO2NPs concentration; we OAT samples (3.04±1.12%, range 1.47-7.82%) (P= 2.50E-6). It was also signifi-
might hypothesize that our results are related to the aggregation states. At high cantly higher in moderate when compared with normal samples (P=0.00067).
concentration, nanoparticles tend to aggregate between them, reducing the The combined analysis of TZI and aneuploidy in the different sperm typologies
surface of interaction with cells. Conversely, at low concentration they can easily showed a significant positive correlation between the frequency of total aneu-
come into contact with cells. ploidy and TZI (R= 0.47, P= 6.72E-9).
Limitations, reasons for caution: These results cannot be extrapolated Limitations, reasons for caution: This study included a restricted number
to in vivo toxicity of CeO2NPs after inhalation, but demonstrate that interac- of samples. Additional data are necessary to corroborate our findings.
tions between CeO2NPs and germ cells induce significant DNA damage and Wider implications of the findings: Morphologically abnormal spermatozoa
oxidative stress. Additional data should be needed to better understand the often have multiple defects. A detailed assessment of their incidence would
mechanism of interaction between cells and NPs by using nano-tomography refine the relevance of morphological defects, single or multiple, on the capacity
imaging analysis. of a sperm cell to support fertilization and embryo development, contributing
Wider implications of the findings: The results obtained give some light to the evaluation of the extent of damage occurring during human
on the complex cellular mechanisms by which ROS generation could exert their spermatogenesis.
biological effects on human spermatozoa after CeO2NPs in vitro exposure. Trial registration number: Not Applicable
Potential impacts of diesel exhaust exposure in couples are a major concern for
public health, highlighting the need for in vivo studies.
Trial registration number: 15447-2018061110211950 P-082 Identification of novel mutations in DPY19L2 responsible
for human fertilization failure and have a favorable clinical
outcomes by artificial oocyte activation
P-080 Air pollution and seminal parameters in subfertile men
X.J. Zhuang1, P. Liu1
“Abstract withdrawn by the authors” 1
Reproductive Medical Center- Peking University Third Hospital, Department of
Obstetrics and Gynecology, Beijing, China
P-081 Correlation between teratozoospermia index (TZI) and
sperm aneuploidy in infertile men Study question: Does artificial oocyte activation (AOA) together with intra-
M. Benincasa1, A. Crippa1, M.C. Magli1, F. Coppola2, G.B. La Sala1, cytoplasmic sperm injection (ICSI) improve clinical outcomes for human fertil-
L. Gianaroli1 ization failure of globozoospermia linked to DPY19L2 mutations patients?
1
S.I.S.Me.R, Reproductive Medicine, Bologna, Italy ; Summary answer: Novel DPY19L2 deletions were revealed by whole-exome
2
Centro Medico Spallanzani, Clinical Center, Parma, Italy sequencing. AOA together with ICSI improved cycles of DPY19L2 mutations
by increasing the fertilization and transferable embryo rates.
Study question: Does the teratozoospermia index (TZI) correlate with the What is known already: In clinical, some infertile couples suffer from recur-
frequency of sperm aneuploidy in a manner that is dependent on the severity rent fertilization failure. Globozoospermia is a rare form of male infertility char-
of the male factor condition? acterised by round-headed sperm and malformation of the acrosome. Although
Summary answer: TZI correlates with the frequency of sperm aneuploidy the fertilization rate was improved by ICSI, about 1–5% of ICSI cycles still display
and increases with the severity of the male factor condition. fertilization failure. The genetic reasons for fertilization failure are largely
What is known already: The contribution of sperm aneuploidy to the embryo unknown. Although pathogenic variants in DPY19L2 are known causes of glo-
chromosome status is generally perceived to be clinically irrelevant in comparison bozoospermia, novel DPY19L2-mutated in infertile patients should be revealed
with the oocyte counterpart. However, increases in sperm aneuploidy have been and the treatment of these DPY19L2 deleted globozoospermic patients by AOA
reported to have an effect on male infertility and on IVF cycle outcomes especially together with ICSI should be evaluate.
in cases of repeated implantation failure and recurrent miscarriages. Infertile Study design, size, duration: Globozoospermic patients were recruited
men typically have significantly higher levels of sperm aneuploidy compared with from the Reproductive Medical Center of Peking University Third Hospital
fertile or normozoospermic males and the level of aneuploidy correlates to the (Beijing China) between January 2016 and June 2019. Informed consent was
severity of the male factor. Sperm aneuploidy is routinely evaluated by FISH obtained from all individual participants included in the study. All procedures
(Fluorescence in Situ Hybridization). performed in studies involving human participants were in accordance with the
Study design, size, duration: Since January 2018, 139 sperm samples from ethical standards of the institutional and national research committees of Peking
patients with a normal karyotype undergoing assisted conception cycles were University Third Hospital and with the 1964 Helsinki declaration and its later
analysed for semen parameters according to WHO 2010. In addition to the amendments or comparable ethical standards.
morphology evaluation, TZI (representing the number of abnormalities per Participants/materials, setting, methods: DPY19L2 were analyzed with
abnormal spermatozoon) was also determined. The same sperm samples were blood and sperm samples from globozoospermic patients. DPY19L2 levels in
also analyzed by FISH to evaluate possible correlations between TZI, aneuploidy human testes were investigated using RT-PCR, western blotting and
and the severity of the male factor. Immunofluorescent analysis, respectively. And its novel mutations using

whole-exome sequencing and sanger sequencing was explored. Human sperm Participants/materials, setting, methods: The study included women
samples from globozoospermic donors with DPY19L2 mutations are subjected younger than 38 years who underwent controlled ovarian stimulation and fer-
to ICSI and AOA. The fertilization rates, cleavage rates and transferable embryo tilization using either ejaculated abnormal sperm (OAT) or surgically extracted
rates of DPY19L2-mutated patients were compared with its previous ART testicular sperm (AS). A comparison between the morphokinetic parameters,
cycles. implantation and clinical pregnancy rates of the two groups was made with
Main results and the role of chance: DPY19L2 mRNA and protein were additional subgroup analysis according to the implantation status. Logistic regres-
abundantly transcribed in male testis in human. Immunohistochemical results sion was conducted to assess the association between sperm origin and embry-
revealed DPY19L2 was localized in the cytoplasm of spermatids and round onic morphokinetic parameters and implantation.
spermatids. Through whole-exome sequencing and sanger sequencing, we iden- Main results and the role of chance: Implantation rate was significantly
tified that five patients carried DPY19L2 deletions and six patients contained higher in the ejaculated sperm group compared to the testicular sperm group
novel DPY19L2 point mutations in couples diagnosed with fertilization failure. (45.8% vs. 33.6%, p=0.02). Embryos from the OAT group reached the later
Expression of these mutations in human testis significantly reduced the levels of morphokinetic milestones - embryonal third cell cycle (ECC3), synchronous
DPY19L2 expression. In addition, the fertilization rate, embryo cleavage rate division (S3) and Morula compaction- faster than embryos obtained from azo-
and transferable embryo rate were significantly higher in the AOA group than ospermic patients. Implanted embryos developed in similar rate in both group.
its previous cycle of group (fertilization rate 50.38% versus 13.86%, respectively, In a multivariate analysis for the entire study population, ejaculated sperm
P < 0.001; embryo cleavage rate 59.16% versus 19.04%, respectively, P < 0.001; (OR=5.88 (CI 95% 2.56-12.50), p<0.001) and time to 8 cell stage (OR=0.90
transferable embryo rate 43.51% versus 16.69%, respectively, P < 0.001). Five (CI 95% 0.82-0.98), p=0.013) were positively associated with successful
live-birth babies were born by AOA together with ICSI. implantation.
Limitations, reasons for caution: Analysis was performed in testicular Limitations, reasons for caution: The two patients’ groups were not homo-
biopsy samples from only a small number of patients with DPY19L2 mutations. geneous in their basic characteristics. A higher rate of nulliparity and increased
In future investigations, a larger sample size should be used and the role of the fragmentation was demonstrated in the AS couples. Information regarding the
other genes involved in the globozoospermic should be analyzed. maximal dose of GT obtained, previous IVF response and ovarian reserve testing
Wider implications of the findings: DPY19L2 variants are highly related was lacking.
to globozoospermia and fertilization failure. AOA could rescue the pheno- Wider implications of the findings: Ejaculated sperm impact on late pre-
type of fertilization failure and help establish pregnancy and lead to live birth. implantation development, especially the third cell cycle and compaction, may
These findings bring us closer to a complete molecular diagnosis for globo- highlight another pathophysiology by which sperm origin effects embryo devel-
zoospermia patients which would help to predict the success of reproductive opmental kinetics. Morphokinetic parameters, with regard to sperm source, may
treatments. assist in predicting implantation.
Trial registration number: This study was supported by the National Natural Trial registration number: Kamin fund
Science Foundation of China (NO.81671513) and Beijing Natural Science
Foundation (NO.7172236). P-084 Sperm fatty acid profile in infertile men.
A. Stendardi1, S.E. Pizzasegale1, F.L. Vellucci1, G. Collodel2,
P-083 Does sperm origin - either ejaculated or testicular – affect E. Moretti2, C. Signorini2, L. Gambera1
early and late embryonic morphkinetic parameters 1
A.G.I. Medica, Reproductive Medicine, Siena, Italy ;
G. Karavani1, N. Schachter - Safrai1, I. Har-Vardi2, A. Buxboim3, 2
Università degli Studi di Siena, Dipartimento di Medicina molecolare e dello
Y. Kantor4, S. Zeev4, O. Yuval4, S. Yoel5, A. Ben-Meir1 sviluppo, Siena, Italy
1
Hebrew University-Hadassah Medical center, Obstetrics and Gynecologyresident,
jerusalem, Israel ; Study question: Are the sperm characteristics correlated with sperm fatty
2
Soroka, The IVF Unit Gyn/Obs- Soroka University Medical Center- Israel- Faculty acid composition?
of Health Sciences- Ben-Gurion University of the Negev- Beer-Sheva- Israel., Beer Summary answer: Our results show a relationship between sperm fatty acid
Sheva, Israel ; composition and sperm characteristics evaluated by light and electron microscopy
3
Hebrew University-Hadassah Medical center, Alexander Grass Center for What is known already: In sperm membrane and semen plasma, lipids and
Bioengineering- School of Computer Science and Engineering- The Hebrew fatty acid (FA) composition are relevant to spermatozoa function and spermato-
University of Jerusalem- Jerusalem- Israel, jerusalem, Israel ; genesis. Mammalian spermatozoa are characterized by a high proportion of
4
The Hebrew University of Jerusalem- Jerusalem- Israel., Alexander Grass Center polyunsaturated fatty acids (PUFA) which play a crucial role in sperm maturation
for Bioengineering- School of Computer Science and Engineering, Jerusalem, Israel ; and motility, in acrosome reaction and fertilization. Due to the high presence of
5
Infertility and IVF Unit, Helen Schneider Hospital for Women, Rabin Medical Center PUFA, sperm are susceptible to lipid peroxidation. It is known that men with
altered seminal parameters had shown different FA composition and different
Study question: Is there an effect of sperm origin on embryo morphokinetics omega-6/omega-3 PUFA ratio.
in couples diagnosed with male factor infertility (MFI)? Study design, size, duration: The proposed investigation is a retrospective
Summary answer: Testicular sperm impacted early developmental stages, study. Twenty-three infertile men (aged 27-38) referred to AGI Medica lab (Siena,
while ejaculated sperm was associated with faster late cell division, faster com- Italy) for semen analysis were enrolled from January 2019 to June 2019.
paction and higher implantation rate. Participants/materials, setting, methods: Semen characteristics were
What is known already: Time-lapse imaging provides a non-invasive tech- evaluated by light and transmission electron microscopy (TEM). TEM data was
nique that enables assessment of embryo quality by morphokinetic parameters. quantified with a mathematical formula which provides numerical scores as the
While embryonic morphokinetic milestones are known to reflect the develop- percentage of apoptosis, necrosis and immaturity and a fertility index. Sperm
mental and implantation potential, it is unclear whether sperm origin, either FA composition was evaluated by a gas chromatography instrumentation
ejaculated or testicular, in MFI has an early or late effect with influence on cleav- (Lipinutragen, Bologna, Italy). The content of FA and the value of the ratios
age, blastulation and implantation potential. There is only scarce data regarding between different FA were correlated with sperm characteristics (Spearman’s
the paternal effect on embryo development, with regard to sperm origin, Rank Correlation Coefficient ).
using TLI. Main results and the role of chance: Semen analysis showed an impaired
Study design, size, duration: The study included a retrospective analysis of semen quality. By TEM investigation, sperm pathologies were increased and
morphokinetic parameters performed by TLI from five medical centers between fertility index reduced as compared to normal values. A positive correlation was
January 2013 and December 2017.The developmental process and kinetics of detected between docosahexaenoic acid (DHA), an omega-3 PUFA, and sperm
424 embryos obtained from ejaculated sperm from couples with MFI attributed concentration, vitality and fertility index. Necrosis was positively correlated with
to Oligoasthenoteratozoospermia (OAT) were compared to 160 embryos oleic, vaccenic, palmitoleic acid content, and negatively with DHA and saturated
derived from surgically extracted testicular sperm from couples diagnosed with fatty acid/monounsaturated fatty acid ratio. The omega-6/omega-3 ratio neg-
azoospermia (AS) atively correlated with sperm concentration, motility and morphology and

positively with sperm necrosis. Sperm immaturity positively correlated with Loss-of- function mutations could lead to male infertility in humans and mice by
eicosatrienic acid. damaging spermatogenesis and making abnormal sperm
With a view to the role of FA in sperm characteristics, these preliminary data Limitations, reasons for caution: Considering the relationship between
show that, i) DHA content appears strictly related to sperm quality, in particular CEP112 and primary ciliarym, we wanted to performed CT and MRI scans on
to sperm vitality; ii) the omega6/omega3 ratio increases in presence of necrosis, the patient that can assess clinical signs of PCD. Unfortunately, the patient
as well as eicosapentaenoic acid/arachidonic acid ratio. refused further examination, therefore we did not assess ciliary beat in the
These results may be implemented by further studies on the relationship patient.
between FA composition and sperm characteristics since a dietary FA supple- Wider implications of the findings: The screening of the deleterious muta-
mentation should improve semen quality. tions of CEP112 could be important for clinical molecular diagnosis of male
Limitations, reasons for caution: The study group is small and selected infertility. Further study is required to elucidate the molecular mechanism of
groups of patients with different reproductive pathological conditions will to be CFAP65 in sperm flagellar development.
compared Trial registration number: 81771642
Wider implications of the findings: A deeper knowledge on sperm FA
profile and its correlation with sperm pathology indicate the potential role of P-086 Microfluidic sperm sorter versus sperm slow prior to ICSI in
personalized nutraceutical treatments to modulate sperm FA composition oligoasthenozoospermia samples with poor DNA Fragmentation
improving male reproductive efficiency. Index (DFI) scores in SCSA testing
Trial registration number: none P. Selvaraj1, K. Selvaraj1, V. Srinivasan2, M. Sivakumar1,
V. Harikrishnan1, S. Balakrishnan1, S. Sudan1
P-085 A novel homozygous CEP112 mutation in humans causes 1
Fertility Research and Women’s Speciality Hospital, G.G. Hospital, Chennai, India ;
male infertility with azoospermia 2
The Tamilnadu Dr.MGR Medical University, Biostatistics, Chennai, India
X. Zhang1, G. Huang2, G. Zhang2, W. Xu2
1
Department of Obstetrics/Gynecology- Joint Laboratory of Reproductive Medicine Study question: Compare the outcomes of sperm preparation by Microfluidics
SCU-CUHK- Key Laboratory of Obstetric- Gynecologic and Pediatric Diseases and and Sperm slow in oligoasthenozoospermia samples with poor DFI and high
Birth Defects of Ministry of Education- West China Second University Hospital- DNA stainability scores in sperm chromatin structural assay(SCSA) prior to ICSI.
Sichuan Universi ; Summary answer: The use of Microfluidics and Sperm slow had no statistically
2
Department of Obstetrics/Gynecology- Joint Laboratory of Reproductive Medicine significant differences on primary outcomes.
SCU-CUHK- Key Laboratory of Obstetric- Gynecologic and Pediatric Diseases and What is known already: Ideal sperm preparation methods for oligoastheno-
Birth Defects of Ministry of Education- West China Second University Hospital- zoospermia samples have long been debated especially if additional testing such
Sichuan Universi as DFI and high DNA stainability (HDS) scores have been added using SCSA.
Micorfluidics has emerged as an atraumatic method of sperm selection, enabling
Study question: To date, several gene mutations have been identified in azo- enhanced retrieval of motile sperm with normal morphology, mimicking nature
ospermia, which can explain the genetic causes of a small number of azoospermia and proving a non static restricted environment. Spermslow on the other hand
cases. Here, we report anovel gene mutations has the advantage of replacing Polyvinylpyrrolidone and providing hyaluronan
Summary answer: Our experimental observations on human subjects sug- and important components for pre-fertilization event of zona-binding and pen-
gested that CEP112 is involved in sperm flagellum structure and that loss-of- etration. This hyaluronan enriched semi viscous medium enhances selection of
function mutations could lead to male infertility low DNA fragmented sperm with better embryo formation rates.
What is known already: Nearly half of male infertility cases are thought to Study design, size, duration: A prospective study was conducted from June
be linked to genetic defects. And 20% of infertile men are diagnosed with azo- 2016 to December 2018 with the sample size of 109 couples undergoing assisted
ospermia. Nonobstructive azoospermia is defined as spermatogenic failure, and reproduction. The mean age of women was 32.26±4.26 years. The mean age of
are often be discovered during the semen analysis showing the absence of sperm men was 36.62±4.5 years. The oligoasthenozoospermic sample had a mean count
in semen. Azoospermia is a heterogeneous disease with many histological phe- of 48.71±13.45 million/ml with motility of 35.13±16.86. The definition for poor
notypes. To date, several gene mutations, including TEX11, SYCP3, have been SCSA score was a DFI of >50% and HDS of >25 %. The outcomes were interpreted .
identified in azoospermia, which can explain the genetic causes of a small number Participants/materials, setting, methods: The study participants were
of azoospermia cases. divided into two groups. Group A (n=56) comprised of sample prepared by
Study design, size, duration: Whole-exome sequencing (WES) was per- Microfludics and Group B comprised of sample prepared by Sperm slow ( n=53).
formed with patient DNA. In Brief, genomic DNA was isolated from peripheral The stimulation protocols were short protocol with antagonist and recombinant
blood samples, and utilized for exon capture using the Agilent SureSelect Human hCG for trigger. All patients underwent ICSI. The outcomes in both groups were
All Exon V6 Kit and sequenced on the Illumina HiSeq X system. Candidate compared as mentioned for Clinical pregnancy, miscarriage rates and live birth rates.
pathogenic variant on the patient was validated by Sanger sequencing in the Main results and the role of chance: There were no statistically significant
patients’ parents as well as the normal controls. Then Knockout mice were used difference between Group A and Group B in patients who have low DFI value
to observe phenotypes. in SCSA test with respect to clinical pregnancy (p=0.112) and miscarriage rates
Participants/materials, setting, methods: A-30-year-old Han Chinese man (p=0.61), live birth (p=0.611), fertilization rates (p=0.185), and implantation
from a consanguineous family was diagnosed with infertility for 2 years, and no rates (p=0.623). This signifies that the MFSS and Spermslow gives better normal
fertility-related diseases were detected in his wife. The parents of the patient embryos fertilization and delivery rates equal as per primary study outcomes.
were also recruited. Whole-exome sequencing (WES) was performed with Limitations, reasons for caution: A larger sample size would add more
patient DNA. Immunofluorescence staining, Scanning electron microscopy (SEM) value to the study.
assay and transmission electron microscopy (TEM) assay were performed Wider implications of the findings: MFSS and SS have proven clinical effi-
according to a protocol described previously. The flow cytometric analysis and cacy in oligoasthenozoospermic males with poor SCSA scoring. We have
cell sorting were performed as previously described. achieved better clinical outcomes when compared to our retrospective results
Main results and the role of chance: To illuminate the genetic cause of using conventional sperm preparation methods in similar groups.
the azoospermia in this study, we performed Whole-exome sequencing Trial registration number: NA
(WES) was performed on the patient and a homozygous mutation
(g.17:64125864_64125941cnv, c.NA, p.NA) was identified in centrosomal pro-
tein 112kDa gene named CEP112, which is primarily expressed in the testis and P-087 Comparison between the sperm retrieval rate by micro-
belongs to the cell division control protein 42 effector protein family. The protein dissection testicular sperm extraction (mTESE) versus convention
encoded by this gene has putative coiled-coil domains and plays a role in sper- TESE (mTESE) combined with stereoscopic dissection
matogenesis. Our experimental observations on human subjects and mice sug- A. AlAhwany1, T. Alglely2, M. Ragab1, H. AlAhwany3
1
gested that CEP112 is involved in sperm flagellum structure and assembly. Cairo University, Department of Andrology, Cairo, Egypt ;

2
Nile IVF Center, Department of Embryology, Cairo, Egypt ; Study question: Does oral antioxidant supplementation for the male partner
3
Nile IVF Center, Department of Gynaecology, Cairo, Egypt improve clinical pregnancy rate in couples undergoing intracytoplasmic sperm
injection (ICSI) for male factor infertility?
Study question: Can conventional TESE (cTESE) combined with stereoscopic Summary answer: Oral antioxidant supplementation for the male partner
dissection achieve a sperm retrieval rate similar to Micro-dissection Testicular resulted in significantly increased clinical pregnancy rate in couples undergoing
Sperm Extraction (mTESE)? ICSI for male factor infertility versus no supplementation
Summary answer: Our study suggests that micro-TESE still has the highest What is known already: Reactive oxygen species (ROS)-mediated damage
recovery rate of sufficient sperms and good quality diatoms for immediate or to sperm contributes significantly to male factor infertility. ROS related injury
potential cryopreservation for ICSI. reduces fertilization potential and adversely affects sperm DNA integrity.
What is known already: Infertile patients with non-obstructive azoospermia Antioxidants act as free radical scavengers to protect spermatozoa against ROS
are treated by testicular sperm harvesting combined with ICSI. Micro-dissection induced damage. During ICSI, use of sperms which have been exposed to ROS
Testicular Sperm Extraction (mTESE) is considered the gold standard procedure mediated damage may affect the treatment outcome. Pre-treatment with anti-ox-
with the highest sperm retrieval rate compared with the conventional TESE. But idants may reduce ROS mediated sperm DNA damage. Currently, there is ambi-
mTESE cannot be widely offered in most infertility centres because of its high guity regarding role of anti-oxidants before ICSI in male factor infertility due to
cost, should be done by an expert well-trained surgeon and requires expensive conflicting results from earlier studies.
equipment. Laboratory stereoscopic testicular tissue dissection helps in identi- Study design, size, duration: This was an open label randomized trial con-
fying the dilated seminiferous tubules. In this study, we tested whether cTESE ducted at a tertiary level infertility clinic between Feb 2013-Jan 2019. The trial
combined by stereoscopic dissection can identify dilated seminiferous tubule and included 200 subfertile couples who were undergoing ICSI treatment for male
subsequent high sperm retrieval rate. factor infertility
Study design, size, duration: This is a prospective comparative study, com- Participants/materials, setting, methods: Couples were randomized into
paring the difference in sperm retrieval rate between Micro-dissection Testicular treatment arm (n = 100) and control arm (n = 100). In the treatment arm, the
Sperm Extraction (mTESE) (n=100) and Convention TESE (cTESE) using male partner received oral anti-oxidants (vitamin C, vitamin E and zinc) for three
Stereomicroscope (n=100), where Two hundred patients with NOA undergoing months just prior to ICSI cycle. In the control arm, no antioxidant was given to
ICSI and TESE were recruited between 2018 and 2019. the male partner. The primary outcome was clinical pregnancy rate and live birth
Participants/materials, setting, methods: Two hundred patients with and miscarriage rates were the secondary outcomes
NOA undergoing ICSI and TESE were recruited. Patients were counselled, Main results and the role of chance: A total of 200 randomized women
consented and randomly randomised. Convention TESE with additional were available for analysis. The clinical pregnancy per transfer was significantly
stereomicroscope dissection was applied to the first group (n=100), while higher following anti-oxidant therapy (54.7% vs.36.2% Odds ratio, OR: 2.1, 95%
micro-TESE was applied to the second group (n=100). Parameters such as Confidence Interval, CI 1.06-4.26) compared to no therapy. There was no sig-
patient’s average age, testicle size, follicle-stimulating hormone (FSH), lutein- nificant difference in live birth rate per transfer (39.1%, vs. 30.4 %; OR: 1.5, 95%
izing hormone (LH), testosterone levels, were analysed as predictors of CI 0.71-3.00) or miscarriage rate per pregnancy (25.7% vs. 8.0%; OR: 3.9, 95%
sperm recovery rate. CI 0.78-20.3) following antioxidant therapy compared to no therapy. However,
Main results and the role of chance: The average age of 100 men in the the intention to treat analysis did not show any significant difference in live birth
micro TESE was 35.74 ± 37.7, the size of the right testicle was 10.5 ± 4.5, the rate per woman randomized (25.0% vs.21.0%; OR: 1.3, CI: 0.65-2.43). The
size of the left testicle was 10.5 ± 4.3, the level of the hormone FSH recorded semen parameters of sperm concentration (18.2 (8.7,37.5) vs. 20.5 (8.1,52.5)
19.3 ± 13.9, and the hormone LH concentration was 17.1 ± 23.8, and the million; P= 0.86) , motility (34 (20,45) vs. 32.0 (18,45) %; P = 0.55) and mor-
testosterone hormone level was 3.4 ± 2.3. In conventional TESE group the phology (2.0 (1.39) vs. 2.2 (1.48); P = 0.63 ) did not show any significant improve-
average age of 100 men 35.17 ± 7.83 and the size of the right testicle 10.9 ± ment before and after anti-oxidant therapy.
4.6 and the size of the left testicle 11.1 ± 4.9 and the level of the hormone FSH Limitations, reasons for caution: Objective assessment of sperm DNA dam-
18.3 ± 10.4 and the hormone LH concentration 21.4 ± 44.1 and testosterone age was not done before and after the anti-oxidant supplementation. The study
level 2.7 ± 0.7. Micro-TESE was successful in obtaining sperm from the testicle duration got prolonged from two to five years due to slow recruitment. However,
in 43% (43/100) of cases. While patients with sperm retrieval in a conventional no major protocol changes were introduced during the study period
TESE manner the success rate was 36% (36/100) showing a statistically significant Wider implications of the findings: The preliminary result of the current
difference (P<0.05). study suggests that while there is an increase in clinical pregnancy rate following
Limitations, reasons for caution: 1) Larger (n) number of patients were anti-oxidant supplementation for the male partner in couples undergoing ICSI
needed. 2) Some of the patients already undergone a partial or conventional for male factor infertility compared to no supplementation, there is no improve-
TESE screening elsewhere. 3) This study was single-blinded and subjective to ment in live birth rate
the surgeon decision during the operation. Trial registration number: CTRI/2013/02/003431
Wider implications of the findings: Even though mTESE shows a higher
recovery rate of sufficient sperms for immediate or potential cryopreservation P-089 Effort to ameliorate sperm profile by folic acid-antioxidant
for ICSI, still, conventional TESE combined with stereoscopic dissection could combination therapy to evade single nucleotide polymorphisms of
be the method of choice of clinics because of its low cost and also giviving better methylenetetrahydrofolate reductase and methionine synthase in
results than partial or conventional TESE screening. idiopathic male infertility
Trial registration number: N/A R. Chattopadhyay1, S. Parvin2, S. Kalapahar2, P. Paladhi3, S. Dutta3,
S. Ghosh3, P. Chakraborty2, B. Chakraborty2
1
Institute of Reproductive Medicine, ART, Calcutta, India ;
P-088 Does oral antioxidant supplementation for the male 2
Institute of Reproductive Medicine, Department of Assisted Reproduction,
partner improve clinical pregnancy rate in couples undergoing
Calcutta, India ;
ICSI treatment for male factor infertility? A randomized 3
University of Calcutta, Department of Zoology, Kolkata, India
controlled trial
T. Joseph1, M. Mascarenhas2, R. Karuppusami3, M. Karthikeyan4, Study question: Does anti-oxidants and folate combination therapy influence
A.T. Kunjummen4, M.S. Kamath4 the association of single-nucleotide-polymorphisms (SNPs) in methylene-
1
Christian Medical College, Reproductive Medicine, Vellore, India ; tetrahydrofolate reductase (MTHFR) and/or methionine synthase (MS) in
2
Leeds Fertility -Leeds Teaching Hospitals NHS Trust-, Leeds Fertility-, Leeds-, hyperhomocysteinemic idiopathic male infertility (IMI)?
United Kingdom ; Summary answer: MTHFR polymorphism with (1298A>C) SNP is
3
Christian Medical College and Hospital, Biostatistics, Vellore, India ; associated with hyperhomocysteinemic IMI. Folate-antioxidant/s combination
4
Christian Medical College and Hospital, Reproductive Medicine, Vellore, therapy confers no significant improvement in sperm parameter/s in
India treated cohort.

What is known already: Abnormal germline DNA methylation has been Summary answer: Microfluidics sperm selection significantly increases the
proposed as possible mechanism compromising spermatogenesis in some men number of blastocysts and improves clinical pregnancy rate in egg-donation (ED)
with IMI. Dietary folate influence epigenetic modification/s during spermato- cycles and high SDF.
genesis. Recently, associations between four SNPs (MTHFR C677T, MTHFR What is known already: Previous studies have described a DNA fragmen-
A1298C, MS A2756G and MTRR A66G) in male infertility are widely studied tation negative effect on early embryo development. Concretely, high levels of
among several ethnicities; however, results remaining contradictory. Folic acid DNA fragmentation have been associated with embryo development arrest, as
supplementation usually been given to infertile men, a time when DNA meth- well as low implantation and pregnancy rates. Conventional methods used to
ylation patterns are being actively maintained and remodeled in male germ select sperm for ICSI, as swim-up or density gradients, require centrifugation. It
cells to prepare the epigenome for embryogenesis. It is therefore crucial to is well known that centrifugation forces increase Reactive Oxygen Species (ROS),
understand the possible impact of antioxidant with folic acid supplementa- one of the main promoter of SDF. Microfluidic systems don’t need centrifugation,
tion in IMI. consequently avoid ROS formation and could allow us to select sperm with
Study design, size, duration: Prospective study; azoospermic/ oligozoosper- better motility, morphology and lower SDF than conventional sperm selection
mic men with IMI (n=102; Group A) served as study cohort. Normozoospermic methods.
men (WHO criteria, 2010) were treated as controls (n=58; Group B). Patients Study design, size, duration: Preliminary retrospective cohort study
are supplemented with folic acid (5mg/day) and a combination of antioxidant/s including until now 63 cycles of ICSI-ED and sperm with high SDF (>30%)
(glutathione: 400 mg/day; vitamin E: 200mg/day) for 3 months. Peripheral blood between January 2017 and October 2019. In all the cycles single fresh blastocyst
samples are collected before and after treatment. The study was conducted transfer were performed. SDF was evaluated by SCD (Sperm Chromatin
from October 2018 to September 2019 and approved by Institutional Ethics Dispersion) test.
Committee of the Institute. Participants/materials, setting, methods: Two groups of cycles were
Participants/materials, setting, methods: Plasma levels of homocysteine, established according to the sperm selection method used for ICSI: microfluidic
vitamin B12, folate are determined by chemiluminescence. PCR-RFLP (poly- chip (G1; n=30) and density gradients (G2; n=33). The groups were homoge-
merase chain reaction- restriction fragment length polymorphism) and Sanger neous in terms of the donor age, mean number of mature oocytes collected
sequencing by MiSeq(Illumina) was performed to detect SNPs in homocysteine and embryos transferred. No differences were found between groups in the
metabolism (MTHFR 677C>T; MTHFR1298A>C; MS2756A>G). The effect of SCD test using T-student statistics (G1: 45.6±14.5%; G2: 39.0±11.7%; p=0.05).
drug supplementation on sperm parameters and apoptosis status was evaluated The results were analyzed using Fisher test.
by Makler’s chamber and flow cytometry respectively. Statistical significance was Main results and the role of chance: The two considered groups of this
set at p < 0.05 as evaluated by student’s T-test. study were diagnosed with pathological percentage of sperm DNA fragmenta-
Main results and the role of chance: In comparison with normozoospermic tion. Fertilization rate was equivalent (p=0.07) between G1 (172/216; 79.6%)
spermatozoa, there was a significant decrease (P < 0.03) in motility and velocity and G2 (169/234; 72.2%). However, good quality blastocyst rate (GQB; ≥3BB
parameters and increase in Caspase+/propidium iodide- (PI-) cells (P < 0.04) Gardner score) of G1 (84/172; 48.8%) was significantly higher (p<0.05) than
in oligospermic patients. Increased (p<0.01) serum homocysteine levels in G2 (64/169; 37.9%), as well as pregnancy rate (G1: 90.0%; G2: 54.55%).
(mmol/L) was documented in Group A with no other changes in blood profile. Additionally, clinical pregnancy rates were higher in G1 (63.3%) than in G2
MTHFR 1298A>C gene polymorphism was documented in 49/102 (48.03%) (48.5%), not being statistically significant. Perhaps, with a higher sample could
in contrast to 11 samples from Group B. 7 participants (14.28%) were homozy- help to asses if the tendency of this increase keeps on the clinical pregnancy
gous for the MTHFR TT variant while 13 (26.53%) and 10 (20.41%) were CC rate. Our results show that processing sperm samples with a microfluidic sperm
and CT genotypes, respectively (Group A). 29 cases exhibited MS 2756A>G sorting device could be a more efficient alternative to density gradients in cycles
polymorphism in contrast to Group B (n=11). Serum folate concentrations where patients were diagnosed with increased SDF. This method allows the
(nmol/L) increased significantly (28 + 2.72 vs. 42 + 3.81; p<0.01) after supple- selection of sperm with a low DNA fragmentation promoting the formation of
mentation with no significant improvements in the three MTHFR genotypes for a higher number of GQB to transfer and/or vitrify.
any of the semen parameters (count, motility). Changes in semen volume, sperm Limitations, reasons for caution: This is a preliminary retrospective study,
motility and sperm chromatin integrity (assessed by sperm chromatin structure so the main limitation is the small sample size achieved until now. These results
assay or SCSA, and reported as % high DNA stainability) was not significant. should also be confirmed by further randomized prospective studies.
Sperm DNA integrity, assessed by SCSA and reported as % DNA fragmentation Wider implications of the findings: Our findings suggest that the application
index (% DFI), demonstrated a trend toward improvement (t-test P = 0.057) in of microfluidic systems could be improving the sperm selection with conserved
oligospermic men after combination regimen. DNA integrity. In this sense, microfluidic systems increase significantly the num-
Limitations, reasons for caution: The global erasure of parental DNA meth- ber of GQB and therefore could potentially improve the cumulative pregnancy
ylation during early stages of embryogenesis is a mechanism for reducing the risk rates and the efficiency of the cycle.
of altered methylation. Thus, recommendations for men with IMI to receive com- Trial registration number: not applicable
bination of antioxidant-folic acid therapy should be taken with caution..
Wider implications of the findings: A personalized approach to combina-
tion of antioxidant-folic acid therapy may be needed, based on factors like genetic P-091 Fertile®ChiP-ZyMōt improves ICSI clinical outcomes in
background of the recipients. Use of combination treatment did not significantly patients with high values of sperm double-strand breaks using
improve semen quality probably because altered methylation may elude global oocytes from both patients and donors.
erasure mechanisms and limit the progress to next stages of mitochondrial S. Camacho Fernandez-Pacheco1, C. Urda-Muñoz1, S. Lara-
development. Cerrillo2, T. Lacruz-Ruiz2, C. Rosado-Iglesias2, M. De la Casa
Trial registration number: NA Heras1, V. Badajoz-Liébano1, J. Gijón de la Santa1, A. García-Peiró2
1
GINEFIV, Assisted Reproduction, Madrid, Spain ;
P-090 Sperm selection using microfluidic sorting chips in 2
CIMAB- Barcelona Male Infertility Centre, Research & Development,
patients with high DNA fragmentation improves clinical outcomes Sant Quirze del Vallés - Barcelona, Spain
in egg-donor cycles
A. Farreras Ayestaran1, A. Munuera1, B. Freijomil1, S. Novo1, Study question: Are clinical outcomes on ICSI cycles improved by the use of
À. Garcia-Faura1, B. Marquès1, F. Garcia1, C. Castelló1, Fertile®ChiP-ZyMōt for sperm selection using oocytes from both patients
M. Lopez-Teijón1 and donors?
1
Institut Marques, Reproductive Medicine Service, Barcelona, Spain Summary answer: Fertile®ChiP-ZyMōt improved biochemical and clinical
pregnancy rates and reduced miscarriage rate in ICSI cycles from patients with
Study question: Can sperm selection using microfluidic sorting chips altered values of dsSDF.
improve reproductive outcomes in patients with high sperm DNA fragmenta- What is known already: Delays in embryo kinetics, implantation failures in
tion (SDF)? ICSI treatments and recurrent miscarriages have been associated to high values

of double-strand breaks (DSB) in sperm. Moreover, a recent study showed that ameliorate SCF. Because the predictability of overall gamete competence through
DSB are not reduced during the sperm selection of an ICSI cycle. semen analysis has been debatable, new bioassays have been proposed. By
The Fertile®ChiP-ZyMōt is a new method for sperm selection based on surveying localization patterns of ganglioside M1, a key regulator of capacitation
microfluidic properties that showed to reduce the presence of DSB in the sperm and acrosomal reaction, we have been able to quantify subtle sperm function
sample. In this sense, the specific reduction of DSB using Fertile®ChiP-ZyMōt and predict probability of generating a pregnancy (PGP).
could improve clinical outcomes after ICSI treatments. Study design, size, duration: In the past 12 months, fresh ejaculates were
Study design, size, duration: This retrospective study included 78 ICSI obtained from 6 consenting men who were initially diagnosed with varicocele
cycles from January 2018 to March 2019. Three groups were classified attending of grade 2 or higher and then treated by varicocelectomy. Ejaculates were
on the origin of the oocytes and the sperm selection method: Control group = obtained again at least 3 months post-surgery to allow a full cycle of spermato-
oocytes from patients and density gradients for sperm selection (n=16); genesis. Semen parameters, SCF, CaP-ScoreTM, and PGP were determined in a
Group 1 = oocytes from patients and Fertile®ChiP-ZyMōt for sperm selection blind fashion and compared among pre- and post-operative specimens.
(n=22); and Group 2 = oocytes from donors and Fertile®ChiP-ZyMōt for sperm Participants/materials, setting, methods: Semen analysis was performed
selection (n=11). All male patients presented high values of DSB. on pre-/post-operative ejaculates. Varicocele was evaluated by physical exam
Participants/materials, setting, methods: Patients included in the study in standing position. Capacitation was measured by CaP-Score™ (Androvia
presented high values of DSB analyzed though the Neutral CometFertility assay LifeSciences) with normal thresholds of >27.6%. A corresponding PGP was
(CIMAB, Spain). Sperm selection was performed using conventional Density previously established by analyzing pregnancy outcomes from fertile and infertile
Gradients (Sperm Grad, Vitrolife, Sweden) (Control group) or the Fertile®ChiP- men who completed 3 IUI cycles with a normal threshold of >32.7%. Total
ZyMōt (DxNow, USA) (Groups 1 and 2). ICSI cycles were performed using capacitated spermatozoa were quantified by volume x concentration x CaP-
oocytes from patients or donors and clinical outcomes were studied. Results ScoreÔ. SCF was assessed using TUNEL with normal threshold of <15%.
were compared between groups, being the statistical significance α = 0,05. Main results and the role of chance: Men (n=6) with grade 2 varicocele
Main results and the role of chance: Women’s age was significantly lower or higher aiming to procreate underwent microsurgical varicocelectomy without
in Group 2 (26.7 ± 4.28) compared to the Control group (35.67 ± 3.43) and any post-operative complication. Semen parameters such as volume and normal
Group 1 (36.17 ± 3.84), p<0.01. morphology that were assessed before surgery did not improve. For all men,
Fecundation rates were slightly higher, even not significantly, in Group 1 sperm concentration was initially 35.0 ± 28.0 x106/ml and became 57.3 ± 29.5
(0.53 ± 0.27) and Group 2 (0.63 ± 0.21) compared to the Control group x106/ml (P < 0.05). Sperm capacitation function, total capacitated spermatozoa,
(0.51 ± 0.29), p=0.80. Biochemical pregnancy was significantly higher in and PGP at baseline were 25.4 ± 3.4%, 18.4 ± 13.3 x106, and 29.3 ± 5.1%,
Group 1 (12/22 (54.5%)) and Group 2 (7/11 (63.6%)), compared to the respectively. After 3 months of post-operative recovery, sperm capacitation,
Control group (3/16 (18.8%), p=0.03. Clinical pregnancy was significantly higher number of total capacitated spermatozoa, and PGP significantly increased to
in Group 1 (10/22 (45.5%)) and Group 2 (7/11 (63.6%)), compared to the 32.0 ± 4.1%, 33.2 ± 16.0 x106, and 41.0 ± 7.2% (P < 0.05), respectively. Genomic
Control group (2/16 (12.5%), p=0.01. Miscarriage rates were significantly higher integrity as measured by SCF was originally above threshold at 17.9 ± 6.6% and
in the Control group (2/2 (100%)) than in Group 1 (2/10 (20%)) and Group 2 significantly decreased to an average of 11.0 ± 4.8% (P < 0.01) after varicocele
(0/7 (0%)), p=0.02. correction.
Even Group 2 showed the best results, there were no significant differences Limitations, reasons for caution: These findings support a beneficial effect
compared to Group 1 for biochemical and clinical pregnancies and miscarriage of varicocele correction. However, the improvement of sperm concentration
rates (p=0.618; p=0.325 and p=0.418, respectively). together with functional assay, as well as the amelioration of genomic integrity,
Compared to density gradients, the use of the Fertile®ChiP-ZyMōt in ICSI needs to be confirmed by testing the embryonic developmental competence of
cycles improved biochemical pregnancy rates x1,65 using oocytes from patients the male gamete.
(p=0,033) and x2,32 using oocytes from donors (p=0,054). Fertile®ChiP-ZyMōt Wider implications of the findings: This analysis confirms the beneficial
also improved clinical pregnancy rates x1,71 using oocytes from patients and effect of varicocelectomy on at least one semen parameter. The utilization of a
x2,68 oocytes from donors (p=0,036). functional assay and the determination of genomic integrity by SCF provide
Limitations, reasons for caution: Despite biochemical and clinical pregnan- information on an individual’s ability to spontaneously reproduce or can help
cies presented significant better results using the Fertile®ChiP-ZyMōt, the guide one toward the preferential method of assisted reproduction.
increase in the fecundation rate was not significant. More studies analysing a Trial registration number: not applicable
larger number of ICSI cycles are needed to confirm these findings.
Wider implications of the findings: The use of the Fertile®ChiP-ZyMōt in
P-093 Andropenia precedes AML diagnosis and is associated with
ICSI cycles to treat high values of DSB in sperm increase biochemical and clinical
fatty marrow
pregnancy rates. These increases are even more important when oocytes are
from a donor. K. Shlush1, N. Zioni2, N. Kaushansky3, G. Oron4, L. Shlush3
Trial registration number: Does not apply. 1
fertility clinic Rabin Medical center, Obstetric and Gynecology, Herzliy, Israel ;
2
Wiezmann institute of science, Immunology, Rehovot, Israel ;
P-092 Varicocelectomy corrects sperm capacitation functions and 3
Weizmann institute of science, Immunology, Rehovot, Israel ; 4Rabin medical
enhances sperm genomic integrity center, Obgyn, Petah Tikva, Israel
M. Haddad1, P. Xie1, A. Parrella1, Z. Rosenwaks1, J. Kashanian2,
G. Palermo1 Study question: What are the effects of andropenia on leukemia and
1
Weill Cornell Medicine, The Ronald O. Perelman and Claudia Cohen Center for bone marrow?
Reproductive Medicine, New York, U.S.A. ; Summary answer: Many years before AML diagnosis testosterone levels are
2
Weill Cornell Medicine, Department of Urology, New York, U.S.A. decreasing. Low testosterone levels after castration cause fatty bone marrow
in mice.
Study question: To determine whether varicocelectomy improves spermato- What is known already: As human age they accumulate somatic mutations.
genesis by enhancing semen parameters, sperm capacitation, sperm genomic These early mutations preleukemic mutations (pLMs) accumulate and eventually
integrity, and overall embryo developmental competence of the male gamete. lead to different myeloid malignancies. Specific pLMs are more common among
Summary answer: Correction of grade 2 or higher varicocele enhanced male elderly males. Furthermore myeloid malignancies are more common among
gamete production, ameliorated sperm chromatin fragmentation (SCF), and males. While large proportion of the population carry pLMs only a small pro-
amplified male gamete capacitation. portion will develop myeloid malignancies. Understanding the mechanisms con-
What is known already: Varicocele is known to induce oxidative damage to tributing to the progression to leukemia is of great importance. One factor
the male reproductive system, affecting semen parameters, sperm chromatin possibly contributing to leukemia evolution might be the accumulation of fat in
integrity, and function. Varicocelectomy has been proposed to enhance typical the bone marrow (BM) with age. A correlation was found between increased
parameters measured in semen analysis, such as concentration and motility, and BM fat and low testosterone levels.

Study design, size, duration: The electronic health records of 4.5 million years. These cycles had a 72.7% fertilization rate (4935/6789), with a 12.0%
Israelis over 15 years were analyzed to identify AML cases (N=987). We iden- implantation rate (112/935) and a 21.7% clinical pregnancy rate (CPR; 92/423).
tified all individuals in this group (pre-AML) and studied their blood counts and Fourteen of these pregnancies resulted in miscarriage (15.2%).
testosterone levels before AML diagnosis and compared it to 500,000 aged A total of 146 couples with an abnormal DSB level were treated by 259 ICSI
match individuals. cycles. The maternal age was comparable between the two groups at 37.3±5
Participants/materials, setting, methods: To study the effect of andro- years, while male partners were much older at 41.4±8 years of age (P<0.01).
penia on BM fat mice were castrated and BM fat was measured after two month This group had a comparable fertilization rate of 70.1% (1739/2481), an implan-
by lipitox staining of CD45 negative cells from the BM. in each group 10 mice tation rate of 10.8% (49/455), and a CPR of 22.9% (40/175). However, these
were used. couples were significantly more likely to experience a miscarriage, which
Main results and the role of chance: Testosterone levels were available occurred at a rate of 30.0% (12/40; P<0.05).
for 76 pre-AML cases on average 4 years before AML diagnosis. Significantly Limitations, reasons for caution: This is a retrospective observation in
reduced testosterone levels was present already four years before AML diagnosis couples with a history of compromised outcomes with assisted reproductive
and remained significantly lower until diagnosis. Low testosterone levels were techniques. It would be ideal to reproduce these findings in a prospective manner
achived after castration. Two month after castration a significant increase in the using solely the neutral Comet assay.
BM fat was noticed with 30% more fat in the castrated mice compared to aged Wider implications of the findings: DSBs in human spermatozoa, which
matched controls. have been associated with structural chromosomal abnormalities, may provide
Limitations, reasons for caution: It is not clear why fatty marrow is pro- information on embryonic developmental potential. Utilizing an assay that
moting leukemia among males. Also the number of pre-AML cases with testos- screens exclusively for DSBs may be more beneficial than total SCF in terms of
terone levels was low and a sampling bias is possible. predicting a couple’s ability to successfully reproduce.
Wider implications of the findings: The effect of low testosterone levels Trial registration number: not applicable
on the BM and leukemia evolution remains unknown.In the current study we
provide evidence that fatty marrow can be induced by castration. Furthermore, P-095 The male factor – impact of semen quality on the
low testosterone levels precede AML. Better understanding the mechanisms insemination result in women with endometriosis
leading to increased leukemia might help in preventing this devastating disease. Z. Kaszás1, A. Nemes1, Á. Anikó1, D. Pócsi1, Á. Murber1,
Trial registration number: NA J. Urbancsek1, P. Fancsovits1
1
Division of Assisted Reproduction Semmelweis University, Department of
Gynecology and Obstetrics, Budapest, Hungary
P-094 The impact of double-stranded DNA breaks in human
spermatozoa on embryo implantation Study question: Do we need higher sperm count to achieve a successful
L. Zimmerman1, D. Keating1, A. Parrella1, Z. Rosenwaks1, insemination cycle with endometriosis patients?
G. Palermo1 Summary answer: Our results suggest that sperm viability and/or its lifetime
1
Weill Cornell Medicine, The Ronald O. Perelman and Claudia Cohen Center for is effected by endometriosis.
Reproductive Medicine, New York, U.S.A. What is known already: Little is known about the effect of endometriosis
on the outcome of insemination (IUI). In the first 6 month after surgery women
Study question: Is there a correlation between double-stranded DNA breaks with minimum to mild endometriosis have the same chance to get pregnant with
(DSBs) in human spermatozoa and ICSI outcome? IUI as couples with unknown infertility. The ESHRE guideline recommend insem-
Summary answer: An increased proportion of DSBs in men with normal ination as treatment for infertility with minimum to mild endometriosis, but this
semen analyses impairs embryo implantation. statement has never been linked to any study with male fertility factors. We
What is known already: Total sperm chromatin fragmentation (SCF) in men don’t know yet whether sperm react to the intrauterine environment effected
with normal semen parameters has been linked to a couple’s ability to successfully by endometriosis, but possibly endometriosis may influence the viability of the
conceive. Recent studies have challenged this, asserting that DSBs, rather than motile sperm.
single-stranded breaks (SSBs), lead to compromised embryo development. Study design, size, duration: This is a retrospective study performed
There are many techniques to assess the genomic integrity of spermatozoa by between 2010 January and 2019 May at university settings. Patients treated with
measuring indistinct SSBs and DSBs, such as the sperm chromatin structure assay, endometriosis and infertility were selected from our institute database such as
terminal deoxynucleotidyl dUTP transferase nick-end labeling (TUNEL), sperm diagnosis of endometriosis, date of previous surgeries, date of insemination and
chromatin dispersion, and alkaline Comet. However, the only method to spe- total motile sperm count (M/ml) after density gradient centrifugation.
cifically detect double-stranded DNA integrity is the neutral Comet test. Participants/materials, setting, methods: Data of IUI treatments were ana-
Study design, size, duration: In the course of 12 months, we carried lyzed in two groups. Endometriosis group: cycles with the diagnosis of previously
out a pilot study to assess exclusively DSBs in spermatozoa by neutral Comet surgically proven endometriosis. Control group: cycles with “unknown infertility”.
assay. There was a linear correlation between Comet and TUNEL (R2=0.96), Endometriosis group was further divided into two subgroups according to
which allowed us to extrapolate the proportion of DSBs in all men who had the time elapsed between surgery and IUI. Subgroup A: IUI ≤6 months after
been screened by TUNEL assay. This value was correlated with ICSI surgery; Subgroup B: IUI in 7-18 months after surgery.
outcome. Total motile sperm count and pregnancy rates were compared in Endometriosis
Participants/materials, setting, methods: A total of 523 men with poor and Control groups.
assisted reproductive outcomes were screened by TUNEL, with ≥500 sperma- Main results and the role of chance: The pregnancy rate of insemination
tozoa assessed per sample and a normal threshold of ≤15%. Neutral comet was cycles of women with endometriosis (23/341; 6,7%) was similar to the Control
performed using a modified in-house protocol with commercially available mate- group (9/171;5,2%) (P=0,513).
rials, with ≥200 spermatozoa assessed, and a normal threshold of ≤3%. All men Subgroup analysis of endometriosis patients showed similar pregnancy rates
underwent ICSI, performed in the standard fashion, with their female partners. in Subgroup A (4/69; 5,8%) and Subgroup B (10/129;7,7%) (P=0,609).
Fertilization, implantation, and pregnancy outcomes were compared according The progressive motile sperm count in the pregnant IUI cycles was 34,9±
to DSBs. 18,4 M/ml, in the Endometriosis group, while 25,8±14,6 M/ml in the Control
Main results and the role of chance: The pilot study yielded an average group. The difference was not significant (P=0,304).
SCF by TUNEL of 11.3±6% and a DSB of 2.2±3% by neutral Comet in a linear The mean of 22,5 ±9,2 M/ml of progressive sperm count was found
relationship (R2=0.96). This allowed us to extrapolate DSB rates in 523 normo- in pregnancy in Subgroup A. That is similar to the Control group’s (25,8±
zoospermic men (2.6±1 mL volume, 42.3±34x106 concentration, 43.1±10% 14,6 M/ml) result.
motility, and 4.1±1% normal morphology) who had undergone TUNEL. However, that of Subgroup B higher sperm number was found in pregnancy
A total of 377 couples with normal DSB rates underwent 736 ICSI cycles. cycles (37,3 ±11,3 M/ml), but the difference was not significant (P=0,327) com-
The average maternal age was 37.0±4 years, and the paternal age was 38.5±6 pared to the Control group.

It seems that within 6 months following the endometriosis surgery patients Wider implications of the findings: Sperm DNA damage independent of
need comparable amount of motile sperm to get pregnant by insemination as its cause, may affect the sperm quality and have implications on patient’s fertility
the patients with unknown infertility. After this half year period, patients need potential. Therefore, we recommended the evaluation of sperm DNA status in
higher sperm count to get pregnant. These differences can be explained with infertile patients with a high clinical varicocele grade in order to optimize sperm
the recurrent appear of endometriosis changing the pelvic environment. quality and pregnancies rates in this population.
Limitations, reasons for caution: The retrospective study design limits our Trial registration number: not applicable
chance to make a strong conclusion. Our small sample number in the subgroups
also limits our chance to find significant differences. P-097 Male lifestyle and pathological conditions associated with
Wider implications of the findings: Our results could also confirm the couple infertility in a referral center in Italy: a cross-sectional
ESHRE guideline regarding the necessity of insemination during the first 6 months study of 13452 male patients
after the endometriosis surgery. We plan to further explore this field, to find M. Dal Canto1, C. Signorini2, L. Mura1, S. Casellato3,
out what kind of biochemical milieu could influence the need for higher sperm L. Carmignani2, M.M. Renzini1, J. Buratini1,4, S. Maruccia3
count to reach pregnancy. 1
Biogenesi - Reproductive Medicine Centre, Istituti Clinici Zucchi, Monza, Italy ;
U.O. Urologia IRCSS Policlinico San Donato, Milano, Italy ;
3
U.O. Urologia Istituti Clinici Zucchi, Monza, Italy ;
P-096 Increased DNA damage in infertile patients with high 4
Sao Paulo State University, Institute of Biosciences, Department of Physiology
grading varicocele
S. Jellad1, F. Hammami1, A. Khalbous2, M. Cbibani3, R. Rachdi3 Study question: Is overall male health associated with male fertility as assessed
1
Miitary Hospital of Tunis- Faculty of Medicine of Tunis, laboratory of by semen analysis in couples undergoing subfertility treatment?
Reproductive Biology- Unit of Assisted Medical Procreation, Tunis, Tunisia ; Summary answer: Oncologic, cardiovascular, metabolic and allergic diseases
2
Miitary Hospital of Tunis, laboratory of Reproductive Biology- Unit of Assisted are more frequent in infertile men with impaired semen analysis than in those
Medical Procreation, Tunis, Tunisia ; with normal semen parameters.
3
Miitary Hospital of Tunis- Faculty of Medicine of Tunis, Unit of Assisted Medical What is known already: Current literature suggests an association between
Procreation, Tunis, Tunisia male infertility and a wide range of other comorbid conditions, ranging from
oncologic, cardiovascular, autoimmune, and metabolic disorders. The exact
Study question: This study aims’ at evaluating the impact of clinical varicocele nature of this putative association remains somewhat unclear, although hypoth-
on sperm nuclear DNA quality and standard semen parameters? esized mechanisms linking men’s health and fertility include genetic, develop-
Summary answer: Clinical varicocele generates a significant increase of sperm mental, and lifestyle-based factors. The male population seeking for fertility
abnormalities and DNA damage and these changes are positively correlated treatment constitutes an extremely valuable opportunity to clarify this issue, but
with varicocele grade. have not been sufficiently explored in previous studies with this aim.
What is known already: Varicocele is thought to progressively reduce sper- Study design, size, duration: Sociodemographic, health, lifestyle and recre-
matogenesis via elevated intratesticular temperature and altered testicular blood ational data from 13452 male patients seeking for couple infertility treatment
flow. The decreased supply of oxygenated blood and nutrients to the sperm were collected at a single referral centre between 01/2013 and 12/2019.
secretion sites reduces sperm quality and quantity, and consequently, their fertility Frequencies of diseases and lifestyle/recreational habits impacting on health
capacity. Although a cause-effect relationship is not established, multiple reviews (smoking, use of alcohol and other drugs of abuse) were compared between
conclude that there is indeed evident association between varicocele and male patients with different fertility as assessed by semen analysis.
increased DNA fragmentation Participants/materials, setting, methods: This is a retrospective study
Study design, size, duration: The study included 45 men assessed by our utilizing data collected from 13452 male partners of couples seeking for infertility
laboratory of reproductive biology of military hospital of Tunis during 6 months. treatment at our clinic. Treatments included 1091 IUI, 7443 ICSI and 1348 IVF.
A prospective study was designed involving one control group of men with The WHO 2010 referral criteria were used to categorized semen parameters
unknown fertility and normal semen parameters (n = 10) and one group of as pathological or normal. Descriptive statistics was applied to describe the
patients with clinically diagnosed varicocele and infertility (n=30). whole cohort. Differences in frequencies were assessed with the Chi Square test.
Participants/materials, setting, methods: Our prospective study involved Main results and the role of chance: The median age was 41.5 years (95%
30 infertile patients with clinical varicocele and 15 control patients referred to CI ±0,09). 8209 (61.0%) men had pathological semen parameters (any type),
our laboratory for routine spermiological exploration). Men with azoospermia, while 5243 (39.0%) individuals presented normal semen parameters. Among
severe oligozoospermia or leucocytospermia were excluded from the study. patients with pathological semen, 3666 (44.6%) had a sperm concentration
Spermograms were performed and analyzed according to WHO guidelines 2010. <15mln/ml, 3689 (44.9%) had a total sperm count <39 mln, 4695 (57.1%) had
The DNA fragmentation was detected by the terminal desoxynucleotidyl trans- a motility <32%, and 6764 (82,3%) men had a normal morphology <4%. In terms
ferase-mediated deoxyuridine triphosphate biotin nick-end labeling of health comorbidities, patients with abnormal semen parameters more fre-
(TUNEL) assay. quently reported a history of hypertension [941 (11.4%) vs. 234 (4.4%)], dia-
Main results and the role of chance: The average age of the patients at betes mellitus [804 (9.7%) vs. 178 (3.3%)] and cardiovascular diseases [613
the time of diagnosis was 26 ± 4 years (19-35) and 35 ± 7 years (27-47) for the (7.4%) vs. 160 (3.0%)], than those with normal semen analysis (p<0.001). Testis
control group. The DNA fragmentation index was significantly higher in patients cancer was more frequently found in men with abnormal semen parameters
with clinical varicocele compared to controls (13.3±3.4 % versus 6.1±2.5%, [140 (1.7%) vs. 24 (0,4%), p<0.001]. Men with impaired semen parameters were
p=0.0001). In addition, the DFI was positively and significantly correlated with more frequently active smokers [3336 (40.6%) vs. 764 (14.5%)] and alcohol
the degree of severity of varicocele thus the DFI was 15.24±1.9% in patients consumers [208 (2.5%) vs. 15 (0.2%)] than those with normal semen parameters
with grade 3 versus 12.92±3.5 % in those with grade 2 (p<0.0001). However, (all p<0.001), while the use of substances of abuse was similar among groups.
an abnormality of at least one of the semen parameters was found in 90% of A history of allergic diseases was more frequently found in men with impaired
varicocele patients, and all semen characteristics such as sperm count, vitality, semen analysis [2031 (24.7%) vs. 477 (9.0%); p<0.001].
mobility and typical forms were decreased compared to the controls. Limitations, reasons for caution: The retrospective nature of the study.
Furthermore, statistically significant negative correlations were noted between Wider implications of the findings: Our data contribute to a better comprehen-
sperm DNA fragmentation index and sperm concentration (p=0.0001), motility sion of the association between men’s health and fertility, while favouring early diagnosis
(p=0.03), and normal sperm morphology (p=0.03). of important male diseases and medical counselling during subfertility treatment.
Limitations, reasons for caution: Although considered as referent technique Trial registration number: Not applicable
the TUNEL assay has certain limits as it doesn’t allow differentiating between
normal and pathological DNA ruptures. Further investigations are also needed P-098 Improved classification of testicular histopathology to
to reveal the cause and effect relationship between the increase of DNA frag- promote uniform diagnosis and discovery of genetic causes for
mentation levels and clinical varicocele, as well as the underlying mechanisms. male infertility

G. Van der Heijden1, M. Wyrwoll2, L. Ramos1, J. Veltman3, 1

Amsterdam UMC- University of Amsterdam, Centre for Reproductive Medicine-
F. Tüttelmann4 Amsterdam Reproduction & Development Institute, Amsterdam, The Netherlands ;
2
1
Radboud University Medical Center, Gynaecology, Nijmegen, The Netherlands ; Monash University, Department of Obstetrics and Gynaecology, Melbourne,
2
University of Münster, Department of Clinical and Surgical Andrology- Centre of Australia ;
3
Reproductive Medicine and Andrology, Münster, Germany ; Nij Barrahûs, Fertility clinic, Wolvega, The Netherlands ;
4
3
Newcastle University, Institute of Genetic Medicine, Newcastle, United Kingdom ; University hospital Brussels, Centre for Reproductive Medicine, Brussels, Belgium ;
5
4
University of Münster, Institute of Human Genetics, Münster, Germany Rijnstate, Department of Clinical chemistry and haematology, Arnhem, The
Netherlands ;
6
Study question: Can a descriptive framework for testicular histopathology be Rijnstate, Department of Obstetrics and Gynaecology, Arnhem, The Netherlands ;
7
designed that identifies phenotypically unique cases and facilitates research on Isala, Department of Obstetrics and Gynaecology, Zwolle, The Netherlands ;
8
the genetic causes of male infertility? Vivaneo Medisch Centrum Kinderwens, Fertility Clinic, Leiderdorp, The
Summary answer: We propose an organized nomenclature for testis histo- Netherlands
pathology that makes use of clearly defined rules and definitions, describing
certain patterns of disruption of spermatogenesis. Study question: Are six cycles of ICI non-inferior to six cycles of IUI in donor
What is known already: With the availability of Next Generation Sequencing sperm treatment in the natural cycle in terms of ongoing pregnancy?
the genetic causes of male infertility are increasingly being chartered. A genetic Summary answer: In donor sperm treatment in the natural cycle in terms of
diagnosis combined with a clear unambiguous description of accompanying testis ongoing pregnancy rate, we could not demonstrate non-inferiority of ICI as
histology will hugely benefit interpretation of the function of identified genes. compared to IUI.
However, a universal ontology for testis histopathology is lacking. The Human What is known already: Both ICI and IUI in the natural cycle are performed as
Phenotype Ontology (HPO) is a formal ontology, which describes phenotypes first line treatments in women who are eligible for donor sperm treatment. IUI is
encountered in human diseases. The HPO allows phenotype driven differential more costly than ICI, due to the involvement of sperm processing.
diagnostics in clinical routine and research. The current hierarchy of terms High quality data on the effectiveness of ICI versus IUI in the natural cycle in terms
regarding azoospermia, is not a systematical one. Moreover, the majority of of ongoing pregnancy are lacking. A large retrospective cohort study performed in
testicular phenotypes are not annotated yet. the Netherlands suggested that ICI and IUI in the natural cycle in donor sperm treat-
Study design, size, duration: We aimed to design a framework that is easy ment resulted in comparable ongoing pregnancy rates of 40% after six cycles.
to understand and straightforward in its application but does contain most rel- Study design, size, duration: We performed a multicenter, non-blinded,
evant information to study the impact of an identified mutation on non-inferiority randomized controlled trial in six fertility clinics in the Netherlands
spermatogenesis. and Belgium. Women were allocated to receive either ICI or IUI in a natural
Participants/materials, setting, methods: The ontology classification is cycle during six cycles. Based on the retrospective cohort study we assumed a
based on qualitative and quantitative histology description of testicular samples live birth rate of 40% after six cycles of IUI. To assess a non-inferiority margin
from azoospermic samples (>1000) in our centers. For the differentiation of 12%, we needed to include 416 women.
between the classification of hypospermatogenesis and complete spermatogen- Participants/materials, setting, methods: All women scheduled for donor
esis, n=117 testicular biopsies post vasectomy cases were used. sperm treatment were eligible, regardless of the indication for treatment. The
Main results and the role of chance: Building on what has been published primary outcome was ongoing pregnancy within eight months after randomisa-
previously we propose a framework for testis histopathology based on the tion leading to a live birth. Secondary outcomes were multiple pregnancy, mis-
presence/absence of germ cells and/or Sertoli cells. Our framework has five carriage, time to ongoing pregnancy and pregnancy complications.
main categories. In addition to Sertoli cell only phenotype we add the classi- We calculated relative risks (RR) and risk difference (RD) and 95% CI. We
fication of tubular shadows, when also Sertoli cells are absent. When germ analysed the data both on an intention to treat and per protocol basis.
cells are present, their stage of arrest, numbers and distribution can inform Main results and the role of chance: Between June 2014 and February
us about the effect of the causal mutation. We therefore propose to distinguish 2019, we included 419 women, of whom 211 women were randomly allocated
Germ Cell Arrest (GCA), in which no spermatozoa are observed, from hypo- to ICI and 208 to IUI.
spermatogenesis (HS). Both categories are sub-divided to obtain information Women’s age was on average 34 years (SD ± 4) in both groups.
on the timing of the arrest (GCA) or the severity of overall loss of spermato- Of the 419 women included, ongoing pregnancy occurred in 52 women (25%)
genesis (HS). To establish a lower threshold for complete spermatogenesis in the ICI group and 77 women (37%) in the IUI group (RR 0.67, 95% CI 0.50
we determined the number of tubules that contain elongating spermatids (ES) to 0.89 and RD -0.12, 95% CI -0.21 to -0.04). Live birth occurred in 51 women
in a collection of samples of men with an obstructive azoospermia due to a (25%) in the ICI group and 76 women (37%) in the IUI group (RR 0.67, 95% CI
previous vasectomy (N=117). The majority of samples contained ES in ≥60% 0.50 to 0.91).
of their tubules, suggesting 50% to be a reasonable cut-off to distinguish In the per protocol analysis we included 193 women in the ICI group and 193
hypospermatogenesis from complete spermatogenesis. We conclude that women in the IUI group. We excluded two spontaneous pregnancies. Ongoing
once this framework is incorporated in the HPO, the standardized vocabulary pregnancy occurred in 51 women (26%) in the ICI group and 76 women (39%)
will facilitate communication and interpretation in both a clinical routine and in the IUI group (RR 0.67,95% CI 0.50 to 0.90 and RD -0.12, 95% CI -0.21 to
research. -0.03). In both the intention to treat and per protocol analysis ICI was inferior
Limitations, reasons for caution: Words describing the world are words to IUI as the left boundary of the 95% confidence interval crossed the pre-set
not the world. Some classifications of histopathology remain arbitrary. absolute difference of 12%.
Discussions should follow to further improve categories and reach a consensus. Limitations, reasons for caution: We did not perform analyses for factors
Wider implications of the findings: Histopathology of the testis is just one possibly affecting ongoing pregnancy rate like quality of frozen-thawed donor
aspect of the diagnosis, treatment and study of male infertility. The use a uniform sperm. These data will be available in July 2020.
ontology for all clinical diagnoses concerning male infertility should be facilitated Wider implications of the findings: In women undergoing donor sperm
and stimulated. treatment in a natural cycle, ICI results in lower ongoing pregnancy rates than
Trial registration number: not applicable IUI. Therefore, IUI should be the preferred treatment.
Trial registration number: NTR4462
P-099 A randomized clinical trial comparing intracervical P-100 Impact of genotype and phenotype on the spermatogenesis
insemination and intrauterine insemination for donor sperm of cystic fibrosis (CF) patients: a cohort study about 57 patients
treatment in the natural cycle. from 1998 to 2019
F. Kop1, M. Van Wely1, A. De Melker1, B.W. Mol2, R. Bernardus3, A. Chargui1, L. Adjiman1, D. Hubert2, C. Jean1, A.S. Gille1,
M. De Brucker4, P. Janssens5, A. Nap6, B. Cohlen7, J. Pieters8, E. Girodon Boulandet3, L. Ferreux1, C. Chalas1, K. Pocate1,
S. Repping1, F. Van der Veen1, M. Mochtar1 J.P. Wolf1, N. Thiounn4, T. Bienvenu3, C. Patrat1

1
Hôpital Cochin - Assistance Publique des Hôpitaux de Paris, Service d’Histologie collection can be proposed only in case of a conceptional project for men in
Embryologie Biologie de la Reproduction, PARIS, France ; couple or before transplantation and immunosuppressive agents
2
Hôpital Cochin - Assistance Publique des Hôpitaux de Paris, Service de Trial registration number: none
Pneumologie - Centre de ressources et de compétences de la mucovisidose, Paris,
France ;
3
Hôpital Cochin - Assistance Publique des Hôpitaux de Paris, Service de Génétique P-101 Male Infertility During the Syrian Crisis: A Case-Control
et Biologie moléculaire, Paris, France ; Study
4
Hôpital européen Georges Pompidou - Assistance Publique des Hôpitaux de Paris, Z. Khrait1, R. AL-Nasser2
Service d’Urologie, Paris, France 1
Fakih Fertility Center, IVF, Dubai, United Arab Emirates ;
2
British-Syrian IVF & Fetal Medicine Center, Fertility, Damascus, Syria
Study question: Is there an impact of CFTR (cystic fibrosis transmem-
brane-conductance regulator) gene mutation type and/or general health con- Study question: To explore the differences in semen analysis parameters
dition and the quality and yield of spermatogenesis? among Syrian males before and during the Syrian crisis to identify the impact of
Summary answer: Neither genetic severity nor clinical severity seems to have war on male fertility.
a negative impact on the characteristics of epididymis and testicular spermatozoa Summary answer: Semen samples obtained during the Syrian war contained
ofn cystic fibrosis (CF) patients. semen in significantly higher quantities but of lower quality. Sperm morphology
What is known already: CFTR protein regulates electrolyte and fluid is subject to aberrations.
transport in many tissues with exocrine function, including male reproductive What is known already: Several studies conducted on veterans and civilians
tract. Mutation of CFTR gene causes CF, which affects the function of several exposed to war-related injurious trauma showed that war operations and trauma
organs, and impairs male fertility. CF is generally associated to an obstructive had no impact on male infertility in terms of fathering a child after exposure to
azoospermia because of bilateral absence of vas deferens and seminal vesicles war incidents.
degeneration. CFTR protein is detected in human fetus at the early develop- Study design, size, duration: Control samples were collected in 2009-2010
mental stages and highly expressed in both testis and epididymis. CFTR pro- for 847 participants (before the war), and case samples were collected in
vides establishment of specific fluid environment for germ cell differentiation 2014-2015 for 1108 participants (during the war). The semen analysis parame-
and maturation. According to literature, it seems likely that CFTR mutations ters of each case were obtained electronically from hospital records. Both verbal
affect the sperm quality. consent and ethical approval were secured. Data analyses were conducted with
Study design, size, duration: This cohort study was conducted in the SPSS version 23.0 with 95% confidence intervals.
Assisted Reproduction Center of an university hospital on 57 patients, for whom Participants/materials, setting, methods: A total of 1108 male cases
a CF has been diagnosed and monitored at Cystic Fibrosis Resource and from the war in Syria (2014-2015) were included in the study and were
Competence Center (CFRCC) in the same hospital. The CF diagnosis was based compared with 847 controls (2009-2010). Exclusion criteria included a BMI of
on CFTR genetic tests and clinical symptoms. All patients were azoospermic and greater than 30, age older than 55 years, confirmed diagnosis of systematic
underwent on a microscopic epididymal sperm aspiration (MESA) and/or a disease, extensive exposure to radiation and chemicals not related to wartime
testicular sperm extraction (TESE) between 1998 and 2019. events (such as exposures among radiologists and chemists), and administration
Participants/materials, setting, methods: Clinical data- one year preceding of medications with a known impact on fertility.
surgical sperm collection - related to the severity of the CF have been collected: Main results and the role of chance: The mean semen volume was
respiratory spirometry data, Pseudomonas aeruginosa colonization, number of significantly higher in cases (3.18 ± 1.68 ml vs 3.45 ± 1.75 ml) than in controls
antibiotic treatment, BMI and Pancreatic insufficiency, as well as CFTR mutations. (p <.001). In contrast, the means of all other parameters (density; motility
Linear regression tests (fisher and χ²) were carried out to establish or not cor- categories A, B, and C; and percentage of normal sperm) were significantly lower
relation between on the one hand the quality of the spermatic sample and on among cases than among controls (all P-values <.001).
the other hand genotype and/ or each clinical parameter Conclusion: Semen samples obtained during the Syrian war contained semen
Main results and the role of chance: The mean age of patients at sperm in significantly higher quantities but of lower quality. Sperm morphology is subject
retrieval was 31.3 years [21-55]. The mean BMI was 21.3 Patients with severe to aberrations that highlight the risk of fertility problems. Motility and normal
genotype represented 67.8% and likely (67.3%) an external pancreatic deficiency. sperm percentage are the most affected parameters.
P.aeroginosa colonization was revealed in 30% of cases and 45.6% patients Limitations, reasons for caution: While the findings of our study are prom-
received at least one IV antibiotic treatment a year before MESA and/or TESE. ising, the extent to which psychological stress affects spermatogenesis remains
Respiratory function has been impaired in 18 cases (31.6%) with a maximal unclear. A large randomized controlled trial is recommended to fully elucidate
expiratory volume per second (MAVS) < 40%. the specific confounding factors implicated in spermatogenesis and to understand
Spermatozoa were founded for all patients and frozen after 58 surgical the effect of stress, particularly war, on the deterioration of sperm quality.
sperm collections (one patient has benefited of 2 TESE): 20 MESA, 32 MESA Wider implications of the findings: It is recommended that fertility health
+ TESE and 6 only TESE. Spermatic parameters results were: presence of should an area of focus for those exposed to wartime events.
epididymal spermatozoa in 89.7%, a mean epididymal sperm numeration at Trial registration number: not applicable
66.0 ±105.5 million, subnormal mean epididymal vitality at 57.8%.
Progressive epididymal motility. Total testicular motility and testicular vitality P-102 Microfluidic sperm selection for couples with a history of
were decreased (respectively: 2.5±4.7%, 6.0±6.8% and 41.1±25.3%). The aneuploid embryos
mean number of epididymal motile progressive spermatozoa was satisfac- A. Parrella1, D. Tavares1, M.S. Wang1, M. Haddad1, Z. Rosenwaks1,
tory (10.2±18.83 million). G.D. Palermo1
The presence and quality of epididymal spermatozoa was not affected by the 1
genetic severity of the disease (p = 0.65). Likewise, and after several univariate Reproductive Medicine, New York, U.S.A.
analyses there was no statistically significant correlation between the parameters
evaluating clinical severity and those relating to epididymal sperm. Study question: Does selecting the highest progressively motile spermatozoa
Limitations, reasons for caution: The lack of standard values for epididymal with optimal genomic integrity enhance the likelihood of generating euploid
and testicular sperm parameters made harder the choice of cut-offs to establish embryos and achieving higher pregnancy rates?
correlation. This leads us to choose the WHO recommendations for sperm Summary answer: Microfluidic sperm selection (MFSS) identified spermato-
count and vitality and the mediane for motility. Our study needs to be completed zoa with the highest chromatin integrity, leading to higher implantation and
by a multivariate analyse. delivery rates.
Wider implications of the findings: Regarding to the better quality of life What is known already: Genomic impairment of the male gamete can hinder
of CF patients nowadays and to our results- with no significant correlation embryo cleavage and implantation. Dysfunction of the male genital tract increases
between genetic and/or clinical severity and spermatogenesis, the surgical sperm both single-strand (ss) and double-strand (ds) DNA nicks and breaks that can

inhibit the developmental competence of embryos. In particular, ds DNA breaks Study question: Are there different outcomes of intracytoplasmic sperm
present in the spermatozoa of fertile donors, at a proportion as high as 40%, injection (ICSI) in NOA patients with fresh and frozen-thawed testicular sperm
may contribute to embryo aneuploidy with consequent implantation impairment. versus AID sperm?
Study design, size, duration: From October 2016 through January 2020, Summary answer: Comparing similar outcomes with fresh and frozen-
26 consenting couples underwent a new ICSI cycle in which spermatozoa were thawed testicular sperm, NOA patients showed best ICSI outcome with
selected by MFSS. Most couples had a history of high sperm chromatin fragmen- AID sperm.
tation (SCF) in their ejaculate, a high proportion of aneuploidy embryos, or What is known already: Approximately 60% of azoospermia cases are due
recurring implantation failure after ICSI. to NOA caused by testicular dysfunction.NOA patients can become fathers
Participants/materials, setting, methods: SCF was measured by TUNEL through micro-TESE which is associated with a high sperm retrieval rate (SRR)
on raw semen specimens as well as after density gradient centrifugation (DGC) of 30-75%. So they have two treatments either synchronous oocyte retrievals
and MFSS. ICSI was carried out with spermatozoa selected by the two different for their partners or testicular sperm cryopreserved when they could gain sperm
methods, and resulting embryos underwent preimplantation genetic testing for by surgery.
aneuploidy (PGT-A). Fertilization and clinical pregnancy outcomes, following Study design, size, duration: Comparing the outcomes of NOA patients
replacement of thawed euploid blastocysts, were recorded and compared both undergoing ICSI with fresh and frozen-thawed testicular sperm versus NOA
between the two sperm selection methods. patients undergoing ICSI with AID sperm was conducted to achieve this goal
Main results and the role of chance: A total of 26 men ( 39±7 years) had after excluding infertility caused by female factors.
a mean sperm concentration of 43.3±41.6x106/mL, 34.6±17 motility, and Participants/materials, setting, methods: We analyzed the outcomes of
2.4±1% morphology. After DGC and MFSS, the sperm concentration was 4.1±5 474 NOA patients undergoing 413 ICSI cycles with fresh testicular sperm, 198
and 3.4±4x106/mL, with 70±32% and 95±15% motility, respectively (P<0.0001). ICSI cycles with frozen-thawed testicular sperm and 170 azoospermic patients
Average SCF decreased from 22% in raw samples to 18% following DGC, and with donor sperm undergoing 197 ICSI cycles between January 2015 and March
became 2.8% after MFSS processing (P<0.0001). 2019. The parameters were fertilization rate (FR), 2PN cleavage rate (2PNCR),
A total of 14 men underwent 29 ICSI cycles with their female partners (37.3±5 blastocyst formation rate (BFR), implantation rate (IR), cumulative pregnancy
years) with DGC-selected spermatozoa, achieving a 56.7% fertilization rate rate (CPR), cumulative live-birth rate (CLBR), miscarriage rate (MR) and no
(177/312), resulting in 63.2% (55/87) morphologically good-quality embryos; embryo available cycle rate (NEACR).
18.4% were euploid at time of transfer in 9 couples, resulting in a 21.4% (3/14) Main results and the role of chance: Comparing fresh and frozen-thawed
CPR and 33% pregnancy loss (1/3). Subsequently, they underwent ICSI with testicular sperm, NOA patients using AID sperm showed the better outcome
MFSS, achieving a 74% fertilization rate (137/186; P<0.001), with 61% (51/84) after ICSI-ET (for FR, BFR, IR, CPR, CLBR and NEACR, all p values were 0.000).
good-quality embryos of which 45.2% (38/84) were euploidy (P<0.001). A total NOA patients had the similar ICSI outcomes with fresh and frozen-thawed tes-
of 10 couples received a replacement, achieving a 75% implantation rate (9/12; ticular sperm and this results were not effected by male female age, female BMI,
P<0.001), resulting in a 90% CPR/cycle (9/10); most delivered (P<0.001) or male age, male BMI, male FSH, male LH and male T after adjusting confounding
are ongoing. factors by linear and logistic regression analyse.
Next, we treated solely with MFSS 12 couples with a history of all Limitations, reasons for caution: The small number of transfer cycles for
aneuploid embryos at an outside center. They underwent ICSI at our center, NOA patients (due to the exclusion criterias) reduced the precision of our
achieving an 85% fertilization rate, a 60% (9/15) implantation rate, and the estimates. However, our results provide a valuable indication of the ICSI outcome
same CPR. for NOA patients using fresh or frozen-thawed testicular sperm.
Limitations, reasons for caution: This study represents a preliminary exper- Wider implications of the findings: Our results provide a valuable infor-
iment on a small number of subjects. While the oocyte contribution to aneu- mation about similar ICSI with fresh or frozen-thawed testicular sperm for coun-
ploidy cannot be discounted, MFSS was able to yield the highest progressively selling of NOA patients. They maybe choice AID sperm if they want a better
motile spermatozoa with optimal genomic integrity capable of enhancing the ICSI outcome.
chances of generating euploid embryos. Trial registration number: 81521002
Wider implications of the findings: The occasional presence of
ds-DNA in the male gamete has been considered responsible for increasing P-104 Does sperm DNA fragmentation influence blastocyst
chromosomal structure abnormality. MFSS of highly motile and genetically com- ploidy?
petent male gametes may enhance the chances of obtaining a euploid conceptus D. Mavrelos1, B. Sowry2, B. Jones3, G. Morris1, S. Lewis4,
for transfer. J. Ben Nagi5
University College London Hospital, Reproductive Medicine Unit, London, United
Kingdom ;
P-103 Comparison the ICSI outcomes of three protocols including 2
Centre for Reprodutive and Genetic Health, Embryology, London, United Kingdom ;
fresh and frozen-thawed testicular sperm and donor semen (AID) 3
Imperial College London, Reproductive Medicine, London, United Kingdom ;
for non-obstructive azoospermia (NOA) patients 4
Queen’s University Belfast, Reproductive Medicine, Belfast, United Kingdom ;
L. Zhang1 5
Centre for Reprodutive and Genetic Health, Reproductive Medicine, London,
1
li.zhang, Department of Obstetrics and Gynecology, Beijing, China United Kingdom
Fresh testicular Frozen testicular P-value P-value P-value

AID sperm
sperm sperm (fresh vs. frozen) (fresh vs. AID) (frozen vs. ADI)
FR% (n) 44.8 (2172/4844) 49.22 (1036/2105) 70.11 (1581/2255) 0.001 p<0.001 p<0.001
BFR% (n) 26.31 (241/916) 22.40 (99/442) 33.79 (197/583) 0.119 0.002 p<0.001
IR% (n) 29.61 (215/726) 26.18 (83/317) 34.92 (161/461) 0.259 0.055 0.010
CPR% (n) 42.51 (176/414) 35.86 (71/198) 60.91 (120/197) 0.117 p<0.001 p<0.001
CLBR& (n) 28.99 (120/414) 25.25 (50/198) 50.76 (100/197) 0.795 p<0.001 p<0.001

Study question: Is high sperm DNA fragmentation associated with reduced the first to the third tertile (88% to 62%, p = 0.08). Although this difference was
blastocyst euploid rate? not statistically significant, we believe that it might have impacted the results of
Summary answer: We show a significantly lower euploid blastocyst rate in the third tertile. The surgical time progressively reduced from the first through
couples with high sperm DNA fragmentation. the third tertile, and the trend curve reached a plateau after 55 cases (203 to
What is known already: Sperm DNA fragmentation has been associated 156 minutes, p < 0.01). Patency rates were high since the first tertile (85%, 90% and
with clinical pregnancy miscarriage. A putative mechanism is karyotypic abnor- 93%, from the first to the third tertile, p = 0.75). There were no statistical differences
malities in the blastocysts resulting from conception with sperm with high DNA regarding the patency rates among the groups, but they seemed to improve through
fragmentation. the tertiles. Likewise, the post-operative total sperm count had a clinically, but not
Study design, size, duration: We included 174 couples undergoing treatment statistically, significant improvement through the tertiles (88%, 93% and 100%, p =
during the study period. Data for each ART cycle was held on IDEAS version 6 0.78). Meanwhile, the post-operative total motile sperm count was similar among
Mellowood Medical and analysed using SPSS version 24.0. the groups (33, 22 and 26 million/mL, consecutively). The overall pregnancy rate for
Female and male partners age was grouped into age ranges <35; 35 – 37; patients with more than 12 months of follow up was 42%.
38 – 39; 40 – 42; >42 years for analysis. Limitations, reasons for caution: The retrospective nature of the results
Participants/materials, setting, methods: We included 174 couples under- presented here should be taken into account before the generalization of our
going treatment during the study period. Data for each ART cycle was held on conclusions. Additionally, the surgeon was submitted to intense microsurgery
IDEAS version 6 Mellowood Medical and analysed using SPSS version 24.0. training before the beginning of this study, which may not be the case for all
Female and male partners age was grouped into age ranges <35; 35 – 37; urologists interested in the infertility field.
38 – 39; 40 – 42; >42 years for analysis. Wider implications of the findings: Vasectomy reversal is a challenging
Main results and the role of chance: Median maternal age was 39 years procedure, and at least 55 procedures are needed to achieve a good surgical
(IQR 37 – 41) and paternal age 40 years (36 – 44). During the study period 797 time. MIM fellowship training provides microsurgical skills that help to accelerate
blastocysts were biopsied and 246 (31%) were reported euploid. Euploidy rate by the procedure learning curve.
maternal age was <35; 78/155 (50% ), 35 – 37; 72/171 (42%), 38 – 40; 65/173 Trial registration number: Not applicable
(27%), 41- 42 27/150 (15%) and >42; 4/56 (7%). Couples with a female partner
under 35 and abnormal sperm DFI had a significant lower euploidy rate compared
to couples with normal DFI (31/76, 41% vs. 47/79, 59%), p=0.0198. There was P-106 Contribution of sperm genomic and extragenomic markers
no significant difference in older age groups, 35 – 37 (45/119, 38% vs. 27/52, 52%) as a critical determinant of embryonic development and embryo
p=0.085, 38 – 40 (38/130, 29% vs. 27/108, 25%) p=0.465, 41 – 42 (18/96, 19% viability in spontaneous and assisted conceptions
vs. 9/72, 11%) p= 0.159 and >42 (4/43, 9% vs. 0/9, 0%), p=0.17 . V. Dhawan1, D. Deka2, N. Malhotra2, V. Dadhwal2, N. Singh2,
Limitations, reasons for caution: This is a small retrospective study with R. Dada1
important clinical implications but prospective studies are needed for 1
All India Institute Of Medical Sciences, Anatomy, New Delhi, India ;
confirmation. 2
All India Institute Of Medical Sciences, Obstetrics & Gynaecology, New Delhi,
Wider implications of the findings: Abnormal sperm DNA fragmentation India
is associated with less euploid blastocysts in couples with a female partner under
35 years of age. Study question: Does the transmission of sperm genomic and extragenomic
Trial registration number: NA cargo hold any relevance in early embryonic development and embryo viability
in successful pregnancy outcomes?
Summary answer: The dynamic intertwined role of sperm DNA and RNA
P-105 Impact of male infertility fellowship training on the elements delivered at fertilization have postfertilization functions and navigate
vasectomy reversal learning curve early embryonic development in successful pregnancy outcomes.
G. Wood1, E. De Paula Miranda2, F. Tenorio Lima Neto3 What is known already: The suite of complex repertoire of sperm RNA
1
Urologist, Urology, Sao Paulo, Brazil ; delivered to the oocyte at fertilisation have the potential to navigate the early
2
Universidade Federal do Ceará, Urology, Fortaleza-CE, Brazil ; embryonic development. The epigenetically marked sperm genome successfully
3
Instituto de Medicina Integral Professor Fernando Figueira, Urology, Recife-PE, Brazil orchestrates in synchrony with the extragenomic cargo. Sperm with disturbed
genomic integrity and dysregulated levels of sperm RNA may be responsible for
Study question: How many vasectomy reversal procedures are needed for a a successful fertilisation but may affect the pregnancy outcomes and even affect
trained microsurgeon to achieve good patency results and reduce operative time? the health of the offspring.
Summary answer: Vasectomy reversal is a challenging procedure, and at least Study design, size, duration: A case control study of male partners of
55 procedures are needed to achieve a good surgical time. couples experiencing RPL (n=75), recurrent implantation failure (n=75) and 30
What is known already: Male infertility microsurgery (MIM) is an area in controls at AIIMS, New Delhi, India. Study duration was 2 years.
intense development, requiring intensive surgeon training and its results are highly Participants/materials, setting, methods: A case control study of male
dependent on surgeon’s skills and domain of microsurgical techniques. Vasectomy partners of couples experiencing RPL (n=75), recurrent implantation failure
reversal (VR) is the most cost-effective option for couples desiring offspring after (n=75) and 30 controls at AIIMS, New Delhi, India.
vasectomy, and is one of the most difficult microsurgical procedures. We report Main results and the role of chance: The transcript levels of SOX3, WNT5A,
the learning curve results for VR performed by a single surgeon after his MIM RPS6, RBM9, RPL10A showed significant difference beyween RPL patients and
fellowship training. controls. While the levels of FOXG1, RPS6, RBM9, RPL10A, RPS17 and TOMM7
Study design, size, duration: In a retrospective cohort fashion, we reviewed showed significant difference between RIF patients and controls. The mean ROS
the charts of all patients who underwent VR from June 2016 to August 2019 by and DFI was seen to be significantly higher (in both RPL and RIF patients as
a single surgeon who finished a MIM fellowship training in May 2016. All proce- compared to controls in both RPL and RIF patients (p <0.001). The odds of
dures were performed using an operating microscope. A multilayer technique occurrence of RPL and RIF was 12.41 and 13.68 times higher, whose ROS>
was used for vasovasostomies (VV), and the LIVE technique for 28 % [(OR 12.41, 95% CI: (6.28-22.29) and OR 13.68 (6.52-28.71)] (p<0.001).
vasoepididymostomies. While the odds of occurrence of RPL and RIF 12.68 and 18.87 times higher,
Participants/materials, setting, methods: At the end of the study dura- whose DFI>30 (OR 12.68, 95% CI: (6.53-23.55) and OR 18.87 (6.52-28.71),
tion, 69 patients were included. They were divided into tertiles by dates. We (p<0.001). The mean sperm telomere length was seen to be lower in RPL
assessed and compared the following variables: surgical time, complications, (p=0.054) and RIF (p=0.0294) patients with respect to controls.
postoperative semen analysis parameters, patency, and pregnancy rates. Limitations, reasons for caution: A potential limitation of this study is the
Main results and the role of chance: 26 patients were included in each smaller sample size recruited for the current study. Also the contribution of
tertile. There were no differences between tertiles regarding baseline charac- maternal factors and the possible role of other environmental factors which may
teristics. The proportion of men who underwent bilateral VV decreased from affect implantation and early embryogenesis cannot be ignored.

Wider implications of the findings: The complex interplay of the various is inconsistent because in the case of spermatozoa absence, only cells from
transcripts have seen to have significant effects in pregnancy outcomes and health the male genital tract and from the compromised geminal epithelium were
trajectory of the future progeny. Any perturbation in this has the potential to available.
affect the early embryonic development and embryo viability. This may further Wider implications of the findings: Our epigenetic analysis demonstrated
alter sperm methylation pattern adversly affecting sperm epigenome. that key genes related to spermatogenesis were unbalanced, suggesting a mat-
Trial registration number: Not Applicable urational arrest as the cause for the azoospermia. The identification of specific
genes in men with failed TESE may provide valuable information on the ability
to successfully predict the likelihood of retrieving spermatozoa with TESE.
P-107 Epigenetic Assessment by RNA Sequencing of Non- Trial registration number: not applicable
Obstructive Azoospermic Men to Predict Successful Testicular
Sperm Retrieval P-108 Can Cytomegalovirus (CMV) infection affect male
K. Hancock1, A. Parrella1, S. Cheung1, B. Barker1, Z. Rosenwaks1, reproductive function? Results of a retrospective single-center
G.D. Palermo1 analysis
1
Weill Cornell Medicine, The Ronald O. Perelman and Claudia Cohen Center for A. Natali1, G. Cito1, R. Picone2, R. Fucci2, E. Micelli2, A. Cocci1,
Reproductive Medicine, New York, U.S.A. M. Di Mauro3, C. Giachini2, F. Rizzello2, A. Minervini1, M. Carini1,
M.E. Coccia2
Study question: Can epigenetic profiling of key genes provide information on 1
Careggi Hospital - University of Florence, Dept. of Urology, Florence, Italy ;
the germinal epithelium function of men with non-obstructive azoospermia 2
Careggi Hospital - University of Florence, ARTs Center, Florence, Italy ;
(NOA) and the likelihood of retrieving spermatozoa? 3
University of Catania, Dept. of Urology, Catania, Italy
Summary answer: By sequencing transcripts in NOA men, we identified genes
related to maturational arrest in men with different sperm retrieval outcomes Study question: To investigate the impact of chronic CMV infection on semen
with testicular sperm extraction (TESE). parameters in men with couple infertility and the influence on the reproductive
What is known already: Azoospermia accounts for about 15% of male outcomes of CMV-seronegative women suffering from tubal factor
infertility cases, and while it is rarely caused by pre-testicular factors, the most Summary answer: CMV did not seem to play a key role in male reproductive
common forms are testicular and post-testicular. The more challenging type is function, as well as in influencing sperm fertility potential in assisted reproductive
secretory azoospermia, in which scattered persistent germinal epithelium strive outcomes.
to support the germline meiotic process, failing to yield spermatozoa via testicular What is known already: Cytomegalovirus (CMV) is mainly investigated for
biopsy in almost 50% of cases. To judge the reproductive potential of these the consequences of prenatally transmission from mother to fetus, that can lead
individuals, we assessed the gene expression profile to predict the ability to to severe disturbances of development. However, the impact of infection on
identify injectable gametes. the male reproductive health has been received little consideration, despite a
Study design, size, duration: Over a 3-month period, we executed a pro- potential negative effect of the virus on the spermatogenesis.
spective study on 3 consenting men who yielded no sperm after extensive semen Study design, size, duration: From February 2016 to January 2019, all the couples
analysis. Differential expression was carried out on the ejaculate and compared referring to our ARTs Centre for infertility due to female tubal factor, were retrospec-
to a fertile donor. Subsequently, they underwent TESE, and gene expression tively reviewed. All the men were divided into two groups: Group 1 included CMV
analysis was carried out both in comparison to a man with obstructive azoosper- IgG-seropositive men, Group 2 comprised CMV IgG-seronegative patients.
mia (OA), as well as their ejaculated specimen, in relation to whether sperma- Participants/materials, setting, methods: On the day of fresh IVF/ICSI
tozoa were retrieved with TESE. treatment, we collected data of the seminal characteristics, as follows: volume,
Participants/materials, setting, methods: Three men presented with no pH, total sperm count/ml, total sperm concentration, viability, progressive motil-
spermatozoa in their ejaculates after an extensive sperm search and were diag- ity (PR), non-progressive motility (NP), immobility, total motility and normal
nosed as NOA. Subsequently, they underwent TESE. RNA expression was morphology. Two-pronuclear (2PN) fertilization rate (FR), 1-2-3PN FR, cleavage
performed on the leftover ejaculate specimen and on the testicular specimens rate (CR), miscarriage rate (MR), pregnancy rate (PR) and live birth rate (LBR)
from these individuals. Purified RNA was sequenced by Illumina HiSeq at were collected.
2x150bp per lane with ~58M reads/sample. A log2fold change of >1 and Main results and the role of chance: 222 men were included: 115 (51.8%)
P<0.001 were considered significant. in the Group 1, 107 (48.2%) in the Group 2. There was reported a low trend
Main results and the role of chance: Three consenting men (36.3±5yrs) towards higher sperm concentration/ml, total sperm count and viability in CMV
were included in this study. All underwent extensive semen analysis of their IgG-seronegative males, compared to CMV IgG-seropositive, but no statistically
ejaculate, and no spermatozoa were found. These men were diagnosed with differences were found among groups. Semen volume, pH, motility and normal
NOA following urological evaluation. Quantitative analysis of RNA extracted sperm morphology were similar between the two groups. Considering the sub-
from these ejaculated samples averaged a concentration of 2.3±1 ng/ul and an group of men, partners of CMV IgG-seronegative females, 65 couples (29.2%)
RNA integrity of 7.0±1. Compared with the ejaculated donor, we found ~3000 were selected. Overall, median 2PN FR was 67%, total FR 83%, CR 100%, PR/
genes differentially expressed for each man, with 1379 in common, most asso- cycle 26.2%, MR 10.8%, LBR/cycle 15.4%. No significant differences were found
ciated with meiotic arrest (TEX11, DMC1, MEOB, MEI1, STX2). regarding the reproductive and pregnancy outcomes between CMV IgG-
These men underwent TESE and a gene expression analysis in comparison seropositive men and those seronegative.
with an OA control, ~3000 genes were identified, with 702 in common, including Limitations, reasons for caution: We did not test the presence of CMV
TSPYL1 and TSPYL2, associated with spermatogenesis. DNA in the seminal samples, due to the time and cost of performing polymerase
Spermatozoa were successfully identified in the TESE specimen of 2 men chain reaction (PCR) and cell culture, compared to the more rapid and inexpen-
(38.5±6yrs) at a 0.08±0.1x103/ml concentration. RNA concentration was sive detection of CMV IgG and IgM assays.
5.57±1 ng/ul with 7.0±1 integrity. Genes associated with cell proliferation Wider implications of the findings: The viral infection did not seem to play
(MAPRE1, RAN), apoptosis (ARL6IP1), embryo development (IL6ST), and sper- a key role in male reproductive function and in influencing sperm fertility potential
matogenesis (AKAP11) were underexpressed in the ejaculate compared to tes- in the assisted reproduction. CMV serology screening could be crucial to identify
ticular biopsy (P<0.001). primary acute infection by detecting IgM seropositivity, in order to prevent virus
In a 35-year-old man with no spermatozoa in testicular biopsy, RNA- transmission by sperm sample.
sequencing revealed several genes that were uniquely overexpressed in the Trial registration number: not applicable
ejaculate compared to the testicular. These were associated with spermatogen-
esis(DAZAP2, SYNJ2BP), cell cycle arrest(CAB39), DNA replication(NFIB), and
cilia formation(IFT57) (P<0.001). P-109 Impact of male body mass index on Invitrofertilisation
Limitations, reasons for caution: This is a preliminary analysis, which outcome-Live birth rate.
needs to be corroborated by further observations. The source of the RNA R. Mushtaq1, C. Yearwood2, Y. Khalaf2, T. El-Toukhy2

1
CARE Fertility United Kingdom, Clinical, Bedford, United Kingdom ; Study design, size, duration: Retrospective analysis was conducted on 96
2
Guy’s and St Thomas foundation trust hospital, Assisted conception unit, London, discarded blastocysts with consent from patients out of embryos collected and
United Kingdom cultured in our clinic from 2013 to 2018. All of these embryos were time-lapse
monitored by EmbryoScope (Vitrolife).
Study question: Does male body mass index effect outcome of assisted repro- Participants/materials, setting, methods: The subject embryos were
ductive technology? biopsied with TE and NGS chromosome analysis was performed. These embryos
Summary answer: Male body mass index does not influence Invitrofertilisation were classified by time-lapse videos, those with 2 cells in the first cleavage and
live birth rate after adjusting for important confounding factors. 4 cells in the second cleavage are the good cleavage groups, those with 3 or
What is known already: Male factor infertility could be contributory in up to 50% more cells in the first cleavage, or those with 5 cells or more in the second
of cases seen in assisted reproduction clinics. Obesity is becoming pandemic and is cleavage are the irregular cleavage groups.
contributory factor in overall fall noted in semen parameters worldwide. There are Main results and the role of chance: As a result of image analysis by time-lapse
many studies on impact of female BMI on outcomes of IVF. There is conflicting evi- monitoring, there were 53 good cleavage groups and 43 irregular cleavage groups.
dence regarding the effect of raised paternal BMI on outcome of assisted reproductive The proportions of euploid embryos, aneuploid embryos and mosaic embryos were
technology. Most of the studies are small and retrospective. 43.4% (23), 45.3% (24) and 11.3% (6) in the good cleavage group, 41.9% (18), 46.5%
Study design, size, duration: It is a prospective observational study, including (20), and 11.6% (5) in the irregular cleavage group, there was no significant difference
438 couples having IVF or ICSI treatment at Guy’s and St Thomas’ hospital, in euploid rate between the two groups (The odds ratio of the euploid rate of the
London between July 2015 and June 2016. irregular cleavage group to the good cleavage group was 0.94).
Participants/materials, setting, methods: Couples undergoing first or Limitations, reasons for caution: Because PGT-A is not approved in Japan,
second IVF cycle between July 2015 to June 2016 where recruited before starting this study was conducted using only discarded embryos that were not used for
the IVF cycle. Both male and female BMI were measured and recorded at the transfer with the consent of the patient.
start of treatment. All IVF/ICSI cycle characteristics were recorded and all preg- Wider implications of the findings: This study showed that Irregular cleav-
nancies followed up till delivery age in early embryogenesis did not reduce the euploid rate after reaching the
Main results and the role of chance: Of the 438 couples included in the blastocyst. Irregular cleavage embryos that have reached the blastocyst need
study, 165 (38%) male partners had a normal BMI (18.4-24.9) and 273 (62%) not be excluded from transfer.
had a raised BMI (25-41.9). There was no significant difference between the two Trial registration number: not applicable
groups in sperm parameters. The clinical pregnancy and live birth rates were
not significantly different between the two groups (33% vs 35% and 25% vs 31%,
respectively, P=0.19). After adjusting for important confounders, including female P-111 MicroRNAs secreted by human embryos are potential
partner age, female BMI, duration of infertility, pre-treatment AMH level, total biomarkers for clinical outcomes of assisted reproductive
dose of gonadotrophins used during ovarian stimulation, number of oocytes techniques
collected, method of oocyte fertilisation (IVF vs ICSI) and number of embryos Z. Li1, F. Fang2, X. Chengliang1
transferred, the likelihood of a live birth outcome was not significantly different 1
Huazhong University of Science and Technology, Institute of Reproductive Health,
in the raised male BMI group compared to normal male BMI group (OR 1.22, WuHan, China ;
95% CI 0.69-2.14, P=0.49). There was no significant difference in the live birth 2
Huazhong University of Science and Technology, Department of Obstetrics and
rate per cycle between overweight (BMI 25-29.9, n=193) and obese (BMI Gynecology- Union Hospital- Tongji Medical College, WuHan, China
30-41.9, n=80) male partners (28% vs 38%, P=0.1).
Limitations, reasons for caution: These results are from prospective but Study question: Investigate whether the miRNAs secreted by human embryos
small study with live birth rate as the end point . in culture medium can be used as embryonic biomarkers.
Wider implications of the findings: Larger proportion of men had high Summary answer: Our results indicate that miRNAs in human embryo culture
BMI in our study. This could be reflective of rising obesity. While our study has media may serve as novel and invasive biomarkers for embryo selection during
not shown a significant impact of male BMI on live birth rate in IVF cycles, the IVF/ICSI-ET cycles.
health implications of high BMI in population should be considered. . What is known already: MicroRNAs (miRNAs) are important regulators of
Trial registration number: not applicable many biological functions, including embryo implantation and development.
Recently, it is reported that miRNAs in biofluids are predictive for physiological
and pathological processes.
Study design, size, duration: The culture media were prospectively collected
POSTER VIEWING SESSION from embryos of patients who underwent routine in vitro fertilization (IVF) or
EMBRYOLOGY intracytoplasmic sperm injection (ICSI) at reproductive medicine center with
informed consent. And
Participants/materials, setting, methods: A high-throughput miRNA
sequencing method was applied to detect the miRNAs profiles in culture media
of embryos with different reproductive outcomes (15 samples were mixed for
P-110 Irregular cleavage in early embryogenesis does not reduce each group). Furthermore, qRT-PCR and droplet digital PCR (ddPCR) to verify
the euploidy after reaching the blastocyst the target miRNAs at single sample level (100 cleavage samples in total, 50 with
S. Watanabe1, K. Yoshikai1, M. Tomida1, S. Suzuki1, Y. Matsuda1, successful pregnancy and 50 with failed pregnancy). Receiver operating charac-
T. Kato2, E. Nakano1, H. Kurahashi2, T. Sawada1 teristic (ROC) analyses were performed for differentially expressed miRNAs.
1 Main results and the role of chance: The miRNA sequencing showed that
Sawada Women’s Clinic, ART Lab., Nagoya, Japan ;
2 embryos with successful pregnancy secreted different miRNA profiles into the
Institute for Comprehensive Medical Science- Fujita Health University, Division of
culture media, compared with embryos with failed pregnancy. These differentially
Molecular Genetics., Toyoake, Japan
expressed miRNAs were predicted to be involved in multiple biological pro-
Study question: Does euploid rate decrease when the irregular first or second cesses, cellular components and molecular functions. And 18 miRNAs were
cleavage embryos reach the blastocyst? selected for validation by qRT-PCR in mixed samples and significantly different
Summary answer: The euploidy of the irregular first or second cleavage expression of these 10 miRNAs between two groups were identified. The
blastocysts was equivalent to the normal cleavage blastocysts. ddPCR results revealed that hsa-miR-26b-5p, hsa-miR-451a and hsa-miR-21-5p
What is known already: There are some reports that aneuploidy is high could be stably detected in the culture medium of one single embryo at cleavage
when the irregular cleavage embryos develop into blastocysts but there are stage. After statistical analysis, we found that the cleavage embryos with suc-
reports that the rate of transfer pregnancy and miscarriage of irregular cleavage cessful pregnancy presented decreased expression of hsa-miR-26b-5p and hsa-
blastocysts is equivalent to that of the normal cleavage blastocysts. miR-21-5p in the culture media. Moreover, the Receiver Operating Characteristic

1
(ROC) curve analysis indicated that hsa-miR-26b-5p and hsa-miR-21-5p could Yanaihara Women’s Clinic, Obstetrics and Gynecology, Kamakura, Japan ;
2
serve as potential biomarkers for reproductive outcomes. Gunma Paz University, Graduate School of Health Sciences, Takasaki, Japan
Limitations, reasons for caution: More clinical trials are needed to deter-
mine the sensitivity and specificity of miRNA biomarkers for embryo selection Study question: Can fertilization be obtained by pressing single sperm onto
and more basic research are also necessary to develop better detection methods an unfertilized oocyte after in vitro fertilization (IVF) without breaking the oocyte
of miRNAs with low input. membrane?
Wider implications of the findings: Together, our findings highlight the Summary answer: Fertilization was obtained by pressing zona pellucida (ZP)-
important predictive potential of miRNAs secreted by human embryos in culture bound sperm directly onto the oocyte membrane, a procedure that we desig-
media, which is meaningful for noninvasive embryo selection during IVF cycles. nated as assisted sperm fusion insemination (ASFI).
Trial registration number: none What is known already: Rescue intracytoplasmic sperm injection (ICSI),
which is ICSI of unfertilized oocytes after conventional IVF, has been reported
to prevent total fertilization failure. However, recent studies have shown that
P-112 An analysis of 2.1PN (2PN+1 micronuclei) formation.
the outcome of rescue ICSI is unsatisfactory, resulting in a poor fertilization rate
N. Okuyama, MSc1, N. Oka1, N. Aono1,2, T. Hashimoto1, and a high degeneration rate. A micromanipulation technique that permits fer-
K. Kyono1,2 tilization without breaking the oocyte membrane would be expected to reduce
1
Kyono ART Clinic Takanawa, Department of Gynecology, Tokyo, Japan ; further the degeneration rate after rescue ICSI.
2
Kyono ART Clinic Sendai, Department of Gynecology, Sendai, Japan Study design, size, duration: This study of the ASFI method was performed
between January 2019 and December 2019 on 49 subjects. The results of ASFI
Study question: What factor causes 2.1 PN zygotes? were compared with those obtained in a previous trial of rescue ICSI that was
Summary answer: High-aged female patients (OR: 1.17, 95%; CI: 1.09-1.25; performed between July 2018 and December 2018. The present study was
P<0.001) and frozen-thawed oocytes (OR: 3.85, 95%; CI: 1.79-8.26; P<0.001) approved by the local ethics committee.
were more likely to produce 2.1PN zygotes. Participants/materials, setting, methods: The acrosome reaction (AR)
What is known already: We confirm fertilization when two pronuclei (PN) rate of 49 motile sperm collected from the ZP was assessed. Sixty-nine motile
and second polar body are seen, and in some cases, 2PN + micronuclei (2.1PN) sperm bound to the ZP were aspirated directly using an injection pipette.
is recognized. It is reported that some 2.1PN zygotes may develop to blastocyst These motile sperm were pressed onto the membrane of 69 unfertilized
and lead to successful clinical pregnancy and live birth; however, there is a risk oocytes at 6 hours after IVF; pressing was performed with the injection pipette
that 2.1PN zygotes have the possibility of triploid (Capalbo et al., 2017). for 30 seconds.
A limited number of studies have reported on 2.1PN, but its mechanism is Main results and the role of chance: The AR rate of sperm collected
still unclear. from the ZP was significantly higher than that of control sperm (98.0% vs.
Study design, size, duration: This retrospective single-center study was 28.6%, p <0.01). The mean and standard deviation of ages of the women in
performed from January 2015 to December 2018 including a total of 659 patients ASFI and rescue ICSI were 36.1 ± 3.9 and 37.7 ± 4.3 years, a difference that
and 1161 cycles. A total of 7939 oocytes were inseminated either by conven- was not significant (p >0.05). All sperm collected from the ZP adhered to the
tional IVF (cIVF) or ICSI. Logistic regression analysis was conducted to investigate oocyte membrane after ASFI. Time-lapse cinematography showed that sperm
the cause of 2.1PN. Inseminated oocytes were cultured in time-lapse monitoring pressed onto the oocyte membrane remained there for approximately 30
system. We defined a 2.1PN when two equal-sized PN, second polar body, and minutes before being incorporated into the oocyte membrane. There was no
less than 1/3 the size of micronuclei were observed. significant difference in fertilization rate (69.6% vs. 70.0%), degeneration rate
Participants/materials, setting, methods: We analyzed the associations (0% vs. 4%), number of good-quality embryos at day 3 (58.3% vs. 51.4%), or
between the number of PN and embryo development; background profiles such blastocyst formation rate (58.5% vs. 42.4%) between the ASFI and rescue ICSI
as female age, serum AMH, and BMI; ICSI or cIVF; ejaculated sperm or TESE groups, respectively.
sperm; fresh oocytes or vitrified-warmed oocytes; and with or without artificial Limitations, reasons for caution: The major limitation of this study was
oocyte activation (AOA). Good quality blastocyst rates were defined according the use of sperm bound to the ZP. For more general application, a method of
to Gardner’s criteria. Chi-square test and logistic regression analysis were con- artificially inducing AR will need to be established. Moreover, our sample size
ducted where appropriate. was relatively small; further studies with a larger number of patients will
Main results and the role of chance: Fertilization results were as follows: be needed.
26.7% (n=2122) for 0PN, 3.2% (n=255) for 1PN, 62.1% (n=4928) for 2PN, Wider implications of the findings: ASFI is expected to improve the survival
1.0% (n=83) for 2.1PN, 4.1% (n=328) for 3PN, and 2.8% (n=223) for >3PN. rate of oocytes and to increase the number of the embryos available for implan-
There were statistically significant differences in the blastocyst formation rates tation. Additionally, ASFI may contribute to the elucidation of the mechanism
and good quality blastocyst rates between 2PN, 2.1PN and 3PN embryos, of fertilization, given that the technique permits observation of the process of
(49.6% vs 30.1% vs 14.3%, P<0.001) (34.0% vs 20.5% vs 7.0%, P<0.001). High- fusion between the oocyte and sperm.
aged female patients (OR: 1.17, 95%; CI: 1.09-1.25; P<0.001) and vitri- Trial registration number: not applicable.
fied-warmed oocytes (OR: 3.85, 95%; CI: 1.79-8.26; P<0.001) were more likely
to obtain 2.1PN zygotes. No association was found between 2.1PN formation P-114 Non-invasive prediction of the blastocyst formation by
and serum AMH (OR: 0.96, 95%; CI: 0.86-1.06; P=0.448), BMI (OR: 0.96, 95%; morphokinetics discriminant analysis
CI: 0.89-1.03; P=0.295), ICSI or cIVF (OR: 1.33, 95%; CI: 0.61-2.11; P=0.702), A. Iaconelli Jr.1, A. Setti2,3, D. Braga2,3, L. Vingris4, E. Borges Jr.1,3
ejaculated or TESE sperm (OR: 1.40, 95%; CI: 0.65-3.00; P=0.392) and with or 1
Fertility Medical Group, Clinical department, São Paulo- SP, Brazil ;
without AOA (OR: 1.18, 95%; CI: 0.28-5.09; P=0.821). 2
Fertility Medical Group, Scientific research, São Paulo- SP, Brazil ;
Limitations, reasons for caution: PGT-A was not analyzed due to the 3
Instituto Sapientiae - Centro de Estudos e Pesquisa em Reprodução Assistida,
regulation of the Japan Society of Obstetrics and Gynecology. Scientific research, São Paulo, Brazil ;
Wider implications of the findings: This study indicated that 2.1PN may 4
Fertility Medical Group, IVF laboratory, São Paulo- SP, Brazil
be derived from oocyte factors such as advanced female age and oocyte cryo-
preservation. 30.1% of 2.1PN zygotes developed to blastocyst; however, 2.1PN Study question: Is the discriminant analysis of embryo morphokinetics
zygotes may be triploid (Capalbo et al., 2017). Hence, a careful choice for able to predict the blastocyst formation chance on the 5th day of embryo
embryo transplantation is needed. development?
Trial registration number: none Summary answer: Early kinetic parameters may predict which embryo is able
to develop into blastocysts and those failing to blastulate.
P-113 Fertilization by direct pressing of zona pellucida-bound What is known already: At the early stage of embryonic development,
human sperm onto the oocyte membrane the transcription of embryonic genes is quiescent, and development is con-
S. Hatakeyama1,2, Y. Araki2, Y. Araki2, S. Ohgi1, A. Yanaihara1 ducted by maternal proteins and RNAs. Subsequently, embryonic genome

activation (EGA) occurs, followed by the developmental control switching non-male factor infertility report heterogeneous results, thereby supporting the
to the nuclear genome. Studies have suggested that morphological grade is need for a meta-analysis.
not accurate enough to predict the developmental potential. Most embryos Study design, size, duration: Systematic review and meta-analysis of pub-
reach the cleavage-stage, regulated by maternal factors, and EGA never lished research articles. Searches were conducted on MEDLINE, EMBASE, and
occurs. Extended embryo culture and transfer at blastocyst stage is an the Cochrane Library from 2004 to June 2019 using medical subject headings
alternative which allows the selection of embryos at more advanced stages, and free text terms for ‘ICSI’, ‘IVF’, ‘non male factor’, ‘unexplained infertility’,
after EGA, increasing the implantation rate and minimizes the multiple preg- ‘fertilisation failure’, ‘pregnancy’, ‘live birth’.
nancies risk. Participants/materials, setting, methods: Studies preselected on title and
Study design, size, duration: Kinetic data were analyzed in 139 patients and abstract were assessed using adapted Newcastle-Ottawa Quality Assessment
1219 zygotes cultured until day 5 in a time-lapse incubator system, between Scales. Only articles describing primary outcome of live birth were included.
March/2019 and November/2019. Timing to specific events from the point of Risks of bias were assessed using ROBIN-1 tools. Study selection, bias assess-
insemination was determined using time-lapse imaging. A stepwise discriminant ment and data extraction were performed by two independent reviewers
function analysis determined which kinetic markers discriminate between according to Cochrane methods. Risk ratio (RR) IVF/ICSI and 95% confidence
embryos that reached the blastocyst stage or not. Moreover, cut-off points were interval (CI) were estimated using random effect model. Pre-specified sensitivity
established halfway between the averages that were significantly different analysis and meta-regression on mean female age were performed.
between the blastocyst and non-blastocyst embryos. Main results and the role of chance: Thirty-nine full-text articles were
Participants/materials, setting, methods: This study was performed in a preselected out of 1,615 references. Of these, 17 were included in the meta-anal-
private university–affiliated IVF center. The analyzed kinetic markers were: pro- ysis, corresponding to 395,414 assisted reproductive technology cycles (136,178
nuclei appearance (tPNa), timing to pronuclei fading (tPNf ), timing to two (t2), IVF and 259,236 ICSI cycles). IVF showed significantly higher LBR per cycle when
three (t3), four (t4), five (t5), six (t6), seven (t7), and eight cells (t8). Durations compared to ICSI (RR=1.13; 95%CI=1.05-1.23). No statistical difference was
of the second (t3-t2) and third (t5-t3) cell cycles (cc2 and cc3, respectively), as found for clinical pregnancy rate per cycle (RR=1.09; 95%CI=0.93-1.29).
well as timing to complete synchronous divisions s1 (t2-tPNf ), s2 (t4-t3) and s3 Fertilization rate was significantly lower in IVF cycles (RR=0.80; 95%CI=0.71-
(t8-t5) were calculated. 0.89), while proportion of cycles with total fertilization failure was not statistically
Main results and the role of chance: A total of 563 embryos reached different (RR=0.97; 95%CI=0.75-1.25).
the blastocyst stage (46.15%). The discriminant function correctly classified Sensitivity analyses, even those after removing studies at high risk of bias or
76.1% of original cases, best predicting blastocyst formation (95.0%). The outlier studies, led to similar results and conclusions. Meta-regression showed
cross-validated classification showed that overall 75.9% were correctly classified. a significant association between RR of live birth and mean female age, in favour
In this model, mean t2 (25.29 ± 3.19 vs. 27.26 ± 5.33), t7 (54.21 ± 8.40 vs. of IVF (p<0.001).
57.34 ± 11.62), s2 (1.43 ± 2.69 vs. 3.24 ± 4.64) and s3 (8.48 ± 7.84 vs. 13.75 Limitations, reasons for caution: The validity of meta-analysis results
± 10.05) were significantly different (p<0.001) between the blastocyst and the depends mainly on the quality and the number of published studies available.
non-blastocyst group, respectively. Cut-off points were established halfway Indeed, this meta-analysis contains only one randomised controlled trial.
between those averages, at 26.27 for t2, 55.78 for t7, 2.34 for s2 and 11.12 for Wider implications of the findings: Conventional IVF should be preferred
s3, for the prediction of blastocyst development. to ICSI in non-male factor infertility. The use of ICSI should be confined to male
Limitations, reasons for caution: Retrospective nature of this study and factor infertility. This evidence will help improve patient counselling.
the small sample size may be a reason for caution. Trial registration number: Prospero CRD42019136383
Wider implications of the findings: The identification of markers of the
blastocyst formation potential may lead to the benefits of the extended embryo
culture without exposing the embryo to the deleterious effects of the in vitro P-116 A prospective randomized sibling-oocyte study of two
culture for an extended period of time (i.e. epigenetic changes in trophectoderm uninterrupted media systems for culturing blastocysts.
cells leading to abnormal implantation and placentation). M. Calonge1, D. Machuca1, B. Morales1, C. Troncoso1,
Trial registration number: Not applicable M. Meseguer2
1
IVI Santiago de Chile, IVF Laboratory, Santiago- RM, Chile ;
2
IVI Valencia, IVF Laboratory, Valencia, Spain
P-115 Intracytoplasmic sperm injection (ICSI) is not superior Study question: There is any difference in the probability of getting a live birth
to conventional in vitro fertilisation (IVF) in non-male factor when the embryos are cultured in two single step media Vitrolife or Lifeglobal ?
infertility: a systematic review and meta-analysis Summary answer: No statistically significant difference in live birth rates
A. Finet1, E. Arbo2, M. Colombani3, B. Darne4, V. Gallot5, between study groups, being uninterrupted embryo culture a feasible strategy
V. Grzegorczyk-Martin1, S. Languille4, T. Freour6 with satisfactory outcomes.
1
Clinique Mathilde, Assisted Reproductive Technologies, Rouen, France ; What is known already: Currently, commercial media are available in different
2
Gedeon Richter France, Medical Affairs, Paris, France ; formulations to support specific stages of embryo development. Culture of
3
Clinique Bouchard-Elsan, Institut de Médecine de la Reproduction, Marseille, embryos is possible under different conditions: whether the media is refreshed,
France ; changed or left undisturbed for the 5-6 days of embryo culture. Continuous
4
Monitoring Force, Statistical Analysis, Maisons-Laffitte, France ; culture can be interrupted or uninterrupted depending on the renewal of the
5
Hôpital Antoine Béclère- Assistance Publique – Hôpitaux de Paris, Service de medium on day 3. Both culture systems have demonstrated excellent clinical
Médecine de la Reproduction et Préservation de la Fertilité, Clamart, France ; outcomes. Very well knows brands like Vitrolife and Lifeglobal have uninterrupted
6
CHU de Nantes, Service de biologie et médecine de la reproduction, Nantes, culture media with different specifics formulations but also are different in prices
France and time of expiration.
Study design, size, duration: We performed a prospective randomized
Study question: Does ICSI perform better than conventional IVF in non-male study between November 2017 and December 2018. A total 137 patients from
factor infertility in terms of live birth rate (LBR)? IVI Santiago were included in the study. All mature oocytes were randomized
Summary answer: ICSI is associated with lower LBR per cycle than conven- before ICSI to be cultured under either one of the two uninterrupted media
tional IVF in non-male factor infertility. formulations tested; Vitrolife or Lifeglobal. Clinical outcomes were compared
What is known already: Over recent decades, the use of ICSI has noticeably between study groups.
increased and is now significantly more frequently used than conventional IVF Participants/materials, setting, methods: Inclusion criteria were patients
in most countries. As the prevalence of male infertility has not increased con- younger than 37 years and recipient of fresh/vitrified oocytes, with a minimum
comitantly, the relevance of ICSI in non-male factor infertility can be questioned, of 7 MII to ICSI . Exclusion criteria was patients with severe pathologies in
especially when its economic and/or psychological consequences are considered. sperm sample. All transfer and vitrification of embryos were performed in
Numerous studies comparing ICSI and conventional IVF in various cases of D5/D6 according to established protocol. Fertilization, blastocyst, pregnancy,

implantation miscarriage and live birth rates were compared between both ranged from 23-88% for autosomal chromosomes and 27-63% for sex chromo-
study groups. Variable significance was analyzed through chi square and logistic somes for samples from laboratories using a single-step culturing system (n=2).
regression. This was lower than the concordance rates of 45-85% for autosomes and 61-93%
Main results and the role of chance: One hundred and thirty seven study for sex chromosomes from laboratories using a two-step culturing system (n=7).
subjects participated in the study, from which 108 underwent an oocyte donation Sex chromosome concordance varied between laboratories regardless of the
cycle. In total, 1884 MII oocytes were randomized to both media formulations. culturing system used, suggesting that embryological processes, such as the tech-
Nine hundred and thirty four oocytes were microinjected and cultured in Vitrolife nique and accuracy of cumulus cell removal, are important influencers on the
medium and nine hundred and fifty in Lifeglobal medium. Fertilization rates were accuracy in NI-PGT-A testing. Media droplets ranged in size from 12-70ul, with
comparable between groups being 74% in Lifeglobal and 78% Vitrolife group. a mean drop volume of 32ul, and a mean collection time of 60hr (4-144hr).
Moreover, no significant differences were obtained in basculation rates (62% Assisted hatching was perform for some but not all protocols.
Lifeglobal and 54% Vitrolife) , pregnancy rates (67% Lifeglobal and 58% Vitrolife ), Limitations, reasons for caution: Limited numbers of media samples were
implantation rates (%62 Lifeglobal and 53%Vitrolife ) and miscarriage rates (15% available for each culturing protocol and researchers performing the data analysis
Lifeglobal and 4%Vitrolife). On the other hand, the best 1 or 2 blastocysts based were not blind to the culturing conditions or PGT-A biopsy results. Maternal
on morphology grading, independent of from which study group coming from DNA contamination of media may be underestimated, as it was only possible
were transferred (average 1.16). More blastocysts were selected to transfer in to detect when the embryo in culture was male.
the Lifeglobal group because of their had a better morphology (58% v/s 42%) Wider implications of the findings: Currently there are still many questions
nevertheless the differences were not statistically significant. Finally in our main on what effect embryology has on the quantity and quality of embryo DNA
objective live birth rate, no statistical differences were found (48% Lifeglobal and found in spent culture media. Successful NI-PGT-A may require specific culturing
53%Vitrolife) conditions and individual laboratories are best placed to further identify key steps
Limitations, reasons for caution: More studies can be conducted to to generate quality concordant results from spent media.
ingress the number of patients to confirm our findings. Trial registration number: Not applicable
Wider implications of the findings: In agreement with other studies our
results confirm that uninterrupted culture media is providing optimal results. As
both brands analyzed had comparable results we might suggest each laboratory P-118 Spontaneous blastocyst collapse during pre-vitrification
can select one of those taking into account other important characteristics of equilibration is related to a lower pregnancy rate: a prospective
management such as prices and durability. cohort study
Trial registration number: 1706-SCL-064-MC M. Murakami1, S. Mizumoto2, K. Tanaka2, H. Otsubo2, Y. Nagao2,
T. Kuramoto3
1
Kuramoto Women’s Clinic, Research laboratory, Fukuoka, Japan ;
P-117 Observations from a multi-centre study using spent culture 2
Kuramoto Women’s Clinic, IVF laboratory, Fukuoka, Japan ;
media for NI-PGT-A 3
Kuramoto Women’s Clinic, President, Fukuoka, Japan
M. Jasper1, F. Bardan1, C. Robinson1, S. Myers1, M. Fraser1
1
PerkinElmer Health Sciences Australia Pty Ltd, PerkinElmer Health Sciences Study question: Is there an association between the spontaneous collapse
Australia Pty Ltd, Adelaide, Australia pattern of a blastocyst in an equilibration solution (ES) before vitrification and
implantation success?
Study question: Does non-invasive preimplantation genetic testing for aneu- Summary answer: Ongoing pregnancy rates (PRs) after warmed ET decreased
ploidy (NI-PGT-A) of spent embryo culture media require specific culturing in blastocysts that collapsed completely during equilibration before
conditions? vitrification.
Summary answer: Individual laboratories rigorously follow defined embryo What is known already: Previous time-lapse studies have found that blasto-
culturing protocols to maximise pregnancy outcome and the implementation of cysts that spontaneously collapse during embryo culture are less likely to hatch
non-invasive testing may be possible with minimal protocol changes. in mice and implant in humans, suggesting a negative impact of the collapse on
What is known already: Several laboratories are testing the feasibility of reproductive outcomes. Conversely, it has been shown that artificial shrinkage
using spent embryo culture media for Non-Invasive Pre-implantation Genetic of human blastocysts before vitrification improves embryo survival and/or preg-
Testing for Aneuploidy (NI-PGT-A). Results published to-date show different nancy rates. However, shrinkage is not induced in some vitrification protocols,
rates of concordance with embryo biopsy between laboratories. Variability in including ours, as blastocyst viability was already high. To date, no study has been
the culturing conditions and methodologies being tested may explain the conducted to examine the effect of spontaneous blastocyst collapse during
observed difference, as culture conditions are likely to impact the accumulation pre-vitrification equilibration on the clinical outcome.
of both embryonic and contaminating maternal and sperm DNA in spent embryo Study design, size, duration: This study included 471 patients undergoing
culture media. As such, optimization of culturing conditions may significantly their first autologous IVF/ICSI cycle and freeze-all strategy at the blastocyst stage
influence the concordance of results for NI-PGT-A and could assist a controlled in our clinic between June 2018 and October 2019. For patients with multiple
evaluation of its use in place of biopsy-based PGT. vitrified blastocysts, embryos for transfer were selected hierarchically based only
Study design, size, duration: NI-PGT using the PG-Seq™ Rapid Non- on morphological scoring (Gardner score) during culture. To minimize bias, only
Invasive PGT kit (PerkinElmer) was performed on spent embryo culture media the data from single ETs with Day 5 blastocysts (Score 4, excluding CC) in the
samples from several laboratories implementing different culturing conditions. first warmed cycle of each patient were analyzed.
The influence of culturing conditions, including culture volume and collection Participants/materials, setting, methods: Blastocysts were vitri-
time on concordance between NI-PGT results and embryo biopsy results was fied-warmed in in-house prepared solutions using Rapid-i carriers. Prior to vit-
analysed and conditions found to have an influence were identified. rification, intact blastocysts were equilibrated in 10% ethylene glycol (12 min,
Participants/materials, setting, methods: Laboratories provided spent 37°C). Their spontaneous collapse patterns were assessed under an inverted
culture media samples for protocol development and/or testing. Once the microscope before being vitrified in 15% DMSO + 15% ethylene glycol + 0.5 M
embryo was removed from the culture droplet, the culture media was collected sucrose. Blastocyst collapse was defined as the separation of the trophectoderm
and stored at -20°C, with ethics approval. Equivalent template volumes of cells from the zona pellucida. Collapsed blastocysts with/without blastocoel
spent embryo culture media were amplified using the PG-Seq™ Rapid Non- cavity were called partially/completely collapsed blastocysts.
Invasive PGT kit protocol, a novel single tube amplification and indexing work- Main results and the role of chance: Non-collapsed (NC), partially col-
flow specifically optimized for NI-PGT-A, and DNA yield was assessed before lapsed (PC), and completely collapsed (CC) blastocyst ETs were 22.9%
sequencing. (108/471), 53.5% (252/471), and 23.6% (111/471) of the ET cycles, respec-
Main results and the role of chance: Whole genome amplification resulted tively. Most embryos (99.6%, 441/443) survived warming. PRs per ET tended
in the amplification of 77-100% of spent embryo culture media samples provided to decrease as blastocysts progress from one collapse pattern to another. In this
by the different laboratories (n=9). Ploidy concordance with the embryo biopsy study, the ET cycles were divided into NC/PC and CC groups to facilitate data

analysis. Maternal age was similar between the groups. Clinical PRs per ET were and SCM (3.4±5.5 vs 2.9±2.3, p=0.075). However, blastulation rate on day 5
61.7% (NC/PC) versus 51.4% (CC) (odds ratio (OR): 1.52, 95% confidence was significantly higher for embryos cultured in CCM (72.2% vs 63.1, p=0.017).
interval (CI): 0.97-2.39, P = 0.061). Ongoing PRs per ET were 55.8% (NC/PC) The usable blastocyst rate was not significantly different between the two
versus 37.8% (CC) (OR: 2.07, 95% CI: 1.31-3.30, P = 0.001). A logistic regression groups compared (76.5% vs 79.3%, p=0.448 for CCM and SCM). From a
analysis was performed on the ongoing PRs to account for the effects of some Poisson regression model adjusted for confounding factors, no differences
possible confounding factors, including the complete blastocyst collapse, blas- were observed in euploidy rate between CCM and SCM (OR=2.4387, CI 95%:
tocyst morphology, MII oocyte number, and maternal characteristics (age, parity 0.93-6.38, p=0.0699 for CCM and OR=1.1791, CI 95%: 0.94-1.48, p=0.1572,
(yes/no), smoking (yes/no), and BMI). In addition to the maternal age (years, for SCM). Only patients age was associated with the euploidy rates, regardless
OR: 0.89, 95% CI: 0.84-0.94, P < 0.001) and blastocyst morphology (good the type of culture medium (OR=0.9494, CI 95%: 0.93-0.97, p<0.001). A
(4AA/4AB/4BA) versus not, OR: 1.60, 95% CI: 1.09-2.36, P = 0.017), the linear mixed model was performed controlling for confounding factors and a
blastocyst collapse (yes versus no) had a significant effect on the results (OR: significant increase in toP-quality blastocysts (ICM= A or B and TE=A) was
0.48, 95% CI: 0.31-0.77, P = 0.002). found when embryos were cultured in SCM (OR=-0.2545, p=0.814 for CCM
Limitations, reasons for caution: Cryoprotectant concentration of our ES and OR=0.5070, p=0.036 for SCM).
is relatively low. Thus, when considering the collapse pattern as a criterion for Limitations, reasons for caution: The current results are based on a ret-
selecting embryos for transfer, caution is needed as ES osmolarity can influence rospective observational study.
blastocyst shrinkage. Further studies with more participants are required to Wider implications of the findings: Under controlled conditions, in vitro
confirm the results. outcomes are comparable when using a CCM or SCM, although quality of blas-
Wider implications of the findings: This is the first study to demonstrate tocysts can be improved when embryos are cultured in SCM. As expected,
a relationship between spontaneous blastocyst collapse during pre-vitrification patient’s age has the most important impact on euploidy rate.
equilibration and a decreased pregnancy rate. Studying the collapse pattern may Trial registration number: None
assist in selecting the most viable blastocysts after warming and increase IVF
success rates. P-120 Embryos that divided chromosome correctly develope
Trial registration number: UMIN000039264 to the morphologically-good blastocyst in spite of abnormal
cytokinesis at 1st mitosis
P-119 Influence of culture media on embryo developmental S. Hashimoto1, Y. Nakaoka2, Y. Morimoto3
pattern and ploidy status: a sibling oocyte study. 1
Osaka City University Graduate School of Medicine, Reproductive Science
A. Abdala1, I. Elkhatib1, A. Bayram1, A. Arnanz1, A. El-Damen1, Institute, Osaka, Japan ;
B. Lawrenz1,2, H.M. Fatemi1, N. De Munck1 2
IVF Namba Clinic, IVF Namba Clinic, Osaka, Japan ;
1 3
IVIRMA Middle East, Fertility Clinic, Abu Dhabi, United Arab Emirates ; HORAC GrandFront Osaka Clinic, HORAC GrandFront Osaka Clinic, Osaka, Japan
2
Women´s University Hospital Tuebingen, Obstetrical Department, Tuebingen,
Germany Study question: Why do human embryos normally develop after abnormal
cytokinesis at 1st mitosis?
Study question: Is there a difference in embryo development and euploidy Summary answer: Only embryos that underwent correctly chromosomal
rate between blastocysts cultured in continuous culture media (CCM) or sequen- division developed to the morphologically-good blastocyst in spite of abnormal
tial culture media (SCM)? cytokinesis at 1st mitosis.
Summary answer: While the blastulation on day 5 is increased in CCM, the What is known already: It has been shown that a live birth from embryos
usable blastocyst and euploidy rates are not significantly different between CCM which underwent abnormal cytokinesis at 1st mitosis. During abnormal cytokinesis,
and SCM. there are two main patterns in chromosomal division, such as dual spindle forma-
What is known already: Culture medium is fundamental for in vitro blastocyst tion without syngamy and multipolar spindle formation with syngamy. It’s unlikely
development. While CCM supplies all nutrients that embryos need to grow (“let to believe these abnormally chromosome-separated embryos could develop to
the embryo choose”), SCM mimics physiological conditions (“back to nature”). babies. There is the potential to include morphologically-abnormal cytokinesis
Many studies evaluated whether CCM improves clinical outcomes as compared without abnormal chromosome partitioning, such as fragmentation.
to SCM, hence, comparisons failed to identify a superiority. Recent data suggest Study design, size, duration: This study was approved by the ethical com-
that euploidy rates may be affected by culture media, however, non-consensus mittee of the Japan Society of Ob/Gy. We assessed chromosomal behavior
has been reached due to the variability of experimental designs, underpowered during 1st mitosis in human zygotes and their development in vitro using 140
studies and heterogenicity that might affect outcomes. The lack of standardiza- donated zygotes. Pronuclear ova were donated from infertility couples after
tion of the comparative studies drives to controversial conclusions of which obtaining their informed consent.
culture media improve euploid outcomes. Participants/materials, setting, methods: Donated pronuclear ova were
Study design, size, duration: A single center observational cohort injected with a mixture of mRNAs encoding EGFP-EB1 and mCherry-his-
study was performed between September 2018 and March 2019, including tone-H2B. Dynamic changes of their chromosomes were monitored continu-
1452 mature oocytes (MII). Patients who underwent preimplantation genetic ously using a confocal microscope inside an incubator every 15 min for 24 h.
testing for aneuploidies (PGT-A) with at least four fresh autologous MII insem- After live imaging, morphological changes of embryos were recorded by a con-
inated by ICSI were included. Severe male factor was excluded. According to ventional time-lapse system using a light microscope.
clinical practice, sibling MII oocytes were randomly split between two media: Main results and the role of chance: To directly examine possible idea for
751 in Global Total LP (CCM) and 701 in Sage cleavage and blastocyst inclusion of embryos without abnormal chromosome partitioning into abnormal
media (SCM). cleavage, we observed chromosomal behavior during 1st mitosis using live cell
Participants/materials, setting, methods: Zygotes were cultured up to imaging system. One hundred-forty zygotes underwent 1st mitosis after RNA
day 7 with both media refreshed on day 3 under 6% CO2, 5% O2 and 89% N2, injection. Abnormal cytokinesis at 1st mitosis was observed in 45 zygotes (32%).
pH=7.2-7.4. Fertilization, cleavage, D5 blastulation, usable blastocyst (total In 12 zygotes, normal chromosome division was observed under a confocal
blastocysts biopsied/total blastocysts formed) and euploidy rates were microscope (27%) despite abnormal cytokinesis. Three zygotes developed to
recorded. Blastocyst expansion, inner cell mass (ICM) and trophectoderm the blastocyst stage from normally chromosome-divided embryos (25%). One
(TE) quality were graded according to Gardner´s scoring system, prior TE of these blastocysts were categorized as morphologically-good (4BB) by
biopsy on day 5, 6 or 7. TE samples were analyzed for PGT-A by next gener- Gardner’s strict criteria. Monitoring of chromosomal behavior at 1st mitosis using
ation sequencing. a confocal microscope enables us to distinguish embryos which underwent cor-
Main results and the role of chance: Patient mean age was 34.3±6.0 rect chromosomal division in spite of abnormal cytokinesis. The data of the
years old. No differences were observed in fertilization and cleavages rates present study suggest that embryos which underwent chromosomal division
(77.4% vs 75.5%, p=0.468, and 97.6% vs 99.1%, p=0.089 for CCM and SCM, correctly could develop to live babies in spite of abnormal cytokinesis at 1st
respectively). Total number of blastocysts formed was similar between CCM mitosis.

Limitations, reasons for caution: It is not ethically permitted in clinical fields 2

University of Health Sciences- Gülhane Training and Research Hospital, IVF
to monitor the chromosomal behavior at 1st mitosis of human embryos under Center, Keçiören, Turkey
a confocal microscope. Further studies are required to detect the difference
between correct and incorrect chromosomal division using a light microscope. Study question: What is the rates of clinical pregnancy after the transfer of
Wider implications of the findings: Live imaging using a confocal micro- vitrified and thawed human embryos on Day 3, Day 4, and Day 5 of embryonic
scope inside an incubator yielded information about chromosome behavior development.
during preimplantation embryo development that were unobservable by a time- Summary answer: Higher rates of pregnancy were determined with the
lapse system using a light microscope. Live imaging is expected to play an import- freezing of human embryos on Day 4 and the transfer of those embryos
ant role in understanding chromosomal aberrations during pre-implantation on Day 5.
embryo development. What is known already: One of the most important stages in clinical infertility
Trial registration number: Japan Society of Obstetrics and Gynecology 112 treatment today is frozen embryo transfer. The cryopreservation of human
embryos allows us to increase the potential of conception for any in vitro fer-
tilization (IVF) cycle to its highest level and to prevent the waste of embryos.
P-121 Prevalence and predictive factors for complete fertilization Vitrification, which is currently used to freeze human oocytes and embryos at
failure in in vitro fertilization treatment cycles: a retrospective the pronucleus, cleavage, morula, and blastocyst stages, is a simple, low-cost,
analysis of a large-scale nationwide database study and effective method.However, there is no consensus regarding which stage of
T. Takahashi1, K. Ota1, H. Mizunuma1 embryonic development is most suitable for vitrification with maximum devel-
1
Fukushima Medical University, Fukushima Medical Center for Children and opment after the embryo is thawed.
Women, Fukushima, Japan Study design, size, duration: 148 embryo transfer cycles, using embryos
frozen with the closed vitrification method, thawed during the three-year period
Study question: What is the prevalence of and what are the risk factors for between January 2016 and December 2018 were investigated. Results of the
complete fertilization failure (CFF) in patients undergoing in vitro fertilization embryos frozen were compared in three groups based on the day they were
(IVF) treatment cycles. frozen Day 3, Day 4, or Day 5. This study is the first one in which pregnancy
Summary answer: The prevalence of CFF was 13.2% and the major predictive results are compared following freezing and thawing of human embryos in the
factor for IVF cycles with CFF was a retrieved oocyte number. cleavage, morula, and blastocyte stages.
What is known already: It is reported that the rate of CFF in IVF treatment Participants/materials, setting, methods: 128 patients, and 148 cycles
cycles ranges from 5 to 15%. A more accurate indicator of prevalence is belonging to this infertile patients, were included in our study. Male majority
unknown, and the predictive factors for CFF in IVF cycles based on real-world factor cases were excluded from the study. Closed vitrification method were
data are unclear. used during the study. 17.4% of patients’ embryos were frozen on Day 3, 55.6%
Study design, size, duration: A retrospective cohort study was performed, on Day 4, and 27% on Day 5.The clinical pregnancy and ongoing pregnancy
based on 1404648 treatment cycles in registered assisted reproductive technol- results were examined, after thawing.
ogy (ART) data from the Japan Society of Obstetrics and Gynecology between Main results and the role of chance: The clinical pregnancy and ongoing
2007 and 2012. pregnancy results were examined, after thawing, in all groups according to
Participants/materials, setting, methods: We analyzed data from 220,065 their infertility reasons, with no significant difference observed in terms of
fresh cycles of IVF when one or more oocytes were retrieved. An instance of clinical pregnancy and ongoing pregnancy rates (p:0.061, p:0.493). 20% of the
CFF was defined as zero fertilized oocytes obtained. The IVF cycles with CFF embryos frozen on Day 3 were also transferred on Day 3; 52% were trans-
or with one or more oocytes fertilized were compared to determine predictive ferred on Day 4; and 28% were transferred on Day 5. 26.2% of the embryos
factors for CFF using multivariate logistic regression analyses. The area under frozen on Day 4 were transferred on Day 4, and 73.8% were transferred on
the receiver operating characteristic (ROC) curve was used to assess the dis- Day 5. All embryos frozen on Day 5 were transferred the same day. Clinical
criminative ability of the logistic models. pregnancy and ongoing pregnancy rates were observed to be significantly
Main results and the role of chance: The total percentage of CFF in IVF higher in the results of embryos frozen on Day 4 and transferred on Day 5,
cycles was 13.2% (29076/220065). On multivariate analysis, patient age (adjusted according to the transfer day (clinical pregnancy rates were 28.5% on day four
odds ratio [aOR]: 1.00, 95% confidence interval [CI]: 1.00-1.01, p<0.01), oocyte and 49.1% on day five)
number (aOR: 0.78, 95% CI: 0.77-0.79, p<0.01), sperm motility (aOR: 0.99, Limitations, reasons for caution: There is a need for comparative studies on
95% CI: 0.98-0.99, p<0.01), and male factor infertility (aOR: 1.43, 95% CI: vitrification of human embryos at different stages with a larger patient population.
1.38-1.49, p<0.001) were independent predictive factors of CFF. The percent- Wider implications of the findings: This study is the first one in which
ages of CFF cycles with one, two, three, four, and five oocytes retrieved were pregnancy results are compared following freezing and thawing of human
34.2%, 15.3%, 8.6%, 6.0%, and 4.6%, respectively. The efficacy of oocyte number embryos in the cleavage, morula, and blastocyte stages. Higher pregnancy rates
and sperm motility for predicting CFF were evaluated using ROC curves. The can be achieved by transferring, in the blastocyst stage, embryos that were frozen
areas under the curves for oocyte number and sperm motility were 0.79 and and thawed in the morula stage.
0.59, respectively. Setting the threshold at 2 for oocyte number offered the best Trial registration number: 19/48
compromise between sensitivity (0.66) and specificity (0.80).
Limitations, reasons for caution: As cases with serious semen abnormalities
may have undergone intracytoplasmic injection, a selection bias cannot be ruled P-123 The expression of Anti-Müllerian hormone receptor
out in the IVF treatment cycles analyzed. type II (AMHRII) in human oocytes from the in vitro fertilization
Wider implications of the findings: This study showed that collecting a programme and their maturation with recombinant AMH
large number of retrieved oocytes may prevent CFF. These results provide J. Bedenk1, E. Vrtačnik-Bokal1, N. Jančar1, K. Geršak1, I. Virant-
information that may be useful for counseling patients before their undergoing Klun1,2
ART treatments. 1
University Medical Centre Ljubljana, Department of Obstetrics and Gynaecology,
Trial registration number: not applicable Ljubljana, Slovenia ;
2
University Medical Centre Ljubljana, Centre for Clinical Research, Ljubljana, Slovenia
P-122 Investigation of transfer results of human embryos which

Study question: Is AMHRII receptor present in the immature and mature
are vitrified and thawed in cleavage, morula and blastocyst stages
non-fertilized oocytes from the IVF programme; can recombinant AMH affect
C. Korkmaz1, U.G. Yıldız2, U. Fidan2, B. Baykal2, S.T. Ceyhan2, oocyte maturation in vitro?
E. Agacayak2 Summary answer: AMHRII receptor is more expressed in immature than
1
University of Health Sciences- Gülhane Training and Research Hospital Ankara- mature oocytes and immature oocytes can be matured in vitro in the presence
Turkey, IVF Center, Ankara, Turkey ; of recombinant AMH only.

What is known already: AMHRII, a key receptor for AMH, is expressed in Study design, size, duration: The present study includes 308 couples
Sertoli and Leydig cells in testes and in granulosa and theca cells in the ovaries. This who underwent assisted reproduction treatment at UMR-centre HERA
receptor is crucial for the function of Anti-Müllerian hormone (AMH) in follicular between October 2009 to April 2019. The couples were counseled for in
cells, especially as a regulator of growing ovarian follicles. Despite this, we could vitro treatment and were proposed to perform ICSI and IVF on sibling
not find any studies on the expression of AMHRII receptor in human oocytes. oocytes. In none of the cases infertility was due to severe male factor or
Therefore, we decided to analyze the oocytes for AMHRII receptor expression previous fertilization failure after in vitro technique. The average age for
and mature the oocytes in vitro with recombinant AMH in the maturation medium female patients was 33.4.
to perhaps demonstrate the direct action of this hormone on oocytes. Participants/materials, setting, methods: The patients underwent ovarian
Study design, size, duration: During two years, we included two groups of stimulation using GnRH agonist or antagonist protocol. After oocytes retrieval,
oocytes – 24 immature (GV) and 24 mature (MII) oocytes to perform immu- oocytes-cumulus complex were randomly divided into two groups. Oocytes
nocytochemistry for expression of AMHRII and elucidate whether the maturity were simultaneously fertilized by ICSI and IVF with the same semen sample.
stage of oocytes has an effect on expression of AMHRII receptor. In the next Fertilization and embryo development were assessed by time-lapse monitoring.
step, 15 immature and 15 mature oocytes were analysed on expression of Embryos were transferred on the same cycle and surplus blastocysts vitrified
AMHRII gene. Immature oocytes (n=15) were matured in vitro in the presence for further transfer. Fertilization rate, embryo kinetic and embryo implantation
of recombinant AMH to test its activity. rates were compared between ICSI and IVF.
Participants/materials, setting, methods: We used the oocytes from Main results and the role of chance: On 3,400 oocytes collected, 1,270
patients included in the in vitro fertilization programme (ICSI) after their written were fertilized by ICSI and 1,035 by IVF. Fertilization rate was statistically higher
informed consent. Immunocytochemistry was performed using the anti-AMHRII in ICSI group compared to IVF group (76.3% in ICSI versus 58.6% in IVF, p
antibody; fluorescence was quantified with Fiji software. Gene expression analysis <0.0001). There were no statistically significant differences in cleavage rate
for AMHRII, ZP1, ZP2, ZP3, ZP4, BMP4 and GDF9 was performed using qPCR (98.7% in ICSI versus 98.7% IVF; p> 0.05), embryonic quality in GII (39.7% versus
after superamplification, as well as in vitro maturation of oocytes with recom- 41.8%) and the rate of usable embryo on total produced embryos per technique
binant AMH (R& DSystems 1737-MS, 100 ng/mL) alone in the maturation (49.4% versus 46.2%) between ICSI and IVF (p>0.05). The rates of clinical
medium of MediCult IVM System. pregnancy (46.6% vs. 46.8%), of abortion (12.7% vs. 13.3%) were also compa-
Main results and the role of chance: The AMHRII receptor was expressed rable (p> 0.05). Implantation rate was statistically higher for embryos from IVF
in both immature and mature ocytes, mostly in a spotted pattern through the compared to the sibling embryos produced in ICSI (42.6% versus 55.2%, p
whole cell membrane, as well as in the zona pellucida, but in a much lower <0.05). Embryo kinetic was faster in ICSI compared to IVF.
quantity. There was also accumulation of AMHRII protein around the genetic Limitations, reasons for caution: Not all vitrified surplus blastocysts have been
material (germinal vesicle) in the immature oocytes, as revealed by immuno- thawed and transferred. Consequently cumulative pregnancy rate is incomplete.
cytochemistry. The qPCR analysis confirmed the expression of gene AMHRII Wider implications of the findings: ICSI may induce adverse event on
as well as the co-acting genes related to zona pellucida (ZP1, ZP2, ZP3, ZP4) embryo competence. In case of absence of indication for ICSI, IVF should be
and development (BMP4 and GDF9) in immature and mature oocytes, which preferred as fertilization method.
may reflect the potential function of AMHRII in the oocytes. We also proved Trial registration number: xx
that AMHRII gene was expressed 10x more in immature oocytes than in mature
oocytes (P=0,006; Student T-test). All immature oocytes matured in vitro in P-125 Identification of the optimal puncture position on oolemma
the presence of recombinant AMH alone (without FSH and HCG) in the without degeneration in Piezo-ICSI using image analysis: a pilot
maturation medium. Our preliminary data show that AMH does not act only study
on the follicular cells but may also act on the oocyte directly. S. Mizuta1, T. Maekawa2, Y. Kinishi3, H. Matsubayashi2, K. Kitaya1,
Limitations, reasons for caution: The limitations of this research may T. Takeuchi2, Y. Hata3, T. Ishikawa1
be: impaired quality of mature oocytes which did not fertilize after ICSI and a 1
Reproduction Clinic Osaka, Reproductive Medicine, Osaka, Japan ;
relatively low number of oocytes. A proportion of them degenerated during 2
Reproduction Clinic Tokyo, Reproductive Medicine, Tokyo, Japan ;
the immunostaining procedure. This research is still ongoing to increase the 3
University of Hyogo, Graduate School of Simulation Studies, Kobe, Japan
number of oocytes.
Wider implications of the findings: Based on our data, we assumed that Study question: Is it possible to identify the optimal puncture position on
AMH, in addition to follicular cells, may directly act on oocytes in vitro. This oolemma which may not cause degeneration in Piezo-ICSI using image analysis?
implies a new understanding of the human oocyte maturation process. Further Summary answer: Visualizing the likelihood of unintentional membrane rup-
research will show whether recombinant AMH can improve the process of ture using image analysis can identify positions that result in lower risk of oocyte
human oocyte maturation in vitro. degeneration following Piezo-ICSI.
Trial registration number: Republic of Slovenia National medical ethics What is known already: Oocyte degeneration may take place in Piezo-ICSI as
committee, trial number 0120-546/2018/6 a result of unintentional membrane rupture in the puncturing process. Identifying
the appropriate puncturing position may decrease the likelihood of membrane
P-124 ICSI versus IVF on sibling oocytes: The real efficacy rupture and thus degeneration, but this cannot be done visually. Image feature
comparing laboratory results and clinical outcomes analysis is generally used to represent the useful features such as color, brightness,
S. Chamayou1, R. Lombardi1, C. Ragolia1, C. Alecci1, G. Storaci1, and contour. Among capturing image features, Local Binary Patterns (LBP) can
R. Sapienza1, S. Romano1, C. Cardea1, A. Liprino1, E. Maglia1, efficiently summarize the local structures of images, and it has been applied in
A. Guglielmino1 texture analysis in various fields including face recognition. There were no studies
1
U.M.R. Soc. Coop. a.r.l., Unità di Medicina della Riproduzione, Sant’Agata li identifying the optimal puncture position using image analysis.
Battiati-Catania, Italy Study design, size, duration: We retrospectively performed the LBP method-
ology to analyze the moving images of 131 oocytes following Piezo-ICSI between
Study question: ICSI versus IVF : which technique is more efficient based on August 2018 and January 2019. These oocytes were categorized as either having
in vitro results and clinical outcomes in absence of severe male factor? Conclusions undergone unintentional rupture (UR: n = 30) or no rupture (NR: n = 101).
from sibling oocytes study. Participants/materials, setting, methods: LBP values were calculated in
Summary answer: Despite of the highest fertilization rate in ICSI, IVF-derived the analysis region centered around the puncture position. Median values for
embryos have a higher implantation rate compared to ICSI-derived embryos. UR and NR were calculated to select an effective pattern for rupture evaluation
What is known already: The ICSI procedure has been invented to treat from the 256 shape patterns acquired by LBP. After choosing the three effective
severe male infertility and full fertilization failure after conventional IVF. Nowadays patterns with hierarchical clustering, we employed Ward’s hierarchical cluster
ICSI tends to be applied in every case to ensure fertilization as reported in the analysis method and calculated the Euclidean distance between the cluster bary-
latest report from the European IVF-monitoring Consortium (2014) where ICSI center and each data point to define an index indicating membrane rupture
resulted applied in 71.3% of the fresh treatments. implausibility.

Main results and the role of chance: Two clusters, the cluster A and B, Logistic regression found the followings to be significant: Age at freezing
were classified from hierarchical clustering. Following ICSI, 2 out of 27 oocytes (p < 0.001, OR 0.939, 95% CI 0.908 - 0.971), Day 5 vs Day 6 transfer (p < 0.001,
from the cluster A and 28 out of 104 from the cluster B resulted in UR, OR 2.060, 95% CI 1.443 - 2.978), trophectoderm score of A upon thawing
indicating that the cluster A strongly represented NR group and the cluster (p =0.01, OR 1.482, 95% CI 1.100 – 1.997), and the expansion score upon
B weakly represented UR group, with the sensitivity as 0.93. A significant thawing (p < 0.001 overall via likelihood ratio Chi-square test). Embryos
difference between the UR and the NR group was reported from the Euclidean with an expansion score of 1 have 75.7% lower odds of live birth than embryos
distance calculations between the barycenter of Cluster A and each data point with grade 6 expansion score (OR 0.243, 95% CI 0.085 – 0.722). The score for
(P = 0.001), where data showed a longer distance from the barycenter inner cell mass was not found to be significant.
amongst the UR group and a shorter distance in the NR group. All degener- Limitations, reasons for caution: Other potential covariates including body
ated oocytes after ICSI procedure were from UR (27.7%, 8/30). Of these, mass index, parity and aetiology of infertility were not included in the analysis.
two were from the cluster A (100%, 2/2) and six were from the cluster B Other morphokinetic factors such as abnormal cleavage may have been included
(21.4%, 6/28), while the 2PN rate was 0% and 67.9% in the clusters A and into selection process for freezing and thawing, but were not included in the
B, respectively. On the other hand, no degeneration was observed from the analysis.
NR group, and the 2PN rate was 84.0% and 81.6% in the clusters A and B, Wider implications of the findings: The study provides a simple decision
respectively. Fertilization and degeneration results in NR were significantly support tool for embryologists to decide on which frozen blastocysts to transfer.
better than UR groups. Morphological features such as trophectoderm grade and expansion score are
Limitations, reasons for caution: This was a retrospective pilot study with predictors of livebirth. Wherever possible, a day 5 blastocyst should be trans-
a small sample size, and this was a single-center study. Moreover, this study did ferred in preference to a day 6 blastocyst.
not include embryonic development and clinical outcomes. Further prospective Trial registration number: not applicable
studies in large samples size are needed.
Wider implications of the findings: We can retrospectively classify UR or P-127 Understand chromosomal self-correction of human embryo
NR from the shape feature of the oolemma by the image analysis with AI. If we with non-invasive genomic test
could recognize optimal puncture position by visualizing shape features of the J. Yan1, M. Li1, L. Xiang1, X. Wu1, Y. Li1
oolemma in real-time, we could reduce degeneration of the oocytes after 1
The First People’s Hospital of Yunnan Province-, Reproductive Medical Center-,
Piezo-ICSI. Kunming, China
Study question: Can mosaic embryo commit self-correction or elimination
P-126 Day of blastocyst freezing is a better prognostic factor process during development? Are there differences between pre-implantation
than morphology for livebirth in single frozen embryo blastocyst (D1~D5) and post-implantation stage (D5~D14)?
transfer Summary answer: Chromosomal self-correction could be observed in both
M.W. Lui1, C. Lamb2, D. Thong3, S. Pickering2, K.J. Thong2 pre- and post-implantation stage. In this research, most of the aneuploidy cells
1
Queen Mary Hospital- Hong Kong, Obstetrics and Gynaegolocy, Hong Kong, decreased before Day 5 post fertilization.
Hong Kong ; What is known already: Chromosomal mosaicism in human pre-implantation
2
Royal Infirmary of Edinburgh, Edinburgh Fertility and Reproductive Endocrine embryos has been described, and is regarded as a significant factor that contrib-
Centre, Edinburgh, United Kingdom ; utes to unsuccessful embryo implantation and spontaneous abortion. It is
3
Statistician, independent researcher, Edinburgh, United Kingdom reported in cleavage stage, the mosaic embryo is up to 70%. and in blastocyst
the ratio reduces to about 30%. Current hypothesis is during the development
Study question: Is the day of blastocyst freezing or blastocyst morphology a embryo excluded the aneuploidy cells for self-correction. However, lack of
better prognostic factor for livebirth in single frozen embryo transfer (FET)? proper approaches, the quantitatively reports are still rare. The post-implantation
Summary answer: Day-5 blastocyst is associated with higher livebirth rate stage situation is also elusive.
than day-6 blastocyst, even with a better morphological scoring at freeze. Study design, size, duration: From 2019~2020, 36 frozen embryos from
What is known already: Assessment of development potential is essential 23 patients (age 22~34 years), who determine to donate their embryos after
in reducing the pregnancy interval in the era of single embryo transfer and successful pregnancy with informed consent in Reproductive medical center, the
optimizing the livebirth rate from treatment. The rate of blastocysts development first people’s hospital of Yunnan province,China.
(day-5 or day-6 blastocysts) (1) and morphological assessment (2, 3) have both Participants/materials, setting, methods: For each cleavage embryo,
been identified as significant prognostic factors for livebirth in blastocyst(s) trans- 2~3 blastomeres are tested and the embryo was continually cultured to blas-
fer. However, we are uncertain which of the parameters is a better prognostic tocyst. and collected the medium In Day5, 1~2 TE and 1 ICM are biopsied.
factor for livebirth rate for women undergoing FET. For mimicing post-implantation embryo in vitro, three-dimensional blasto-
Study design, size, duration: A retrospective study was carried out for all cyst-culture system(Nature, 2010) is adopted. This method allowed the devel-
single blastocyst frozen transfer from January 2015 to December 2018 in a single opment of 3D structure of embryo and mimic the in vivo situation better. The
fertility centre. Cases requiring extended culture after thawing and incomplete culture medium are collected in Day8/10/2. Medium and embryo are
data were excluded. The cases were identified from the centre’s computerized sequenced with NGS (Yikon).
data. Morphological assessment was carried out using the Gardner and Main results and the role of chance: In our study, 92% (297/323) samples
Schoolcraft Grading System (4). Good quality embryos were defined as 1-6AA, are successfully amplified. Data from ICM are considered as represented result
3-6AB or 3-6BA. for the embryo. In pre-implantation stage, mosaicism portion over 20% is
Participants/materials, setting, methods: A total of 1541 single blastocyst included, and 24% (6/25) of the embryos carrying at least one mosaic chromo-
transfers were carried out during the study period and 1162 transfers were some. 72% (17/25) results from mediums are at least consistent with TE or ICM.
included in the analysis. Chi-square testing and binary logistic regression of live In pre-implantation stage, the consistent of each blastomeres are 83% (55/66)
birth rate against: age of women at freezing, interval of freezing, embryo quality TE and ICM are 86.3%(19/22). 16%(4/25) self-correction (CNV in blastomeres
after thaw and change in quality from freezing to thawing were performed. but not in TE/ICM or the mosaic ratio decreased) and 4%(1/25) novel chro-
Women with morulas after thaw were excluded in the analysis using logistic mosomal aneuploidy (CNV in TE but not in blastomeres) are observed.
regression. 8%(2/25)aneuploidy is detected in medium but not in either blastomere
Main results and the role of chance: The mean age of women at freezing or TE/ICM.
and duration of freezing were 34.0 +/- 3.9 years and 0.9 +/- 1.3 years respec- In post-implantation stage, 81.8%(9/11) are developed embryos carrying
tively. The overall livebirth rate was 31.4%. The livebirth rate was the highest consistent chromosomal situation. One duplication of chromosome 14 is
for good quality day-5 blastocysts of 39.4%, followed by 25.2% in poor quality observed in Day8 but not in Day10 and Day12(1/11) .
day-5 blastocysts. It was lower for good quality day-6 blastocysts of 22.5% and Limitations, reasons for caution: The size of the research is still small; the
15.9% in poor quality day-6 blastocysts (p <0.001 for differences and trend). post-implantation and pre-implantation embryos are not the same batch; some

of the donated embryos are not from the ICSI cycle, potentially maternal con- atmospheric fluctuations, accumulation of volatile organic compounds or ammo-
tamination could not be excluded. nium built-up may negatively affect the culture environment. Future research is
Wider implications of the findings: Non-invasive chromosomal test provides needed to compare reproductive outcomes in RCT performed with patient’s
us powerful tool to study the genomic change. We find more mosaicism in medium own oocytes
but not in biopsy, and bsed on current study size, most self-correction could be Wider implications of the findings: The present study suggests that commercial
observed before Day5, for post-implantation stage, more evidence are required. single culture media when used in a continuous (one-step) approach may result in
Trial registration number: NSFC 31700798 different outcomes in terms of proportion of suitable blastocysts for clinical use. Single
media formulation, laboratory conditions and the culture environment should be
P-128 Prospective comparative study between two commercial properly validated independently on each laboratory conditions
single culture media using sibling donor oocytes collected from Trial registration number: Not applicable
randomized ovaries
C. MIRET LUCIO1, M. Benavent1, M. Escriba1, A. Garcia1, P-129 pregnancy is significantly correlated with the blastocyst
D. González1, N. Borges2, G. Calderon2, J. Crespo3, J. Teruel1 width and area: a time-lapse study.
1
Equipo Médico Crespo, IVF Lab, Valencia, Spain ; R. Sciorio1, D. Thong2, K.J. Thong3, S. Pickering3
2 1
Embryotools, IVF Lab, Barcelona, Spain ; Royal Infirmary of Edinburgh, Edinburgh Assisted Conception Programme- EFREC,
3
Equipo Médico Crespo, Reproductive Medicine, Valencia, Spain Edinburgh- Scotland, United Kingdom ;
2
RIE, Efrec, Einburgh, United Kingdom ;
Study question: To compare embryo development and clinical outcomes 3
RIE, Efrec, Edinburgh, United Kingdom
between two commercial single culture media using sibling donor oocytes col-
lected from randomized ovaries Study question: Does blastocyst width and area affect pregnancy outcome in
Summary answer: Our study suggests that different commercial single media IVF/ICSI cycles?
used uninterruptedly can yield different blastocyst developmental rates, depend- Summary answer: Clinical pregnancy rate (CPR) is significantly higher for blas-
ing on their formulation and/or environmental culture conditions tocysts with a larger width and area, which was confirmed by logistic regression.
What is known already: In recent years, there has been a renewed interest What is known already: In order to maintain pregnancy rates following single
in the use of single media to support human embryo culture in vitro. These media embryo transfer, optimisation of embryo culture and selection is vital. Time-lapse
are based on a single formulation designed following the principle of allowing monitoring (TLM) has the potential to play a crucial role by providing sequential
developing embryos to choose the nutrients according to their needs. An increas- images of embryo development and minimal disturbance, thereby increasing the
ing number of single media have now become commercially available and have probability of selecting embryos with high implantation potential. However,
been swiftly introduced in IVF laboratories worldwide. Whether all single media transfer of good quality embryos does not always lead to successful pregnancy.
are equally suitable for the continuous (one-step) culture of human embryos Therefore, in this study morphometric evaluation was performed. We used a
remains unclear, as recent studies suggest that the formulation, laboratory envi- single assessment of blastocyst area and maximum width to evaluate if these
ronment and culture conditions may result in different outcomes parameters correlated with pregnancy outcomes.
Study design, size, duration: This is a single-center prospective study per- Study design, size, duration: A retrospective analysis of 664 patients who
formed between February and December 2019 that included 84 donors and 94 had eSET was carried out between April 2014 and August 2019. Embryos were
recipients. Ovaries were randomized using a computer-generated randomization cultured individually in 6.0% CO2, 5.0% O2, 89.0% N2, using single step medium
list. Sibling oocytes collected from each randomized ovary were processed and (G-TL™ Vitrolife, Göteborg, Sweden) in the time-lapse incubator (Embryoscope,
cultured in two commercial single media (A and B). The corresponding series ES-D, Vitrolife).
media (fertilization, handling or culture) offered by each brand was used to Participants/materials, setting, methods: All embryos were evaluated using
perform all procedures involving oocytes and sperm samples of the two exper- calibrated annotation tools of the EmbryoViewer. Drawing tools were used to
imental arms measure specific variables such as the maximum blastocyst width and blastocyst
Participants/materials, setting, methods: Oocytes were injected by ICSI area. Data obtained was assessed in terms of CPR at 7 weeks gestation. Statistical
and then cultured individually in miniGPS dishes (LifeGlobal) prepared with either analysis was performed with Mann-Whitney U-test and Logistic Regression.
single medium A or B (25ul medium/well) at 37ºC, in an atmosphere of 6-7% Main results and the role of chance: Variables analysed in this study were
CO2/5% O2/88-89% N2 of a dry atmosphere incubator (IVFcube, Astec). correlated to clinical outcomes. The age [median (range) years old] of women
Laboratory conditions, such as temperature, humidity and volatile organic com- was 34 (22-44). Our results show women who were pregnant had a significantly
pounds (VOC) levels were monitored continuously (Octax Log&Guard, Vitrolife) (P<0.01) larger blastocyst width [median (range) µm] 184 (125-239) versus
during the study period and pH measured in a weekly basis in the two single those who were not pregnant 160 (120-230). A significantly (P <0.01) larger
media used [median (range) µm2] blastocyst area 26099 (12101-45280) was seen in pregnant
Main results and the role of chance: The pH average values were similar versus non pregnant women, 22251 (10992-37931).
in both media (medium A 7.28±0.06 vs medium B-7.27±0.05), and the mean A univariate logistic regression performed showed that blastocyst width
value of VOCs 0.032 ppm. A total of 1103 MII oocytes were injected by ICSI [(OR = 1.026, 95% CI = (1.019, 1.033)] was found to be significant with P-value
(A, n=558 and B, n=545). The proportion of fertilized oocytes was identical <0.0001. This shows that for every µm increase of blastocyst width, the odds
between the two media (A:83.9% and B:79.4%), but oocyte degeneration rate of clinical pregnancy increase by 2.6%.
post-ICSI was significantly lower in group A (3.6% vs 7.3%; p=0.0059). The mean A univariate logistic regression performed also showed that blastocyst area
number of embryos that reached the blastocyst stage cultured in the single [(OR = 1.00008, 95% CI = (1.00006, 1.00011)] was found to be significant with
medium A (64.3%) was significantly lower (p=0.004) than that obtained in P-value <0.0001. The lower odds ratio is explained by a larger range in blastocyst
medium B (73,2%). Similarly, a higher proportion (p=0.031) of blastocysts suit- area. This shows that, for every µm2 increase of blastocyst area, the odds of
able for clinical use (transferred or cryopreserved) was obtained in medium B clinical pregnancy increase by 0.008%. Hosmer-Lemeshow tests of calibrations
(61%) compared to medium A (53.8%). were performed to verify calibration.
Ninety-four patients had an embryo transfer on day 5/6 with either fresh or Limitations, reasons for caution: 6.6% of patients (44/664) were excluded
cryopreserved blastocysts cultured in single medium A (n=39) or B (n=56), with from the study because dimensions could not be measured accurately. Although
a mean number of 1.2 blastocysts transferred/patient in the two groups. No our findings show a clear effect of blastocyst dimensions on clinical pregnancy
differences were found in terms of clinical pregnancy (69.2% and 64.3%, respec- rate, further studies are necessary to confirm these observations.
tively) or implantation rates (67.4% and 56.1%, respectively) between both Wider implications of the findings: Confirmation of our findings could
groups. Miscarriage rates were similar between group A (18.5%) and group B result in establishment of a new algorithm to improve embryo selection at the
(12.5%). Three mixed transfers were excluded from the interpretation of results blastocyst stage. However, this may not exclude the importance of other mor-
Limitations, reasons for caution: Continuous embryo culture in single media phological features/grades in embryo selection.
largely relies on optimal laboratory conditions and a series of osmolality or Trial registration number: not applicabale

P-130 A prospective randomised control study comparing underlie molecular and cellular alterations in either oocytes or sperm, which may
reproductive outcome of day 5 Quarter laser zona thinning partially affect also the embryos that continue their development. By comparing cycles
assisted hatching (qLZT-AH) in frozen thawed embryo transfers with the same number of generated embryos, but different FR, we test whether FR
H. Sharma1, K.D. Nayar1, G. Kant1, S. Draboo1, M. Singh1, per se affects pregnancy and LB rates after fresh embryo transfer.
R. Bhattachrya1, S. Gupta1, K.D. Nayar1 Study design, size, duration: Retrospective cohort study of 7,782 oocyte
1 donation cycles performed in one fertility clinic between 2011 and 2019. Between
Akanksha IVF Centre- Mata Chanan Devi Hospital, Reproductive Medicine, New
5-14 metaphase II oocytes (MII) were assigned to each oocyte recipient. Each
Delhi, India
cycle gave rise to 4, 5 or 6 D+1 embryos after elective ICSI. Reproductive results
Study question: Can quarter zona laser thinning assisted hatching (qLZT-AH) were analyzed per groups with the same number of embryos (4-5-6), regardless
improves reproductive outcome in day 5 frozen embryo transfer cycles? of the number of MII needed to obtain those embryos (varying FR within groups).
Summary answer: Quarter laser thinning assisted hatching (qLZT-AH) on Participants/materials, setting, methods: A minimum of 5 MII were
day 5 frozen thawed embryos is associated with improved implantation rate, assigned to each recipient hence, cycles with 4 embryos presented 33-80% FR,
clinical pregnancy rate over no assisted hatching. cycles with 5 embryos 38-100% FR, and cycles with 6 embryos 43-100% FR. For
What is known already: Hatching is a process where blastocyst escape the each group of embryos (4-5-6), the FR range was divided in categories containing
Zona pellucida (ZP) membrane prior to implantation. This is accomplished >160 cycles each. Mean embryo morphological score, pregnancy and LB rates
in-vivo by secretion of hatching factors and lysine production by trophectoderm after fresh ET, were compared within groups and along FR available categories
of embryo but in in-vitro fertilisation when embryos are frozen under ultra low using ANOVA, Chi2 and linear-by-linear tests.
temperature, may lead to zona hardening. This may inhibit or reduce the chances Main results and the role of chance: Oocytes were inseminated with
of spontaneous hatching. partner sperm in 84.1% of cycles (FR: 77.3%; LB rate: 35.6%) or donor sperm
With the advent of laser assisted hatching (LAH), this complication could be in 15.9% of cycles (FR: 77.9%; LB rate: 39.8%), equally distributed within the
overcome with focussed laser light to produce opening in ZP with a single pulse three groups. Fresh transfer of 1 (9.1% of cycles), 2 (90.7%) or 3 (0.2%) embryos
of few millisecond, with no mechanical, thermal or mutagenic side effects. was performed on D+2 (28.1%) or D+3 (71.9%) post-ICSI. Cycles with 4, 5
Study design, size, duration: A prospective randomised control study was con- and 6 D+1 embryos were 2,873, 2,962 and 1,947, respectively. When consid-
ducted from 1st January to 31st December 2019. All patients whose frozen embryos ering all cycles that yielded 5 embryos, the most common occurrence, clinical
were thawed on day 5 were included. Two hundred day 5 FET cycles were ran- pregnancy and LB rates were comparable across FR range (38.5%-100%), varying
domised by computer generated list and divided into 2 groups. Group A (n=100), in between 42.8% and 47.4% for clinical pregnancy (p=0.46), and between 35.7%
which Quarter laser zona thinning assisted hatching (qLZT-AH) was done after thaw- and 39.9% for LB rate (p=0.56). Similarly, no significant differences were
ing while in group B (n=100) no laser assisted hatching was done after thawing. observed when considering mean embryo morphological score, pregnancy or
Participants/materials, setting, methods: All normoresponder patients LB rates in cycles that yielded 4 or 6 embryos (p>0.05 in all cases). Although
whose embryos were frozen on day 5 were included in this study and patients clinical pregnancy rates and LB rates predictably increased with the number of
with endometrium pathologies were excluded. Quarter laser zona thinning embryos obtained overall (32.0%, 36.6% and 38.0%, for 4, 5 and 6 embryos
assisted hatching (qLZT-AH) was performed after thawing in group A, where respectively, p<0.001), we did not find an effect of FR per se on the performance
25% of surface area and 50% thickness is removed by using laser while in group of the cohort of embryos generated.
B no laser assisted hatching was done. Groups were compared on the basis of Limitations, reasons for caution: A minimum of 5 MII were assigned to
implantation rate, clinical pregnancy rate and miscarriage rate. recipients, therefore it was not possible to perform the analysis for the whole
Main results and the role of chance: None of the Frozen embryo transfer range of FR. Our results should not be extended to cycles with FR <30%. Studies
cycle was cancelled and no loss of embryo was reported during thawing process. specifically focusing on low and very low FR are needed to clarify this further.
The cryosurvival rate was 96% in group A and 95% in group B which is in the Wider implications of the findings: FR per se does not affect reproductive
range of cryopreservation key performance indicators. No significant difference outcomes. Cycles with lower FR after ICSI result in similar LB rate than cycles with
in female age, BMI and AMH was observed between the two groups. higher FR if the number of generated embryos is the same. These results can
There was a statistically significant increase in implantation rate (35% vs. 23.07%, inform patient counseling, and the management of oocyte donation programs.
p=0.004) and clinical pregnancy rate (50% vs. 35%, p=0.031 ) when day 5 frozen Trial registration number: NA
thawed transfers assisted with qLZT-AH was done, while no difference in mis-
carriage rate (6% vs. 5.70%, p=0.906) was noted.
Limitations, reasons for caution: Larger randomised control studies are P-132 Morphokinetic analysis of embryos originated from vitrified
needed to strengthen these results. oocytes in infertile patients
Wider implications of the findings: We have demonstrated that day5 frozen A. Coello1, M. Meseguer1, M. Nohales1, L. Alegre1, J. Remohi2, A. Cobo1
embryo transfers assisted with quarter laser zona thinning assisted hatching 1
IVIRMA Valencia, IVF laboratory, Valencia, Spain ;
(qLZT-AH) results in better reproductive outcome than if no hatching is done. 2
IVIRMA Valencia, Ob/Gyn, Valencia, Spain
This study strengthen the current trend of freezing more embryos at blastocyst
stage and then assisting with qLZT-AH can further improve results. Study question: Does oocyte vitrification alter embryo morphokinetic param-
Trial registration number: MCDH/2019/15 eters in infertile patients?
Summary answer: Morphokinetics of embryos generated from vitrified
P-131 Is there a cohort-effect of low fertilization in human oocytes is similar to that observed in fresh oocytes in infertile patients, except
embryos? Analysis of live birth rate in 7,782 oocyte donation for time to first cell división.
cycles. What is known already: Although oocyte vitrification seems not to alter
embryo morphology and clinical outcomes, some differences have been found
M. Torra1, D. Garcia1, R. Vassena1, A. Rodriguez1
in kinetic parameters of embryos originated from vitrified oocytes compared to
1
EUGIN, Clinica Eugin, Barcelona, Spain those of embryos coming from fresh oocytes in donor cycles. These differences
have revealed a delay of 1 hour in each cell division until early blastocyst stage in
Study question: Does fertilization rate per se, rather than having few embryos the vitrification group. Currently, there is lack of information about embryo mor-
available for embryo transfer, affect live birth rates in ICSI cycles? phokinetic parameters after oocyte vitrification in autologous cycles.
Summary answer: Fertilization rate per se does not affect clinical pregnancy Study design, size, duration: This was a retrospective study including 2617
and live birth rates after ICSI embryos originated from fresh oocytes (475 cycles) and 716 embryos derived
What is known already: Low fertilization rates (FR) after ICSI reduce the embryos from vitrified oocytes (118 cycles). All embryos were cultured in a time-lapse
available for transfer. The number of embryos is a predictor of pregnancy and live monitoring system (Embryoscope, Vitrolife and Geri, Genea). Embryo develop-
birth (LB) rates, and controlling for this variable is imperative in order to understand mental events were annotated together with the corresponding timing of events
the cohort effect of FR. A high percentage of non-fertilized oocytes at D+1 could in hours after intracytoplasmic sperm injection (ICSI).

Participants/materials, setting, methods: Kinetics parameters analyzed cf-mtDNA content did not differ among morphological inner cell mass (A: 11.9
included: time to pronuclear appearance and fading (PNa, PNf ), cleavage divisions ± 8.0, B: 14.2 ± 6.8, C: 13.2 ± 5.0, P = 0.31) or trophectoderm (A: 12.1 ± 6.4,
until 8 cells (t2-t8), early, expanded and hatching blastocyst (tB, tBE, tBHi), as well B: 12.1 ± 8.2, C: 15.9 ± 5.8, P = 0.20) grades. The analysis of MPs by TLM
as second cell cycle duration (cc2) and the synchrony in the division (s2). Embryos showed that the time taken from starting of blastulation to the expanded blas-
were classified according to the hierarchic tree model by Meseguer tocyst (tEB - tSB) and from the blastocyst stage to the expanded blastocyst (tEB
et al. 2011. Implantation and clinical pregnancy rates were also evaluated. - tB) significantly correlated with the amount of cf-mtDNA in the corresponding
The analyzed variables were compared using chi-square test and 95% SCM (tEB - tSB: r = 0.46, P < 0.01, tEB - tB: r = 0.47, P < 0.01). Furthermore,
confident intervals. during the blastocyst stage, the mean cf-mtDNA content was higher for blasto-
Main results and the role of chance: No significant differences in morphokinetic cysts that experienced BC than in those that did not experience BC (BC group:
parameters were found between embryos originated from fresh and vitrified oocytes, 14.9 ± 7.7 vs. non-BC group: 7.67 ± 3.9, P <0.01).
except for t2. The proportions of embryos allocated to categories A–E in the hier- Limitations, reasons for caution: The main limitation of this study was the
archical tree were similar between groups. The blastocyst formation rate was 52.0% small sample size; hence, we were unable to expand our hypothesis to clinical
(95% CI: 49.7%-54.3%) in the fresh oocytes group and 45.3% (95% CI: 40.7%-49.9%) pregnancy outcomes at this point.
in the vitrified oocyte group. No differences on implantation rate [41.5% (95% CI: Wider implications of the findings: This is the first report showing an
34.2%–48.8%) vs. 48.9% (95% CI: 34.9%–62.9%)] and clinical pregnancy rate [39.8% association between the amount of cf-mtDNA in SCM and MPs of blastocysts.
(95% CI: 31.6%-48.0%) vs. 55.0% (95% CI: 39.6%-70.4%)] were found between the It has been reported that the duration of expansion and presence of BC is
fresh and vitrified group respectively. associated with clinical outcomes. Thus, cf-mtDNA may be a useful non-invasive
Limitations, reasons for caution: The retrospective nature of this study is the marker for blastocyst selection.
main limitation. Additionally, the smaller sample size compared to previous morphoki- Trial registration number: not applicable
netic analysis in donor cycles makes it difficult to drawn definitive conclusions.
Wider implications of the findings: The delay observed in donor cycles P-134 Original autologous partial oocyte vitrification program
has not been confirmed in this study, most probably because patient’s infertility “ICSI-Vit-program”: evaluation of the cumulative pregnancy rate
condition is affecting the morphokinetics of embryos originated from fresh and impact on the number of supernumerary cryopreserved
oocytes and, therefore, the effect of vitrification is not reflected in the analysis. embryos
Trial registration number: 1511-VLC-062-AC P. Boyer1, M. Gervoise-Boyer1, C. Siraudin1, A. Amar-Hoffet1,
D. Montjean1
P-133 Amount of cell-free mitochondrial DNA in spent culture 1
Hôpital Saint-Joseph, Laboratoire de Biologie de la Reproduction, Marseille Cedex
medium correlates with morphokinetic parameters of human
08, France
blastocysts
M. Kobayashi1, R. Suzuki1, K. Tsukamoto1, H. Hasegawa1, T. Kyoya1, Study question: Does “ICSI-Vit-program” consisting in partial vitrification of
A. Kikumoto1, T. Sakurai2, S. Saito2, J. Kobayashi2, H. Iwata3 MII-oocytes and delayed ICSI on thawed oocytes affect cumulative pregnancy
1
Kanagawa Ladies Clinic, Embryology Labolatory, Yokohama, Japan ; rate after fresh and frozen embryo transfer ?
2
Kanagawa Ladies Clinic, Department of Gynecology, Yokohama, Japan ; Summary answer: ICSI-Vit program during autologous ICSI cycle increases
3
Tokyo university of agriculture, Faculty of Agriculture Department of Animal cumulative early pregnancy rate lowers the number of cryopreserved embryos
Science, Atsugi, Japan and helps to promote single embryo transfer policy
What is known already: Partial vitrification of MII oocytes was introduced
Study question: Does the amount of cell-free mitochondrial DNA (cf-mtDNA) and validated in our laboratory as a tool to limit embryo freezing to meet legal
in spent culture medium (SCM) correlate with the morphokinetic parameters requirements. Post-warming oocyte survival rate was reported to average 85%
(MPs) of human blastocysts? in IVF-treated patients. This implies a loss of 15% of oocytes and therefore a
Summary answer: The duration of blastocyst expansion (tEB - tB and tEB - potential decrease in the number of embryos available. However, the top quality
tSB) and presence of blastocyst collapse correlated with the amount of embryo rate is not higher than 50% with fresh oocytes and not all the supernu-
cf-mtDNA in SCM. merary embryos can be cryopreserved. Data do not answer the question: could
What is known already: The mitochondrion is an important organelle for a non-survival oocyte after vitrification/thawing lead to a transferable embryo
embryonic development. cf-mtDNA can be detected in SCM, and it has been if it had been freshly fertilized
reported that the amount of cf-mtDNA in SCM of early cleavage-stage embryos Study design, size, duration: To study cumulative pregnancy rate (the sum
(day 3) reflects subsequent embryonic development and pregnancy outcomes. of pregnancies obtained in fresh oocytes, frozen embryos and when applicable
Accumulating evidence shows that MPs of human blastocysts obtained from vitrified/warmed oocytes cycles)
time-lapse monitoring (TLM) facilitates the selection of competent blastocysts in couples included in ICSI-Vit program, we conducted a retrospective analysis
for clinical pregnancy. However, there have been no reports regarding the rela- of 378 ICSI-Vit cycles from September 2011 to December 2018 including 363
tionship between the amount of cf-mtDNA in SCM and MPs of embryos. frozen embryo transfers compared to 420 conventional ICSI cycle without
Study design, size, duration: This was a retrospective cohort study. oocyte vitrification and fertilization of the entire MII oocyte cohort performed
We targeted 20 couples who underwent in vitro fertilization (IVF)/ intracyto- between January 2003 and July 2011including 353 frozen embryo transfers.
plasmic sperm injection (ICSI) treatment between January and December 2019. Participants/materials, setting, methods: Cycles with MII oocyte number
We measured the cf-mtDNA copy number in the SCM (20 μL) of embryos ≥12 at pick-up were included, cycles without embryo transferred were not
(total: 53) that were individually cultured to the expanded blastocyst stage for included. Data collection: Patient age, numbers of collected oocytes, vitrified
5 or 6 days. Incubation was performed using a time-lapse incubator oocyte after pick-up; fresh embryos obtained after fresh oocytes ICSI, Vit-fresh
(EmbryoScope; Vitrolife, Sweden). oocytes ICSI or frozen transferred embryos, early pregnancies (hCG blood test
Participants/materials, setting, methods: Amount of cf-mtDNA, maternal >100 IU) and deliveries were collected. Statistical analysis was performed.
age, type of insemination method, total embryo culture hours, morphological Cumulative rates were calculated including: fresh embryos transferred, frozen
blastocyst quality (according to Gardner–Schoolcraft grading), certain MPs, and embryos and embryos from Vit-fresh oocytes.
presence of blastocyst collapse (BC) were analyzed. DNA extracted from the Main results and the role of chance: We showed that vitrifying a part of
SCM was quantified by real-time PCR (CFX Connect™; Bio-Rad, USA) using a Metaphase II oocytes during ICSI-Vit program allowed superior cumulative preg-
specific primer set. Statistical analysis was performed using Spearman’s correla- nancy rate when comparing with conventional ICSI (67,9%, vs 49,7% p<0,05).
tion or Mann-Whitney-U-test. Besides, patients enrolled in ICSI-Vit program had fewer embryos transferred
Main results and the role of chance: The amount of cf-mtDNA did not (1,2±0,5 vs 1,6±0,5 in conventional ICSI cycles [p<0,05]) and a mean of 1,5±0,7
correlate with maternal age (r = - 0.15, P = 0.28), the total culture hours (r = embryo was transferred in vitrified/warmed oocytes cycles ([p<0,05]).
0.18, P = 0.19), or the type of insemination method (mean cf-mtDNA ± standard Moreover we observed lower mean number of cryopreserved embryos in ICSI-
deviation; IVF: 12.0 ± 7.1, ICSI: 13.4 ± 7.8, P = 0.56). In addition, the mean Vit program during fresh embryo transfer cycles (1,2±1,4 vs 3,3±2,9 in

conventional ICSI cycles [p<0,05]) and 0,6±0,6 embryos were cryopreserved Considering the number of embryos included in this analysis, the LBR obtained
in vitrified/warmed oocytes cycles. Women included in both programs were of from these slow developing embryos is unlikely to be a chance finding.
same mean age and had equal mean number of collected oocytes. However, Limitations, reasons for caution: This is a single-center study, which was
fertilization rate of fresh oocyte was lower in ICSI-Vit program (65,7% vs 69,9% performed with a modified microdrop vitrification/warming protocol where all
in conventional ICSI [p<0,05]), although no difference was found between fresh steps were carried out at 37oC and some steps did include an oil overlay. Thus,
oocytes and sibling vitrified/warmed in ICSI-Vit program (65,8% vs 65,4%). our results should be tested with a vitrification/warming protocol that uses room
These encouraging results may bring a tremendous change in IVF strategy by temperature.
providing an opportunity for better oocyte use. Wider implications of the findings: Slow developing embryos on day 5
Limitations, reasons for caution: Oocyte vitrification can be proposed with that reach blastocyst stage by day 6 substantially contribute to LBR in a vitrifica-
oocyte storage instead of embryos when necessary. The compared groups were tion/warming program. This holds even true for day 6 morphologically poor
treated at different periods and cryopreservation methods were different (slow quality blastocysts. These findings, if confirmed, may challenge current practice
freezing in ICSI group and vitrification ICSI-Vit). This inconsistency may partially to discard slow developing embryos on day 5 from vitrification.
explain the benefit described. That point is being investigated. Trial registration number: not applicable
Wider implications of the findings: Our results validate the vitrification of
oocytes as an alternative to freezing embryos which may remain a real difficulty P-136 Can monozygotic twinning in ART be explained by absence
for various legal, ethical or religious reasons. ICSI-Vit program gives couples the of zygotic polarization?
opportunity to have multiple ICSI cycles with fresh embryo transfers in a context A. Mikich1, G. Collar1, M.I. Kulmann2, M. Ferreira3, S. Mattiello3,
free of ovarian hyper-stimulation N. Frantz3
Federal University of Rio Grande do Sul, Morphological Sciences, Porto Alegre, Brazil ;
2
Nilo Frantz Reproductive Medicine, Clinical embryology, Porto Alegre, Brazil ;
P-135 Slow developing embryos undergoing compaction or 3
Nilo Frantz Reproductive Medicine, Human Reproduction, Porto Alegre, Brazil
cavitation on day 5 substantially contribute to live birth rates after
single day 6 vitrified-warmed blastocyst transfer Study question: Is the absence of zygotic polarity associated with Monozygotic
J. Gunst1, M. Vynck2, A. Van de Vijver3 twinning (MZT) in ART cycles?
1
AZ Sint Jan Brugge-Oostende, Laboratory Medicine - Centre for Reproductive Summary answer: Yes, zygotic polarity was not observed in any of the zygotes
Medicine, Brugge, Belgium ; that resulted in MZT. Enlarged periviteline space (PVS) and low Z-score were
2
AZ Sint Jan Brugge-Oostende, Laboratory Medicine, Brugge, Belgium ; also detected.
3
AZ Sint Jan Brugge-Oostende, Centre for Reproductive Medicine, Brugge, Belgium What is known already: The incidence of MZT is increased after ART pro-
cedures. The origin of MZT associated with IVF is still unclear. Blastocyst culture
Study question: Does the status of slow developing embryos on day 5 and and ICM splitting during hatching, ICSI and assisted hatching (AH) are some of
subsequent vitrification on day 6 impact clinical outcome in a vitrified-warmed the factors possibly associated with MZT. Little attention has been given to
transfer cycle? zygotic events that may contribute to monozygosity. Following sperm-oocyte
Summary answer: Slow developing embryos that reach blastocyst stage by day interaction, pronuclei (PNs) develop and migrate toward a central position in
6 have a potential for live birth which is independent of the status on day 5. the zygote. Alignment of PNs onto the polar axis is a fundamental event for
What is known already: Transfer of a single blastocyst either in a fresh cycle normal first cleavage and development. Non-polarization leads to random ICM
or after vitrification / warming is becoming widely applied. An important aspect location in the blastocyst and anomalies in development.
is the selection of the embryos that are eligible for transfer and/or for cryopres- Study design, size, duration: We retrospectively analyzed zygote images taken
ervation. Slow developing embryos on day 5, showing signs of compaction or 16-17 hrs post-ICSI, from 14 frozen embryo transfer cycles that resulted in MZT
cavitation, are either discarded or left in culture until day 6 for re-evaluation gestations, between 2017 and 2019. The zygotes analyzed were the ones that gave
using scoring systems that assess blastocyst expansion and morphology of inner rise to the blastocysts transferred in the 14 MZT gestations. Morphology of the
cell mass (ICM) and trophectoderm (TE). We investigated the potential of transferred blastocysts and cycle characteristics were also examined.
embryos that reached blastocyst grade >3 by day 6 in regard to clinical outcome Participants/materials, setting, methods: Observations of images and sta-
after vitrification / warming. tistical analysis were carried out to investigate putative origin of MZT. The pattern
Study design, size, duration: This single-center retrospective cohort study of alignment of the PNs toward the polar bodies (PBs), angle between the PBs,
was undertaken between 2011/01 and 2018/12. Day 5 embryos with slow devel- periviteline space (PVS) and Z-score were assessed. Inner cell mass and expansion
opment, expressed as compaction or cavitation (blastocyst grade 1/2), were /hatching were assessed in day-5 blastocysts. Ovarian stimulation, oocyte number,
cultured to day 6. Blastocysts that had reached stage >3 were vitrified. We included fertilization rates, assisted hatching and cytogenetic analysis were considered. MZT
433 patients with 601 day 6 vitrified warmed blastocyst cycles resulting in 567 was assessed by ultrasound at 6-7 and 8-9 weeks of gestation.
(94.3%) single vitrified warmed blastocyst transfers (SVBTs). Cycles with PGT, Main results and the role of chance: The transfer of a single blastocyst (7
donor oocytes, re-vitrification and an aberrant day 5 status were excluded. cases) and of a single day-3 embryo (1 case) resulted in two GSs. One single
Participants/materials, setting, methods: Microdrop vitrification was per- blastocyst transfer resulted in three GSs, each with an embryo; another single
formed using RapidVit Blast Kit/RapidVit Omni Kit (Vitrolife). Blastocysts were transfer resulted in one GS with two embryos. When two blastocysts were trans-
exposed to vitrification solutions 1&2 in microdroplets under oil, followed by solution ferred, two cases presented a single GS with two embryos and one case showed
3 without oil and subsequent vitrification using a closed device (Rapid-i; Vitrolife). three GS with one embryo each. One double blastocyst transfer resulted in three
Warming was performed using RapidWarm kits, with no oil in Warm 1 solution GSs. 16 out of 18 zygotes did not present their PNs in alignment with the PBs.
followed by microdroplet warming in solutions 2, 3 and 4 under oil. All procedures Two zygotes presented peripherally positioned PNs with their longitudinal axis
were carried out at 37oC. Clinical outcome was assessed after SVBT. aligned with one PB and the meridional axis aligned with the other PB. Majority of
Main results and the role of chance: For the 567 SVBTs that were performed zygotes (15/18) presented a great angle separating the two PBs and an enlarged
on day 6, the status of development on day 5 was characterized as compaction PVS (14/18). 12 zygotes scored as Z3-Z4 and 6 as Z2. ICM of eight blastocysts
(n=171; including embryos showing starting, partial and completed compaction; was large and tightly compacted, of seven embryos it formed two distinct cell
C), early blastocyst grade 1 (n=218; BL1) and blastocyst grade 2 (n=178; BL2). masses. Blastocysts were expanded and 11 were herniating at transfer. There was
The percentage of day 6 SVBT with good quality blastocysts was 69.6%, 68.3% no significant differences between the number of follicles, collected oocytes, fer-
and 77.5% for status C, BL1 and BL2, and 30.4%, 31.7% and 22.5% for poor quality tilization rate, assisted hachting and cytogenetic analysis between the cycles with
blastocysts, respectively. The overall resulting live birth rates (LBR) were 28.1% / MZT and contemporary cycles that resulted in single gestation.
28.0% and 27.5% for day 6 SVBT from C, BL1 and BL2 derived embryos. LBR Limitations, reasons for caution: Results were based on observations made
were higher for SVBT of good (BB or better) quality blastocysts (C: 29.4% / BL1: on zygotes and blastocysts of 14 IVF cycles. The occurrence of GSs derived
30.2% / BL2: 31.9%) compared to those performed with poor (lower than BB) from natural conception cannot be eliminated. A larger cohort of cases from
quality blastocysts (25.0% / 23.2% / 12.5%, respectively). different IVF centers should be studied to confirm our findings.

Wider implications of the findings: MZT represents an undesired IVF P-138 Insights into early human development from abnormal
outcome, due to the possible adverse health consequences for the babies and fertilisation
their mothers. A careful zygotic assessment should be performed and taken into G. Coticchio, Ph.D.1, F. Gurioli1, R. Sciajno1, C. Lagalla1,
account, together with blastocyst ICM morphology, before elective single N. Tarozzi1, M. Nadalini1, C. Zacà1, A. Borini1
embryo transfer or double embryo transfer. 1
9.baby, Family and Fertility Center, Bologna, Italy
Study question: Does morphokinetic analysis of human abnormal fertilisation,
P-137 The impact of single blastomere cell cycle duration of early one- (1PN) and three-pronuclear (3PN) fertilisation, offer cues to better under-
cell division on blastocyst formation and blastocyst quality. stand the very early steps of development?
M. Kljajic1, T. Krebs2, D.H.A. Maas2, N. Saymé2 Summary answer: 1PN/3PN morphokinetic analysis gives insights into
1
Saarland University Medical Center, IVF Laboratory, Homburg, Germany ; unknown aspects of fertilisation, concerning for example the role of the polar
2
Team Kinderwunsch Hannover, Department of Obstetrics- Gynecology & Assisted body, origin of pronuclei and pronuclear chromatin.
Reproduction, Hannover, Germany What is known already: The basic morphological attributes of human fertili-
sation have been known since the dawn of human IVF. For decades, such a knowl-
Study question: Does the time duration of the second and third cell cycle for edge has derived from static observation at a single time point. Initial application
single blastomere affect blastocyst development and blastocyst quality? of Time-Lapse Microscopy (TLM) in the late 1990’s revealed the beauty and mor-
Summary answer: The time duration of the second and third cell cycle phokinetic complexity of fertilisation. However, in-depth dynamic observation of
strongly affects blastocyst development as well as blastocyst quality. fertilisation has been pursued only in the last few years, with findings that have
What is known already: In the last decade, the introduction of time-lapse shed new light on cytoplasmic phenomena (e.g. the cytoplasmic wave and halo)
technology enables almost continuous monitoring of embryo development. This and the importance of cell symmetry for embryo development. Unfortunately,
technology generates comprehensive information regarding morphology and abnormal fertilization has been largely neglected, despite potential for research.
kinetics of embryo development and facilitates observation of dynamic, and Study design, size, duration: This retrospective study involved TLM obser-
often transient, events occurring between static observation periods. Together, vation of 378 normally (2PN) and abnormally (1PN and 3PN) fertilised oocytes
these have been defined as ‘morphokinetic’ variables. Strong correlations utilised in ICSI cycles carried out between 2017 and 2019. Maximum three sibling
between embryo kinetics and positive outcomes have been demonstrated in fertilised oocytes (one normal and one or two abnormal) per patients were
various studies. Some of the studies which involved cleavage time, cleavage analysed to reduce possible patient-specific biases. Oocytes of patients with
interval, and cleavage synchrony reported that embryos with high developmental different diagnoses of infertility were included in the analysis, while cases involving
potential could be effectively identified using these parameters. cryopreserved gametes or surgically retrieved sperm were excluded.
Study design, size, duration: Cell cycle duration is calculated using time-lapse, Participants/materials, setting, methods: Microinjected oocytes were
according to a single cell division. The injection time of ICSI was designated as assessed by a combined TLM-culture system (Embryoscope). Oocytes not
“time zero” (t0), and computer software was used to calculate the time duration amenable to TLM assessment, due to excess of residual corona cells or inade-
of the first (cc1a, t2-tPNf ), second (cc2a, t3-t2; cc2b, t4-t2) and the third cell cycle quate orientation for observation of polar body II (PBII) emission, were not
(cc3d, t8-t4). Obtained results of cell cycle duration were later associated with analysed. Fifteen parameters were identified and monitored, relevant to meiotic
the embryo capability to forming a blastocyst as well as with blastocyst quality. resumption, pronuclear dynamics, chromatin organization, cytoplasmic/cortical
Participants/materials, setting, methods: A total of 87 blastocysts from modifications and embryo quality. Diverse statistical tests were used depending
20 patients undergoing an antagonist cycle for ICSI treatment between November on the numerical category of parameters compared between groups.
2019 and December 2019 were evaluated. All blastocysts were cultured in Main results and the role of chance: All major fertilisation phenomena – e.g.
Embryoscope™ according to the manufacturer’s specifications (Vitrolife, polar body II (PBII) extrusion, cytoplasmic wave, cytoplasmic halo presence and
Sweden). The Gardner and Schoolcraft scoring system was used to describe disappearance, PN migration, PN juxtaposition (where applicable) and nucleolar
blastocyst quality. Correlations between the data were calculated using logistic precursor bodies (NPB) redistribution – were discerned in the three PN categories.
regression analysis. Statistical significance was defined as p<0.05. All statistical As a general trend, both 1PNs and 3PNs extruded the PBII with a delay of more
analyses were performed using SAS software. than 0.6 hours (P=0.0007), but only 1PNs accumulated a developmental delay,
Main results and the role of chance: Morphokinetic data showed that the starting from disappearance of the cytoplasmic halo, that amounted to almost 6
time duration of first (cc1a), and second (cc2a; cc2b) cycle were significantly hours at the time of cleavage (T=26.5, 32.6 and 27.3 in 2PNs, 1PNs and 3PNs,
different between embryos that reached the blastocyst stage and those embryos respectively; P<0.0001). More intriguing finding, with significant developmental
that did not. The first cell cycle was calculated as a time difference between implications, derived from the observation of specific fertilisation events. For
pronuclei disappearance and the first cell cleavage, where time duration was example: a) PBII extrusion was observed in the majority (45/75, 60%), but not all,
significantly different (p<0.05) in successfully formed blastocysts versus arrested of 1PNs and in a large proportion (61/136, 44.8%) of 3PNs; b) both the cyto-
or non-blastulating embryos. What is more this parameter as well significantly plasmic wave and cytoplasmic halo were observed, not only in 2PN and 3PN, but
affects blastocyst quality (p<0.05). Almost the same results were obtained for also in all but one 1PNs; c) NPB clustering and localization in a confined nuclear
the second cell cycle where was confirmed that duration of single blastomere region (NPB polarization) occurred in 2PNs (100%), 3PNs (96%) and, astonishingly,
division (cc2a: p< 0.001) and (cc2b: p<0.01) significantly correlate with blasto- in most 1PNs (73%). Finally, on day 2/3, 1PNs were relatively more affected by
cyst formation potential. Therefore, blastocyst quality was also affected by the developmental delay and blastomere fragmentation (data not shown).
time duration of a single blastomere division cycle (cc2a, p<0.01; cc2b, p< 0.05). Limitations, reasons for caution: This is a pilot study requiring extension
On the other hand, the duration of the third cell cycle (cc3d) between embryos and refinement of data, as well as independent verification.
which successfully reached the blastocyst stage and those embryos that failed Wider implications of the findings: These observations suggest a reap-
were not statistically significant. What is more, blastocyst quality neither was praisal of fertilisation. For example: 1PNs, with/without PBII emission, might be
affected by this morphokinetic parameter. mono- or digynic, respectively; 3PNs not always derive by failed PBII extrusion;
Limitations, reasons for caution: The disadvantage of the relatively small the cytoplasmic wave, generated by the microtubular aster, can form in the
patient cohort was balanced with a wider range of patients’ age (25-37). Further absence of the male pronucleus; NPBs polarize in isolated pronuclei.
research should link these morphokinetic parameters with pregnancy rate and Trial registration number: Not applicable
live birth rate as well.
Wider implications of the findings: The potential of the present findings is P-139 The impact of Vitrification in the outcome of IVF in an
considerable as we believe that these results are helpful for a better understanding oocyte donation program.
of the association between embryo morphokinetic parameters and blastocyst devel- P. Tatsi1, T. Chartomatsidou1, E.G. Papanikolaou2, R. Najdecki2,
opment. Also, the present work illustrates the possibility of additional information F. Chouliara2, I. Asuchidou2, C. Ioakeimidou2, E. Timotheou1
that can potentially be incorporated into an embryo classification model. 1
Assisting Nature, Embryology Laboratory, Thessaloniki, Greece ;
Trial registration number: ‘not applicable’ 2
Assisting Nature, Clinical Department, Thessaloniki, Greece

Study question: How does Vitrification and double Vitrification (cryopre- What is known already: Vitrification is becoming the most widely adopted
served embryos produced from cryopreserved oocytes) affect the result of an embryo cryopreservation technique for higher embryo survival rate and live
oocyte donation program? birth rate than other methods.In order to avoid the risk of multiple pregnancies,
Summary answer: Oocyte and embryo Vitrification does not affect the out- most countries recommend reproductive centers to transfer only one blastocyst
come of IVF with donor oocytes. However, cryopreserved embryos from vit- per cycle, causing surplus of surviving embryos might occasionally be used in
rified donor oocytes exhibit reduced implantation potential. FET cycles. But to our knowledge, limited data was available for FET outcome
What is known already: Vitrification of oocytes, embryos and blastocysts is with twice cryopreserved human embryos, and the sample size was small .With
successfully performed worldwide in IVF Treatments and Oocyte Donation the increase number of repeated cryopreserved embryo transfer cycles, the
Programs. Vitrification of Oocytes and Embryos does not impair oocyte and impact of repeated-frozen-thawed manipulate to clinical pregnancy outcomes
embryo viability or implantation potential. No statistically significant difference needs further to be elucidated.
is observed in the outcome after blastocyst embryotransfer originated from fresh Study design, size, duration: The purpose of this retrospective cohort study
versus vitrified donor oocytes. In addition, embryos derived from fresh donor was to evaluate the pregnancy and perinatal outcome of revitrified human
oocytes show an extremely high implantation potential and exceptional clinical day5/6 blastocysts derived from surplus of surviving blastocysts in the last FET
pregnancy and live birth rates. On the other hand, few is known regarding cycles. Cohorts of 320 transfered blastocysts with either twice-frozen-thawed
cryopreserved embryos that were produced from cryopreserved donor oocytes. and once-frozen-thawed between January 2016 and December 2018 were
Study design, size, duration: This study was conducted between January included in the study. Only IVF/ICSI cycles were included,PGT cycles were
2015 and December 2018 to examine the impact of Vitrification and compare excluded. Match-pair analysis was used to control for some patient basic char-
the pregnancy and live birth rate after embryotransfer of day 5 embryos of fresh acteristics,then compared two groups.
oocytes-fresh embryotransfer (FROFRE) versus vitrified donor oocytes-embry- Participants/materials, setting, methods: During the match-pair analysis,
otransfer (CROFRE) and cryopreserved embryos derived from fresh oocytes we paired twice-frozen-thawed and once-frozen-thawed groups’ patients data
(FROCRE) and cryopreserved embryos originated from cryopreserved oocytes through maternal Age,body mass index, endometrial thickness. The two groups’
(CROCRE). A total of 425 donations were included in the study. paired ratio was 1 (80 cycles):3 (240 cycles). The total 320 transfered blastocysts
Participants/materials, setting, methods: A total of 425 oocyte recipient cycles gave live birth to 105 infants, of which twice-frozen-thawed (22 infants) and
were included in the study. 111 FROFRE cycles, 92 CROFRE cycles, 184 FROCRE once-frozen-thawed groups (83 infants). Finally,we compared clinical pregnancy
and 38 CROCRE cycles. 896 fresh oocytes were utilized in FROFRE cycles and 736 outcome parameters between the two groups,included implantation,clinical
vitrified oocytes were thawed and used in CROFRE cycles. ICSI was implemented in pregnancies,live deliveries, miscarriage rate, infants’ birth body length, birth
all cases and embryotransfer and vitrification was performed on day 5 on blastocyst weight and so on.
stage. Positive hCG, clinical pregnancy and live birth rates were monitored. Main results and the role of chance: We regarded once-frozen-thawed
Main results and the role of chance: Positive β-hCG, clinical pregnancy rate FET as the control group to compare with twice-frozen-thawed FET group. By
and live-birth rate per embryotransfer were compared between our groups, statistical comparison(twice-frozen-thawed versus control ),we found maternal
FROFRE-CROFRE-FROCRE-CROCRE. Furthermore, embryo quality and blas- age(31.60±4.24 versus 32.33± 4.34, P >0.05,), body mass index (21.62±3.34
tulation rate was similar among fresh donated oocytes and from vitrified oocytes. versus 22.53± 4.74kg/m², P >0.05), endometrial thickness (9.78±1.54 versus
β-hCG was 69%, 59% and 54% between FROFRE, FROCRE and CROFRE groups 9.53± 1.64, P >0.05), Embryo transfer cycles(2.11±1.34 versus 2.83± 1.55,
respectively. No statistically significant difference was observed in the clinical preg- P >0.05) and number of embryos transferred (1.0 versus 1.0, P >0.05) were all not
nancy rate, which was 55% (FROFRE), 52%(FROCRE) and 47%(CROFRE) and significantly different between two groups. While clinical pregnancy rate(36.25% ver-
the live birth rate 46% (FROFRE), 44% (FROCRE) 42% (CROFRE). The CROCRE sus 46.25%,P >0.05) , implantation rate (36.25% versus 46.25%, P >0.05) and live
cycles exhibited the lowest rates 35% (β-hCG), 19% (clinical pregnancy) and 15% birth rate (27.50% versus 34.58%, P >0.05) were all lower in twice-frozen-thawed
(live birth). Oocyte vitrification and warming is common and effective method of group,but the difference was not significant by the Chi-square test .Unexpectedly ,
delivering donor oocytes to patients and success rates all over the world are as miscarriage rate (7/29-24.14% versus 29/111-26,13%,P >0.05) was lower in twice-
high and similar to results in fresh donor egg cycles. One remaining question is if frozen-thawed group but not significantly. As for the perinatal data, no significantly
these previously frozen oocytes, once warmed, inseminated and cultured to later differences were observed in gestational age(38 w±3.4d versus 38 w±5.3d, P >0.05),
embryonic stages can be again re-vitrified and warmed with similar results as in sex ratio(11/11 versus 48/35,Boy/Girl), birth body length (49.15±4.12cm versus
fresh oocyte donation cycles. According to our findings there is a significant impact 49.09±3.88 cm, P>0.05) and birthweights (3220.00±422.3 g versus 3116.99±453.6,
of double-vitrification in pregnancy and live birth rates. P>0.05) between groups. Likewise, in the live birth individuals,there were no significant
Limitations, reasons for caution: Although our primary results show that differences between the two groups on the incidence of pre-term birth (3/22 versus
clinical impact in success rates of double-vitrification is significantly impaired in 11/83 ), low birthweight(2/22 versus 7/83 ), small for gestational age, large for
our oocyte donation program, this needs further investigation in larger random- gestational age and macrosomia.
ized studies. Few CROCRE cases were included and the embryos studied were Limitations, reasons for caution: Because the refrozen group’s sample size
chosen for second, third or final embryotransfer after previous CROFRE cycle. was limited,we did not distinguish some confounding factors associated with clin-
Wider implications of the findings: It is important to study and perceive ical pregnancy outcomes, included type of infertility,cause of infertility, different
if embryos derived from vitrified donor oocytes perform similarly to fresh development stage,and the embryo morphological score,insemination method,en-
oocytes after double vitrification and warming. The couples interested in oocyte dometrial preparation protocol , embryo cryopreservation duration and the long-
donation and IVF treatments should be thoroughly informed about their options term effects of re-vitrification method on offspring’s health outcomes.
and success rates. Wider implications of the findings: The results provide feasibility to use
Trial registration number: Not Applicable refrozen blastocysts in FET which can obtain an acceptable clinical outcome
expectation ,when there is no other primary cryopreservation blastocyst to
P-140 Clinical pregnancy and perinatal outcomes of twice-frozen- choose. Meanwhile, patients should be fully informed of possible reduction in
thawed embryo transfers: a retrospective comparative study clinical outcome parameters.
B. Cai1, W. Yali1, X. Yanwen1, Z. Canquan1 Trial registration number: not applicable
1
First Affiliated Hospital of SunYat-sen University, reproductive medicine center,
P-141 Comparison of clinical results of embryo culturing using last
Guangzhou-Guangdong, China
generation incubators: is there a better option?
Study question: Dose repeated-frozen-thawed human embryos can achieve M.D.L.Á. Valera Cerdá1, C. García Gimeno1, L. Bori Arnal1,
comparable clinical pregnancy and perinatal outcomes to conventional once J.M. De los Santos Molina2, A. Pellicer Martínez3,
frozen-thawed embryos ? M. Meseguer Escrivá2
Summary answer: Acceptable clinical outcomes could be expected from 1
IVIRMA, Research Laboratory, Valencia, Spain ;
transfer of recryopreserved blastocysts, while perinatal outcome was not 2
IVIRMA, IVF Laboratory, Valencia, Spain ;
affected and clinical pregnancy parameter was lower but not significant. 3
IVIRMA, President- Reproductive Medicine, Valencia, Spain

Study question: Is there a significant difference in clinical results using different surrounding micro-environment and cell-free microRNAs reflect on the quality
last generation time-lapse incubators (TLM) for embryo culturing, using benchtop and ploidy status of embryos. It is known that miR-290 members are regulators
incubators (CI) as a reference? of apoptosis in cells. In pre-implantation embryos, miR-294 levels in media
Summary answer: We did not find significant differences in pregnancy and directly correlate with apoptosis in the blastomeres. However, there is a lack of
implantation rates between last generation incubator systems. When grouped, mechanistic explanation behind this observation. It is not clear whether high
TLM showed better outcome compared to CI. miR-294 in cells causes DNA fragmentation or increased apoptosis results in
What is known already: Time-lapse systems (TLM) bring to IVF laboratories intense release of miR-294, which could serve a secretory function.
a clear advantage, providing new information of embryo development without Study design, size, duration: Mouse zygotes (N=28) were cultured to the
perturbing their culture. However, their performance as incubators must be small blastocyst stage. At this stage, UV radiation was used to induce DNA
tested versus those conventional benchtop incubators (CI), to assure they pro- damage in order to mimic the final stages of apoptosis. The culture was extended
vide a proper environment for embryo culturing. To date, there is insufficient to allow for cellular responses to DNA damage. The blastocysts were stained
evidence of actual significant differences in clinical outcomes varying on the type at Day 5 of development to verify DNA damage. Media samples were individually
of incubator used, although some studies show a slight improve with time-lapse processed and analysed for miR-294. UV-treated and control samples were
systems. Previously published studies have found a significant increase in compared for microRNA levels (t-test).
good-quality blastocyst and euploidy rates using time-lapse incubators. However, Participants/materials, setting, methods: Frozen mouse zygotes (B6C3F-1 x
comparative studies on the matter are usually incomplete. B6D2F-1) were cultured at 37oC, 5% CO2 to the early blastocyst stage. The blasto-
Study design, size, duration: We present a retrospective study including cysts (N=15) were exposed to UV radiation (5 s, 302 nm) and cultured for 16 more
8446 ICSI cycles performed in IVIRMA Valencia from two consecutive years, hours. Control embryos were cultured in parallel (N = 13). The embryos were stained
both with own and donated oocytes. Fertilized oocytes were cultured until with the TUNEL method to assess DNA damage. Media samples (20µl) were indi-
blastocyst stage using different systems: CI (Astec, Japan) or incubators with vidually analysed for miR-294 expression using PCR technology.
TLM, including Embryoscope, Embryoscope Plus (Vitrolife, Denmark) and Geri Main results and the role of chance: UV radiation can be used to induce
(Genea, Australia). Out of 26100 viable embryos, 9817 were selected for trans- DNA damage and study apoptosis-triggered mechanisms. For embryos, expo-
fer, 7574 in single embryo transfer (SET), with clinical outcome data of 9103 sure to 302 nm for 5 s at the early blastocyst stage causes increased DNA
transferred embryos available. breakage but maintains blastocyst viability for enough time to investigate molec-
Participants/materials, setting, methods: All incubators included in the ular changes. The direct effects of UV radiation on DNA breakage was verified,
study are well stablished and routinely used in the clinic for embryo culturing. with the UV-treated group showing higher extent of apoptosis (and reduced
Scoring and selection for transfer or freezing were performed according to the cell count) compared to the control embryos. The average miR-294 Ct in the
ASEBIR criteria, by morphological assessment combined with morphokinetic SCM of the UV group was 25.1 ± 1.1 (ΔCt 6.3 ± 1) and in the control group
selection methods. A statistical analysis was performed for comparing clinical 27.1 ± 1.5 (ΔCt 8.3 ± 1.5). The results from the t-test showed that the blasto-
results between incubators, using the Statistical Package for Social Sciences cysts with UV-induced DNA damage released significantly more miR-294 in the
(SPSS). Results are presented as pregnancy, ongoing pregnancy and implantation media compared to controls (p<0.01).
rates per incubator, separately by oocyte origin. Limitations, reasons for caution: These results are limited to mouse
Main results and the role of chance: The statistical analysis shows that embryos and further research is needed for applications in other species.
there is no significant difference in the clinical results of cycles using the different Wider implications of the findings: UV radiation is a useful approach to
types of TLM. For autologous cycles, the pregnancy rate of fresh transferred mimic the last stages of apoptosis and study related cellular responses. Apoptosis
embryos (N=744) was 59.4% for Embryoscope (ESD), 64.1% for Embryoscope in pre-implantation embryos triggers the intense packaging and release of miR-
Plus (ESD+) and 61.7% for GERI (P=0.305). For Egg donation cycles, having a 294 in the extra-cellular environment with possible actions as a singaling molecule
total of 2384 transferred embryos, pregnancy rates resulted in 72.9% for ESD, for communication with the endometrium.
74.1% for ESD+ and 69.3% for GERI (P=0.173). Regarding implantation rates, Trial registration number: Not applicable
in procedures with own oocytes resulted in 48.14% using ESD, 50.35% for ESD+
and 50.79% using GERI incubators (P=0.855). As results for cycles with donated P-143 Oocyte central granularity is associated with reduced
oocytes, the implantation rate was 61.42% for ESD incubators, 61.99% for ESD+ fertilization rates and altered fertilization dynamics
and 58.78% with GERI, not having a statistically significant difference (P=0.288). C. Moutier1, M.C. Guglielmo1, P.V. Novara1, F. Brambillasca1,
When comparing CI with TLM for autologous cycles, the pregnancy rate of fresh A. Bartolacci1, M. Mignini Renzini1, M. Dal Canto1, J. Buratini1
transferred embryos was 69.0% for TLM and 66.1% for CI (P=0.046), while 1
Biogenesi Reproductive Medicine Centre, Istituti Clinici Zucchi, Monza, Italy
ongoing pregnancy also resulted significantly higher in the standard cycles when
compared TLM 60.9% vs CI 54.5% (p=0.047). Study question: Is cytoplasmic central granularity (CCG) in oocytes associated
Limitations, reasons for caution: Other variables should be assessed as with alterations in fertilization rates and dynamics?
well, such as embryo quality, good-quality embryo rate and life-birth rate per Summary answer: Oocyte CCG is associated with reduced fertilization rates,
incubator. Furthermore, this study only compares four types of incubator but as well as delayed pronuclei disappearance and first cell division after ICSI.
there are many more available. What is known already: Oocyte quality is a major determinant of ICSI
Wider implications of the findings: The results of this study can serve as success. Oocytes are routinely selected according to morphological criteria
a quality check of the procedures in the clinic. It reassures that all TLM used in before ICSI, but the relationship between different oocyte morphological fea-
the clinic yield similar results and there is no bias in the clinical results depending tures and developmental competence is not entirely known. Oocyte CCG is
on which TLM is used. associated with alterations in the actin cytoskeleton and meiotic spindle, both
Trial registration number: Not applicable playing critical roles in meiosis completion, pronuclei formation and all steps of
P-142 Increased DNA fragmentation in blastocysts triggers the embryo mitotic divisions. Faster fertilization morphokinetics is associated with
release of miR-294 in spent culture medium higher rates of embryonic development.
Study design, size, duration: We performed a retrospective analysis com-
D. Makri1, W. Maalouf1
paring fertilization rates and fertilization morphokinetic parameters of control
1
University of Nottingham, Child Health- Obstetrics- and Gynaecology, (morphologically normal) and CCG oocytes, obtained from patients under 40
Nottingham, United Kingdom years of age subjected to ICSI in our Fertility Centre from January 2016 to July
Study question: To study whether extended apoptosis in embryonic blasto- 2019. ICSI cycles were selected in order to equally distribute infertility factors
meres enhance the packaging and release of miR-294 by blastocysts. in both experimental groups. Morphokinetic parameters were assessed by time-
Summary answer: Induced DNA fragmentation at the early blastocyst stage lapse technology.
causes increased release of miR-294 in the culture medium. Participants/materials, setting, methods: A total of 1075 metaphase II
What is known already: MicroRNAs are important regulators of cell signaling, oocytes were injected by ICSI obtaining an overall fertilization rate of 76,5%
growth, and cell death. Pre-implantation embryos release microRNAs in the (823/1075). Morphokinetic parameters until the first division were compared

between CCG and control oocytes (n=141/group). Maternal age and body Limitations, reasons for caution: The study was retrospective but a pro-
mass index (BMI) did not differ between experimental groups (CGC: age=34,44 spective clinical trial is underway. In addition, all the samples were collected in
± 3,13; BMI=23,35 ± 4,13. Control: age=34,40 ± 3,14; BMI=22,64 ± 3,42). the same clinic using the same culture media. The test needs to be validated in
Main results and the role of chance: Fertilization rate was significantly lower multiple clinics and culture media.
in CCG oocytes compared to control [69,1% (141/204) vs. 78,3% (682/871); p Wider implications of the findings: 90% aneuploidy positive identification
< 0,01]. Time of pronuclei fading was longer (p = 0,02) for CGC oocytes com- without biopsy is as high as the highest reported NI-PGT results. Further advan-
pared to control oocytes (24,52h ± 3,39 and 23,72h ± 4,13 respectively). The tages of metabolomics is that is cheaper than NI-PGT (NGS) and does not require
first mitotic division was delayed in embryos obtained from CCG oocytes com- change in embryology protocols. Combined with markers for embryo viability
pared to control embryos (27,43h ± 3,57 vs. 26,53h ± 3,88 respectively, p = 0,02). other than euploidy might yield further improvements in embryo selection.
Limitations, reasons for caution: The retrospective nature of the study Trial registration number: NA
and the need of further studies to assess if CCG is also associated with subse-
quent embryo competence to implant and provide a live birth. P-145 A new role of Repulsive Guidance Molecule C as a ligand
Wider implications of the findings: Our study contributes to a better of neogenin enhanced in follicular development of ovary in the
understanding of the relationship between oocyte morphology and quality, while mouse
providing valuable references for the improvement of ICSI outcomes. J.H. Lee, Ph.D.1,2, Y.J. Kim1, K.H. Choi3, K.Y. Kang3, H.S. Kim3,
Trial registration number: Not Applicable H. Jeong3, H. Kang4, Y.R. Park3, S.J. Kim5, M.K. Kwon5, H.O. Kim5,
M.K. Koong5, Y.S. Kim5, T.K. Yoon5, J.J. Ko2
P-144 A metabolomics approach to identify aneuploid embryos to 1
CHA Fertility Center Seoul Station, Reproductive Molecular Medicine, Seoul,
increase the effectiveness of ART cycles Korea- South ;
S. Cabello-Pinedo1, H. Abdulla2, M.L. Seth-Smith1, M. Escriba3, 2
CHA University, Biomedical Science, Seoul, Korea- South ;
J. Crespo3, S. Munné1, J.A. Horcajadas Almansa1 3
CHA Fertility Center Seoul Station, Embryology Lab, Seoul, Korea- South ;
1 4
Overture Life SL, Scientific department, Madrid, Spain ; CHA Fertility Center Seoul Station, Andrology Lab, Seoul, Korea- South ;
2 5
Texas A&M University Corpus Christi, Chemistry, Corpus Christi, U.S.A. ; CHA Fertility Center Seoul Station, IVF Clinic, Seoul, Korea- South
3
Juana Crespo Clinic, Embryology, Valencia, Spain
Study question: In the ovary, Repulsive Guidance Molecule c (RGMc) stimu-
Study question: Can euploid embryos be identified in a non-invasive manner lated neogenin signal has a specific function for follicle development and oocyte
by measuring the concentration of specific metabolomic biomarkers in spent maturation?
culture media? Summary answer: Neogenin is specifically localized at the follicle and RGMc as
Summary answer: Specific metabolites in embryo spent culture media are ligand promotes follicle development and the maturation of the oocyte in the ovary.
correlated with euploidy status of blastocyst stage embryos. What is known already: Neogenin has reported a gene level expression in the
What is known already: Embryo Implantation potential and euploidy rates mammalian ovary. However, neogenin as a multiple function receptor remains
decrease with advancing maternal age. Preimplantation Genetic Testing (PGT) unknown as the main role and ligand for folliculogenesis in the ovary of mammals.
has been used to avoid the transfer of aneuploid embryos, which would not Study design, size, duration: This is an animal model study for the role of and
result in a successful pregnancy. However, embryo biopsy is invasive and oper- RGMc as a ligand of neogenin during the follicular development phase in the ovary
ator-dependent resulting in wide variability in pregnancy rates. Although DNA of the mouse. We compared the study between controls (n=25 head) versus
can be detected in spent media, correlation with trophectoderm biopsy varies RGMc treatment (n=25 head) groups. We analyzed the follicular
widely between different studies. Recently, metabolomics studies on spent cul- development ratios of mouse ovary with RGMc + PMSG induction. And we study
ture media have shown promising results to predict implantation potential. In uP- and down-regulation whole transcriptome clusters of RGMc treated ovary.
this study we aim to determine if euploidy status can also be ascertained by Participants/materials, setting, methods: We identified the gene, the protein
non-invasive metabolomics analysis. expression of neogenin and the localization in the ovary. Then we studied the role of
Study design, size, duration: This study includes spent media samples col- the neogenin through superovulation by PMSG with RGMc treatment as a ligand of
lected before trophectoderm biopsy from embryos that were later analyzed by neogenin for in vivo follicle development activation. And afterwards, we performed an
PGT (NGS). Among them, 60 out of 116 were classified as aneuploid, 42 as entire transcriptome next generation sequencing process for the identification of the
euploid and 14 as mosaic. Samples were analyzed to find metabolites differentially RGMc effect, which linked the specific factors to the follicle development in the ovary.
abundant in the different embryo groups, leading to the definition of a method Main results and the role of chance: Neogenin is abundantly expressed in
that allows selection of euploid embryos for transfer without the need for biopsy the ovary and located at the developmental oocyte phase from primordial and
avoiding possible damage to viable embryos. antral follicles. Therefore, the RGMc treated ovary exhibits a 40% increase in
Participants/materials, setting, methods: Patients undergoing PGT were the number of follicles compared with the control ovary. And RGMc treated
included in this study. Spent media samples were collected just before biopsy, ovary present Oct3/4, Nanog, p63, and Neogenin significantly higher gene
ultrafiltered to remove molecules >3KDa and run through a UPLC- Fusion expressed than control ovary. The especially experimental group, Nanog and
Orbitrap MS/MS system at 500,000 FWHM mass resolution. Different machine p63 three times higher expression than the control group. In the western blot
learning techniques were applied to reduce the huge number of metabolites to data also present similar pattern like RT-PCR. To identify the transcriptomic
a limited number of informative biomarkers for euploidy. signature between RGMc-treated and control mice, RNA-seq was performed.
Main results and the role of chance: The analysis of mass spectrometry As a result, total of 275 genes was differed by more than 4-fold change between
(MS) led to the identification of a sub-set of biomarkers whose concentration the RGMc-treated and control groups. Among the total 275 genes, 197 and 78
differed between aneuploid, euploid and mosaic embryos. The concentration genes were relatively uP-regulated in RGMc-treated and control group, respec-
of this subset of biomarkers was increased in aneuploid embryos and reduced tively (Figure 4A and 4B). The 197 genes overexpressed in the RGMc group
in euploid and mosaic embryos. Fifty-four out of the 60 aneuploid embryos were significantly enriched to cilium organization, axonemal dynamin complex
(90%) showed a reduced concentration of aneuploid profile specific metabolites assembly, microtubule bundle formation, icosanoid metabolic process, prosta-
and different from euploidy. glandin metabolic process, and prostanoid metabolic process. The 78 genes
The challenge of this study relies on the wide range aneuploidy profiles found down-regulated in the RGMc group were significantly enriched to ovulation cycle
in blastocysts, from monosomies to trisomies for each possible chromosome, process, hormone biosynthetic process. Specifically, RGMc treated ovary shows
alone or in combination with other abnormalities. Each aneuploidy may produce specific uP-regulation factors as a prostaglandin linked signal.
different metabolic alterations, resulting in very diverse and heterogenous met- Limitations, reasons for caution: This study did not the analysis of the
abolic profiles. Given the diversity of the aneuploid group, and the positive human samples because of animal experiments for follicle development and
identification of such profiles into the aneuploid category, the sub-set of iden- oogenesis. Therefore, further investigations aimed specific analysis of RGMc in
tified biomarkers are probably definitory of a particular\specific metabolic profile human at improving controlled ovarian hyperstimulation study with RGMc of
and could hardly be found by chance. poor ovarian response.

Wider implications of the findings: RGMc as a ligand of neogenin involved Study question: Does the hazardous atmospheric air can impact embryo
follicle development and maturation of oocytes. RGMc stimulated the PSDN development in the IVF laboratory even after standard air quality management
signaling pathway for follicular development. This platform can apply to poor and use of air filters?
ovarian responses as a specific stimulator for enhanced follicle development for Summary answer: There was a decrease in key performance indicators of
the in vitro fertilization patients of the poor ovarian responder. IVF lab with increased fragmentation, poor embryo development and reduced
Trial registration number: 2018R1D1A1B07050138, 2018R1C1B5045516, reproductive outcome.
2018R1D1A1B07044016 and 2019R1A2C1086882. What is known already: According to WHO survey amongst 1650 cities in
the world, Delhi the capital has worst air quality. With the air quality index falling
P-146 Soluble protein Cripto-1 promotes aberrant cell divisions in drastically from Moderate (101-200) level between January to September to
human embryonic cells severe or hazardous (500+) level from October to December. The factors for
V. Porokh1, P. Vanhara1, Z. Holubcova2, A. Hampl1 poor air quality is stubble burning, road dust, cold weather and vehicle pollution.
1
Masaryk University- Faculty of Medicine, Histology and Embryology, Brno, Czech Studies have supported that air quality is critical to embryo development and
Republic ; for overall success of IVF. Both animal and human studies have suggested an
2
Reprofit International- Clinic of Reproductive Medicine, Research&Development, association between poor air quality conditions and impaired embryo develop-
Brno, Czech Republic ment, resulting in decreased implantation and pregnancy rates.
Study design, size, duration: A retrospective study was conducted from 1st
Study question: Do secretory factors present in culture media affect the January to 31st December 2019. Patients were divided in 2 groups. Group (A)
fidelity of mitotic divisions during preimplantation embryo development? from October to December, when atmospheric air quality was hazardous and
Summary answer: Cripto-1 signaling induces the formation of multipolar mito- Group (B) patients from January to September, when atmospheric air quality
ses hence increasing the risk of aneuploidy in early human embryogenesis. was within normal range. Both groups were compared on the basis of fertilisation
What is known already: Mitotic errors are common in preimplantation devel- rate, fragmentation rate, Day 3 grade A embryo development rate, implantation
opment. Abnormal segregation of genetic material results in aneuploidies, spon- rate and clinical pregnancy rate.
taneous abortions, and birth defects. Despite its major importance for assisted Participants/materials, setting, methods: All patients undergoing fresh
reproduction, our knowledge of molecular mechanisms governing cell division day 3 embryo transfers from the month of January to December were included.
in early human embryos remains rudimentary. Out of the total 276 patients, 60 patients had their embryo transfer in the month
Study design, size, duration: A basic research study involving low-passaged of October to December (study group), while 216 were those from month of
human embryonic stem cells (hESCs; lines CCTL12, CCTL14, MUES1) and spent January to September (control group). Same culture media was used throughout
cultivation media from 30 IVF cycles. Media from successful cultures with >80% the period. There was no change in clinical or embryology team.
fertilized eggs developing into blastocysts (n=15) were compared with cultiva- Main results and the role of chance: The average AQI in Delhi was recorded
tions in which all embryos arrested in cleavage stage (n=15). A combination of around 500 between October to December 2019, while the maximum was recorded
microscopic and immunoanalytical assays was used to investigate the link more than 1200. The quality of atmospheric air was correlating with the quality of
between embryonic secretome and cell division errors. embryo development. In group A (Oct-Dec). Fragmentation rate was significantly
Participants/materials, setting, methods: The pluripotent stem cells were higher in Group A than Group B.( Fragmentation <10% : 53.03% vs 70.6%; p=
derived from the inner cell mass of human blastocysts donated for research. 0.00001, Fragmentation 10-20% :30.80% vs. 18.78%; p=0.0002, Fragmentation >20%:
The single-step medium (CSCM-C, Irvine) from each group culture was collected 16.16% vs 10.6%; p=0.029). There was also a statistically significant decline in fertil-
on the day-5/day-6 following ICSI and frozen in -20oC prior to analysis. The ization rate (62.5% vs. 70.07%, p=0.008), Day 3 A grade embryo formation rate
immunoaffinity proteomic array and ELISA were performed to assess soluble (53.03% vs 70.6%; p=0.00001), Implantation rate (11.6% vs 25%; p=0.011), Clinical
factors in the stem cell/human embryo microenvironment. Pregnancy rate (15.7% vs 43.1%; p=0.025).
Main results and the role of chance: Immunofluorescent staining of cultured Limitations, reasons for caution: Multi centric studies are needed to
hESCs revealed that 9-15% of mitotic cells exhibited supernumerary centrosomes strengthen these results.
and aberrant mitotic spindles. The presence of more than 2 centrosomes and the Wider implications of the findings: We have demonstrated that poor
formation of multipolar division apparatus was found to promote the incidence of atmospheric air during October to December in Delhi INDIA has a negative
aneuploidy in later stem cell generations. Immunoaffinity assay of 120 soluble proteins impact on embryo development which also decreases reproductive outcome
detected 18 molecules that were specifically produced by low passage hESCs with a even after standard air quality management. During this period either case can
high incidence of multipolar mitoses. Functional experiments led to the identification be avoided or more stringent air quality should be maintained.
of the soluble form of Cripto-1protein as a factor capable to induce multicentrosomal Trial registration number: MCDH/2019/09
phenotype. To complement results from the hESCs model, the analysis was under-
taken on the spent media from human embryo cultures. Our finding that the soluble P-148 Does the number of oocytes affect the cumulative clinical
Cripto-1 was enriched in the microenvironment of human embryos which failed to pregnancy rate following the transfer of all frozen–thawed embryos
reach the blastocyst stage supports the notion that this secretory molecule adversely L. Kedar1,2, A. Ben-Haroush1,2, A. Wertheimer1,2, O. Sapir1,2,
affects genomic stability during preimplantation development. E. Shlush1,2, Y. Shufaro1,2, G. Oron1,2
Limitations, reasons for caution: The limitation of this preliminary 1
Belinson, Belinson Infertility and IVF Unit- Helen Schneider Hospital for
study must be interpreted with caution due to the small study size. Further research
Women- Rabin Medical Center- Beilinson Hospital, Petach Tikva, Israel ;
is needed to evaluate whether the secretion of Cripto can be used as a negative 2
Tel-Aviv University, Sackler Faculty of Medicine, Tel-Aviv, Israel
indicator of embryo´s developmental competence in clinical practice.
Wider implications of the findings: Here, we present soluble Cripto-1 as a Study question: Does the number of oocytes affect the cumulative clinical
potential inducer of multipolar mitosis that may compromise the developmental pregnancy rate from the transfer of all frozen-thawed embryos.
potential of human embryos. The future investigation of soluble factors in the spent Summary answer: The cumulative pregnancy rate from frozen-thawed
embryo culture medium holds the potential to discover novel non-invasive biomarkers embryo transfers is strongly related to the number of oocytes retrieved, regard-
of human embryo quality and developmental potential. less of the fresh transfer outcome.
Trial registration number: not applicable What is known already: Recent studies show that the number of oocytes
retrieved is correlated with the live birth rate of the fresh embryo transfer. While
most studies have addressed the outcome of the fresh embryo transfer, only
P-147 Effect of Hazardous Air quality index on embryo limited data is available on the cumulative pregnancy rate following the transfer
development in an IVF laboratory in New Delhi, India of all fresh and frozen-thawed embryos in patients that utilized all their embryos
G. Kant1, K.D. Nayar1, M. Singh1, S. Draboo1, H. Sharma1, F. Khan1, which is the most significant outcome.
S. Gupta1, R. Bhattacharya1, D. Nayar1 Study design, size, duration: The study included all oocyte collection cycles
1
Akanksha IVF Centre, Reproductive Medicine, New Delhi, India between January 2009 and September 2019 in a tertiary medical center that had

a fresh embryo transfer and utilized all their frozen-thawed embryos originating The network was then trained to classify embryos based on their known implan-
from the same cohort of retrieved oocytes. tation outcomes. UBar predictions were compared to embryologists’ grades
Participants/materials, setting, methods: Cumulative pregnancy rate was and implantation outcomes.
calculated as any pregnancy from a transfer of frozen-thawed embryos originating Main results and the role of chance: An encoder was trained on over 2.5
from the same cohort of retrieved oocytes. The number of oocytes was divided million images from unlabeled (unknown outcomes) time-lapse videos, maxi-
into quartiles and the cumulative pregnancy rate from the frozen-thawed mizing the information available in the visual data, and “translating” it into feature
embryos was calculated in each group. Logistic regression analysis was utilized vectors. A memory-based network was built to create initial predictions based
to adjust for potential confounders including maternal age, treatment cycle num- on embryologists’ grades. Because grades somewhat correlate with implantation
ber, primary or secondary infertility, fresh embryo transfer outcome and the success, the network used the results from graded videos to set initial feature
number of oocytes retrieved. weights. Videos from KID embryos were then used as input to train the network
Main results and the role of chance: During the study period, there were to predict implantation probability.
1,306 oocyte retrieval cycles that had a fresh embryo transfer and utilized all Performance of the UBar predictions exceeded that of the expert panel as
their frozen-thawed embryos originating from the same cohort of retrieved calculated by the area under the receiver operating characteristic curve (AUC
oocytes. In the first quartile (Q1) 1- 8 oocytes were retrieved (n=369), of the ROC): 0.82 and 0.58, respectively. For a clinically relevant performance
Q2 (25-50%) 9-11 oocytes (n=263), Q3 (50-75%) 12-16 oocytes (n=399) and assessment, the positive and negative predictive values (PPV and NPV) were
Q4 (>75%) >17 oocytes (n=275). The cumulative pregnancy rate from fro- calculated for UBar predictions and compared to those of the expert panel.
zen-thawed embryo transfers originating from the same cohort of retrieved Both the PPV (93%) and the NPV (58%) of UBar significantly surpassed the
oocytes increased with the number of oocytes retrieved Q1 76/369 (20.6%); corresponding values of the expert panel (PPV=81% and NPV=23%), implying
Q2 87/263 (33.1%); Q3 137/399 (34.3%); Q4 106/275 (38.5%) p<0.001. that application of UBar in a clinical setting could potentially improve embryo
This increase in the cumulative clinical pregnancy rate was significant with the transfer outcomes. The increased NPV is particularly meaningful as an indication
number of oocytes retrieved among patients who conceived in the fresh transfer that UBar more accurately identifies embryos that would likely not implant.
and ( p=0.002) of those who did not (p=0.01). On a logistic regression model, Limitations, reasons for caution: In order to further improve results, the
maternal age and the number of oocytes were significant independent predictors model will be applied to a larger amount of samples originating from multiple
for a cumulative clinical pregnancy rate from the frozen-thawed embryo transfers IVF clinics. New ML models will also be developed, and clinical parameters will
regardless if a pregnancy was achieved in the fresh transfer. be explored in future experiments to account for external factors that may
Limitations, reasons for caution: The limitation of the study is in the ret- influence implantation aptitude.
rospective nature of the study. Wider implications of the findings: Our ML-based embryo outcome pre-
Wider implications of the findings: Our findings might challenge the con- diction model predicts implantation probability more effectively than a panel of
cept of optimal stimulation when looking at the cumulative pregnancies arising expert embryologists. Adjunct use of UBar in IVF clinics could aid in selection
from transfer of all frozen-thawed embryos originating from the same cohort of embryos with the highest implantation competence for transfer, as well as
of retrieved oocytes. With agonist triggering and freeze all strategy, targeting deselection of potentially non-implantable embryos.
for a higher stimulation is not only safe but also quite effective. Trial registration number: Not applicable
P-149 Deep learning model for improving embryo selection and P-150 The impact of advanced maternal age on early and late
deselection morphokinetic parameters
A. Polsky1, D.H. Silver1, M. Feder1, Y. Gold-Zamir1, S. Rosentraub1, O. Lebovitz1, Y. Atzmon1, N. Aslih1, M. Michaeli1, D. Paltov1,
E. Shachor1, A. Weinberger1, P. Mazur2, V.D. Zukin2, A.M. Bronstein3 E. Shalom-Paz1
1
1
Embryonics- Ltd., Research and development, Tel Aviv, Israel ; Hillel Yaffe Medical Center Affiliated with Rappaport Faculty of Medicine-
2
Clinic of Reproductive Medicine ‘Nadiya’- Kyiv- Ukraine, Embryology, Kyiv, Ukraine ; Technion- Haifa, IVF Unit- Department of Obstetrics and Gynecology, Hadera, Israel
3
Technion -- Israel Institute of Technology, Computer Science, Haifa, Israel
Study question: Are there any differences between morphokinetic parameters
Study question: Can time-lapse imaging files of developing embryos be used as (MP) and embryo quality (EQ) of embryos derived from young versus advanced
input for a deep learning algorithm to improve implantation probability predictions? maternal age (AMA) women?
Summary answer: Application of time-lapse imaging files to a deep learning Summary answer: Embryos derived from younger women presented a sig-
algorithm produced higher positive and negative predictive values than an exter- nificantly different MP in younger versus AMA women. These differences could
nal panel of embryologists. later cause differences in EQ.
What is known already: Although manual annotation and quality assessment What is known already: The introduction of TLI has provided a more precise
of embryos fertilized in vitro remains the gold standard for embryo selection, understanding of early embryo developmental stages by assigning unique quan-
efforts to improve embryo outcome prediction have become increasingly titative identifiers to each embryo. Data regarding possible association between
computational with research progressively turning toward artificial intelligence embryo morphokinetics and maternal age is limited and conflicting.
(AI). The majority of AI-based algorithms for embryo selection either require Study design, size, duration: Using a time-lapse incubator we retrospectively
user-defined input parameters, or use little of the available embryo information compared between MP and quality of embryos derived from young (<30 years) and
(e.g., still images as opposed to time-lapse). Moreover, published algorithms AMA women (41-44 years). This study included 364 ICSI cycles (173 cycles from
often do not report clinically relevant performance metrics, making it difficult to young and 191 from AMA) performed between January 2016 and December 2018.
transition the results from research to the clinic. Participants/materials, setting, methods: Embryo development was ana-
Study design, size, duration: The dataset consisted of 8,789 retrospectively lyzed with TLI system (EmbryoScope, Unisense FertiliTech). The following kinetic
collected time-lapse videos from embryos cultured to the blastocyst stage. Initial markers were assessed: time to pronuclei fading (tPNf ), and appearance of two
training of a machine learning (ML) model, UBar, was executed on 4,087 of the to eight cells (t2-t8). For embryos cultured to blastocyst, time to morula (tM),
embryos, which were graded by an external panel of embryologists from various start of blastulation (tSB) cavitated and expanded blastocyst (tB, tEB) were also
countries. Time-lapse videos were the input for the model, which was cross-val- recorded. Alpha-ESHRE and KIDScore algorithm were used to determine the
idated on 272 embryos with known implantation data (KID). Prediction of transferred embryo quality. Embryos derived from young women were com-
implantation probability came from the cross-validation data only. pared with those derived from AMA.
Participants/materials, setting, methods: Embryos in this study came Main results and the role of chance: A total of 2021 oocytes were evalu-
from patients who underwent fertility treatment between 2012 and 2019 at the ated. 60.5% (n=1223) of oocytes were derived from patients under 30 years
Clinic of Reproductive Medicine “Nadiya” in Kiev. Resulting time-lapse videos and 39.5% (n=798) from women 41 years old and above. A trend towards faster
were used for developing, training, and 10-fold cross-validation of the model. polar body disappearance observed among patients <30 years compared to
Initial model training used a recurrent neural network to recover panel grades. patient ≥41 years old. (24.7hr vs 25.2hrs respectively, p=0.05). The mean time

points for younger patients were significantly shorter at t3, t4, t5, and t6 com- completion of the dashboard provided peace of mind when leaving work to 82.6
pared to AMA women (37.2, 39.8, 50.1 and 53.6 hrs. versus 38.4, 40.4, 51.2 (68.6, 92.2)% of respondents, and 84.8 (71.1, 93.7)% felt more confident knowing
and 54.7 hrs. respectively) (P<0.05). For embryos cultured to the blastocyst that procedures were completed according to the EWS. Most respondents (73.9
stage tSB, tB and tEB were significantly shorter for embryos derived from younger [58.9, 85.7]%) agreed that EWS allowed double witnessing to be conducted quickly
patients compared to embryos derived from AMA group (100.7, 108.9, 116.8 and accurately, and that labelling standardization increased efficiency (84.8 [71.1,
hrs. versus 104.7, 115.0, 126.6 hrs. respectively, p<0.001). There were no sig- 93.7]%). The majority of respondents considered EWS training easy (78.3 [63.6,
nificant differences between the two groups at t2, t7, t8 and tM. 89.1]%) and would recommend the EWS to another embryologist (88.9 [75.9,
The proportion of toP-quality embryos (KID 4-5) was significantly higher 96.3]%), while 26.1 (14.3, 41.1)% had concerns with transitioning from manual to
among transferred embryos of younger women compared with AMA women EWS. Managers agreed (93.3 [68.1, 99.8]%) that sample chain of custody was
(87.1% versus 76.7%; p=0.007, respectively). Similar results obtained for alpha easily followed using EWS audit reports.
scoring (alpha 2-3) (83.3% vs 72%; p=0.001 respectively). Limitations, reasons for caution: The survey was sent to one network of
Limitations, reasons for caution: Limitations of the present study are those embryology laboratories as it relates to a single EWS. Embryologists’ perceptions
inherent to the TLI technique which is a static system and might be operator may be influenced by length of experience with a particular system and by their
dependent. Since the embryos cannot be rotated once entered to the embryo- general embryology experience and knowledge.
scope, situations like blastomeres overlap or irregular divisions might be difficult Wider implications of the findings: This study indicates that the implemen-
to observe by the embryologist. tation of an EWS was well perceived by most embryologists. Sample identifica-
Wider implications of the findings: This is the first study to compare the tion, traceability, mismatch prevention, laboratory efficiency and embryologist
time-lapse analysis of younger with AMA women up to the blastocyst stages. confidence was perceived to be improved. For the first time, these data captured
Since increasing women’s age was associated with slower embryo developmental embryologists’ opinion on the value of an EWS.
kinetics, these parameters can be potentially utilized to improve embryo selec- Trial registration number: NA
tion and cycle outcome in AMA women undergoing IVF treatment.
Trial registration number: N/A P-152 Multinucleation has no impact on an embryos ability to
develop, implant or sustain a viable pregnancy to live birth
P-151 Embryologists’ perceptions following implementation of an V. Songkhunawej1, L. Burmeister2, K. Sorby1
electronic witnessing system in an in vitro fertilisation laboratory 1
Number 1 Fertility, Embryology, East Melbourne, Australia ;
network 2
Number 1 Fertility, Clinician, East Melbourne, Australia
J. Swain1, K. Wirka2, B. Hayward2, A. Catherino2, R. Holmes3
1
CCRM IVF Network, IVF Laboratory, Lone Tree, U.S.A. ; Study question: The aim of our study was to investigate the impact of mul-
2
EMD Serono, Fertility & Endocrinology, Rockland, U.S.A. ; tinucleation on ART outcomes including embryo development, utilisation, preg-
3
CCRM Boston, IVF Laboratory, Newton, U.S.A. nancy loss and live birth.
Summary answer: Presence of multinucleation appears to have no negative
Study question: How do embryologists perceive the value of an electronic impact on embryo development nor the embryo’s ability to implant and sustain
witnessing system (EWS) with regard to sample traceability, mismatch preven- a viable pregnancy.
tion, workload interruption, laboratory efficiency, system implementation and What is known already: Detected as early as 1987 by Tesarik et al, multinu-
overall embryologists’ confidence? cleation in embryos is thought to arise from abnormalities in DNA synthesis during
Summary answer: The EWS was well perceived by embryologists in all evaluated cleavage. Previous literature demonstrated that multinucleation in embryos was
domains; it enhanced confidence and peace of mind at the end of a work day. linked to increased aneuploidy rate (Kligman et al, 1996) and lower implantation
What is known already: Double witnessing has become an important addi- and live birth rate (Hardarson et al., 2001). However, development of time-lapse
tional task to prevent potential sample misidentification within the in vitro fertil- technology has dramatically improved embryo assessment from just a static time-
isation (IVF) laboratory setting. Best practice dictates the use of double point to continuous monitoring, enabling more robust detection of multinucleation.
witnessing, but while necessary, this can be time consuming and implementation Recent studies suggest that ploidy (Balakier et al., 2016) in addition to perinatal
may vary between laboratories. Electronic witnessing, although not mandated outcomes (Seikkula et al., 2018) are not impacted by multinucleation.
in most parts of the world, has emerged as a potential way to mitigate current Study design, size, duration: In this retrospective study, data was collected
manual double witnessing challenges; however, embryologists’ perspectives from cycles dating from 1st February 2018 to 31st December 2019. In this period,
regarding the value of such technology has not been evaluated. there were a total of 8409 embryos assessed. To date, pregnancy and live birth
Study design, size, duration: This study involved administration of a web- data were available for 363 embryos.
based survey to embryology staff at eight fertility laboratories across the United Participants/materials, setting, methods: Embryos were cultured in the
States and Canada to gather attitudes and perception regarding an EWS (Gidget® Embryoscope+ to day 5 or 6 and assessed for routine morphokinetic parameters.
[Genea BIOMEDX]). The survey remained open for 2 months. There were 73 Multinucleation assessment included: presence or absence at the 2-cell and 4-cell
questions related to: demographics and work experience, sample traceability, stages, number of blastomeres affected, number of nuclei seen. Utilisation, clinical
mismatch prevention, workload and interruptions, laboratory efficiencies, EWS pregnancy and live birth rates were assessed. Transfers were single day 5 embryos.
implementation, managerial tasks and embryologist confidence. To simulate observation of multinucleation in laboratories not using time-lapse,
Participants/materials, setting, methods: The questionnaire was sent to muLtinucleation was assessed in eleven planes at the static timepoint of 42 hours.
embryologists, managers and non-managers in all laboratories. Participants con- Main results and the role of chance: Multinucleation was seen in 31.77%
sented to take the anonymous survey that was composed of Likert scale state- of embryos under time-lapse. However, when assessing at 42 hours, only 8.1%
ments, open-ended questions and ranking questions related to EWS. The values of embryos had visible multinucleation. The occurrence of multinucleation was
corresponded to categorical variables (strongly disagree, disagree, neutral, agree, more likely at the two cell stage in comparison to the four cell stage (20.60% vs
strongly agree), with an option for ‘no viewpoint’ or ‘choose not to answer’ for 6.20%; p = 0.001) with 4.98% of embryos having multinucleation persisting
some questions. Survey results are shown as proportion agreeing with state- through both the two and four cell stages. Prevalance of multinucleation was
ments and 95% confidence intervals (CI). not impacted by age, presence of smooth endoplasmic reticulum discs, uneven
Main results and the role of chance: The survey response rate was 96.2% pronuclei, uneven two cell blastomeres or ploidy. Time to cavitation (both abso-
(50/52). Mean (±standard deviation) experience in a clinical IVF setting was 12.94 lute and relative to syngamy) which was used as a marker of embryo develop-
(±8.67) years for all embryologists (n=45) and 18.67 (±6.79) years for the subset ment, did not appear to be impacted by the presence of multinucleation (78.10h
of managers (n=15); experience with the EWS was 10.9 (±6.30) months and 12.4 vs 78.34h; p = 0.0585) as well as embryo utilisation (51.03% v 47.05%;
[±5.47] months, respectively. Overall (% [95% CI]), 78.3 (63.6, 89.1)% of respon- p = 0.8731). The clinical pregnancy rate was no different when comparing
dents agreed that EWS improved sample traceability and 80.4 (66.1, 90.6)% agreed embryos with or without multinucleation (30.12% v 26.77% respectively; p =
that EWS reduced labelling errors. Just 60.9 (45.4, 74.9)% agreed that EWS 0.4848). Reassuringly, the same was seen in the pregnancy loss rate where
reduced the risk of sample mismatch errors by minimising disruptions. Visual non-multinucleated embryos had a rate of 23.61% versus those with

multinucleation at 16.00% (p = 0.5761). Furthermore, birth weight and gender P-154 Piezo-ICSI can produce significantly higher fertilization
ratios were not significantly different between groups. and blastocyst rates without increasing the risk of birth defects as
Limitations, reasons for caution: Despite having a large sample size for our compared to the IVF
main analysis, after filtering down to embryos that were transferred and had live K. Hiraoka1, M. Ibayashi1, A. Kuga1, N. Uchida1, T. Sujino1,
birth data available, a larger sample size would be more desirable for birth K. Kawai1
outcomes. Interobserver variability is also likely, due to multiple embryologists 1
Kameda IVF Clinic Makuhari, ART Laboratory, Chiba, Japan
making embryo assessments.
Wider implications of the findings: Multinucleation does not appear to Study question: Can Piezo-ICSI increase fertilization and blastocyst rates
have an impact on the capacity of the embryo to develop to blastocyst, implant without increasing the risk of birth defects as compared to the IVF?
or sustain a viable pregnancy to live birth. Multinucleation appears to be an Summary answer: Piezo-ICSI can significantly increase fertilization and blas-
insignificant factor in the developing embryo and should not be used as a criteria tocyst rates without increasing the risk of birth defects as compared to the IVF
for stratification of embryos. What is known already: Recently, the clinical usefulness of Piezo-ICSI has
Trial registration number: Not Applicable been reported by some investigators. All papers reported significantly higher
survival or fertilization rates and embryo development in Piezo-ICSI as compared
P-153 Improving the performance of deep convolutional neural to conventional-ICSI. On the other hand, some papers reported that the com-
networks (CNN) in embryology using synthetic machine- parison of conventional-ICSI and IVF children taking part in an identical follow-up
generated images study did not show any increased risk of birth defects and neonatal complications
in the conventional-ICSI group. To the best of our knowledge, there is no paper
M. Kanakasabapathy1, C. Bormann2, P. Thirumalaraju1, R.
comparing the ICSI results, embryo development and delivery outcome between
Banerjee1, H. Shafiee3
Piezo-ICSI and IVF.
1
Brigham and Women’s Hospital, Division of Engineering in Medicine, Boston, U.S.A. ; Study design, size, duration:
2
Massachusetts General Hospital, Obstetrics and Gynecology, Boston, U.S.A. ;
3
Brigham and Women’s Hospital, Department of Medicine- Harvard Medical
School, Boston, U.S.A. 1. We retrospectively investigated 6,671 mature oocytes between May
2016 and December 2019. Of these, 3,156 mature oocytes were insem-
inated by IVF and 3,516 mature oocytes were inseminated by Piezo-ICSI
Study question: Can we employ synthetic machine-generated images in
and the ICSI results and embryo development were compared.
empowering AI-based morphological assessment of cells with applications in
2. We retrospectively investigated 204 infants born after our IVF program
embryology?
since May 2016. Of these, 82 infants were derived from IVF and 122 infants
Summary answer: The addition of synthetic oocyte images significantly
were derived from Piezo-ICSI and the delivery outcomes were compared.
improved the performance of a CNN in oocyte assessment compared to a CNN
trained on clinical oocyte images.
What is known already: CNNs have shown an enormous potential in embry- Participants/materials, setting, methods:
ology. These neural networks, in general, require large amounts of expertly
annotated data to perform sufficiently well. However, the availability of datasets 1. The fertilization, blastocyst and good quality blastocyst rates of IVF and
is limited to very few fertility centers worldwide. Generative adversarial networks Piezo-ICSI were compared.
(GANs), which excel in generating life-like images, may help in mitigating such 2. The number of embryos transferred, monozygotic twin rates, dizygotic
data requirements. It has been shown previously that oocytes can be differen- twin rates, gestational week, birth weight, the ratio of boy and girl and
tiated based on their potential for fertilization by CNNs through morphological birth defects rates were compared.
analysis (Thirumalaraju et al., 2019). Here, we evaluate whether such a CNN
when trained with additional data generated by a GAN can improve its
The good quality blastocyst was defined as better than grade BB by Gardner’s
performance.
criteria. The data were analyzed by Unpaired T Student, chi-square test or
Study design, size, duration: Using a retrospective dataset of clinical oocyte
Fisher’s exact test.
images with known fertilization outcomes (KFO) and synthetic oocyte images
Main results and the role of chance:
generated by a pretrained GAN, a CNN, henceforth called synthetic CNN
(s-CNN), was trained to classify between oocytes that eventually fertilized nor-
mally, with 2 pronuclei (2PN), and abnormally (non-2PN). The network perfor- 1. Among the IVF and Piezo-ICSI, there were no significant differences
mance was compared to the performance of a CNN, henceforth called the when comparing the average age of women at the insemination (34.3 ±
original CNN (o-CNN), trained using clinical oocyte images only. 4.1 vs. 34.7 ± 4.1). The fertilization rates of IVF and Piezo-ICSI were
Participants/materials, setting, methods: A GAN was trained to generate 73.0% and 83.8%. A significantly higher fertilization rate was observed in
life-like oocyte images from scratch. The s-CNN was trained using a dataset of Piezo-ICSI (P < 0.01). The blastocyst rates of IVF and Piezo-ICSI were
1411 clinical oocytes images with KFO and 1340 synthetic oocyte images gen- 41.3% and 46.2%. A significantly higher blastocyst rate was observed in
erated by the GAN. The o-CNNs were trained as reported previously using Piezo-ICSI (P < 0.01). The good quality blastocyst rates of IVF and
only the 1411 clinical oocyte images with KFO. The trained networks were tested Piezo-ICSI were 19.1% and 22.4%. A significantly higher blastocyst rate
with a common set of 712 clinical oocyte images with KFO. was observed in Piezo-ICSI (P < 0.01).
Main results and the role of chance: The o-CNN performed with an 2. Among the IVF and Piezo-ICSI, there were no significant differences
accuracy of 67.0% (95% CI: 63.4% to 70.4%) and an AUC of 0.6133 in classifying when comparing the average age of women at the embryo transfer (34.6
oocytes based on their eventual fertilization outcome, as reported previously ± 4.0 vs. 35.1 ± 4.1), average number of embryos transferred (1.0 ± 0.3
(n=712). In contrast, the s-CNN performed with an improved accuracy of vs. 1.1 ± 0.3), monozygotic twin rates (1.3% vs. 1.7%), dizygotic twin
82.58% (CI: 79.59% to 85.30%) and an AUC of 0.8123 (n=712) in differentiating rates (1.3% vs. 0.8%), gestational week (38.8 ± 2.8 vs. 38.7 ± 2.7), birth
the oocyte images based on their eventual fertilization outcomes. weight (3036 ± 1751g vs. 3005 ± 1662g), the ratio of boy and girl (51:49
Limitations, reasons for caution: Images were obtained using a single vs. 49:51) and birth defect rates (5.0% vs. 3.3%).
imaging platform (EmbryoScope) at a single timepoint. Fertilization assessments
(2PN evaluations) we performed by individual embryologists from a single center. Data are expressed as mean ± SD.
Wider implications of the findings: The training with GAN-generated data Limitations, reasons for caution: The number of matured oocytes of IVF
helped s-CNN to outperform the conventionally trained o-CNN when evalu- was calculated as the total number of 0PN (with no pronucleus and with1 or 2
ating real oocytes with KFO. Thus, GANs may hold the potential to improve the polar bodies), 1PN (mono-nucleus), 2PN (two-pronucleus), ≥3PN (more than
currently utilized CNNs in embryology. three-pronucleus) at the time of fertilization assessment. So, the 0PN oocytes
Trial registration number: not applicable might be immature at the time of insemination.

Wider implications of the findings: Piezo-ICSI can produce significantly deficient patients (Estimate:1.64, p=0.031). Moreover, in the deficient group, age
higher fertilization and blastocyst rates without increasing the risk of birth defects was significantly higher for aneuploid blastocysts (33.3±5.5 vs 29.2±5.8; p=0.03).
as compared to the IVF. Therefore, Piezo-ICSI could be the main insemination Limitations, reasons for caution: This study is based on a good prognosis
method for human assisted reproductive technology to improve the effective Middle Eastern population, who are mainly VitD deficient or insufficient (<30
utilization rate of precious oocytes. ng/ml), therefore these results can not be extrapolated to other patient groups.
Trial registration number: not applicable Wider implications of the findings: In a deficient VitD population, intra-
follicular concentration of VitD is a marker of embryo competence. The mech-
anism of the impact of VitD on chromosomal status of the blastocysts needs to
P-155 Vitamin D (VitD) in follicular fluid (FF) correlates with the be further investigated. Preliminary data in the literature suggest that telomere
euploid status of the embryo in a VitD deficient Middle Eastern length may be involved.
population. Trial registration number: NCT03073720
A. Arnanz1, N. De Munck1, I. El Khatib1, A. Bayram1, A. El-
Damen1, A. Abdala1, L. Melado2, B. Lawrenz2, A. Pacheco3, H. P-156 Piezo-ICSI can generate significantly higher survival,
Fatemi2 fertilization and blastocyst rates without increasing the risk of
1
IVIRMA Middle East Fertility Clinic LLC, IVF lab, abu dhabi, United Arab Emirates ; malformation as compared to the Conventional-ICSI
2
IVIRMA Middle East Fertility Clinic LLC, Gynaecology, abu dhabi, United Arab A. Kuga1, K. Hiraoka1, M. Ibayashi1, N. Uchida1, T. Suhara2, T. Kaji2,
Emirates ; Y. Kamada2, T. Sujino1, K. Ohuchi3, M. Hayashi3, K. Kawai4
3
IVIRMA Madrid, Andrology, Madrid, Spain 1
Kameda IVF Clinic Makuhari, laboratory, Chiba, Japan ;
2
Kameda Medical Center, laboratory, Chiba, Japan ;
Study question: Is there a correlation between VitD levels in individual FF and 3
Kameda Medical Center, ART center, Chiba, Japan ;
embryo ploidy status among Middle Eastern patients undergoing in vitro fertil- 4
Kameda IVF Clinic Makuhari, ART center, Chiba, Japan
ization (IVF) treatments?
Summary answer: In a Vitamin D deficient population, euploid blastocysts Study question: Can Piezo-ICSI generate higher survival, fertilization and blas-
showed significantly higher levels of 25-hydroxyvitamin D[25(OH)D] and bio- tocyst rates without increasing the risk of malformation as compared with the
available 25(OH)D in FF compared to the aneuploid ones. Conventional-ICSI?
What is known already: The widespread distribution of the VitD receptor Summary answer: Piezo-ICSI can generate significantly higher survival, fer-
in reproductive tissues suggests an important role for VitD in human reproduction. tilization and blastocyst rates without increasing the risk of malformation as
Despite the abundance of sunlight, due to socio cultural/religious habits, the compared with the Conventional-ICSI.
prevalence of VitD deficiency/insufficiency in Middle Eastern women is known What is known already: ICSI has been widely performed by using micropipette
to be among the highest in the world. with spiking tip and usually, the cytoplasm is aspirated to break the membrane
The assessment of patient´s VitD is based on the 25(OH)D metabolite mea- (Conventional-ICSI). Recently, the clinical usefulness of Piezo-ICSI has been
surement. However, most of the circulating 25(OH)D is bound to either vitamin reported by several investigators. In Piezo-ICSI, micropipette with flat tip is used
D-binding protein (VDBP) (88%) or albumin (12%) and less than 1% circulates and the membrane breakage is performed by applying piezo without aspirating
free. The albumin-bound fraction of 25(OH)D plus free fraction equals the the cytoplasm into the micropipette. When comparing the survival and fertilization
bioavailable fraction of 25(OH)D. rates between Conventional-ICSI and Piezo-ICSI, favorable results of Piezo-ICSI
Study design, size, duration: A prospective observational study was per- has been reported. However, little information is available comparing the ICSI
formed between 2017 and 2019, including Middle Eastern couples planned for results and delivery outcome between Conventional-ICSI and Piezo-ICSI.
preimplantation genetic testing for aneuploidies (PGT-A). Trophectoderm biopsy Study design, size, duration:
was performed on 115 blastocysts from 37 couples and subjected to Next
Generation Sequencing. Serum (S) samples were withdrawn on the day of the 1. We retrospectively investigated 5,748 mature oocytes between January
final oocyte maturation and 115 FF samples from follicles ≥14mm were individ- 2012 and December 2019. Of these, 1,538 mature oocytes were insemi-
ually collected during the oocyte retrieval for 25(OH)D, bioavailable 25(OH)D nated by Conventional-ICSI and 4,210 mature oocytes were inseminated by
and free 25(OH)D measurement. Piezo-ICSI and the ICSI results and embryo development were compared.
Participants/materials, setting, methods: Patients were classified into two 2. We retrospectively investigated 178 infants born after our IVF program
groups according to the serum 25(OH)D levels; VitD deficient <20 ng/ml and since January 2012. Of these, 89 infants were derived from Conventional-
insufficient/replete ≥20 ng/ml. IVF and 89 infants were derived from Piezo-ICSI and the delivery out-
Quantitative measurement of total 25(OH)D was performed using the Roche comes were compared.
total 25(OH)D assay with competition principle in an electrochemiluminescence
(ECLIA) binding assay. VDBP was measured using an ELISA kit (R&D system).
Participants/materials, setting, methods:
Free and bioavailable 25(OH)D were determined using the method reported
by Bikle et al (1986) applying a modified Vermeulen formula.
Main results and the role of chance: The average maternal age in both 1. The survival, fertilization and blastocyst rates of Conventional-ICSI and
groups was 31.1±6.0 vs 32.1±5.0; p=0.369, body mass index 26.1±2.9 vs Piezo-ICSI were compared.
25.6±3.8; p=0.499, Antimullerian Hormone 3.8±2.2 vs 3.5±1.4; p=0.400, 2. The number of embryos transferred, monozygotic twin rates, dizygotic
mature oocytes 14.2±6.9 vs 13.0±4.9; p=0.561, respectively in <20 ng/ml and twin rates, gestational week, birth weight, the ratio of boy and girl and
≥20 ng/ml group. Serum values were 13.2±4.0 vs 32.3±9.2; p<.0001, respec- malformation rates were compared.
tively. FF samples from a total of 40 and 75 blastocysts were analyzed of which
52.5% (21/40) and 61.3% (46/75) were euploid, respectively. The data were analyzed by Unpaired T Student, chi-square test or Fisher’s
In <20 ng/ml group, the logistic regression showed that the average values exact test.
of 25(OH)D and bioavailable 25(OH)D were significantly higher in follicles Main results and the role of chance:
resulting in euploid vs aneuploid blastocysts (18.3 vs 13.9; p=0.029) vs (18.1 vs
13.7; p=0.029). However, in ≥20 ng/ml group, no significant differences were 1. The survival rates of Conventional-ICSI and Piezo-ICSI were 92.9% and
found (38.1 vs 40.6; p=0.417) and (37.7 vs 40.1; p=0.441), respectively. 95.2%. A significantly higher survival rate was observed in Piezo-ICSI
A Pearson test (ρ) indicated a strong correlation between S and FF 25(OH) (P < 0.01). The fertilization rates of Conventional-ICSI and Piezo-ICSI
D, bioavailable 25(OH)D and free 25(OH)D values (r= 0.805 vs r= 0.805 vs were 69.9% and 81.1%. A significantly higher fertilization rate was
r=0.834; p<.001 respectively). observed in Piezo-ICSI (P < 0.01). The blastocyst rates of Conventional-
In a multivariate model, confounding for age, BMI and AMH, sufficient/replete ICSI and Piezo-ICSI were 47.1% and 54.4%. A significantly higher blasto-
have a significantly higher probability to have a euploid embryo compared to cyst rate was observed in Piezo-ICSI (P < 0.01).

2. Among the Conventional-ICSI and Piezo-ICSI, there were no significant Limitations, reasons for caution: In the case of biopsy after warming, it is
differences when comparing the average age of women at the embryo difficult to determine the number of warming embryos and re-cryopreservation
transfer (38.4 ± 4.3 vs. 38.7 ± 4.0), number of embryos transferred (1.2 of surplus euploid embryo could increase embryo damage and miscarriage rate.
± 0.4 vs. 1.2 ± 0.4), monozygotic twin rates (4.7% vs. 0%), dizygotic twin Wider implications of the findings: Using recently available data, when
rates (0% vs. 4.7%), gestational week (38.2 ± 1.7 vs. 38.3 ± 2.1), birth faced with the option of fresh embryos, before or after trophectoderm biopsy
weight (2913 ± 462g vs. 2947 ± 489g), the ratio of boy and girl (38:62 for PGT-A, our result supported performing TE biopsy of blastocyst for PGT-A
vs. 53:47) and malformation rates (6.7% vs. 6.7%). before or after vitrification and warming may be the optimal timing for better
clinical outcomes.
Data are expressed as mean ± SD. Trial registration number: Not applicable
Limitations, reasons for caution: In our IVF program, day 2 or 3 after
oocyte retrieval, 1-3 good quality cleaved embryos were cryopreserved or P-158 A machine-learning (ml) based decision support framework
transferred, so the blastocyst culture was performed excluding these embryos. with a case study in frozen-embryo-transfer (fet) in-vitro-
Wider implications of the findings: Piezo-ICSI can generate significantly fertilization (ivf ) for patients with polycystic ovarian syndrome
higher survival, fertilization and blastocyst rates without increasing the risk of (pcos)
malformation as compared to the Conventional-ICSI. Therefore, Piezo-ICSI T.J Ramnath Babu1, V.S. Roy2, Y. Jambunath1, D. Modi3,
could be an alternative ICSI method to the Conventional-ICSI without losing S.B. Bhuvarahamurthi1, N. T s4
valuable oocytes for human assisted reproductive technology. 1
SpOvum Technologies Pvt. Ltd., Product Development, Bangalore, India ;
Manipal Fertility - JP Nagar, Scientific Director & Chief Embryologist, Bangalore,
India ;
3
National Institute For Research In Reproductive Health ICMR-NIRRH, Molecular
P-157 Optimum timing of trophectoderm biopsy with and Cellular Biology Lab, Bangalore, India ;
4
cryopreservation for preimplantation genetic testing for Manipal Fertility - JP Nagar, Clinical Director, Bangalore, India
aneuploidies (PGT-A) cycle
Study question: Can a data-driven framework not just predict, but also aid
E.J. Yu1, E.A. Park1, S.A. Choe1, M.J. Kim1, Y.S. Kim1, J.Y. Lee1,
embryologists/ clinicians in decision-making during a FET-IVF treatment for
E.B. Joe1, H.S. Kim1, J.H. Lee1, H.O. Kim1, Y.S. Kim1, M.K. Koong1,
patients with PCOS?
I.S. Kang1, T.K. Yoon1
1
Summary answer: We show that one can augment the capability of predic-
CHA Fertility Center- Seoul Station, Department of Obstetrics and Gynecology, tion-tools and controllability of parameters using a ML-based framework towards
Seoul, Korea- South improving ‘clinical pregnancy rate’ (CPR).
What is known already: Existing literature primarily focus on prediction, and
Study question: Is the timing of trophectoderm(TE) biopsy before or after seldom aid in the IVF treatment process. Recent decision-support systems in the
vitrification-warming associated with successful clinical outcomes after the IVF domain consider non-clinical parameters like demographic details. However,
embryo transfer(ET) of blastocyst subjected to preimplantation genetic testing the efficacy of the decisions taken during the treatment are not considered. It
for aneuploidies (PGT-A)? is also noted that earlier case studies cover diverse population including several
Summary answer: TE biopsy(PGT-A) with cryopreservation may be the male and female causes of infertility, to predict IVF outcomes. One downside
optimal timing for better clinical outcomes compared to the fresh TE biopsy of including miscellaneous collection of infertility factors is that the model
without cryopreservation. becomes highly uncertain. Specifically, no study has covered PCOS condition,
What is known already: Embryo biopsy and fresh embryo transfer are tra- the most common cause of infertility.
ditionally performed in the PGT-A cycle. However, before ET, the time allowed Study design, size, duration: We developed a supervised machine learning
for genetic analysis of the specimens is restricted, particularly after blastocyst model covering only PCOS in FET-IVF cycle. The purpose of the study is to
biopsy. Cryopreservation of blastocysts after biopsy instead of fresh transfer classify the CPR as either Yes/No. Anonymized clinical data along with CPR
permits more sufficient time for performance of molecular diagnosis. The effect outcomes of 330 PCOS patients who underwent FET-IVF treatment at an IVF
of cryopreservation and warming procedures on clinical outcomes in PGT-A center between 2015 and 2019 were retrospectively analyzed. Patients with
cycle has not been effectively studied. donor samples (donor oocyte/ sperm/ embryo), severe male infertility factors
Study design, size, duration: Retrospective analysis of patients with PGT-A were excluded from this dataset, resulting in 187 (~56%) patient records
cycles was carried out from January 2016 to November 2019. 3684 blastocysts Participants/materials, setting, methods: We avoided sampling to reduce
from cycles were subjected to TE biopsy for performing array comparative genomic uncertainty. Feature engineering was performed to label the details including
hybridization test. Embryos were cultured to expanded blastocyst and performed embryo details (quantity and grade of embryos before freezing, post thawing
TE biopsy on day 5 or day 6, depending on the timing of embryo expansion. Cycles and before embryo transfer) as good or moderate or poor. Then we used naïve
with complete PGT-A diagnosis were 655, among them 349 of which had ET. Bayes algorithm to classify CPR as Yes or No. Here, CPR as ‘Yes’ is defined as
Participants/materials, setting, methods: The performance of different the ultrasound visualization of a pregnancy sac with fetal heart activity from six
groups of PGT-A patients was evaluated. The groups were divided into three; weeks after frozen embryo transfer.
first group (n=118 transfer/677 cases) contained fresh blastocysts that biopsied Main results and the role of chance: The f-value statistical test results
for PGT-A without cryopreservation followed by embryo transfer. In the second suggested that all the three parameters (good (~1) / moderate (~1.5)/ poor
group (n=348 transfer/355 cases), the blastocysts that were biopsied prior to (~7)) related to the embryo grades are more correlated with CPR compared
vitrification; and subsequently warmed euploid blastocysts for ET. The last group to other parameters. Our embryo-grades representation is intuitive and generic
(n=189 transfer/248 cases), the vitrified blastocysts were warmed and then and can be performed across all clinics. For decision making, the embryologist’s
biopsied before proceeding with ET. inputs are included as parameters (features) to the model.
Main results and the role of chance: The total pregnancy and implantation A naïve Bayes classifier model was developed using 95% training dataset with
rates of fresh blastocyst biopsied group was 42.4% (50/118), 35.7%(56/157); seven features, and prediction performance were evaluated using macro f1-score
the cryopreserved-warmed-biopsied blastocyst group showed 56.6% (197/348), and accuracy on the remaining 5%. Macro f1-score was calculated as the har-
51.2% (227/443); and finally biopsied and cryopreserved-warmed group showed monic mean of precision and recall of each class label (Yes, No), hence reducing
54.0% (102/189), 48.6% (123/253); respectively. Second group and third group skewness in prediction.
are significant higher pregnancy rates than first group. Also, The second(biopsied Model predictions using test data resulted in 90% accuracy and 90% f1 score
and cryopreserved-warmed) group and third (cryopreserved-warmed–biopsied) at best.
group are significant higher implantation rates than first group. Also, The second To demonstrate the role of the ML framework, we chose ‘Day of freezing’,
group(36/197, 18.3%) showed higher numerical miscarriage rate than the first ‘Culture type’ and ‘Number of embryos transferred’ as the controllable param-
group(6/50, 12.0%) and third group(16/102, 15.7%). eters for decision-making. The model calculates all scientifically possible

combinations of these parameters and ranks the suggestions with confidence might improve oocyte quality. Further, the differences between transcription and
scores. An embryologist can decide the pathway of the treatment using this mitochondrial activity may be used to develop reliable predictive test for mito-
rank list. chondrial genetic screening
Limitations, reasons for caution: A limitation in this study was the small Trial registration number: not applicable
size of dataset collected from a clinic. This limitation could be overcome by using
more data from multiple clinics, thus improving the model’s performance. P-160 Remote embryo assessment: shifting the paradigm from
Wider implications of the findings: The ML framework shall be able to traditional embryo grading
support and guide embryologists on the different paths for them to plan and A. Picou1, K.A. Wirka2, A.B. Catherino2, B. Hayward2, M.D.
adhere-to in order to achieve better success rates. The framework also sensitizes VerMilyea1,3,4
the criticality of certain data-points which will further impact the perspective of 1
Ovation Fertility, Laboratory and Operations, Austin, U.S.A. ;
an IVF treatment. 2
EMD Serono, Fertility and Endocrinology, Rockland, U.S.A. ;
Ovation Fertility, IVF Lab, Austin, U.S.A. ;
4
Texas Fertility Center, IVF Laboratory, Austin, U.S.A.
P-159 Oocyte maturation changes mitochondrial gene expression
patterns and the localization of polarized mitochondria but does Study question: How confident are embryologists with remote embryo assess-
not affect mitochondrial potential ment (REA) using time-lapse (TL) technology as an alternative to traditional
F. Zambelli1, M. Barragan1, R. Vassena1 embryo assessment (TEA) when performing morphology grading?
1
Clinica Eugin, Research and Development, Barcelona, Spain Summary answer: REA enables morphology grading with high confidence at
fertilization assessment (Day 1) and Days 3, 5, 6 and 7 of embryo development.
Study question: Are there changes in the oocyte mitochondrial transcriptome What is known already: Despite other advances in assisted reproductive
and membrane potential during meiotic maturation? technology, morphology is the stalwart used by embryologists to grade embryos
Summary answer: We identified a decreased mitochondrial gene expression for transfer. To be able to assess and select embryos with minimal culture dis-
and a more compact distribution of polarized mitochondria in MII compared to turbance is highly desirable. The impact of REA enabled by TL technology in the
GV; mitochondrial membrane potential was unaffected in vitro fertilization (IVF) laboratory remains unknown, including its potential
What is known already: Mitochondria localization and activity in oocytes impact on embryo grading and selection compared with TEA (routinely per-
from several species has been associated with the acquisition of developmental formed with a light microscope). Most importantly, the level of confidence in
potential. Several studies have attempted to associate mitochondrial measure- embryo assessments performed remotely by embryologists using this novel
ments such as DNA copy number and gene expression in human oocytes with technology has not been previously evaluated. Here we describe preliminary
reproductive outcomes after IVF. These tests rely on the assumption that a results of a feasibility study.
reduction in the abundance of certain transcripts directly translates to a modi- Study design, size, duration: Non-interventional, longitudinal study in a
fication in mitochondrial activity. However, this correspondence has not yet been single IVF centre over 5 months. Seven embryologists performed Day 1–7 mor-
proven. Moreover, a clear causation between an altered mitochondrial activity phology gradings using TEA and REA (Geri Connect and Assess® [Genea
and the failure to reach MII in human oocytes hasn’t been demonstrated yet. BIOMEDX]). Embryologists rated their level of confidence (high/low) for each
Study design, size, duration: We analyzed the mitochondrial membrane embryo grading and stated reasons for low confidence, when applicable. TEA
potential and the localization of active mitochondria in human oocytes at GV, was performed using standard procedures and REA performed after a ≥1week
GV after 30 hours in culture (FTM-GV), and MII from GV after 30 hours in culture washout period. This study was determined exempt from review by Sterling IRB,
(IVM-MII). Further, we performed a transcriptomic-wide analysis via microarray Atlanta, GA, USA.
of GV and in vivo matured MIIs (IVO-MII) to correlate functional and transcrip- Participants/materials, setting, methods: Data included 5458 morphology
tional measurements. gradings from 72 embryo cohorts evaluated on Days 1,3,5,6 and 7. TL imaging
Participants/materials, setting, methods: 24 oocyte donors (ages 20 was initiated on Day 0. TEA/REA were timed and performed by the same
to 35) provided 63 oocytes (24 GV, 15 FTM-GV, 15 IVM-MII, 9 IVO-MII) for the embryologist; elapsed recording time was used to match timing of TEA (per-
study. Oocytes were stained with JC-1 to identify both polarized and non-po- formed in real time) to the retrospective REA (performed elsewhere at their
larized mitochondria and imaged by confocal microscopy. Intensities of the convenience). REA Block I had no time restrictions; Block II simulated TEA’s time
staining and spatial measurement of the polarized mitochondria layer were per- sensitivity. Confidence in gradings and video usage were reported.
formed using ImageJ. Transcriptome-wide analysis via microarray (HTA 2.0 Main results and the role of chance: Mean durations (minutes ± standard
Affymetrix) was performed using 4 GVs and 9 in vivo matured MII deviation) of cohort assessments were TEA 2.14 (±1.30) and REA 2.02 (±1.18)
Main results and the role of chance: GV and IVO-MII oocytes presented with an average of 10.1 embryos per cohort. Embryologists reported high con-
543 transcripts differentially expressed (516 more abundant in GV, 27 more fidence in 94.4% (636/674) of Day 1 TEA-graded zygotes. REA matched high
abundant in MII). Oxidative phosphorylation and mitochondrial function were confidence in 88.1% (560/636). High confidence was reported for 98.5%
the most affected pathways by Gene Ontology analysis, with a significant (585/594) of Day 3 and 96.0% (1308/1363) of Day 5–7 TEA gradings; REA
decrease of mitochondrial transcripts like ATP5, PDHA, and POLG in MIIs. JC-1 matched in 94.4% (552/585) and 95.0% (1242/1308), respectively. Among
staining indicated in all groups a rather thin layer of polarized mitochondria at embryos graded with low confidence using TEA, 44.7% (17/38) of Day 1, 66.7%
the periphery of the oocyte (within 20 µm from the oolemma), while the inner (6/9) of Day 3 and 72.7% (40/55) of Day 5–7 were graded with high confidence
part of the oocyte (>20 µm from the oolemma) only contained non-polarized using REA. During REA Block I and II, video review was used for 5.1% (53/1033)
mitochondria. We found no differences in the membrane potential of the periph- and 0.5% (8/1666) of embryos, respectively, with 91.8% (56/61) of embryol-
eral mitochondria across groups, (39% increase in polarized mitochondria in ogists having improvement in confidence. The main low confidence reasons
FTM-GV, 15% increase in polarized mitochondria in IVM-MII compared to GV; reported for TEA were “not able to roll embryo around” (56.0% [61/109];
p>0.05). We did observed differences in the distribution of mitochondria mostly for Day 1, 59.0% [36/61]) and “need for a second opinion” (25.7%
between GV, FTM-GV and IVM-MII. Both GV and FTM-GV had a wider distri- [28/109], all Day 5–7). Within the REA group, “poor focus or focal range”
bution of the polarized mitochondria (9.7 ± 1.7 µm and 7.9 ± 1.5 µm, respec- (55.9% [123/220], caused by device malfunction in 25.2% [31/123]) and “not
tively), while in IVM-MII we observed a thinner layer (6.2 ± 1.7 µm), placed in able to roll embryo around” (23.2% [51/220]; mostly for Day 1, 54.9% [28/51]).
the outermost part of the ooplasm (p<0.001). Limitations, reasons for caution: This study was conducted in a single clinic
Limitations, reasons for caution: The origin of MII (both in vitro and in using a specific TL device and REA system; clinics using different devices and
vivo matured) may affect some of the comparative observations; GVs were systems may not observe the same results.
collected after controlled ovarian stimulation and their transcriptome may be Wider implications of the findings: This is the first study to evaluate embry-
different from GVs from unstimulated ovaries. ologist confidence in embryo grading using a remote system. REA embryo grad-
Wider implications of the findings: Our results might inform the develop- ings can be performed with high confidence compared with traditional
ment of better culture media, where the presence of mitochondrial modulators assessment. Improved embryologist confidence for embryos previously graded

with low confidence may be associated with image replay and review effect of obesity on oocyte/embryo quality from the effect of endometrial
enabled by REA. receptivity allowing to study if a higher donor BMI could affect oocytes quality
Trial registration number: N/A and biological outcomes.
Study design, size, duration: We retrospectively analyzed 85 egg donation
cycles (October 2014-January 2020) with ≥ 50% survival rate of vitrified/
P-161 The effect of high humidity by using single step culture warmed oocyte. Only cycles with male age ≤ 45 years old were considered.
media on continuous embryo monitoring incubator. Cycles were divided in two groups: grouP-1 included oocyte-donors with
C. Albert Rodriguez1, M.A. Valera1, S. Pérez-Albalá1, L. Alegre BMI≤23.9 whereas group 2 included donors with BMI≥24. All oocytes were
Ferri1, B. Aparicio-Ruiz1, J. Remohí2, M. Meseguer Escrivá1 injected with fresh ejaculated normozospermic semen (WHO, 2010). Data,
1
IVIRMA, Lab FIV, Valencia, Spain ; shown as avarage±SD, were analyzed with Chi square or Student-t test.
2
IVIRMA, Ob/Gym, Valencia, Spain Participants/materials, setting, methods: GrouP-1 included 57 cycles,
donor BMI (21.28±2.76) and 437 oocytes; grouP-2 included 28 cycles, donor
Study question: Is there any effect of high humidity culture conditions by using BMI (26.03±2.78) and 232 oocytes. Respectively, in grouP-1 and in grouP-2,
single step Media and Continiuous Embryo Monitoring (CEM) on reproductive donor age was 25.39±3.87 and 25.20±3.84, female recipient age was 42.3±3.69
outcome? and 41.9±4.18 (NS). The recipient BMI in grouP-1 and in grouP-2 was respec-
Summary answer: By using single step media and CEM incubator Geri, cul- tively (22.9±3.80) and (23.1±3.60) (NS), the male average BMI were respectively
turing embryos under high humidity does increase reproductive outcome 25.6±3.43 and 25.8±3.46 (NS). Embryos were culture in a time-lapse incubator
What is known already: The first goal of an IVF laboratory is to achieve until day-3 or day-5.
stable conditions to allow the best embryo development. An incubator has Main results and the role of chance: Oocytes survival rate in grouP-1 and
control different variables, like temperature, gas. Usually we don´t need to use in grouP-2 was respectively 88.1% (385/437) and 84.9% (197/232)(NS).
the humid conditions, as oil overlay has supported successful use of a dry incu- Fertilization rates in groups 1 and 2 were respectively 77.4% (297/385) and 67.5%
bator for culture human embryos, preventing changes in the pH, temperature. (133/197) (p<0.05). In grouP-1, a total of 224 embryos (transferred and vitrified)
The evaporation of culture media may affect the osmolality. Therefore, the use were obtained whereas in grouP-2 total embryos were 91. When the efficiency
of humid conditions avoid osmolality changes. of treatments was calculated as the ration between total embryos obtained per
Study design, size, duration: A total of 5755 embryos from ovum donation received oocytes, in grouP-1 a rate was obtained (51.1%, 224/437) higher than
programme and own oocytes and that were culture on a CEM system (Geri in grouP-2 (39.2%, 91/232, p<0.05). No statistical differences were found in beta-
genea biomedix, Australia) were included in a retrospective and multicentric HCG positive (62.2%, 33/53, in grouP-1; 57.7%, 15/26, in grouP-2), clinical preg-
study from three consecutive years. nancy rates (CPR) (52.8%, 28/53, in grouP-1; 53.8%, 14/26, in grouP-2) and
Participants/materials, setting, methods: This incubator has 6 separated implantation rate (IR) (40.9%, 35/86, in grouP-1; 45.4%, 20/44, in grouP-2).
incubators. Three of them works in a dry atmosphere (DC) and the other 3 in Limitations, reasons for caution: In clinical outcomes, only fresh transfers
humid conditions (HC). In DC, 2630 embryos were cultured and a total of 3125 were considered (cumulative data not fully available). Donors was stimulated
embryos were cultured under HC. Retrospectively, blastocyst, good morphology with protocols according to their history and hormonal levels. The sample size
blastocyst rate, pregnancy, and euplody rate. of our population was reduced by selecting male partners <45 year old, a choice
Main results and the role of chance: We found differences in embryo made to reduce the possible male-age-factor effect.
development. We had similar blastocyst rate when the embryos were culture Wider implications of the findings: Increasing donor BMI could affect fer-
under high humidity; 52.8% vs DC 53.1%. When we focused on those blastocyst tilization rate and the amount of embryos obtained suggesting that an ideal BMI
with good morphology we had significantly high quality blastocyst in HC; 1.6% value could be set as cut-off in donor screening. Since clinical outcomes were
vs 9.9%. We also found differences in pregnancy rates between HC and DC, similar in the groups we can hypothesize that endometrial receptivity could adjust
62.4 vs 55.0% respectively. These differences disappeared in thawed procedures the donor BMI effects on oocytes quality.
46.7 % in HC and 50.0% in DC, but the embryos were cultured in normal dry Trial registration number: not available
incubator until transfer. In the group of PGT embryos, we had similar embryo
P-163 Robust embryo scoring model based on artificial
ploidy proportional in both groups 39.6%vs 39.2 % in DC
intelligence (AI) applied to a large time-lapse dataset
Limitations, reasons for caution: The retrospective nature of this abstract
maybe reduce the quality of the evidence presented although sample size is J. Rimestad1, M.F. Kragh2, J.T. Lassen1, A. Tran3, J. Berntsen1
1
outstanding and embryos were cultured with the type of incubator that included Vitrolife A/S, Product Development, Viby J, Denmark ;
2
a time-lapse system. Vitrolife A/S & Aarhus University, Product Development, Aarhus, Denmark ;
3
Wider implications of the findings: Culturing embryos with single step Harrison AI, Co-founder, Sydney, Australia
media may include high humidity as a preference to optimize pregnancy results.
Trial registration number: none Study question: Does an AI-based automatic embryo scoring system perform
consistently independent of patient age, fertilization method, year of treatment,
incubation time, culture media and fertility clinic?
P-162 Effect of higher donor BMI on clinical pregnancy and live Summary answer: The trained AI model has shown to perform consistently
birth rates in IVF cycles. regardless of patient age, fertilization method, year of treatment, incubation
E. Iovine1, V. Casciani1, A. Pristera1, V. Zazzaro1, F. Scarselli1, time, culture media and fertility clinic.
G. Pirastu1, D. Uva1, K. Litwicka1, A. Greco1, P.F. Greco1, What is known already: Use of AI for robust selection of embryos requires
M.C. Greco1, M.T. Varricchio1, A. Ruberti1, M.G. Minasi1, E. Greco1 models trained on large datasets that encompass a wide variation in patient
1
European Hospital, Reproductive medicine, Roma, Italy cohorts and clinical practices. Training with a limited dataset would only be
directly applicable to the clinical setting where it was developed and would
Study question: Could higher donor BMI affect the IVF success rate in egg require retraining should clinical practice change.
donation cycles? Previously, an AI model was reported to sort embryos with positive fetal
Summary answer: Increasing donor BMI reduces fertilization rate and embryo heartbeat (FHB) from all remaining embryos with areas under the receiver oper-
development but not clinical outcomes. ating curves (AUC) from 0.90 to 0.95 (IVY v1.0). However, training AI models
What is known already: Excess weight could be deleterious for women can introduce unintentional biases that should be examined using an independent
causing cardiometabolic health risks and adverse reproductive outcomes. It was test set.
known that women with an overweight or obese BMI, experienced worse out- Study design, size, duration: An AI model was trained for binary classifi-
comes in terms of pregnancy and live birth rates that those with normal BMI. It cation of positive FHB from all remaining embryos, given time-lapse videos of
remains unclear whether those effects are exerted at level of the oocyte quality 98583 embryos from 18 clinics across three continents. The following strati-
or at the endometrial. The oocyte donor model is a good tool to separate the fications were analyzed on a test dataset with 17249 embryos not seen during

training: patient age (for the groups <30, 30-34, 35-39, >39), fertilization between November 2018 and November 2019. The study population consisted
method (IVF, ICSI), year of treatment (2011-2019), incubation time (96-115, of stimulated cycles with minimum 10 oocytes during oocyte pick up with own
115-117, 117-119, 119-130 hours post insemination), culture media and fer- or donor oocytes which were fertilized with partner’s sperm. The sperm was
tility clinic. either from ejaculation with 99% abnormal morphology or from testicular biopsy.
Participants/materials, setting, methods: A total of 115832 embryo vid- Participants/materials, setting, methods: 44 cycles were analyzed in order
eos generated by IVF or ICSI between 2011 and 2019 were included in the study. to evaluate their morphokinetic pattern. Mature oocytes from each patient were
Patient age ranged from 18 to 52. Embryos were cultured using the EmbryoScope divided in two groups: AOA group included oocytes that were incubated in Ca2+
or EmbryoScope+ time-lapse system for at least 4 days. 101188 embryos were ionophore (A23187, CultActive, Gynemed) after ICSI for 15 minutes and NO
deemed non-usable due to PGT results or manual deselection by embryologists. AOA group was the control. All oocytes were cultured in Embryoscope time-
14644 embryos were transferred (fresh or frozen) resulting in 4337 positive FHB lapse incubator and the key time parameters and dynamic events were analyzed
and 10307 negative FHB outcomes. using t-test and Mann–Whitney U test.
Main results and the role of chance: The AUC on all 17249 embryos in Main results and the role of chance: There was no significant difference
the test dataset was 0.95 [95% CI: 0.943-0.955], showing great overall in fertilization rate between 2 groups (AOA-68.41% vs. NO AOA-65.88%,
performance. p>0.05). Furthermore, blastocyst rate was similar between the 2 groups (AOA-
For the four age groups, the n and AUC were <30: 2277, 0.97; 30-34: 4302, 61.48% vs NO AOA-65.74%, p>0.05). The comparison for key time parameters
0.95; 35-39: 4702, 0.95 and >39: 3857, 0.94. For fertilization methods, the AUC and dynamic events (tPB2, tPNa, tPNf, t2, t3,t4, t5, t6, t7, t8, t9+, tSC, tM, tSB,
was 0.96 for IVF (n=4681) and 0.95 for ICSI (n=5073). For years of treatment tB, tEB, tHB, S2, S3, S5, CC2, CC3, Blastulation) showed no significant difference
with more than 250 embryos, the mean AUC across all years was 0.95 [0.91 to between the AOA group and NO AOA group in total (p>0.05). In addition,
0.98]. For the four incubation time groups, the n and AUC were 96-115: 1971, when AOA groups were divided into 2 subgroups a) ejaculated sperm with 99%
0.92; 115-117: 3718, 0.94; 117-119: 2579, 0.94; 119-130: 1871, 0.92. For culture abnormal morphology and b) testicular sperm, the morphokinetic comparison
media, one clinic using two different brands (n=456 and n=536) was investigated of these 2 separated groups to their control did not reach any significant differ-
and had AUCs of 0.92 and 0.91. All AUCs above showed only minor biases, ences (p>0.05).
indicating generalization across subsets. Limitations, reasons for caution: Only a limited number of cases were
Mean AUC across clinics was 0.94 [0.89 to 0.97] for the 12 clinics with more included in this study. Apart from severe male infertility other maternal derived
than 250 embryos in the test dataset. To evaluate model generalization to new factors may also be involved. The results for pregnancy, clinical pregnancy and
clinics, leave-one-clinic-out cross validation was performed on these 12 clinics. live birth were not available for all cases.
Each clinic was evaluated on a model trained on the remaining 11 clinics, resulting Wider implications of the findings: Artificial oocyte activation method
in a mean AUC of 0.94 [0.91 to 0.97]. The similar AUCs indicate generalization does not seem to have an effect on the morphokinetic pattern and embryonic
across clinics. development. Additionally, a benefit on fertilization rate in cases of ejaculated
Limitations, reasons for caution: The different clinics from which the data sperm with 99% abnormal morphology or testicular sperm was not observed.
originated did not use a common strategy for embryo selection, which might Trial registration number: N/A
have introduced biases in model performance. To further validate the AI for
embryo selection power, a prospective study will be performed. P-165 KIDScore helps in selecting the best euploid embryo: a
Wider implications of the findings: Automatic embryo scoring by AI shows retrospective cohort study
convincing results for all provided subgroups of data and can therefore be L. Pellegrini1, L. Parmegiani1, A. Rastellini1, A. Arnone1,
expected to generalize well on new clinics, whose data have not been used for S. Bernardi1, S. Lanzilotti1, A.M. Maccarini1, E. Troilo1,
training the AI. G.E. Cognigni1, M. Filicori1
GynePro Medical Centers- NextClinics International, Reproductive Medicine Unit,
Bologna, Italy
P-164 The effect of artificial oocyte activation (AOA) on Study question: Does KIDScore D5v2 represent an additional tool for the
morphokinetic pattern of human embryos in cases with severe selection of euploid blastocysts and for predicting their implantation?
male infertility Summary answer: KIDScore D5v2 appears to be predictive to discriminate
M. KARAGIANNI1, M.I. Papadopoulou1, C. Oraiopoulou1, between euploid and aneuploid embryos and euploid blastocysts with KIDScore
S. Sgoura1, A. Chatzianastasiou2, P. Moisidou Tsiorva2, E. Taki2, 6.0-10.0 show a significantly higher implantation rate.
A. Vorniotaki2, M. Moysidou1, A. Papatheodorou1, What is known already: Time-lapse microscopy (TLM) offers continuous
N. Christoforidis3, A. Chatziparasidou4 monitoring of embryo development and KIDScore is the software used by the
1
Embryolab, Embryology lab, Thessaloniki, Greece ; TLM incubator Embryoscope (Vitrolife, Sweden) to score embryo morphoki-
2
Embryolab, Andrology lab, Thessaloniki, Greece ; netics, but the prediction of chromosomal abnormalities using morphokinetics
3
Embryolab academy - Embryolab, Clinical, Thessaloniki, Greece ; alone is still insufficient. Preimplantation Genetic Testing for Aneuploidy (PGT-A)
4
Embryolab academy - Embryolab, Embryology lab, Thessaloniki, Greece discriminates between euploid and aneuploid blastocysts; KIDScore could be
used to prioritize the transfer of euploid embryos identified by PGT-A diagnosis.
Study question: Does artificial oocyte activation (AOA) in cases with severe Study design, size, duration: Retrospective cohort study, from June 2016
male infertility alter the morphokinetic pattern of human embryos? to December 2019 on 508 embryos from 156 patients, cultured to blastocyst
Summary answer: This study suggests that there is no significant difference stage (DAY 5-7) and screened by PGT-A. Based on KIDScore, blastocysts were
in morhpokinetics using AOA in cases with 99% abnormal sperm morphology divided in two groups: Group 1 (KS 0 – 5.9; n=320), Group 2 (6.0 – 10; n=188)
or testicular biopsy. Main outcome measures: euploidy rate (number of euploid blastocysts/total
What is known already: Oocyte activation is the result of a cascade of events blastocysts), implantation rate (number intrauterine gestational sacs observed
triggered by sperm. The moment sperm enters oocyte’s cytoplasm it diffuses by transvaginal ultrasonography/nº of transferred blastocysts).
PLCz which in-turn triggers calcium oscillations followed by oocyte activation Participants/materials, setting, methods: Mean female age ±SD was
and fertilization. In cases were low fertilization rates is observed after ICSI for 36.0±4.4 (25-46). Blastocysts were cultured in Embryoscope, assessed by
patients with low sperm count, motility and morphology or patients after tes- KIDScore model D5v2 and biopsied to perform PGT-A (Next Generation
ticular sperm extraction (TESE), sperm related activation deficiency may be Sequencing Veriseq PGT protocol - Illumina, USA). Subsequently, blastocysts
suspected. To overcome low or no fertilization after ICSI, AOA has been used were vitrified and euploid blastocysts were warmed and transferred (100 blas-
with varying success rate. At present the available published data is limited and tocysts; mean 1.14± 0.35).
the value of AOA has not been yet determined. Main results and the role of chance: Female and male mean age
Study design, size, duration: This prospective cohort study included 44 ICSI was comparable between the groups. Euploidy rate was statistically different:
treatments performed at Embryolab Fertility Clinic, in Thessaloniki, Greece 24.6% (79/320) in Group 1 (KS 0 – 5.9) versus 36.7% (69/188) in Group 2

(KS 6.0 – 10; P=0.0040 Chi-squared test), suggesting KIDScore seems to incubator were: 12/20, 16/20, 15/22, 22/30, 27/35 and 31/42 mOsm/kg.
be predictive for selecting euploid blastocysts. Implantation rate was calculated Two dish types could be cultured in their corresponding time-lapse dry incuba-
over the 100 blastocysts transferred; the mean number of transferred tors, as well. In these cases, the osmolality increase was comparable to that
embryos was comparable in each group (Group 1: 1.13±0.34; Group 2: observed in the dry benchtop suggesting that a direct dish fit in the heated plate
1.16±0.37). Implantation rate was significantly lower at 28.2% (13/46) in within time-lapse incubators does not significantly increase evaporation.
Group 1 compared to 51.8% (28/54) in Group 2 (P=0.0288 Yates’s Chi-squared Limitations, reasons for caution: The humidity level in benchtop incubators
test). This result suggests a threshold value based on morphokinetic features may vary and is not usually monitored. In this study, time-lapse dishes were
that could help embryologists to prioritize euploid blastocysts prior to transfer. cultured in a dry benchtop incubator. However, some time-lapse incubators
Selecting euploid blastocysts with KIDScore > 6 seems to promote increased allow for a humidified atmosphere, and its effects on osmolality must be studied
chances of implantation. in each specific dish incubator.
Limitations, reasons for caution: This study is limited to patients from Wider implications of the findings: The starting osmolality of culture media,
only one centre with a relatively low number of blastocysts. Further and larger incubator humidity and time-lapse dish design are important factors to consider
studies are needed to validate these results and to permit efficient clinical during in vitro embryo culture. An excessive evaporation during uninterrupted
implications. culture will result in osmolality increase and may have a detrimental effect on
Wider implications of the findings: Blastocyst selection through embryo development.
time-lapse technology alone should not be considered as a replacement for Trial registration number: N/A
PGT. According to our findings, KIDScore seems to be a potentially
valuable tool for identifying the euploid blastocysts having a higher probability
P-167 Antagomir-Mediated Silencing of let-7a-5p in Mouse
of implantation.
Embryo can improve blastocyst Implantation Rate.
J. Shakerzadeh1, M. Movahedin2, A. Eidi1, N. Hayati Roudbari1,
P-166 The impact of incubator humidity, protein supplementation K. Parivar1
1
and time-lapse dish design on media osmolality during culture. Science and Research Branch- Islamic Azad University, Department of Biology,
M. Garcia-Jimenez1, E. Mestres1, A. Casals1, I. Vanrell1, Tehran, Iran ;
2
A. Villamar1, M. Acacio1, Q. Matia1, J. Cohen2, N. Costa-Borges1, Faculty of Medical Sciences- Tarbiat Modares University- Tehran- Iran.,
G. Calderon1 Department of Anatomical Sciences, Tehran, Iran
1
Embryotools S.L., Research & development, Barcelona, Spain ;
2
ART Institute of Washington, IVF Laboratory, New York- NY, U.S.A. Study question: To evaluate the role of murine let-7a-5p blastocysts in implan-
tation failure and the possibility of let-7a-5p suppression in improvement of
Study question: How do incubator humidity, protein supplementation and implantation in vivo.
time-lapse dish design affect the osmolality of culture media over time? Summary answer: Our results suggested that suppression of embryonic
Summary answer: Incubator humidity and time-lapse dish design play murine let-7a-5p could improve embryonic implantation through the
an important role in the stability of osmolality during culture. Protein upregulation of Igf1, Itgb3 and Tgfb1.
supplementation does not alter osmolality over time. What is known already: Blastocyst implantation needs appropriate molecular
What is known already: Uninterrupted embryo culture is a widespread and cellular communications between blastocyst. microRNAs-mediated gene
practice, but certain key conditions must be considered to avoid osmolality expression of both endometrial tissues and/or blastocyst have impact in the
increasing above detrimental thresholds. Humidified incubation may protect process of implantation and fetal development.
against evaporation, but humidity is a non-binary condition and can vary between Study design, size, duration: This is a case-control study and totally
incubators. 100 adult female mice and 10 adult male mice during 2018-2019 were included
Proteins (like albumin) can have emulsifying properties and may have a (Strain NMRI). We analyzed the expression of murine let-7a-5p in the
potential effect on osmolality during culture under oil. Time-lapse systems control, sham, and treatment embryos (8-cells stage) by quantitative reverse
are usually dry incubators and embryos are cultured uninterruptedly. In transcription PCR (qRT-PCR).
addition, the preparation of time-lapse dishes is time-consuming and uses Participants/materials, setting, methods: In the treatment group,
reduced volumes of media and oil, which may result in some evaporation. IVF drops were transfected with mmu-let-7a-5p antagomir (ABM, Inc., Richmond,
Study design, size, duration: Medium osmolality was measured daily BC, Canada) using ViaFect™ Transfection Reagent (Promega, Madison, WI,
between 0-168h in Petri dishes with 20µl drops overlaid with oil, comparing USA), while the sham group receive only transfection reagent, and control
incubation in a humid vs. dry atmosphere and media with 0 vs. 20mg/ml HSA. group receive no treatment. Expression levels of mRNA targets of let-7a-5p
Osmolality was also measured in six different types of time-lapse dishes after (Igf1, Il1, Itgb3 and Tgfb1) were analyzed by qRT-PCR. The blastocysts were
0h (set-up), 72h (Day 3), 120h (Day 5) and 168h of culture (Day 7). All readings transfer to the pseudopregnant mice to evaluate the effect of let-7a-5p supres-
were performed with a last-generation vapour pressure osmometer (VAPRO sion on embryo implantation.
5600, Wescor). Main results and the role of chance: : Our results showed that there were
Participants/materials, setting, methods: For the humid vs. dry and pro- not significant changes in the developed embryo to the blastocyst stage between
tein vs. no-protein experiments, 35mm Petri dishes were prepared following an the study groups (P> 0.05). Expressions level of let-7a-5p in the treatment group
underlay method by placing the medium droplets below the oil to avoid any were significantly downregulated compared with the controls (FC=0.352,
evaporation during setup. Each type of time-lapse dish was prepared according P=0.039). In contrast, no significant changes were observed between the sham
to its manufacturer’s instructions. All six types of time-lapse dishes were cultured and control groups (P> 0.05). Expression level of the four mRNA targets was
in a joint dry benchtop incubator. For two of those models, culture and mea- not significantly different between the sham and control group (P> 0.05).
surements were replicated in their time-lapse incubator. Expression level of Igf1 (FC=1.8, 0.021), Itgb3 (FC=1.3. P=0.039) and Tgfb1
Main results and the role of chance: Results are shown as the difference (FC=1.4, P=0.032) were significantly upregulated in the anatgomir treated group
between the initial osmolality (once the dish was set-up) and after 120/168h of compared to the control group. The implantation rates were significantly higher
culture. When assessing the effect of protein supplementation, osmolality in the treatment group compared to the control (P< 0.05).
increased by 28/41 and 30/45 mOsm/kg in the 0 and 20 mg/ml HSA groups, Limitations, reasons for caution: Since each microRNA can target many
respectively. In a humid atmosphere, a rise of 15/19 mOsm/kg was observed, of transcripts, we should explore the other potential mRNAs of let-7a-5p in the
compared with 34/42 mOsm/kg in a dry incubator. process of implantation.
The set-up of all the studied time-lapse dishes lead to medium evaporation Wider implications of the findings: A higher implantation rate following
by 5 mOsm/kg in relation to the medium vial. An additional increase of osmo- the suppression of murine let-7a-5p embryo suggested that microRNAs have
lality was detected during culture, notably varying depending on the dish design. important roles in the process of implantation.
For the six studied dish models, the observed increases in a dry benchtop Trial registration number: not applicable

P-168 A prospective study of the effect of laser-assisted P-169 Early recombinant-LH supplementation improves oocyte
hatching on the clinical outcome of single frozen/thawed embryos competence acting on oxidative stress in young poor responder
transfer cycles IVF-patients.
V. Torres Monserrat1, M.C. Delpiccolo1, M.N. Lardizabal1, M. Santonastaso1, F. Caprio2, A. Natella2, F. Mottola3, L. Rocco3,
N. Carizza2, T. Leiva2, E. Navall2, S. Dominici2, R. Sol Ariadna2, R. Ianniello2, D. Mele2, N. Colacurci1
C. Carizza3 1
Università degli Studi della Campania L. Vanvitelli, Dpt. of Woman-Child and
1
Fertya-Grupo Oroño, Embryology Laboratory, Rosario, Argentina ; General and Special Surgery, Naples, Italy ;
2 2
Fertya-Grupo Oroño, Gynecology, Rosario, Argentina ; Azienda Ospedaliera Universitaria L. Vanvitelli, Gynecological-obstetric clinic,
3
Fertya-Grupo Oroño, Medical Director, Rosario, Argentina Naples, Italy ;
3
Università degli Studi della Campania L. Vanvitelli, Dpt. of Environmental-
Study question: Does laser-assisted hatching (AH) improve clinical pregnancy Biological and Pharmaceutical Sciences and Technologies, Caserta, Italy
outcome of frozen/thawed embryo transfer cycles taking into consideration the
day of embryo development and patients´ prognosis? Study question: Does early recombinant (rec)-LH supplementation in ovarian
Summary answer: Laser-assisted hatching does not improve clinical pregnancy stimulation protocol affect oxidative stress in cumulus cells as well as IVF outcome
rate in frozen/thawed embryo transfer. However, good prognosis patients that in poor responder patients?
transfer single day-5 embryos are affected with AH. Summary answer: Early rec-LH supplementation enhances ovarian response
What is known already: Although assisted hatching (AH) is routinely per- to stimulation and embryological parameters reducing oxidative stress markers
formed in frozen/thawed embryo transfer cycles, there is not enough data to in young women with poor ovarian reserve in ICSI cycles.
conclude the benefit of this technique. Despite its widespread use, there is a What is known already: Management of women with poor ovarian response
limited number of prospective studies that have examined the effect of AH on (POR) to stimulation is a major challenge in reproductive medicine. The POR
clinical pregnancy and multiple pregnancy. Most of the investigations with good primary causes remain unknown and oxidative stress was recommended as one
scientific evidence conclude that AH can benefit clinical pregnancy rate in patients of causes. Multiple interventions have been proposed to improve reproductive
with poor prognosis, but it could also increase multiple pregnancy. However, outcomes in POR but showed conflicting results. Ovarian stimulation may have
there are not studies that evaluate the effect of AH and the day of embryo a direct impact on oxidative stress markers with Reactive Oxygen Species (ROS)
development in frozen/thawed embryo transfer production and perturbation of oxidant-antioxidant balance. IVF outcome is
Study design, size, duration: A prospective analysis of the effect of laser-as- adversely affected if imbalance exists between ROS and antioxidants in oocyte
sisted hatching on clinical and multiple pregnancy rates in 331 patients that microenvironment. Early rec-LH supplementation impact in POR ovarian stim-
underwent frozen/thawed single embryo transfer in our clinic from April to ulation protocol to improve IVF outcome are still unclear.
December 2019. Before treatment, patients were explained about actual evi- Study design, size, duration: This prospective randomized controlled study
dence of the technique for them to decide to do it or not. We compare Control included 132 POR patients stratified according to POSEIDON classification
Group, 120 transfer without AH, and Treatment Group, 211 patients with AH. group 3(age < 35, poor ovarian reserve parameters)from March 2015 to April
Embryo development day and patient’s prognosis were analysed 2018. All patients enrolled underwent controlled ovarian stimulation according
Participants/materials, setting, methods: Embryos were incubated in to GnRH antagonist protocol. Patients were randomized to rec-LH supple-
K-System® G-185 incubator with Sage® Medium. Kitazato® Medium Kits were mentation(A group) or no rec-LH supplementation(B group) from second day
used to vitrify and thaw. Single embryo transfer was performed the same day of the cycle. Oxidative stress markers in cumulus cells,oocyte number and
embryo was thawed. AH was performed immediately after thawing with Lykos® quality,fertilization rate, embryo quality, pregnancy rates are the outcome
laser set in the multipulse mode (pulse duration 200us). The chi square inde- measures.
pendence test and Fisher’s exact test were used to compare groups about Participants/materials, setting, methods: Exclusion criteria: PCOs, endo-
clinical pregnancy and multiple pregnancies rates, significance represented by metriosis,metabolic and endocrinology diseases, severe oligoasthenoteratozo-
a p<0.05 ospermia. On the day of oocyte retrieval, cumulus cell samples were mechanically
Main results and the role of chance: Laser-assisted hatching does not removed from oocyte. The intracellular ROS were assessed by H2DCF-DA
improve clinical pregnancy rate (Control Group: 31% vs Treatment Group: 27%, (2’,7’-dichlorofluorescein diacetate) fluorescent probe. DNA fragmentation
P-value=0.404) nor affect multiple pregnancy rate (Control Group: 5.4% vs index (DFI) and apoptotic gene (BAX and BCL-2) expression was assessed by
Treatment Group: 1.8%, P-value=0.561) when considering all patients under- TUNEL Test and qPCR-Real Time respectively. The data were analyzed using
going frozen/thawed single embryo transfer. When analysing only patients with the unpaired Student’s t-test by IBM SPSS-STATISTICS versions 20.0 and con-
bad prognosis, Control Group: 9% vs Treatment Group: 20%, P-value=0.083, sidered significant if P-value ≤ 0.05.
this technique does not improve clinical results either. The same result was Main results and the role of chance: There was no significant
obtained when studying patients with good prognosis, Control Group: 45% vs difference in the baseline clinical characteristics and the number of oocyte
Treatment Group: 33%, P-value=0.088. In addition, we analysed the same groups retrieval in both groups. The mature oocytes number and number of fertilized
and divided them considering the day of embryo development when vitrified. oocytes were higher in patients treated with rec-FSH + rec-LH (A group). The
When considering all patients undergoing frozen/thawed day-5 embryo transfer, average number of high-quality embryos on day 3 in patients treated with rec-
clinical pregnancy was not benefited by AH (Control Group: 39% vs Treatment FSH + rec-LH and rec-FSH was 1.70±0.75 and 0.66±0.47 respectively (p≤ 0.05).
Group: 28%, P-value=0.125). However, when this sub-group was divided accord- After the two ovarian stimulation protocols the differences of number of trans-
ing to prognosis, good prognosis patients were affected by laser-assisted hatching ferred embryos was statistically significant. The pregnancy rate was statistically
(Control Group: 53% vs Treatment Group: 32%, P-value=0.021), whereas, bad higher in patients treated with rec-FSH + rec-LHcompared to patients treated
prognosis patients were not affected (Control Group: 13% vs Treatment Group: with rec-FSH (59.37% versus 34.30%, p≤ 0.05). Intracellular ROS percentage
24%, P-value=0.257). Surprisingly, these results were not seen in the sub-group was significantly lower in POR patient treated with rec-FSH plus rec-LH com-
of day-6 embryo transfer where clinical pregnancy is not affected by performing pared POR patients no rec-LH supplementation (23±5.80 vs. 43±7.50, p≤ 0.05).
AH in the general population (P-value=0.617), nor in good prognosis Cumulus cell DNA fragmentation was significantly lower in in patients treated
(P-value=0.759) or bad prognosis (P-value=0.255) patients with rec-FSH + rec-LH. Cumulus cells surrounding oocytes retrieved from
Limitations, reasons for caution: The main limitation of this study is the patient treated with rec-FSH + rec-LH expressed significantly higher BCL-2
lack of randomness between control group and treatment group mRNA levels than those from patient treated with rec-FSH (p≤0.05) whereas
Wider implications of the findings: This prospective study shows that BAX mRNA levels resulted to decrease significantly (p≤0.05) in the patients
assisted hatching does not benefit clinical pregnancy rates of frozen/thawed treated with rec-FSH + rec-LH compared to patients treated with r-FSH.
single embryo transfer cycles. This is the first prospective study that analyse the Limitations, reasons for caution: Lifestyle factors (i.e. smoking, diet) are
effect of AH on frozen/thawed single embryo transfer in relation to embryo not considered. The cumulus cell samples are pooled.
development day and patients prognosis. Wider implications of the findings: This study showed that rec-LH sup-
Trial registration number: - plementation reduces oxidative stress markers in term of intracellular ROS

production, DNA fragmentation and apoptosis as well as influences oocyte protocols, may result in an improvement in pregnancy outcomes in women
competence in POR patients. Early rec-LH supplementation in controlled ovarian undergoing ART.
stimulation protocol act for strategy to improve IVF reproductive outcomes in Trial registration number: NA
young POR patients.
Trial registration number: N° 48 (30/01/2015)
P-171 Is the frequency of aneuploidy and mosaicism at blastocyst
stage associated to the embryo sex?
P-170 Non-invasive detection of metabolically impaired euploid
blastocysts with low implantation potential P. Villanueva1, J. Huayhua2, L. Noriega-Hoces3, F. Vizcarra1,
J. Noriega-Portella3, L. Guzman Masias1
S. Cabello-Pinedo1, H. Abdulla2, M.L. Seth-Smith1, M. Escriba3,
1
J. Crespo3, J.A. Horcajadas1, S. Munne1 Laboratorios PRANOR, Embriologia, Lima, Peru ;
2
1 ADN Diagnostico, Genetics, Lima, Peru ;
Overture Life SL, Scientific Department, Madrid, Spain ; 3
2 Clinica Concebir, Departamento Medico, Lima, Peru
Texas A&M University Corpus Christi, Chemistry, Corpus Christi, U.S.A. ;
3
Juana Crespo Clinic, Embryology, Valencia, Spain
Study question: Is the chromosome constitution associated to the embryo
Study question: As not all euploid embryos implant, is it possible to sex?
identify non-implanting euploid blastocysts from their spent media metabolomics Summary answer: Male blastocysts are less likely to be euploid considering
profile? confounding variables. Additionally, they have a higher chromosomal unbalances
Summary answer: Analysis of the spent media metabolomics profile could (aneuploidy+mosaicism) in the same embryo.
predict about 30% implantation failure of euploid embryos, coinciding with the What is known already: There is very limited information about embryo
upper limit of PGT-A success. euploidy and mosaicism in relation to embryo sex. In our study, chromo-
What is known already: Embryo implantation potential and euploidy rates some constitution at blastocyst stage was evaluated by next-generation
decrease with advancing maternal age. Preimplantation Genetic Testing (PGT) sequencing (NGS) and confounding factors were controlled. Previous studies
is been used in routine clinical practice to avoid the transfer of aneuploid embryos with lower data found that embryo sex was not significantly associated with
with minimal chance of resulting in a viable pregnancy. However, even the best ploidy status.
PGT-A published results do not exceed 70% ongoing pregnancy rates. Little is Study design, size, duration: Data was collected retrospectively from
understood of the causes of failure of euploid embryos to implant, and methods June 2016 to October 2019. In total, 1212 PGT-A cycles were included.
to discriminate between viable euploid embryos and non-viable euploid embryos, Participants/materials, setting, methods: In total, 4434 blastocysts
that will not result in a successful pregnancy, have not been developed yet. analysed with PGT-A were generated from 1212 cycles. The chromosome
Study design, size, duration: This study includes spent media samples col- constitution was classified: euploid, single-aneuploid, double-aneuploid,
lected before trophectoderm biopsy from embryos that were later analyzed by complex-aneuploid, single mosaic, double mosaic, complex mosaic, mix
PGT and transferred. To develop the algorithm, this study used a dataset that (aneuploid and mosaic) and complex mix.
included 37 samples collected using Vitrolife media that were classified as preg- A multilevel model was made and associations between variables by logistic
nancy (P) or non-pregnancy (NP) according to embryo implantation outcome regression adjusted according to maternal age, blastocyst grade, biopsy day,
after transfer. The algorithm was then tested on spent media of 42 euploid inner cell mass and trophectoderm grade, fertilization method, and
blastocysts. embryologist.
Participants/materials, setting, methods: For the training subset (n=37), Main results and the role of chance: The mean maternal age was 36.2
patients undergoing infertility treatments were included. For the validation stage ± 4.2. The genetic results of 4434 blastocyst were as follow: euploid 46.4%
(n=42), patients whose embryos were analyzed through PGT were recruited. (N= 2058); single aneuploid 14.4% (N=640); double aneuploid 4.7%
In both cases, spent media samples (20-40ml) were collected after incubation (N= 208), complex aneuploid 2.1% (N=94); single mosaic 13.7% (N=607),
between days 3 and 5 and just before biopsy. Before the analysis, samples were double mosaic 4.7% (N=210); complex mosaic 6.6% (N=291); mix 2.8%
diluted with distilled water, ultrafiltered to remove molecules >3KDa and run (N=123) and complex mix 4.6%(N=203). When results were classified
through a UPLC-Fusion Orbitrap MS/MS system to determine metabolite according to the embryo sex, 49.1% (N=1010) were euploid female blastocyst
concentration. and 50.9% (N=1048) were male euploid blastocyst. For single aneuploidy
Main results and the role of chance: Samples from the training subset were was 47.7% (N=305) and 52.3% (N=335); double aneuploid was 46.2%
processed, detecting more than 5,550 metabolites in spent media samples. (N=96) and 53.8% (N=112); complex aneuploidy was 46.8% (N=44) and
Several statistical techniques were applied to reduce this large number of metab- 53.2% (N=50); single mosaic was 44.3% (N=269) and 55.7% (N=338); double
olites to the most informative ones for pregnancy outcome. This led to the mosaic was 52.9% (N=111) and 47.1% (N=99); complex mosaic was 46%
definition of a Vitrolife specific MPI (Metabolite Pregnancy Index), which has (N=134) and 54% (N=157) and complex mix 47.8% (N=97) and 52.2%
proved to be highly predictive of both P and NP embryos. This technique was (N=106) in female and male blastocyst respectively (p≥0.05). Male embryos
tested in a completely different data subset showing an ability to identify 78% of are more likely to have a mix chromosome constitution (aneuploid and
P samples and 61% of NP samples. mosaic) than female embryos (OR=1.63, 95% CI (1.12-2.36), p=0.010).
For the validation part of the study, metabolic profile of culture media coming Furthermore, they are less likely to be euploid (OR=0.88, 95% CI (0.78-0.99),
from euploid embryos (n=42) was studied in order to assess whether the implan- p=0.037) adjusted for all variables.
tation failure of some euploid embryos may be due to metabolic impairment. Limitations, reasons for caution: The results observed in this study should
A 30.95% of euploid embryos in the validation subset showed a metabolomic be confirmed using a larger number of samples.
profile predicting poor implantation potential despite being euploid. Wider implications of the findings: Our data suggest that male blastocyst
Although further research is necessary, these results open the possibility of are associated with a lower euploidy and a higher mixture of chromosomal
applying metabolomics to differentiate, within euploid embryos, those that are unbalanced.
viable from the ones that will not result in a successful pregnancy. Trial registration number: none
Limitations, reasons for caution: The study was retrospective but a pro-
spective clinical trial is underway. In addition, all the samples were collected using
P-172 The transcriptome of paired human oocytes related to
the same culture media and the metabolomic algorithm is media specific. The
progesterone primed ovarian stimulation
study needs to be validated in multiple culture media and test its outcome com-
paring it with more clinical data. F. Martinez1, S. Mateo1, E. Clua1, N. Coca2, J. Rodriguez-
Wider implications of the findings: This study represents an initial step Lumbiarres2, L. Armengol2, M. Roca1, S. Garcia1, N. Polyzos1
1
in the development of non-invasive methods based on metabolomics approach, Hospital Universitari Dexeus, Obstetrics- Gynecology and Reproduction,
that allow identification of viable embryos, not only based on ploidy status. Barcelona, Spain ;
2
The optimization of embryo selection techniques, without altering embryology qGenomics, qGenomics, Barcelona, Spain

Study question: Is there any difference in the human oocyte transcriptome Summary answer: In good-prognosis patients and those with recurrent
under two specific Controlled Ovarian Stimulation (COS)/LH suppression implantation failure, implantation rates (IR) of GQB were similar whether
protocols: progesterone-primed cycles (PP) and GnRH-antagonist cycles (ANT). laser-AH was performed or not before transfer.
Summary answer: Different modes of controlling the endogenous LH secre- What is known already: In assisted reproductive techniques, embryo devel-
tion do not translate in a significantly different oocyte transcriptome. opment conditions are deeply modified. Indeed, the increasing use of extended
What is known already: Although the use of and oral progesterone (PP) instead culture and vitrification may induce a zona hardening, possibly resulting in a
of (ANT) has been effective in controlling LH surge during ovarian stimulation, defective hatching process and implantation failure. Assisted Hatching, defined
evidence from a randomized trial reported lower live birth rates from PP-derived as the artificial breakage of the zona pellucida (ZP), appears to be a tool to
oocytes, suggesting a detrimental effect on oocyte competence. A novel potential overcome this issue and improve IVF/ICSI outcomes. To date, several studies
method to assess an effect of stimulation on the oocyte level is single-cell tran- have focused on the outcomes of AH performed at the cleavage-stage, con-
scriptome, though powerful genomic techniques allow this even in a single-cell. In cluding to an increase in clinical pregnancy rates, especially in poor-prognosis
the current study we aimed to evaluate the effect of PP stimulation on oocyte patients. However, investigations on the interest of AH at the blastocyst stage
developmental competence, through transcriptome analysis of In-vivo-matured are still controversial.
oocytes (transcriptome remarkably stable across ages and ovarian reserve). Study design, size, duration: This prospective randomized study has been
Study design, size, duration: Retrospective study comparing single-cell in progress since September 2017. Inclusion criteria were: female age ≤37 years,
transcriptome of 24 mature paired oocytes from four oocyte donors (OD) who fresh/vitrified day-5 GQB (defined as: blastocoel expansion B3/B4, inner cell
underwent two consecutive COS/IVF cycles at private, university-based IVF mass and/or trophectoderm graded A/B, according to the grading system
center between January 2017 and March 2018 were included. described by Gardner and Schoolcraft), absence of risk factors for implantation
OD underwent two COS/IVF cycles, under two LH suppression protocols failure. So far, 260 fresh/vitrified GQB transfers have been randomly allocated
(PP-Group: 75mg daily oral Desogestrel; ANT-Group: 0.25mg daily s.c. to AH group (n=122): laser-AH performed prior to transfer; or no-AH group
Ganirelix) until GnRH agonist trigger. (n=138): no hatching.
Participants/materials, setting, methods: Four healthy oocyte donors Participants/materials, setting, methods: In AH group, a single hole com-
underwent two consecutive COS/IVF cycles, under two LH suppression pro- pletely through the ZP was performed using a 200-µsecond laser pulse (Lykos,
tocols (PP-Group: 75mg/day Desogestrel; ANT-Group: 0.25mg/day Ganirelix) Hamilton Thorne).
until trigger. Mature MII oocytes were vitrified according to standard procedure. In the preliminary analysis, the included population was stratified by number
Three oocytes from each cycle (PP/antagonist) from the same donor were of good-quality (GQ) embryos previously transferred, leading to “good prog-
thawed, total-RNA was extracted, cDNA was prepared and labeled prior to nosis” (GP) and “recurrent implantation failure” (RIF) subgroups (respectively
hybridization on PrimeView Human Gene Expression arrays(Affymetrix). Gene defined as ≤4 and >4 GQ cleavage-stage embryos, or ≤2 and >2 GQB trans-
expression levels were normalized (RMA) and compared between treatments ferred without proven implantation). Clinical outcomes were compared between
after data normalization (LIMMA). AH and no-AH groups.
Main results and the role of chance: Overall, we analyzed the transcrip- Main results and the role of chance: In the “GP” analysis, AH (n=108) and
tome from 24 oocytes derived from 4 oocytes donors who underwent no-AH (n=117) groups were comparable considering female age, body mass index,
2 consecutive ovarian stimulation cycles, 1 with GnRH antagonist and 1 with PP type and cause of infertility, and attempt rank. On average, 1,13±0,34 and
(12 oocytes from ANT and 12 oocytes from PP). 1,08±0,28 GQB were transferred in AH and no-AH groups, respectively (p=0,29).
Comparisons in the gene expression profile from total single-cell RNA Quality of the transferred blastocysts was similar as well: 57,4% of toP-blastocysts
extracted from oocytes derived from ANT or PP ovarian stimulation did not [B4AA/AB/BA] in AH group vs. 59,1% in no-AH group, respectively (p=0,60).
demonstrate any significant difference. Following, adjusting P-values for multiple IR and clinical pregnancy rates (CPR) were not significantly different whether
testing, no statistically significant differences in gene expression levels were laser-AH was performed or not prior to transfer (IR: 39,4% in AH group vs. 43,6%
observed between oocytes obtained from the two evaluated Controlled Ovarian in no-AH group; p=0,51, and CPR: 40,7% vs. 45,3%, respectively; p=0,17).
Stimulation (COS)/LH suppression protocols. Despite that single-cell transcrip- Considering the “RIF” subgroups, patients’ characteristics and the mean num-
tome profile did not differ when comparing different LH suppression protocols ber of GQB transferred (1,57±0,51 vs. 1,46±0,51, respectively; p=0,51) were
( ANT vs. PP), of interest, other experimental conditions have been have sig- similar between AH (n=13) and no-AH (n=23) groups. There was no difference
nificantly influenced oocyte transcriptome (P<0.05). in the proportion of toP-blastocysts transferred (66,7% vs. 69,7%, p=0,82).
Limitations, reasons for caution: The study was restricted to healthy Interestingly, GQB transferred after laser-AH yielded similar IR and CPR com-
OD and thereby other conditions related to infertility or older age were pared to GQB transferred without laser-AH (IR: 42,9% in the two groups; CPR:
not considered. This is a descriptive study with a limited number of samples 50,0% vs. 54,5%, p=0,79).
reflecting the difficulty to recruit human oocytes, especially from women Limitations, reasons for caution: These preliminary results need to be
with infertility. confirmed on a larger series, especially in the “RIF” subgroup. Moreover, a poten-
Wider implications of the findings: To our knowledge this is the first study tial impact of AH on twin pregnancy rates should be examined.
utilizing singe-cell transcriptome analysis in order to evaluate the effect PP sup- Wider implications of the findings: If confirmed, these findings would
pression of LH on oocyte developmental competence. Our results add reassur- suggest that assisted hatching might not be beneficial to young patients, with or
ing information to the clinical available data regarding the safety of the PP ovarian without history of implantation failure, as long as good-quality blastocysts are
stimulation protocols transferred. Finally, the hypothesis of zona hardening due to exposure to culture
Trial registration number: Not Applicable and/or vitrification media might be questioned.
Trial registration number: 2018-A02395-50
P-173 Interest of laser assisted hatching prior to good-quality
blastocyst transfer: intermediate results of a prospective P-174 The effect of ovine whole ovary cryopreservation on the
randomized study antral follicle oocyte and its developmental potential
C. Herbemont1, Y. Boumerdassi1, B. Bennani Smires1, S. Sarandi1, A. Dadson1, D. Wang1, J. Hernandez-Medrano1, W. Maalouf1
O. Selmi1, C. Vinolas2, C. Lethielleux2, F. Krief2, I. Cedrin- 1
University of Nottingham, Department of Obstetrics and Gynaecology, Nottingham,
Durnerin2, M. Peigné2, M. Grynberg2, C. Sifer1 United Kingdom
1
Hôpital Jean Verdier, Histologie-Embryologie-Cytogénétique-Biologie de la
Reproduction-CECOS, Bondy, France ; Study question: Are oocytes obtained from antral follicles of sheep viable
2
Hôpital Jean Verdier, Médecine de la Reproduction, Bondy, France with the capacity to mature following whole ovarian cryopreservation (WOCP)
using the slow freezing method?
Study question: The aim was to assess the impact of laser assisted hatching Summary answer: Oocytes isolated from whole ovary cryopreserved and
(AH) on the implantation potential of good-quality blastocysts (GQB) trans- thawed antral follicles fail to mature in vitro and record high level of induced
ferred in a selected population. apoptosis.

What is known already: Whole ovary cryopreservation using slow freezing 6.8% (118/1737), respectively, while the ANA (+)/ACA (+) group had rates
and transplantation restores ovarian function and natural fertility resulting in of 60.2% (80/133) and 26.3% (35/133), respectively. The ANA (+)/ACA (+)
multiple live births in sheep. This is as a result of the preservation and survival group showed a significantly higher multiple nucleation rate (p<0.05). Although
of primordial and preantral follicles. there were no significant differences in cleavage rates among the groups (ANA
Study design, size, duration: A total of 540 antral follicle oocytes (control, (+)/ACA (-): 75.4%, 1310/1737 and ANA (+)/ACA (+): 58.6%, 78/133), the
CT, n=304 and WOCP, n=236) were aspirated from abattoir-sourced sheep occurrence of fragmentation in cleaved embryos was significantly higher in the
ovaries. After collection, oocytes were classified according to cumulus oocyte ANA (+)/ACA (+) group (ANA (+)/ACA (-):42.2%, 553/1310, and ANA (+)/
complex (COC) morphology into normal (A&B) or abnormal (C&D). Oocytes ACA (+):57.7%, 45/78, p<0.05). The good-quality blastocyst formation rates
were matured in vitro and nuclear maturation assessed after 24 hours. The via- were significantly lower in the ANA (+)/ACA (+) group than in the ANA (+)/
bility and apoptosis of antral follicle oocytes was also assessed immediately ACA (-) group (ANA (+)/ACA (−): 32.7%, 429/1310); ANA (+)/ACA (+):
following cryopreservation and subsequent thawing. 14.1%, 11/78), p<0.05). A total of 268 and 11 cycles of single vitrified-warmed
Participants/materials, setting, methods: Ovaries were randomly allocated blastocyst transfer were performed in the ANA (+)/ACA (-) and ANA (+)/
into fresh (control, CT) and WOCP. WOCP and thawing was as described by ACA (+) groups, respectively. No significant differences were observed between
Onions et al 2008. COCs were then retrieved and grouped into the various grades the groups in clinical pregnancy and ongoing pregnancy (ANA (+)/ACA
based on the cytoplasm and presence or absence of cumulus cells. The oocytes (−):53.4%, 143/268 and 44.0%, 118/268; ANA (+)/ACA (+):72.7%, 8/11 and
were then assessed for viability and apoptosis using propodium iodide (PI) and 54.5%, 6/11).
terminal deoxynucleotidyl fluorescein-dUTP nick end labelling (TUNEL) respec- Limitations, reasons for caution: The main limitation of the present study
tively. Nuclear maturation was assessed by DAPI following in vitro maturation (IVM). was its retrospective design and small sample size. The rate of ANA (+)/ACA
Main results and the role of chance: Percentage of COCs classified as normal (+)was small (0.8%, 10/1,227) in all IVF patients during the study period.
was similar (P>0.05) between WOCP (54.5±12.7%) and CT (54.3±8.0%). Therefore, these were the only cases of ANA (+)/ACA (+) available.
Moreover, PI positive oocyte percentages were comparable (P>0.05) between Wider implications of the findings: Oocytes in ANA (+)/ACA (+) patients
WOCP and CT: 32.6% (n=23/70) and 19.8% (n=15/76), respectively. However, showed a significantly higher incidence of multiple nucleation, which was asso-
nuclear maturation was lower (p<0.05) in WOCP (8%, n=1/64) compared to ciated with a significantly lower good-quality blastocyst formation rate. However,
CT oocytes (67%, n=53/65), whereas TUNEL apoptotic index was higher in acceptable pregnancy outcomes in those patients would be achievable if
WOCP oocytes than controls (75.4%, n=52/69 vs. 26.6%, n=17/76). good-quality blastocysts are obtained and transferred.
Limitations, reasons for caution: Whole ovary cryopreservation caused a Trial registration number: None.
significant loss of cumulus cells in the oocytes obtained from antral follicles.
These results depict clearly shows the inability of the oocytes to develop further, P-176 Effects of maternal ageing on trophectoderm transcriptome
however does not conclude on the quality of the oocyte. dynamics and the potential impact on embryo-endometrial
Wider implications of the findings: Isolating oocytes from antral follicles interactions.
of freeze-thawed sheep ovary for in vitro maturation and fertilisation may not P. Ntostis1, G. Kokkali2, D. Iles1, J. Huntriss1, A. Pantou2,
be a feasible path for clinical application. K. Pantos2, H. Picton1, D. Miller1
LICAMM Institute, Discovery and Translational Science, Leeds, United Kingdom ;
2
Genesis Athens Clinic, Reproductive Medicine Unit, Athens, Greece
P-175 The impact of anti-centromere antibodies (ACAs) on Study question: What differences are there in the human trophectoderm
embryonic and pregnancy outcomes following IVF and single transcriptome among young, intermediately aged and older women with respect
vitrified-warmed blastocyst transfer to the receptive endometrium?
T. Okubo1, K. Omi1, T. Hayashi1, N. Onda1, T. Segawa1 Summary answer: The most significant gene ontologies from young/inter-
1
Shimbashi Yume Clinic, Advanced medical research institute of fertility, Tokyo, Japan mediate to advanced maternal age trophectoderm involved extracellular exo-
somes and, when receptive endometria were considered, the regulation of
Study question: Do anti-centromere antibodies (ACAs) negatively influence apoptosis.
embryonic and pregnancy outcomes following IVF and single embryo transfer? What is known already: Chromosomal nondisjunction is the most frequent
Summary answer: ACA negatively influences embryonic outcomes following cause of recurrent miscarriages, with women over 35 years of age also experi-
IVF, while pregnancy is not affected once expanded blastocysts are produced encing lower pregnancy rates. Euploid embryo transfer reported decreased live
and transferred. birth rates in women over 35 years, with women over the age of 40 experiencing
What is known already: ACA is a member of the anti-nuclear antibody significantly lower implantation and pregnancy rates. Several factors involved in
(ANA) family. It has been considered that ACA may be the major antibody of embryo-endometrial communication may contribute to lower success rates,
ANA group, which has adverse effects on clinical outcomes following IVF-ET. with blastocyst-secreted exosomes potentially affecting endometrial receptivity
Since the population of ANA positive (+) and ACA positive (+) infertile patients and gene expression, regulating embryo implantation. To date, no study has
is low, studies regarding the significance of ACA, among the types of ANA, in specifically explored the trophectoderm transcriptome with respect to the
the possible interference with embryonic and pregnancy outcomes are limited. maternal age and endometrial receptivity.
Study design, size, duration: A retrospective cohort study of 601 cycles Study design, size, duration: Fifteen women treated for infertility in a single
(223 patients, mean age: 39.9±4.1 years), including 560 cycles (213 patients, IVF unit agreed to participate in this study. Trophectoderm biopsies (Day 5
mean age: 40.0±4.0 years) involving ANA (+)/ACA (-) and 41 cycles (10 blastocysts) were obtained from 4 women below 30 years (young cohort), 8
patients, mean age: 39.0±4.3 years) involving ANA (+)/ACA (+), was conducted women aged 30-39 years (intermediate cohort) and 3 women over 40 years old
between May 2017 and March 2019 in a single centre. Embryonic and pregnancy (AMA cohort). Samples were collected over a 6-month period and RNA was
outcomes following single ET were compared among the groups. isolated. Our study design also included the analysis of transcriptome data from
Participants/materials, setting, methods: A total of 1,419 oocytes (ANA receptive endometria.
(+)/ACA (-): 1,224 and ANA (+)/ACA (+): 195) were retrieved. Oocyte Participants/materials, setting, methods: Library construction and deep
maturation status was defined by first polar body visualization or meiotic spindle RNA sequencing included 10 trophectoderm biopsies derived from young
confirmation. Oocytes were inseminated using IVF or ICSI. The primary out- women (24.4±2.0 years), 16 from intermediate aged women (34.3±2.6 years)
comes were normal fertilization, multiple nucleation (3PN<), cleavage, and and 6 from women of advanced maternal age (AMA) (42.0±1.1 year)
good-quality blastocyst formation rates (Gardner criteria: grade >3). Secondary (mean±SD). RNA expression profiles were obtained from all 32 trophectoderm
outcomes were clinical pregnancy (gestational sac observation) and ongoing biopsies (Day 5 blastocysts). Trophectoderm characterisation and ontological
pregnancy rates (foetal heartbeat observation). analysis were followed by the investigation of the complementarity between
Main results and the role of chance: The ANA (+)/ACA (-) group had young/AMA trophectoderm and receptive/non-receptive endometrial tran-
normal fertilization and multiple nucleation rates of 80.0% (1389/1737) and scriptomes. Chromosome aneuploidy analysis was also performed.

Main results and the role of chance: RNA sequencing revealed 233 sig- couple (22.6% per couple, 451/1890 total embryos). Clinical pregnancy following
nificantly differentially expressed genes (DGE) (580 transcripts), between young the day 4 biopsy was 44.5% (53/119, 1.1 embryos per ET) for con-current
and AMA trophectoderm transcriptomes, with FDR ranging between 5.59 x transfers and 36.2% (17/44, 1.1 embryos per ET) for follow-up FET cycles
Ε-08 and 0.049 in the young and 7.29 x Ε-06 to 0.049 in the AMA cohorts. resulting in overall clinical pregnancy of 42.9% (70/163) and 66 live births.
These results correspond to log fold changes of between 0.66 and 9.97 in the Neonatal outcomes included five preterm deliveries (5/63, 7.9%) with zero
young and between -0.98 and 9.24 in the AMA cohort, respectively. No signifi- babies born with small for gestational age weight and zero days in NICU for
cantly differentially expressed genes were revealed between young and inter- deliveries ≥37 weeks of gestation (0.03 ± 0.2 days). A high proportion of biop-
mediate cohorts. sied embryos per patient with no genetic abnormalities reached the blastocyst
Cellular component analysis of the differentially expressed genes between stage (79.2%) when assessed on day 5. However, a large proportion of utilised
young and AMA trophectoderm, revealed factors potentially participating in embryos per patient did not reach the expanded blastocyst stage on day 5
molecular trophectoderm-endometrial communication, including 42 genes (45.4%). A total of 11 live births resulted from day 4 biopsy method from
strongly associated with extracellular exosomes (FDR 8.2E-06). Considered embryos that would otherwise not have met the requirements for undergoing
alone, the AMA cohort did not reveal any significant ontologies, however a trend a day 5 blastocyst biopsy.
for higher representation of mitotic spindle and cytosol ontologies was noted. Limitations, reasons for caution: The results represent the experience
These results may suggest the role of extracellular exosomes in the communi- gained from current practice and not of a prospective controlled study. No
cation between trophectoderm and endometrium in human. When additional embryos were cultured to day 6, therefore the developmental potential of cer-
endometrial gene expression data were included in our analysis, the regulation tain embryos classified as not expanded blastocysts at day 5 of embryo devel-
of apoptosis was strongly represented in both embryonic and endometrial tran- opment is unknown.
scripts, suggesting that this could be an important process linked to successful Wider implications of the findings: Day 4 embryo biopsy can be a safe and
implantation. effective stage for obtaining genetic material for PGT. Therefore this method
Limitations, reasons for caution: The relatively high variation in trophec- can be used as an alternative approach to current embryo biopsy practices or
toderm RNA expression presents a reason for caution. However, the relatively used in combination with blastocyst biopsy when there is a small cohort of
low variation within the young and AMA cohorts and the fact that a minimum available embryos for biopsy
of 6 samples were analysed (from at least 3 different women per cohort), min- Trial registration number: Not applicable
imized the potential impact on our analysis.
Wider implications of the findings: Understanding the effects of maternal P-178 Does assisted hatching of vitrified blastocysts increase
ageing on the trophectoderm RNA, could identify processes that delineate nor- clinical pregnancy rates? A randomised controlled trial
mal embryo development. Considering also the receptive endometrial transcrip- C. Cordero Rosales1, S. Cortés1, C. Andrés1, R. Pandolfi1,
tome, further insights into the mechanisms involved in successful implantation C. Rodriguez1, M. Saladino1, J. Harper2
may be revealed. These findings have the potential to increase the low implan- 1
Clínica Tambre, FIV Labotary, Madrid, Spain ;
tation rates of AMA women. 2
Institute for womwn`s health, Universyty College, London, United Kingdom
Study question: Does assisted hatching (AH) of vitrified blastocysts increase
P-177 Clinical application, safety and efficacy of Day 4 biopsy for clinical pregnancy rates?
preimplantation genetic testing. A retrospective cohort analysis. Summary answer: Our study found that AH of warmed vitrified blastocysts
G. Liperis1, H. Swearman1, J. Crittenden1, C. Sjoblom1 had no effect on clinical pregnancy rates.
1
Institute of Reproductive Medicine- University of Sydney, Westmead Fertility What is known already: The use of AH has been controversial. Some studies
Centre, Sydney, Australia have shown a benefit of AH on clinical pregnancy rates for various groups of
patients, including those with cryopreserved embryos, but these effects are not
Study question: To evaluate the safety and efficacy of Day 4 embryo biopsy seen on live birth rates. Other studies have shown no difference at all. Few
Summary answer: The developmental potential of embryos and cycle out- studies have looked at the effects of AH on vitrified blastocysts.
comes suggest that day 4 biopsy is a safe and effective stage at which to perform Study design, size, duration: This randomised controlled trial was performed
embryo biopsy. between September 2018 and December 2019. 210 patients undergoing blastocyst
What is known already: Embryo biopsy routinely takes place on day 3 (cleav- warming were randomly allocated into a control or AH group. All patients had a
age stage) or day 5-6 (blastocyst stage) of embryonic development; with the single blastocyst transfer (sET). Patients undergoing PGT-A, day 3 frozen embryo
latter preferred due to the ability to biopsy more cells, providing increased transfers and double embryo transfers (dET) were excluded. T-test was performed
genetic material for analysis. A limitation of blastocyst biopsy is that several as statistical analysis, and p<0.05 were considered statistically significant.
embryos capable of generating a pregnancy may not be biopsied if they fail to Participants/materials, setting, methods: The embryos were frozen at
reach the blastocyst stage. In a recent study, transfer of euploid embryos fol- the blastocyst stage using the open system, Kitazato vitrification and thawing
lowing morula biopsy on day 6 has shown significantly lower implantation rate protocol. Only blastocysts of good quality were frozen (AA,AB,BA,BB) using
and birth rate compared to transfer of euploid blastocysts. the Gardner scoring system. Blastocysts were frozen for between 4 days and 4
Study design, size, duration: A retrospective analysis was undertaken of years. After warming, the AH was performed immediately with the Octax-Laser
patients having day 4 embryo biopsy at a single Centre between 2014 and 2017. at 2.6ms performing in between 2-3 pulses. Blastocysts were cultured for a
The reasons for PGT-A included implantation failure, history of recurrent mis- minimum of 2 hours before transfer. A routine embryo transfer was performed.
carriages and advanced maternal age, whilst PGT-M was employed for specific Main results and the role of chance: 132 patients were allocated into the
gene defects and PGT-SR for structural rearrangements. Average patient age AH group and 126 to the control group. The age of the woman’s egg whose
was 36.4 ± 4.8 years. A total of 152 cycles of PGT from 148 couples were blastocyst was frozen ranged from 18-45 and there was no difference between
biopsied on day 4 (1890 embryos). the AH and control group.
Participants/materials, setting, methods: Participants were consented For the AH group, 70 patients had a positive hCG (53.03%), and 55 had a
patients undergoing IVF/ICSI cycles combined with PGT. Following fertilisation, confirmation of clinical pregnancy by ultrasound scan by a positive heart beat
fresh or frozen-thawed embryos, cryo-preserved at the 2PN stage for banking (41,67%). 9 of the clinical pregnancies had a miscarriage (13%). To date, from the
purposes, were cultured to day 4. On day 4, all embryos suitable for biopsy were ongoing pregnancies there are 16 live births. For the control group, 60 patients had
decompacted in Ca2+/Mg2+ free medium and 2-6 cells were removed for analysis. a positive hCG (47,61%), and 57 had a confirmation of clinical pregnancy by ultra-
The genetic analysis was performed on the same day and euploid/non affected sound scan by a positive heart beat (45,23%). 15 of the clinical pregnancies had a
embryo(s) were either transferred on day 5 or vitrified. miscarriage (25%). To date, from the ongoing pregnancies there are 10 live births.
Main results and the role of chance: Couples had an average of 12.4 ± There were no statistical differences after comparing positive B-HCG from
4.1 embryos biopsied with an average of 2.7 ± 1.3 cells per embryo aspirated. patients undergoing AH and those that were not (p:0.45; 95% CI). There were
Following genetic analysis, 2.5 ± 1.9 euploid/normal embryos available per no significant differences in the outcomes between groups (p:0.16 95% CI).

Limitations, reasons for caution: Currently we do not have all the live birth Summary answer: In a series of 31017 oocyte retrievals, the 22.7% delivery
rate for this study. rate after fresh embryo transfers (ET) increased to 25.7% when including also
Wider implications of the findings: There has been much ongoing debate first frozen ETs
about the use of AH in clinical embryology. In our clinic, we no longer perform What is known already: Based on 12732 fresh IVF cycles, we presented in
AH routinely after blastocyst warming. Clinics need to carefully consider if they 2010 a “totally inclusive and mutually exclusive” clinically relevant classification
are charging for this procedure. system for reporting IVF results, imitating Robson’s 10-group model for classifying
Trial registration number: Not applicable cesarean sections. The most important factors for predicting delivery after IVF
were female age, number of aspirated oocytes, cumulative gonadotropin dose,
P-179 Cell-free mitochondrial DNA facilitates granulosa cells number of previously failed IVF cycles, and previous IVF delivery. The system
apoptosis and reduces developmental competence of oocytes has since then been used in Sweden for open and fair comparisons of delivery
from aged mice rates between IVF clinics, and for revealing temporal changes. However, after
Y. Liu1, Q. Shen1, W. Xiang1, L. Zhang1 introduction of the freeze-all strategy the system is no longer “totally inclusive
1 Study design, size, duration: The material was obtained from the Swedish
Tongji Medical College- Huazhong University of Science and Technology, Institute
national quality register for assisted reproduction (Q-IVF) during 30 months 2015-
of Reproductive Health and Center for Reproductive Medicine, Wuhan, China
2017. The register includes six public and 12 private clinics. The delivery rate per
Study question: Is the addition of cell-free mitochondrial DNA(cf-mtDNA) oocyte retrieval and ET, respectively,was calculated separately for fresh ETs and
to the culture medium affects mouse granulosa cells function and developmental when including first frozen ETs (FFET) within 6 months when freeze-all was done.
competence of oocytes matured in vitro? Participants/materials, setting, methods: 31017 oocyte retrievals resulted
Summary answer: Cf-mtDNA may bind to TLR9 and activate NF-κB /MAPK in 25987 ETs and 7967 deliveries. The revised model included 10 groups: Female
signal pathways, subsequently induces granulosa apoptosis and reduces devel- age (1) >42 years; (2) 40-41; (3) 36-39, previously ≥3 failed fresh IVF cycles; (4)
opmental competence of oocytes from aged mice. <36, ≥3 failed fresh IVF cycles; (5) 36-39, previous IVF child; (6) < 36, previous
What is known already: Quantitative analysis of cf-mtDNA in human follicular IVF child; (7) 38-39, no previous IVF child; (8) 36-37, no previous IVF child; (9)
fluid showed that the relative content of cf-mtDNA was positively correlated with 31-35, no previous IVF child; (10) ≤30, no previous IVF child.
patient age, and the relative content of cf-mtDNA may be closely related to oocyte Main results and the role of chance: By adding FFETs performed within 6
developmental competence. MtDNA, like bacterial DNA containing a large num- months after oocyte retrieval to fresh ETs, i.e. including FFETs when cycle can-
ber of unmethylated CpG sequences, can causes inflammation as a damage-asso- cellation was done due to freeze-all, the rate of delivery per oocyte retrieval
ciated molecular pattern molecule when it is released out of the cells. increased from in mean 22.7% (95% CI 22.3-23.2; range 7.5-35.8%) to 25.7%
Study design, size, duration: C57BL/6J mouse ovarian granulosa cells and (95% CI 25.2-26.2; range 8.4-39.1%). The mean increase was 2.4% with the
cumulus-oocytes complex were cultured accompanied with cf-mtDNA in vitro. lowest increase in group 2 (0.6%) and the largest in group10 (5.0%). The cor-
Mitochondrial function, expression levels of genes and proteins related apoptosis responding figures for delivery per ET were 27.1% (95% CI 26.6-27.7; range
and signal transduction pathway in granulosa cells, estrogen and progesterone 9.6-39.4%) and 30.7% (95% CI 30.1-31.2; range 10.8-43.1%). For both delivery
levels in the media were tested. 2-cell embryos and blastocysts were also counted per oocyte retrieval and delivery per ET, the lowest success rates were found
after maturation and fertilization in vitro of oocytes from young and aged mice. in group 1 (Fresh ET 7.5%, FFET after cancelled cycle 8.4% per oocyte retrieval;
Participants/materials, setting, methods: Mitochondrial function was 9.6% respectively 10.8% per ET) and the highest in group 6 (Fresh ET 35.8%,
assessed by measuring mitochondrial membrane potential, ATP levels, mtDNA FFET 39.1% per oocyte retrieval; fresh ET 39.4%, FFET 43.1% per ET).
copy numbers, and the apoptosis genes and apoptosis pathway proteins expres- Limitations, reasons for caution: The classification system should include
sion were evaluated by Western-blot assay, the mRNA expression of the apop- deliveries after freh ETs and FFETs, but not both in an individual, and exclude
tosis genes were also tested. Chemiluminescence immunoassay (CLIA) was used multiple ETs of frozen/thawed embryos. A Swedish consensus decided the 6
to detect the levels of estrogen and progesterone. month limit, since later FFETs may happen very occasionally.
Main results and the role of chance: Granulosa cells in the cf-mtDNA treatment Wider implications of the findings: Our 10-group classification system will
group had lower ATP content (P<0.05),higher apoptotic cells percentage (P<0.01), reflect variations in IVF results over time in a clinic, and between clinics on
and higher mRNA and protein levels of apoptosis-related factors than the control. national and international levels. The system can reveal inferior results in specific
Also, the expression levels of TLR9, NF-κB and MAPK proteins in granulosa cells groups, to be scrutinized and improved. Further sub-classifications relative to
were significantly increased in the cf-mtDNA treatment group (P<0.05), suggesting ovarian sensitivity indices are optional.
that cf-mtDNA induced apoptosis of granulosa cells was associated with the binding Trial registration number: Not applicable
of cf-mtDNA to TLR9 receptors and then activation of NF-κB and MAPK pathway.
Blastocyst formation rate of oocytes matured in vitro from aged mice decreased P-181 Embryo Biopsy on Morphologically Poor Quality
significantly (P<0.05), however, developmental competence of oocytes from young Blastocyst – will it help improve the Reproductive Outcomes?
mice had not been affected when cf-mtDNA was supplemented into in vitro matu- S.T. Mathew1, K. Mantravadi2, D.G. Rao3
ration medium, suggesting that increase of cf-mtDNA in media for aged mouse 1
Oasis Fertility, Embryology, Vishakapatanam, India ;
oocytes could be a factor to affect oocyte quality. 2
Oasis Fertility, Embryology, Hyderabad, India ;
Limitations, reasons for caution: Findings in vitro might not be suitable for 3
Oasis Fertility, Medical, Hyderabad, India
in vivo.
Wider implications of the findings: This study suggests that extracellular Study question: Embryo biopsy on morphologically poor quality blastocyst –
mtDNA could cause to damage to granulosa cell function and subsequently will it help improve the reproductive outcomes?
decrease of oocyte developmental competence. Summary answer: Transfer of PGT-A screened euploid, morphologically
Trial registration number: no poor quality blastocyst (MPB) can improve the overall pregnancy outcome com-
pared to non-screened MPBs.
P-180 An unbiased classification system including both fresh and What is known already: Traditionally embryo quality based on morphological
first freeze-all cycles for reporting and comparing IVF results parameters is considered as a major predictive factor for the pregnancy outcome
P. Saldeen1, K. Källén2 in ART. Particularly, a strong association has been reported between the blas-
1
Institution of Clinical Sciences Malmö- Lund University, Nordic IVF Malmö, tocyst morphology and pregnancy outcomes. However, morphological assess-
Malmö, Sweden ; ment is still not an ideal way to select and de-select embryos to optimize
2
Institution of Clinical Sciences Lund- Lund University, Center of Reproductive reproductive outcomes. Currently, the focus is to find a method, which enables
Epidemiology- Lund University, Lund, Sweden us to pick the best embryo for optimal results.
Study design, size, duration: All autologous frozen blastocyst embryo trans-
Study question: Is it more justified to include not only fresh, but also first fers (FET) with known pregnancy outcomes from January 2016 to March 2019
freeze-all cycles in a classification system for reporting IVF result were analyzed. Patients who had a day 5/6 transfer with one or two MPBs were

considered in this study. A total of 86 patients underwent FET with non PGT-A Live imaging by time-lapse monitoring began immediately after oocyte transfer
MPB and 18 patients with PGT-A MPB. Statistical analysis of the obtained data to the incubator, by the use of the Primo Vision. All data analysis was performed
was performed to find an association between the PGT-A of MPB and repro- using ImageJ/Fiji.
ductive outcomes. Main results and the role of chance:
Participants/materials, setting, methods: A total of 104 patients were
recruited for the retrospective study. Group I had 86 patients who underwent • Lower concentrations of Etoposide cause DNA damage but most oocytes
FET with non PGT-A MPB and group II had 18 patients with PGT-A MPB. progress through meiosis-I and subsequently extrude the first polar body
Among the clinical outcomes clinical pregnancy rate (CPR), miscarriage rate (PB1) to form a metaphase-II egg, revealing the absence of a DNA-damage-
(MR), implantation rate (IR) and live birth rate (LBR) were included and perinatal induced SAC response.
outcomes includes average birth weight (BW), preterm delivery and delivery • DNA damage accumulated in meiosis-I, such as could occur during in vitro
complications. maturation procedures, does not prevent polar body extrusion and therefore
Main results and the role of chance: The CPR, MR, IR and LBR between could persist in morphologically normal metaphase-II oocytes.
group I and group II were found to be 43.02% vs. 66.67%, 15.12% vs. 16.67%,
26.53% vs. 66.67% and 16.46% vs. 50% respectively. Among the perinatal out- Limitations, reasons for caution: no limitations or reasons for caution
comes there was no incidence of low birth weight (LBW) or any congenital Wider implications of the findings: Multiple aspects of the clinical IVF
anomalies. Average birth weights and preterm delivery rates were 2.8 vs. 2.5kgs environment could potentially damage the DNA of cultured oocytes.
and 1.27% vs. 0% respectively. Clinical outcomes seem to be better with PGT-A This may explain poor embryo quality and repetitive implantation failures in
group. Perinatal outcomes were comparable between both the groups. Results many of the cases.
obtained in this study re-assure that embryo biopsy technique on poor mor- Trial registration number: not applicable
phology embryos doesn’t seem to affect the perinatal and neo-natal outcomes.
P-183 Oxygen concentration in culture media equilibration: does
PGT-A on MPB can be used as an additional tool to pick the best embryo.
it have an impact on pregnancy rate?
Limitations, reasons for caution: Retrospective study, Small sample size,
Relative subjectivity of embryo scoring. E. De Martino1, P. Filardi1, G. Terrado2, N. Passi2, M. Gomez Peña1,
Wider implications of the findings: Our findings may help to reassure M. Papayannis1, J. Maidana1, I. De Zúñiga2, L. Kopcow2, M. Horton2,
women that morphologically poor quality embryos can be screened for PGT-A A. Oubiña2, F. Sobral2, C. Bisioli1
1
and if euploid, can offer the best reproductive outcomes. Apart from morphol- Pregna Reproductive Medicine, Embryology Laboratory, Buenos Aires, Argentina ;
2
ogy, PGT-A can be an additional tool for selecting and de-selecting the embryos Pregna Reproductive Medicine, Reproductive Medicine, Buenos Aires, Argentina
for optimizing pregnancy outcomes.
Trial registration number: not applicable Study question: Can we culture human embryos at low oxygen in media that
has been equilibrated overnight at atmospheric oxygen without affecting preg-
nancy rates?
P-182 Acute exposure to Etoposide does not activate DNA Summary answer: Pregnancy rates are not affected when embryos are cul-
damage checkpoints in Meiosis I of human oocytes. tured at low oxygen in media that has been equilibrated at atmospheric oxygen
M. Botzaki1, K. Agapitou1, E. Nikitos1, S. Kallergi1, K. Niaka2, P. concentration.
Marangos2 What is known already: Low oxygen has been shown to be crucial to culture
1 embryos up to the blastocyst stage. Despite this, many laboratories still culture
INSTITUTE OF LIFE-IASO Maternity hospital, IVFunit, Athens, Greece ;
2 embryos in atmospheric conditions. Others use the old incubators for media
University of Ioannina, Department of Applications and Technology-, Ioannina,
equilibration overnight in order to save space in the low oxygen ones. When
Greece
oocytes or embryos are placed into these high oxygen dishes they will be
Study question: Can spindle disruptions induced by Etoposide, a topoisom- exposed to a temporary atmospheric state while low concentration is finally
erase II inhibitor, activate DNA damage checkpoints in Meiosis I of human reached. Although it is generally pointed out that embryo culture must be entirely
oocytes? done at low oxygen, there are few studies proving that this is true.
Summary answer: DNA damage accumulated in meiosis-I, does not prevent Study design, size, duration: This prospective cohort study included 146
polar body extrusion and therefore could persist in morphologically normal women performing fresh ART treatment between May 2018 and July 2019.
metaphase-II oocytes. Exclusion criteria included couples with severe male factor and donor cycles.
What is known already: Severe spindle disruption activated the checkpoint Participants/materials, setting, methods: According to the culture media
to prevent polar body extrusion, moderate spindle insults fail to prevent polar equilibration methods used in our laboratory (overnight dishes at atmospheric
body extrusion, causing defective chromosome segregation that likely leads to or low oxygen) and the incubators space availability, patients were randomly
uneuploidy. Studies showed that mouse oocytes with damaged DNA can resume divided into two groups:
meiosis and undergo Germinal Vesicle Breakdown (GVBD), but then arrest in
metaphase of meiosis-I in a process involving Spindle Assembly Checkpoint • Group 1: Atmospheric oxygen (n=94)
(SAC) signalling.Such a mechanism could help prevent the generation of meta- • Group 2: Low oxygen (n=52)
phase-II (Met-II) eggs with damaged DNA.Little has been known about the
spindle checkpoint in human oocytes. Primary outcome was clinical pregnancy rate. Secondary outcome was implan-
Study design, size, duration: Donor immature oocytes aged <35years in a tation rate. No differences in patient age, insemination method and infertility diag-
time frame of two years. nosis were observed. Chi-square and Student’s t test were used as appropriate.
Experiments performed: 8 experiments involving 4 control and 8 oocytes Main results and the role of chance: No significant differences were found
treated with 100μg/ml Etoposide for 1h, from 8 individual donors. in clinical pregnancy rate (23.4% vs. 21.1%) and implantation rate (15.4% vs
26 experiments involving 54 control and 57 oocytes treated with 20μg/ml 14.5%) between groups. Treatment outcome seems not to be affected by an
Etoposide for 1h, from 16 individual donors. exposition of around 4 hours to an atmospheric oxygen concentration in the
Histone H2AX and is used as an immunofluorescence marker for the detec- beginning of the culture. Although several studies suggest a negative impact of
tion of double-strand breaks. Draq7 is used for DNA staining. GV-stage oocytes oxidative stress in embryo development, there are still few regarding the effect
are fixed immediately after treatment. of a brief exposition to atmospheric oxygen. Nevertheless it is generally recom-
Participants/materials, setting, methods: Human oocytes were recovered mended by personal communications that embryo culture must be done at low
from donor ovaries. Germinal Vesicle (GV)-stage oocytes were treated with oxygen since the very beginning.
high (100μg/ml) and lower (20μg/ml) concentrations of Etoposide for 1h. Limitations, reasons for caution: Our study is limited by its sample size
Following Etoposide treatment, treated and control oocytes were placed in the and the need to go beyond pregnancy and implantation rates. In this sense it
incubator at 37.5oC under 7.1% CO2. would be important to also study the possible effect that these two different

oxygen concentrations may have on the embryo physiology or on the new- P-185 The non invasive embryoscope morphokinetic kidscore
born health. grading system and PGT-A ploidy status
Wider implications of the findings: To the best of our knowledge, this is P. Sopaboon1, M. Tuntipoj1, P. Suanbooncharoen1, S.
the first analysis comparing two media equilibration conditions using atmospheric Chanchamroen1, W. Quangkananurug2, S. Seweewanlop1, P.
oxygen incubators as a possible viable option in cases of new technologies access Assawakittananon1, T. Thongsook1
limitations. More studies are necessary to elucidate the possible effects of tran- 1
Safe fertility Center, Laboratory, Bangkok, Thailand ;
sient oxygen concentration variation on human embryo development. 2
Safe fertility Center, Medical Director, Bangkok, Thailand
Study question: Could the KIDscore number indicate the risk of aneuploid?
Does the euploid and aneuploid embryos differ in KIDscore number?
P-184 The usefulness of metaphase I oocytes for intracytoplasmic
Summary answer: There is no particular KIDscore number in indicating
sperm injection
euploid embryo. The euploid and complex aneuploid embryos were statistical
J.P. Bilibio1, A.J.C. Meireles2, Y. Maciel3, P. Sales2, F. Nascimento2, significant difference in KIDscore number.
P.L. Lorenzzoni-4 What is known already: It has been shown that euploid embryo tend to
1
Universidade Federal do Pará, Ginecologia e obstetrícia, Belém ; implant. The invasive biopsy followed by PGT-A is widely use to identify the
2
Clínica de Reprodução Assistida Pronatus-, Reprodução Assistida, Belém, Brazil ; ploidy status of embryo. The non-invasive embryo selection is expected to
3
Universidade Federal do Pará, Ginecologia e obstetrícia, Belém, Brazil ; predict ploidy status to enhance the possibility of implantation. The time-lapse
4
Grupo de Pesquisa Bilibio, Reprodução Assistida, Belém, Brazil incubator recently play a role in culturing and selecting the best embryo to
transfer. EmbryoScope(Vitrolife) has launched the KIDscore system to be the
Study question: The aim was to evaluate the fertilization rate and blastocyst evaluation tool for scoring the embryos according to their statistical viability. The
formation rate of oocytes in metaphase I (MI) in women who underwent con- time-lapse incubator combined with the software is costly and not much data
trolled ovarian hyperstimulation for intracytoplasmic injection. have been published about the use of this algorithm.
Summary answer: MI oocytes collected at stage MI after controlled ovarian Study design, size, duration: Randomized prospective study. 205 blastocysts
hyperstimulation that did not mature in vitro to MII have a blastocyst formation in IVF/PGT-A cycle from couples attending an independent IVF clinic (Safe
rate of 0.6%. Fertility Center, Bangkok, Thailand) between April to May 2019 were enrolled
What is known already: It is known that an increased pregnancy rate is in this study.
directly related to the number of embryos in the blastocyst stage obtained by Participants/materials, setting, methods: All embryos were cultured in
the in vitro fertilization (IVF) cycle, and this number depends on the quantity and time-lapse incubators (EmbryoScope, Vitrolife). Embryo grading was done by
quality of MII oocytes collected. As the number of these oocytes is inversely both morphology grading (ACE/NEQAS system) and morphokinetic grading
proportional to the age of the patient, the use of MI oocytes for IVF, especially (KIDscore, Vitrolife). The KIDscore morphokinetic parameters observed were
in couples with advanced age, has been discussed as an alternative. PN assessment, PN fading, time (t) to 2, 3, 4, 5 and 8 cells, tSB and tB followed
Study design, size, duration: A prospective cohort study was performed by morphology grading of ICM and TE. The blastocysts were biopsied on Day
among women from whom at least 1 MI and 1 MII oocyte was obtained after 5/6 and analysed by PGT-A(NGS, Illumina).
controlled ovarian hyperstimulation for intracytoplasmic sperm injection during Main results and the role of chance: The blastocyst formation rate, blas-
the study period,June 2018 to June 2019. tocyst analyzed for PGT-A rate were 64.50% and 86.13% respectively. Of the
We collected 1907 oocytes from 164 women (1291 MII, 352 MI and 258 205 blastocysts, 134 were euploid, 14 were mosaic, 33 were single chromosome
prophase I or atresic) aneuploid, 19 were complex chromosome aneuploid and 5 were DNA degra-
Participants/materials, setting, methods: After oocyte classification, the dation. The overall KIDscore average range was 6.0±2.25. The euploid embryos
MII and MI oocytes were incubated for 4 hours. had 6.27±2.24 on average in KIDscore number with 95% Confident Interval
After 4 hours in the single step balanced embryonic culture medium (Global (CI):5.89-6.66. While the mosaic, single chromosome aneuploid, complex chro-
Total), 1649 oocytes were fertilized by intracytoplasmic injection (ICSI), of which mosome aneuploid and DNA degradation were 6.04±2.29 (95% CI:4.72-7.37),
1291 were in the MII stage (Group MII); of the 358 oocytes in the MI stage, 205 5.75±2.17 (95% CI:4.98-6.52), 4.99±2.34 (95% CI:3.86-6.12) and 5.42±2.01
matured to MII oocytes (Group MI-MII), and 153 remained in stage MI (Group MI). (95% CI:2.92-7.92).There are statistically significant difference in euploid and
Main results and the role of chance: After 4 hours, the maturation rate complex chromosome aneuploid group (P=0.021)in KIDscore number. The
was 57.2%; 205 matured in vitro to MII (Group MI-MII) and 153 remained in KIDscore number 8.0-9.9 bound to be euploid, mosaic, single chromosome
stage MI (Group MI). aneuploid and complex chromosome aneuploid as 76.74%, 9.30%,13.95% and
The normal fertilization rate was directly associated with oocyte maturation, 9.30% respectively. Whereas the mid-grade KIDscore number 4.0-6.0 had
with rates of 79.1%, 60.2%, and 31.9% in MII, MI-MII and MI oocytes, respectively 64.58%,10.41%, 27.08% and 14.58% in euploid, mosaic, single chromosome
(P <0.001). aneuploid and complex chromosome aneuploid respectively. The data could
The MI group had an odds ratio (OR) of 7.6 (IC 5.2 - 11.2, P <0.001) for imply that the high grade KIDscore number has more chance to be euploid than
abnormal fertilization compared with the MII group. the mid-grade and low-grade score.
The risk of embryo polyploidy was higher in the MI group, with an OR of 2.6 Limitations, reasons for caution: The current sample size is small and data
(1.28-5.41,P = 0.008), than in the MII group. were collected from a single clinic which has a limited generalizability and poten-
The blastocyst formation rate was directly associated with oocyte maturation: tial bias. As routinely use is running, we will report on a larger sample size to
36.4% for MII, 11.4% for MI-MII and 0.6% for MI. provide more reliable effectiveness.
MI oocytes had an OR of 86.9 (CI 12.1 - 623.4, P <0.001) for not forming a Wider implications of the findings: When selecting the embryo to transfer,
blastocyst compared with MII oocytes. considering not only the morphology scoring, the other scoring may benefit to
Limitations, reasons for caution: We found a decrease in the blastocyst help select the best embryo to get the best chance to implant. The non invasive
formation rate from MII oocytes but no difference in MI-MII oocyte in patients evaluation tools could be consider to use as it less harming to the embryo.
over 40 years of age. Maternal age does not seem to influence the rate of Trial registration number: not applicable
blastocyst formation from MI-MII oocytes, but these results need to be con-
firmed in other studies. P-186 Effect of different equilibration times on clinical and
Wider implications of the findings: Oocytes collected at stage MI after neonatal outcomes in human blastocyst vitrification
controlled ovarian hyperstimulatio that do not mature to MII in vitro have a S. Mitsuhata1, Y. Fujii1, Y. Endou1, M. Hayashi1, H. Motoyama1
blastocyst formation rate of only 0.6%, so their use should be discouraged. On 1
Kurashiki Medical Clinic, IVF center, Kurashiki, Japan
the other hand, the use of MI-MII oocytes may increase the amount of blasto-
cysts available for embryo transfer. Study question: What are the effects of shortened or prolonged equilibration
Trial registration number: 55625115.0.0000.0018 time during human blastocyst vitrification?

Summary answer: A shorter (5–11 min) equilibrium time is sufficient for dilemma of how to proceed when only one embryo is available, as frequently
vitrifying smaller, non-expanded blastocysts. observed in POR. Therefore, the implantation rate and pregnancy outcomes in
What is known already: In human embryo vitrification, the duration of relation to an individual embryo remain unclear.
embryo’s suspension in the vitrification solution is a critical consideration. This Study design, size, duration: This is a retrospective cohort study including
process should be strictly controlled within 1 min. However, exposure to the 622 patients with POR who underwent IVF/ICSI cycles in two reproductive
equilibration solution is flexible. A prolonged exposure to the equilibration solu- centers (from Belgium and Chile) between January 1st 2011 and December
tion may be harmful and may influence the embryonic developmental potential, 1st 2018.
whereas a shortened exposure may impair the penetration of cryoprotectants Participants/materials, setting, methods: All women who fulfilled the
into the blastomeres. With regard to human blastocysts, few studies examine Bologna criteria POR and underwent IVF/ICSI cycles with only one available
the effects of different equilibration times on clinical and neonatal outcomes. embryo on day 3, were included. The decision to transfer one embryo on day
Study design, size, duration: This was a retrospective study based on data 3 or day 5 was made by the treating clinician and only Grade I-III were trans-
collected between November 2008 and November 2015. A total of 225 blas- ferred. Embryo quality was similar in both groups. We excluded patients who
tocysts, 105 non-expanded blastocysts and 120 expanded blastocysts, obtained did not have an available embryo on day 5.
from 173 patients were analyzed. Non-expanded and expanded blastocysts were Main results and the role of chance: 622 Bologna criteria POR patients
divided into 2 groups according to the equilibration time: 5–11 min and 12–15 with only one available embryo on day 3 were included in the analysis: 498 were
min. Clinical and neonatal outcomes of warmed blastocysts were compared in categorized to the “day 3 group” and 124 to the “day 5 group”. There were no
each group. baseline differences between the groups. In the day 5 group, higher clinical preg-
Participants/materials, setting, methods: The blastocysts were vitrified nancy rates per started cycle (28% versus 19%, P <0.001) and higher live birth
and warmed following the Cryotop method. Blastocysts with blastocoels occu- rates (18% versus 11%, P = 0.043) were obtained compared to the day 3 group.
pying less than half of the embryo volume and showing no signs of zonae thinning Multivariate logistic regression analysis for relevant confounders, showed that
or expansion were categorized as non-expanded blastocysts. Blastocysts with the only independent variable that significantly associated to higher LBR was the
thinning zonae and blastocoels that occupied greater than half of the embryo day of the transfer (OR 1.97, 95% CI 1.12-3.44, P = 0.02).
volume were identified as expanded blastocysts. Hatching blastocysts were clas- Limitations, reasons for caution: The main limitation in our study is the
sified as expanded blastocysts. retrospective design. In addition, the decision to transfer the embryo on day 3
Main results and the role of chance: Both equilibration times (5–11 min or day 5 was made by the treating clinician during their IVF cycle and according
and 12–15 min) in the non-expanded blastocyst group resulted in effective vit- to the progress in embryo development, which may be a potential source of bias.
rification. There were no differences in the survival (96.9% vs. 92.5%, 95% CI: Wider implications of the findings: In POR with only one viable embryo
0.48–13.4), implantation (20.0% vs. 20.0%, 95% CI: 0.38–2.62) and live birth on day 3 of embryonic development, performing the transfer on day 5 was found
rates (12.3% vs. 17.5%, 95% CI: 0.23–1.92) between the groups. While the to achieve higher pregnancy and LBR compared with day 3 transfer. This chal-
results for the expanded blastocysts, the survival (86.7% vs. 96.7%, 95% CI: lenges current practices to be modified to improve fertility outcomes in this
0.05–0.99, P<0.05 ), implantation (23.3% vs. 45.0%, 95% CI: 0.17–0.81, P<0.05) specific group of patients.
and live birth rates (16.7% vs. 38.3%, 95% CI: 0.14–0.75, P<0.01) were signifi- Trial registration number: not applicable
cantly decreased in the 5-11 min group compared with 12-15 min group. There
were no differences in gestational age, birthweight, proportion of male babies,
rates of cesarean section, and congenital abnormalities. P-188 Sorbitol accumulation decreases oocyte quality of aging
Limitations, reasons for caution: This study is limited by sample size. There mice through increasing oxidative stress
is possible dispersion of the embryo quality and cause of infertility. Further Y. Zhang1, H. Liu1, L. Li1, Z. Yan1, Y. Mao1, L. Gao1, J. Liu1, Y. Cui1
prospective randomized studies will be needed to confirm the findings of 1
State Key Laboratory of Reproductive Medicine- Clinical Center of Reproductive
this study. Medicine, First Affiliated Hospital- Nanjing Medical University, Nanjing, China
Wider implications of the findings: Prolonged exposure to cryoprotectants
may incur harmful effects on embryonic development; thus, shortening expo- Study question: Whether could the sorbitol accumulation in oocytes decrease
sures to equilibration solution are desirable. This study shows that outcomes the quality of aging oocytes ?
are satisfactory if unexpanded blastocysts are vitrified with shorter exposure Summary answer: The excessive sorbitol decreases the oocyte quality of
times. Therefore, reducing the cytotoxicity of cryoprotectants and vitrification aging mice, and the sorbinil supplementation, inhibiting theproduction of sorbitol,
process is completed faster. can improve the quality of aging oocytes.
Trial registration number: not applicable What is known already: Sorbitol is a by-product of glucose metabolism
through the polyol pathway. Many studies have demonstrated that the excessive
P-187 Cleavage stage versus blastocyst stage embryo transfer in sorbitol can decrease the cellular activation due to the increased ROS.
Bologna poor responders: a retrospective analysis. Study design, size, duration: This is an in vitro study with COCs obtained
J. Errazuriz1,2, F. Pais1, A. Racca2, G. Verheyen2, I. Filselcker1, from 9 month-old female mice, to explore the sorbitol accumulation related
P. Gonzalez1, H. Tournaye2, C. Blockeel2 with oocyte quality.
1
Centre for Reproductive Medicine, Departamento de Ginecología y Obstetricia- Participants/materials, setting, methods: The COCs obtained from 9
Clínica Alemana- Facultad de Medicina- Universidad del Desarrollo., Santiago, month-old female mice were in vitro cultured with or without sorbinil, a kind of
Chile ; aldose reductase inhibitor as the control and the inhibitor group. In the young
2
Centre for Reproductive Medicine, Universitair Ziekenhuis Brussel- Vrije group, the COCs obtained from 6-8 week-old female mice were cultured the
Universiteit Brussel., Brussels, Belgium same as the control. MII oocytes were collected. The ROS and GSH were tested.
The level of Aldose reductase, SOD1 were measured. The contents of sorbitol
Study question: Do poor ovarian responders (POR) with only one available were measured by HPLC/MS.
embryo on day 3, achieve higher pregnancy rates by transferring cleavage or Main results and the role of chance: The maturity rate of oocytes in the
blastocyst stage embryos? inhibitor group was significantly higher than that in the control group (increased
Summary answer: In POR with only one embryo available, transferring the 16%, P<0.05). The ROS florescence intensity in the inhibitor group was
embryo on day 5 was found to achieve higher live birth rates compared decreased by 42% comparing with the control group (p<0.0001). The GSH
with day 3. florescence intensity in the inhibitor group was increased by 17% comparing with
What is known already: IVF/ICSI cycles in POR remain challenging regarding the control group (P<0.05). Interestingly, SOD1 was upregulated in the inhibitor
ovarian stimulation, and are characterized by low live birth rates (LBR) and high group (P<0.05). The content of sorbitol in the control group was drastically
cancellation rates. Traditionally, embryo transfer was carried out on day 3; how- increased comparing with the other groups (P<0.001). The level of Aldose
ever, in recent years there has been a shift in practice towards embryo transfer reductase was increased in the in vitro aging oocytes of 9 month-old female mice
at the blastocyst stage. Nevertheless, these studies have not addressed the and the in vitro maturation oocytes of 6-8 week-old female mice (both P<0.05).

The sorbitol was accumulated in the in vitro aging oocytes comparing with the Summary answer: It is safe to transfer poor quality embryos when they are
MII oocytes matured in vivo of 9 month-old female mice (P<0.05). the only option for fresh ET.
Limitations, reasons for caution: Sorbitol accumulation decreases the What is known already: Implantation potential of poor morphology embryos
oocyte quality of aging mice, while whether the content of sorbitol in human is much lower than of those of good and fair quality. At the same time, there
oocytes increases during in vitro maturation is sparsely understood. are indications that it might be safe to transfer embryos of poor quality as such
Wider implications of the findings: The excessive sorbitol may decreases transfers do not increase the rate of chromosomal abnormalities, congenital
the oocyte quality of aging mice, and the inhibition of sorbitol production during malformations of children born, or perinatal complications and mortality.
IVM may improve the oocyte quality by decreasing the oxidative stress. However, the data available on the outcomes of IVF cycles in which poor quality
Trial registration number: not applicable embryos are transferred is limited. It is also uncertain which developmental
stage: cleavage or blastocyst is more favorable for poor quality embryos to
P-189 Slowly developing blastocysts: fresh embryo transfer on day transfer.
5, on day 6 or frozen embryo transfer? Study design, size, duration: A retrospective analysis using individual patient
data with positive controls. All patients undergoing fresh embryo transfers of
A. Reignier1, A. Inquel1, S. Loubersac1, J. Lammers1, P. Barriere1,
poor quality embryos between 2012 and 2016. 738 poor quality embryos from
T. Lefebvre1, T. Freour1
1
488 patients were assessed. The control group consisted of 9893 fair and good
CHU Nantes, Biologie et médecine de la reproduction, Nantes, France quality embryos from 5994 patients.
Participants/materials, setting, methods: IVF cycles with or without ICSI
Study question: Should slowly developing blastocysts be transferred on day
involving fresh embryo transfers of poor quality embryos on day 3 or on day 5.
5, when expanded on day 6, or be frozen in order to optimize live birth
The main outcomes of day3 and day 5 poor quality embryo transfers were
rate (LBR)?
compared. We also compared the main outcomes of cycles with poor quality
Summary answer: Fresh day 5 early cavitating blastocyst transfer resulted in
embryo transfers with good and fair quality embryo transfers.
higher LBR than fresh expanded day 6 blastocyst transfer or frozen-thawed day
Main results and the role of chance: No significant differences in the
6 embryo transfer.
biochemical pregnancy rate, implantation rate, clinical pregnancy rate, miscarriage
What is known already: LBR are higher after fully expanded blastocyst fresh
rate and live birth rate were found between the groups of poor quality embryos
transfer on day 5 than after slow-growing expanded blastocysts fresh transfer
transferred on day 3 or on day 5. Although the biochemical pregnancy rate and
on day 6. Embryo-endometrial asynchrony could explain this decrease. In this
implantation rate were lower for the group of poor quality embryos than for
respect, some studies suggested day 6 blastocysts should systematically be fro-
the control (13.9% vs 37.2%, p<0.001 and 6.9% vs 29.4%, p<0.001 respectively),
zen, but this still remains unclear. Moreover, there is no consensus about the
statistically significant differences between the portion of the biochemical preg-
best strategy to adopt with slowly developing blastocysts which are stil at the
nancies which resulted in clinical pregnancies and the portion of clinical preg-
early cavitating stage (B1 or B2 according to Gardner & Schoolcraft criteria) on
nancies which resulted in live births in both groups were not observed (72% vs
day 5, as to whether they should be transferred on day 5, 6 or vitrified on day 6.
78.3%, p=0.2313 and 79.6% vs 79.3%, p= 0.9636 respectively).
Study design, size, duration: This was a retrospective monocentric cohort
Limitations, reasons for caution: This was a single-center retrospective
follow-up study including 306 cycles performed between 2016 and 2017 with
study. Embryo morphology grading was assessed by 5 experienced embryolo-
at least 1 early cavitating blastocyst available on day 5 which was either trans-
gists, but it still remains a subjective type of evaluation.
ferred fresh on day 5 or cultured up to day 6 and transferred fresh or vitrified.
Wider implications of the findings: Unlike the common belief, our data
Participants/materials, setting, methods: All patients with early cavitating
suggest that transfers of poor quality embryos do not increase the rate of mis-
blastocyst on day 5 (Gardner stage B1 or B2) were included and allocated to 3
carriages or stillbirths and should be considered safe. Transfer of embryos of
groups. Group 1 had fresh unexpanded blastocyst transfer on day 5 (n=68),
poor morphology results in lower but still acceptable live birth rates.
group 2 had fresh full expanded blastocyst transfer on day 6 (n=57) and group
3 had frozen embryo transfer after vitrification on day 6 (n=181). LBR was the
main outcome and was compared between the 3 groups.
P-191 Pre-maturation culture with CNP before IVM does not
Main results and the role of chance: Female age (33.8, 32.9, 34.1 years
improve oocyte competence and embryo development in women
old) and smoking status (19.1, 14.0, 13.3%) were comparable between the
with PCOS
groups 1, 2 and 3 respectively. Female BMI was significantly higher in group 1
than in groups 2 and 3 (26.1 versus 24.7 and 23.7 kg/m² respectively). Serum Y. Lin1, X. Zheng1, J. Yan1, X. Song1, R. Li1, J. Qiao1, C. Ma1
1
AMH level was significantly higher in group 3 than in groups 1 and 2 (3.1 versus Peking University Third Hospital, Center for Reproductive Medicine, Beijing, China
2.3 and 2.4 ng/mL respectively). LBR was significantly higher in group 1 (fresh
transfer on day 5) than in group 3 (frozen day 6 transfer): (26.4%) versus (13.8%); Study question: Whether a pre-culture with C-type natriuretic peptide (CNP)
p < 0.05. LBR was 14.0% for group 2 (fresh transfer on day 6) without significant before in vitro maturation (IVM) improves oocyte competence and embryo
difference with groups 1 and 3. developmental potential or not?
Limitations, reasons for caution: The relatively limited numbers of cycles Summary answer: Pre-maturation culture (PMC) with CNP before IVM does
included as well as the retrospective design of the study are significant limitations. not improve oocyte competence and embryo development in women with
Vitrification of early blastocysts on day 5 has not been evaluated in this study. polycystic ovarian syndrome (PCOS).
Wider implications of the findings: Fresh day 5 transfer should be pro- What is known already: Numerous studies in animal models have indicated
moted for early cavitating blastocyst rather than fresh day 6 transfer or vitrifi- the benefit of a pre-culture with CNP on IVM of immature oocytes. In 2017,
cation. These preliminary results should be integrated when the embryo culture Sánchez et al reported that a new IVM culture system enhances human oocyte
and transfer strategy is considered by the clinicobiological IVF staff, and should competence and embryo yield and thought that the improvements were mainly
be used to improve patients’ counselling. attributed to the PMC step in presence of CNP. However the positive effects
Trial registration number: none of CNP on IVM has not yet been repeated by other research teams.
Study design, size, duration: A prospective self-control study involved 192
cumulus–oocyte complexes (COCs) from 15 women with PCOS to evaluate
P-190 Should we transfer poor quality embryos? effects of PMC with CNP on oocyte competence and subsequent embryo devel-
A. Kirillova1 opmental potential. This study was conducted from March 2019 to
1
Federal State Budget Institution “National Medical Research Center for Obstetrics- November 2019.
Gynecology and Perinatology named after Academician V.I. Kulakov of the Ministry Participants/materials, setting, methods: PCOS women undergoing lap-
of Healthcare of Russian Federation”, Department of preservation and restoration aroscopic salpingectomy for hydrosalpinx in a university-affiliated reproductive
of the, center were recruited and COCs were retrieved while surgery. COCs obtained
were randomly allocated to control group (conventional IVM for 30h) or PMC
Study question: Should poor quality embryos be transferred? group (PMC with CNP for 24h + conventional IVM for 30h). Oocyte maturation

and embryo development after ICSI were compared between groups. Cumulus Wider implications of the findings: This study found that rescue ICSI must
cells (CCs) were collected for assessments of the expressions of NPR2 and GJA1 be performed after 8 hours to eliminate the risk of artificial polyspermy.
mRNA and cGMP level. However, delaying fertilization may reduce the quality of the oocytes. Therefore,
Main results and the role of chance: Fifteen patients were included in the further studies are needed to determine new criteria for performing rescue ICSI
study. Total 192 COCs were divided randomly into PMC group (n=93) and as soon as possible.
control group (n=99). Oocyte maturation rates were similar between PMC Trial registration number: not applicable
group (53.8%) and control group (47.5%, p=0.384). Subsequent fertilization rate
(54.0% vs 63.8%) and available embryo rate (40.0% vs 53.2%) per mature oocyte
P-193 Effect of Time Elapsed Between Thaw and Embryo Transfer
also showed no significant difference (p=0.326 and 0.193, respectively) between
on pregnancies rates in Patients with Cryopreserved-thawed
groups. Six blastocysts were acquired in control group (12.8%) while no blasto-
embryo transfer: A Systematic Review and Meta-Analysis
cyst formation in PMC group (0%, p=0.029). Expressions of NPR2 and GJA1
mRNA of CCs denuded from COCs were similar between two groups. No L.H. Sordia-Hernandez1, F.A. Morales Martinez2, E. Gutierrez
significant difference was found in cGMP level of CCs between groups too. Orozco1, O.H. Valdes Martínez1, A. Flores Rodriguez3, P. Leyva
Limitations, reasons for caution: The sample size was not large as previous Camacho4, N. Alvarez Villalobos3, J.A. Zuñiga Hernandez1
1
studies. As no blastocyst was acquired in PMC group, the planned analyses of Universidad Autonoma de Nuevo Leon, Centro Universitario de Medicina
morphology, cell number, aneuploidy rate, DNA methylation and imprinted Reproductiva, Monterrey, Mexico ;
2
genes of blastocyst were not undergone. Because of the discouraging results, Universidad Autonoma de Nuevo León, Centro Universitario de Medicina
imaging of cumulus-oocyte transzonal connection was not taken. Reproductiva, Monterrey, Mexico ;
3
Wider implications of the findings: The results of this study found that Universidad Autonoma de Nuevo Leon, Plataforma INVEST Medicina UANL –
pre-cultured with CNP before IVM did not improve oocyte competence and KER Unit Mayo Clinic KER Unit México-, Monterrey, Mexico ;
4
embryo developmental potential. Further studies exploring a better IVM culture Universidad Autonoma de Nuevo Leon, Plataforma INVEST Medicina UANL –
system are still needed. KER Unit Mayo Clinic KER Unit México, Monterrey, Mexico
Trial registration number: 2018SZ-032
Study question: Does short or long culture time between thawing and transfer
P-192 What are the best decision criteria for rescue of cryopreserved embryos have an impact on reproductive outcomes after in
intracytoplasmic sperm injection (ICSI), reducing the risk of vitro fertilization (IVF)?
artificial polyspermy? Summary answer: There are no differences in reproductive outcomes if
T. Kishida1, H. Kikuchi1, K. Yamada1, H. Kanno1, R. Kishida1, T. cryopreserved embryos are transferred after overnight culture or after two
Sato1, M. Yuki1, A. Kitsu1, S. Fumiya1, M. Katagiri1, M. Shinagawa1, hours of culture after thawing
Y. Hiroaki1 What is known already: There is currently no clear consensus on the optimal
1
time for the thawing of embryos in freezing protocols, despite the fact that the
Sendai ART Clinic, ART Lab., miyagi, Japan
identification of an adequate culture time would improve embryo culture envi-
ronments allowing for better reproductive results
Study question: To establish the decision criteria (DC) for rescue ICSI, reduc-
Study design, size, duration: A systematic review of studies evaluating the
ing the risk of artificial polyspermy using time-lapse imaging systems.
association of two different culture times after thawing, and before transfer, of
Summary answer: Postponing the observation time of the second polar body
cryopreserved embryos with the incidence of pregnancy, implantation, abortion,
(PB) extrusion is most effective for reducing the risk of artificial polyspermy after
live birth or ongoing pregnancy, and ectopic pregnancy in individuals after IVF
rescue ICSI.
fertilization
What is known already: The decision to perform rescue ICSI is determined
Participants/materials, setting, methods: A literature search was performed
by combining the observation time of the second PB extrusion following the
in PubMed, EmBase, and the Cochrane library (from January 2000 to august 2019).
conventional IVF and the second PB extrusion rate at that time. Artificial poly-
A cumulative meta-analysis and evaluation of heterogeneity was performed for
spermy, created by performing rescue ICSI to normally fertilized oocytes after
the clinical pregnancy rate. The quality of the included studies was assessed using
conventional IVF, must be avoided. However, the optimal DC for rescue ICSI
Cochrane’s Risk of Bias tool and ROBINS I for observational studies
for reducing the risk of artificial polyspermy has not been established.
Main results and the role of chance: Five studies were included, two were
Study design, size, duration: This was a retrospective study conducted at
randomized controlled trials and three were retrospective published between
Sendai ART clinic in Japan from 2017 to 2019. A total of 1480 mature oocytes,
2010 and 2019. One study included exclusive transfers of day 5 embryos; four
taken from 437 cycles, were cultured using a time-lapse incubator after short
studies included only transfers of day 3 embryos. The clinical pregnancy rate
insemination (4 hours). These oocytes were checked from the time of insemi-
had a high degree of heterogeneity and no difference was found between short
nation to the second PB extrusion and we determined the number of pronuclei.
and long embryo culture
Participants/materials, setting, methods: The target oocytes for rescue
Limitations, reasons for caution: One limitation was the quality of studies
ICSI were extracted on the basis of various DC, using time-lapse images. DC
reported and included. Most were retrospective in design and only two were
were set by combining the extrusion time of second PB (<4, 4, 5, 6, or 7 hours
RCTs. The risk of bias was deficient, with moderate being the best quality. The
after IVF) and the ratio of the oocytes extruding the second PB per mature
small number of studies reported on this subject was a very important limitation
oocytes (0%, 0-30%, or 0-99%). The normally fertilized oocytes rate following
Wider implications of the findings: No differences were found if cryopre-
IVF among the extracted target oocytes were examined as the risk rate of
served embryos were transferred after overnight culture or after two hours of
artificial polyspermy.
culture after thawing. Large-scale randomized clinical trials should be conducted
Main results and the role of chance: The risk rates of artificial polyspermy,
to definitively clarify if there is a difference in clinical outcomes after different
when setting the extrusion time of the second PB at <4 hours, were significantly
culture time between thawing and transfer of cryopreserved embryos
higher than those at 4, 5, 6, and 7 hours (<4 hours: 37.0-40.8%, 4 hours: 15.0-
Trial registration number: CRD42019137136
25.8%, 5 hours: 6.3-9.0%, 6 hours: 2.0-4.3%, and 7 hours: 0-1.0%, P<0.05). The
risk rates tended to decrease gradually as the extrusion time was delayed.
Conversely, there was no significant difference in risk rates between the ratios P-194 Feasibility of distance-based scoring in the Well-of-the-Well
of the oocytes extruding the second PB. The risk rates decreased to 0% when (WOW) culture system to improve blastocyst development
the extrusion time was set at 7 hours, and the ratios of the oocytes that extruded R. Minekawa1, T. Yoshida1, H. Ogata1, S. Ogata1
second PB were 0% or 0-30%. 1
Ogata Family Clinic, Obstetrics and Gynecology, Hyogo, Japan
Limitations, reasons for caution: This study is a single-center retrospective
study. We expect the extrusion time of the second PB to change depending on Study question: To provide quantitative assessment of optimal arrangement
the ART protocol (e.g., controlled ovarian stimulation protocol and start time of embryos in the Well-of-the-Well (WOW) culture system to improve blasto-
of insemination). cyst development and quality.

Summary answer: Scorings that were based on numbers of surrounding affect TE quality/integrity, hence implantation, studies are needed in order to
embryos and distance between adjacent embryos were associated with blasto- evaluate the effect of different biopsy techniques
cyst formation. Study design, size, duration: This was a prospective observational study
What is known already: It is known in bovine and mouse embryos that performed between 2018 and 2019. 286 PGT-A treatments with 990 TE biopsies
culturing embryos in groups is superior to single culture for blastocyst develop- were included. Of these, 145 euploid embryos were individually transferred, 84
ment due to secretion of autocrine and paracrine factors to neighboring biopsied using Pulling and 61 via Flicking. When using Pulling, laser pulses debil-
embryos. The WOW system provides group culture environment in single drop itate cell junctions and applying moderate tension the TE sample is detached.
of medium that could stimulate diffusion of autocrine and paracrine factors while However, when using Flicking this tension is replaced by a cutting motion gen-
observing individual embryo development simultaneously. It has been reported erated between the holding and biopsy pipettes.
in human embryos that density may affect embryo quality in microwell group Participants/materials, setting, methods: In every single embryo biopsy,
culture dish. the number (1-10) and intensity (ms) of laser pulses was registered, as well as
Study design, size, duration: This retrospective analysis of 476 embryos the operator and the method used for the biopsy (Pulling or Flicking). Blastocyst
was conducted over a 24-month period between 2017 and 2019. Embryos area (µm2) before and after TE biopsy as well as just before embryo vitrification
originated from women <40 years old were enrolled in the study when nine and was measured for each transferred embryo.
more embryos were obtained in each treatment cycle. The WOW system named The main outcomes were implantation rates and embryo collapse-expansion
LinKID®micro25 (DNP, Japan), which is consist of 5x5 microwells, were used. dynamics.
Embryos were cultured in single-step medium in LinKID®micro25 up to blasto- Main results and the role of chance: No differences were found between
cyst stage without refreshment. groups regarding egg age (Pulling: 33.9 vs Flicking: 32.97; p=0.885), maternal
Participants/materials, setting, methods: Embryos were firstly compared age (Pulling: 39.69 vs Flicking: 39.68; p=0.94), PGT indication (p=0.422), embryo
between two groups according to number of adjacent embryos; Group A (1-4) expansion (p=0.205) and quality (p=0.63), and laser intensity (Pulling: 416.4ms
and B (5-8) respectively. Secondly, considering different intra-microwell distance, vs Flicking: 438.3ms; p=0.184). Otherwise, the number of laser pulses was higher
cumulative scoring was set up as follows; 2 points were given when embryos when using pulling (Pulling: 5.4 pulses vs Flicking: 3.49 pulses; p<0.05). This
were allocated side-to-side or up and down, and 1 point was given when difference was expected and intrinsically attributed to the technique. To ensure
embryos were situated diagonally. Then embryos were grouped based on total that this was not a confounding factor we analysed implantation rate based on
scores; Low (2-6), Intermediate (7-9), and High (10-12) points (P) groups. the number of pulses and no significant differences were detected (1-3 pulses:
Main results and the role of chance: Blastocyst formation rate and per- 63.6%; 3-6 pulses: 58.1%; >6 pulses: 76.5%).
centage of good quality blastocysts (≥3BB by Gardner grading scheme) were Clinical outcomes, such as implantation rate (Pulling: 65.0% vs Flicking: 58.2%),
main outcome measures. In the first analysis the blastocyst formation rate was miscarriage rate (Pulling: 10.0% vs Flicking: 9.1%), and ongoing pregnancy rate
significantly higher in Group B (A: 50.3% vs B: 59.7%, P=0.048) but no differences (Pulling: 55.0% vs Flicking: 49.1%), were equivalent between groups (p>0.05).
were found in average age (A: 34.0 vs B: 34.2 years) and the percentage of good Regarding collapse-expansion dynamic, we found that embryos from Flicking
quality blastocyst (A: 67.0% vs B: 69.8%). In the second part of study, there was group showed the highest collapse percentage ((µm2 Pre-µm2 Post)/µm2 Pre)
no difference among three groups in terms of average age; Low P (N=167) 34.1, after biopsy (Pulling: 47.4% vs Flicking: 56.1%; p<0.05). Nevertheless, we did
Intermediate P (N=217) 34.0, High P (N=92) 34.6 years. Blastocyst formation not found any difference in the speed of expansion (%expansion/hour) after
rates were 50.3%, 56.7%, and 64.1% in Low P, Intermediate P and High P group biopsy (Pulling: 11.3%/h vs Flicking: 11.0%/h; p>0.05). Neither the percentage
respectively, and it was significantly higher in High P group compared to Low P of collapse nor the speed of expansion seem to affect implantation rates
group (P=0.037). The more points it gained, the higher percentage of good (p=0.325).
quality blastocyst (65.5% vs 69.9% vs 71.2%) was observed although there was Limitations, reasons for caution: These results are preliminary. In order to
no statistical significance. ensure the safety of both techniques more data should be analysed. If possible,
Limitations, reasons for caution: This is a retrospective analysis of database, the impact on live birth rates would be the ideal outcome to assess if any of
and sample size is limited as cases were restricted to patients obtained more these two techniques proves to be superior.
than nine embryos for equalizing background conditions. Our study focused on Wider implications of the findings: Our results show that the type of
blastocyst formation and quality, hence there is no enough data to evaluate technique used to perform TE biopsy does not affect clinical outcomes on
pregnancy rate and birth outcomes. euploid embryos. Although Flicking proved to elicit a stronger contraction post
Wider implications of the findings: This distance-based scoring approved biopsy, probably by generating more mechanical stress to the trophectoderm,
possible autocrine/paracrine effect for human embryo development. It is the this did not seem to impair reproductive potential.
first quantitative method to propose optimal arrangement of embryos in the Trial registration number: Not applicable
WOW culture, which could lead to better chance for clinical pregnancy. Cases
with fewer embryos could also have benefit if included in the future study. P-196 Impact of cell exclusion and extrusion during the peri-
Trial registration number: Not applicable compaction period on blastocyst formation and subsequent live
birth
P-195 Pulling vs Flicking. Does biopsy technique affect clinical K. Shimazaki1, G. Coticchio2, K. Ezoe3, K. Ohata3, M. Ninomiya1,
outcomes on euploid embryos? N. Wakabayashi1, T. Okimura1, K. Uchiyama1, T. Kobayashi4,
V. Montalvo1, S. Novo1, A. Garcia-Faura1, B. Marques1, F. Garcia1, C. Lagalla2, A. Borini5, K. Kato4
C. Castelló1, M. López-Teijón1 1
Kato Ladies Clinic, IVF Laboratory, Tokyo, Japan ;
1 2
Institut Marques, Reproductive Medicine Service, Barcelona, Spain 9.Baby- Family and Fertility Center, IVF laboratory Unit, Bologna, Italy ;
3
Kato Ladies Clinic, R&D Division, Tokyo, Japan ;
Study question: Has the technique used during trophectoderm biopsy any 4
Kato Ladies Clinic, Gynecology, Tokyo, Japan ;
effect on euploid embryos? 5
9.Baby- Family and Fertility Center, Reproductive Medicine IVF, Bologna, Italy
Summary answer: Pulling and Flicking techniques provide, when applied prop-
erly, optimal and similar results. Therefore, both techniques can be used indis- Study question: Are cell exclusion and extrusion during the embryo peri-com-
tinctly to perform trophectoderm biopsy. paction period associated with embryo development, pregnancy outcomes and
What is known already: Embryo biopsy for Preimplantational Genetic Testing patient characteristics after blastocyst transfer?
(PGT) has to accomplish two main goals: the biopsied material has to mirror Summary answer: Cell exclusion and extrusion during peri-compaction are
the genetic constitution of the embryo, and embryo viability cannot be impacted. associated with impaired blastocyst expansion, reduced live birth rates, male
The strategy that comes closer to guarantee these two aspects is trophectoderm age and low sperm-motility.
(TE) biopsy. Since there is no standardized TE biopsy procedure, two techniques What is known already: The biological and clinical significance of morpho-
are coexisting (Pulling and Flicking). The main difference between both techniques logical alterations in human morula stage embryos is poorly understood, in part
is how cells are detached from the embryo. Considering that TE biopsy could because most assessments of morula morphology have been performed

statically. With the flexibility offered by time-lapse technology, different patterns abnormalities, but there are few reports regarding the timing of the first cleavage
have recently been identified in compacted embryos: fully-compacted morula in DC embryos.
(FCM), partially-compacted morula with cells excluded before compaction Study design, size, duration: This was a retrospective study of data acquired
(Excluded-PCM), and partially-compacted morula with cells extruded after com- by time-lapse imaging. The study included 679 embryos from 353 IVF cycles in
paction (Extruded-PCM). Excluded-PCM and extruded PCM have been associ- 286 patients that underwent intracytoplasmic sperm injection (ICSI) treatment
ated with decreased blastocyst formation, and increased degeneration rates, for blastocyst cryopreservation between May 2018 and August 2019. A total of
respectively. At present, our overall knowledge of possible associations between 373 blastocysts were cryopreserved, and of these, 190 were used for a single
compaction patterns and pregnancy outcomes, and patient/embryonic charac- vitrified-thawed blastocyst transfer by December 2019. Embryo selection for
teristics is rather limited. transfer was based on blastocyst grade and diameter.
Study design, size, duration: Time-lapse videos from 384 morula stage Participants/materials, setting, methods: Embryos with DC at the first
embryos in 291 patients who underwent ICSI with minimal stimulation and single cleavage were recorded as DC(+), and embryos with normal cleavage were
vitrified-warmed blastocyst transfer (SVBT) from April 2017 to March 2018 recorded as DC(-). We compared the time from ICSI to the beginning of the
were retrospectively analysed. Compaction patterns were assessed and com- first cleavage (t2), and the time from the beginning to the end of the first cleavage
pared with embryonic and pregnancy outcomes after SVBT. Other factors pos- (d2) between groups. In addition, we compared implantation rates, miscarriage
sibly associated with compaction patterns, such as patient- and cycle-characteristics rates and live birth rates in single vitrified-thawed blastocyst transfer cycles
and early cleavage-stage development, were investigated. between groups.
Participants/materials, setting, methods: Following ICSI, time-lapse vid- Main results and the role of chance: T2 and d2 of DC(+) embryos (27.9
eos spanning fertilization to blastocyst stage were analysed. The number of cells hours and 5.5 hours, respectively) were significantly longer than those of DC(-)
excluded before compaction, and cells extruded after compaction (if any) were embryos (25.6 hours and 3.3 hours, respectively, p<.0001), even when the
annotated. Possible correlations of compaction patterns with blastocyst expan- embryos developed into blastocysts on Day 5 (t2: 27.0 hours in DC(+) and 25.2
sion rates, blastocyst morphological grade, clinical and ongoing pregnancy rate, hours in DC(-), p<.0001, d2: 5.9 hours in DC(+) and 3.2 hours in DC(-),
and live birth rate were evaluated. Other possible relationships between com- p<.0001). T2 of embryos that reached high-quality blastocysts or that were
paction patterns and patient characteristics, including serum hormone levels, used in single vitrified-thawed blastocyst transfer were similar in DC(+) (25.7
semen parameters, and incidence of abnormal cleavage patterns were also hours and 26.1 hours, respectively) and DC(-) embryos (24.8 hours and 25.2
assessed. hours, respectively). However, d2 was significantly longer in DC(+) embryos
Main results and the role of chance: The incidences of FCM, Excluded- (5.8 hours and 5.7 hours, respectively) compared to DC(-) embryos (3.0 hours
PCM, and Extruded-PCM were 51.8%, 29.7%, and 18.5%, respectively. and 3.1 hours, respectively, p<.0001). In addition, in DC(+) embryos, the rate
Blastocyst formation and cryopreservation rates in Excluded-PCM and of blastocyst formation on Day 5 was higher when t2 was <26 hours than when
Extruded-PCM groups were significantly lower than that in the FCM group, t2 was >=26 hours (80.3% vs. 22.9%, p<.0001). No significant differences were
particularly when the numbers of cells excluded and extruded were more than detected in implantation rates, pregnancy loss rates and live birth rates between
3 and 2, respectively. Lower morphological grades of inner cell mass and DC(+) and DC(-) embryos (29.2% vs. 40.1%, p=0.303, 28.6% vs. 29.7%,
trophectoderm were observed in both Excluded-PCM and Extruded-PCM p=0.390, 20.8% vs. 27.4%, p=0.494, respectively). Congenital abnormalities
groups compared with the FCM group. Cochran-Armitage trend testing were not detected in either group.
demonstrated that the ongoing pregnancy and live birth rates were significantly Limitations, reasons for caution: Small sample size is the primary limitation.
lower in Excluded-PCM and Extruded-PCM groups than in the FCM group In this study, we focused only on the first cleavage, and therefore events after
(P = 0.0147, P = 0.0348, respectively). Increases in incidence of cell exclusion the second cleavage were not evaluated. Because chromosomal analysis is strictly
and extrusion were significantly associated with increases in male age and prohibited in Japan, this study had no available data regarding the chromosomal
decreases in sperm-motility (P = 0.0108, P = 0.0019, respectively). ploidy of the blastocysts.
Furthermore, the embryos in the Excluded-PCM group exhibited significant Wider implications of the findings: The time of the first cleavage was
correlations with higher incidence of direct cleavage, rapid cleavage, and asym- prolonged in DC embryos, which should be investigated in future studies.
metrical division (P = 0.0145, P < 0.0001, P = 0.0232, respectively). Embryos However, our results suggest that the timing of the beginning of the first cleavage
in the Extruded-PCM group did not correlate with irregular division in early could be a useful indicator for embryo selection when the only transferrable
stage cleavage. embryos are DC embryos.
Limitations, reasons for caution: The number of analysed embryos is rel- Trial registration number: not applicable
atively limited; therefore, further studies are required to more reliably assess
the clinical significance of these alternative patterns of compaction. P-198 Detailed quality assessment of direct cleavage embryos
Wider implications of the findings: These results support the possibility during the first division using a combined early cleavage with an
that compaction patterns can serve as a predictive parameter for pregnancy embryo morphological grading method
outcomes. In addition, they shed new light on the phenomenology and impor- H. Tomari1, K. Honjo1, Y. Nagata1
tance of a poorly investigated stage of human preimplantation development. 1
IVF NAGATA Clinic, ART Lab, Fukuoka, Japan
Study question: Is it possible to perform a detailed quality assessment of
P-197 The timing of the first cleavage of direct cleavage embryos direct cleavage (DC) embryos at the early embryonic stage (day2)?
was delayed compared to normal cleavage embryos. Summary answer: The quality of DC embryos can be assessed in detail by
H. Takahashi1, R. Hirata1, T. Habara1, N. Hayashi1 combining the timing of the first cleavage and the morphological evaluation
1
Okayama couple’s clinic, laboratory, Okayama, Japan on day2.
What is known already: Several studies have reported a clear correlation
Study question: Are the early dynamics of direct cleavage (DC) embryos between the occurrence of DC (divided into three or more cells) during the
different from normal cleavage embryos? first division of an embryo and impaired embryo development potential in
Summary answer: In DC embryos, blastocyst transfer led to live birth, but humans. In addition, it has also been reported that the pregnancy rate due to
the timing of the first cleavage was delayed compared to normal cleavage transfer of DC embryos that developed to blastocysts is similar to that of normal
embryos. cleavage embryo transfer. However, only few studies have reported on methods
What is known already: DC embryos exhibit lower blastocyst formation for assessing DC embryo quality.
rates. However, we previously reported that DC embryos exhibit a similar Study design, size, duration: This retrospective observational study was
implantation potential to that of normal cleavage embryos if the blastocyst stage performed in a single in vitro fertilization (IVF) center. This study included patients
is reached, in which embryo selection for transfer is based on blastocyst grade undergoing IVF or intracytoplasmic sperm injection. We analyzed 1,242 DC
and diameter. Prior reports suggest that the timing of the first cleavage is asso- embryos with normal fertilization using a time-lapse incubator. All study partic-
ciated with embryo development and higher incidence of chromosomal ipants provided informed consent and the study design was approved by the

ethics committee of the IVF Nagata Clinic, Fukuoka, Japan. DC embryos were donors’ age we found significant differences: 33.9% for the youngest group,
classified into 3- and ≥4-cell (4 cells or more) groups. 39.1% for the group of 20-25 years and 42.5% for donors over 25 years
Participants/materials, setting, methods: Ex.1: We compared blastocyst (p=0.022). When adjusting by confounding factors (recipient age, number of
formation rates between the two groups. Ex.2: The two groups were further transferred embryos, and day of embryo transfer), LBR was lower to the <20
classified into six grades by the following method. Embryos were evaluated for years’ group (OR: 0.70; CI 95%: 0.50-0.99) and to the 20-25 years’ group (OR:
early cleavage (EC) or late cleavage (LC) at 27 hours after insemination and 0.85; CI 95%: 0.74-0.98) compared to the ≥26 years’ group. No significant dif-
morphology was scored on day2 (poor, ≥4-cells with ≥50% frag.; fair, ≥4-cells ferences were observed neither in fertilisation rates (74.2%, 76.1% and 77.5%)
with <50% and ≥20% frag.; good, ≥4-cells with <20% frag. and equal blasto- nor in embryo development rates (57%, 61.4% and 62%). The number of
mere). The blastocyst formation rates of each group were compared. good-quality embryos transferred was significantly lower in the <20 years’ group
Main results and the role of chance: Ex. 1: Among the 1,242 DC embryos, (1.03±0.71; 1.18±0.69; 1.19±0.67; p=0.015).
669 were in the 3-cell group and 573 were in the ≥4-cell group. The blastocyst Limitations, reasons for caution: The group of donors <20 years included
and high-quality blastocyst formation rates were significantly higher (p<0.01) in in our study was relatively small and smaller as compared with the groups of
the 3-cell group than in the ≥4-cell group (53.5% vs. 32.7%, 28.0% vs. 14.1%, 20-25 years and ≥26 years. Nonetheless, the sample size had adequate statistical
respectively). Ex. 2: Among the 669 embryos in the 3-cell group, 211 were in power in order to identify significant differences in live birth rate.
the EC-fair embryos, 141 were in the EC-poor embryos, 75 were in the LC-fair Wider implications of the findings: Oocyte donor age <20 years as com-
embryos, and 242 were in the LC-poor embryos. Among the 573 ≥4-cell group, pared with older oocyte donors is associated with significantly lower live birth
102 were in the EC-fair embryos, 127 were in the EC-poor embryos, 90 were rates among oocyte recipients. Further studies are needed to confirm our find-
in the LC-fair embryos, and 254 were in the LC-poor embryos. The blastomeres ings, in order to determine whether in the future oocyte donation programs
of DC embryos were unequal and no embryo was evaluated as good. The should also have a lower age limit.
blastocyst and high-quality blastocyst formation rates were significantly higher Trial registration number: -
(p<0.05) in the EC-fair embryos of the 3-cell group than in the other groups
(71.6% vs. 18.5%–57.8%, 44.1% vs. 5.1%–33.3%, respectively).
Limitations, reasons for caution: This study lacked data about implantation P-200 Blastocyst utilization rate – is it a matter of culture media?
rates after embryo transfer. To investigate the implantation ability of DC M. Schenk1, N. Reinschissler2, C. Fritz3, M. Hörmann-Kröpfl3,
embryos, it is necessary to increase the number of cases where data were G. Weiss4
analyzed after embryo transfer. 1
Das Kinderwunsch Institut Schenk GmbH, Research & Development, Dobl,
Wider implications of the findings: The EC-fair embryos of the 3-cell group Austria ;
had high blastocyst development ability for DC embryos. The results suggest 2
Das Kinderwunsch Institut Schenk GmbH, Research and Development, Dobl,
that the detailed evaluation of DC embryos at the early embryonic stage is not Austria ;
only predictive of embryogenic potential, but also useful for the selection of 3
Das Kinderwunsch Institut Schenk GmbH, Embryology, Dobl, Austria ;
embryos for early embryo transfer. 4
Das Kinderwunsch Institut Schenk GmbH, Research and Development, Dobl bei
Trial registration number: Not applicable Graz, Austria
P-199 The impact of the oocyte donor’s age on the recipient’s Study question: Is there a difference in blastocyst formation rates in embryos
outcomes: Should we exclude very young women from oocyte cultured with G-TLTM (Vitrolife) versus GM501 (Gynemed)?
donation? Summary answer: G-TLTM reveals a higher blastocyst formation rate com-
M. Roca Feliu1, E. Clua1, M. Tresánchez1, S. García1, N.P. Polyzos1, pared to GM501.
F. Martínez1 What is known already: A variety of studies have been carried out to com-
1
Hospital Universitari Quirón Dexeus, Obstetricia- Ginecología y Reproducción, pare the effects of different culture media types on embryo development, show-
Barcelona, Spain ing controversial results. A recent Cochrane review suggested that available data
are insufficient to conclude the best culture medium for embryo quality, preg-
Study question: Does the oocyte donor’s age have an impact on the live birth nancy and implantation. Besides the use of the medium, the day of embryo
rate (LBR) in the recipients? transfer is also under debate. A current meta-analysis showed no differences in
Summary answer: The LBR is significantly lower when donors are younger reproductive outcomes comparing blastocyst (d5) and cleavage-stage (d3)
than 25 years and, especially, when they are younger than 20 years. embryo transfer in clinical practice, however, the trend for a blastocyst transfer
What is known already: Donor’s age becomes one determining factor for is increasing since it resembles the natural process of implantation.
successful in oocyte donation programs (OD). It has been demonstrated that Study design, size, duration: The prospective study included 641 embryos
success is determined by embryo quality which depends on the oocyte quality from patients undergoing IVF (in vitro fertilization) and ICSI (intracytoplasmic
which in turn depends on the donor’s age. In most countries where OD is sperm injection) treatment. Patients with ≥ 6 MII oocytes were included in the
anonymous, law or guidelines establish that age must be between 18 and 35. study. Oocytes were splitted equally into the two media types GM501 and
Previous studies have shown that donor age < 25 was not associated with better G-TLTM, both overlaid with mineral-oil (Gynemed).
outcomes. However, no studies have separately analysed LBR in very young Participants/materials, setting, methods: The study was designed and
donors (<20 years) despite previous reports suggesting a higher aneuploidy rate conducted at the Kinderwunsch Institut Schenk GmbH (Dobl, Austria) in 2019.
in women between 18 and 25 years. Fertilized occytes from women aged 18 - 41, cultured in Embryoscope time-lapse
Study design, size, duration: A retrospective study of 3766 oocyte reception system (Vitrolife) were included while oocytes/embryos with polar body biopsy/
cycles performed between January 2009 and December 2018 was carried out. trophectoderm biopsy were excluded. The utilization rates of day 3 and day 5
Cycles using vitrified oocytes, PGD or TESE were excluded. embryos were analyzed.
Participants/materials, setting, methods: The cycles were categorized Main results and the role of chance: The number of achieved fertilized
considering the donors’ age groups: < 20, 20-25 and ≥26 years. The statistical oocytes was 308 in GM501 media and 333 with G-TLTM, respectively. Overall
tests used were Chi-square test for evaluating the differences in LBR and ANOVA the utilization rate was 45.7% using G-TLTM media compared to 36% in the
to test differences in fertilisation and embryo development rates as well as for GM501 group. A subgroup analysis of day 3 revealed a utilization rate of 13.6%
testing embryo quality. A generalized linear mixed model was applied to estimate with GM501, compared to 10.5% in the G-TLTM group. Additionally, the utiliza-
the odds for every endpoint. Patient was treated as random factor to avoid tion rate of day 5 embryos was 22.4% using GM501 versus 35.1% with G-TLTM.
repeated observations effect. Limitations, reasons for caution: The sample size may be seen as study
Main results and the role of chance: A total of 3766 oocyte reception limitation. However, statistically significant results were obtained. Nonetheless,
cycles were analysed. The pregnancy rate was 51.4% and the LBR was 40.7%. results should be confirmed with a higher sample size.
In 4.7% of the cycles the age of the donor was <20 years, in 41.1% was between Wider implications of the findings: The data show a significant better rate
20-25 years and in 54.2% was ≥26 years. When we analysed LBR according the of blastocyst utilization with G-TLTM media. Since around 5 days after fertilization

the uterus is particularly receptive, it is tempting to speculate that transferring P-202 Synchronizing blastocyst at hatching status in frozen
blastocysts in general is more favorable for a successful treatment outcome. embryo transfer cycles: A randomized controlled trial
Trial registration number: Not Applicable Y. Magdi Abd-Elkreem1, M.H. Zidan2, M.A. Ibrahim3, Y. Edris3,
A. El-Gohary3, A. Elattar4
P-201 supplementation of culture medium with melatonin 1
Al-Yasmeen Fertility and Gynecology Centerv, IVF Laboratory, Banha, Egypt ;
improves blastocyst development and reverses glucose 2
Gannah IVF Hospital, IVF Lab. director, Cairo, Egypt ;
intolerance in IVF mice 3
Faculty of Medicine- Benha University, Department of Obstetrics and Gynecology,
Y. Jia1, W. Liu1, D. Bai1, K. Li1, M. Chen1 Benha, Egypt ;
1 4
Center for Reproductive Medicine- Shanghai First Maternity and Infant Hospital, Al-Yasmeen Fertility and Gynecology center, . Department of Obstetrics and
Tongji University, Shanghai, China Gynecology, Benha, Egypt
Study question: Whether the adverse effects of in vitro fertilization (IVF) on Study question: Is culture of vitrified-warmed blastocysts for another day
glucose metabolism could be reversed by supplementation of culture medium after warming and selection of only blastocysts that could reach hatching status
with melatonin in mice? associated with improved outcomes?
Summary answer: Supplementation of culture medium with melatonin Summary answer: Culture blastocyst with ≤ 3grade of expansion for another
improved IVF blastocyst differentiation, reduced excessive weight gain after birth day after warming may be associated with improved implantation and ongoing
and normalized glucose intolerance in IVF male mice. implantation rates.
What is known already: Approximately more than 8 million children have What is known already: Failure of blastocyst hatching has been identified as
been conceived by assisted reproductive technologies worldwide. Increasing a potential obstacle limiting human implantation efficiency. On the other hand,
evidence suggests that IVF treatment may be associated with an increased risk discordance between the stage of transferred blastocyst and the receptive endo-
of developing obesity and metabolic diseases in adulthood. IVF mouse models metrium. We aimed in the present study to identify whether transferring frozen/
confirmed adult IVF offspring showed excessive weight gain on a high fat diet, thawed blastocysts at hatching status could improve the ongoing implantation
impaired glucose metabolism and cardiovascular dysfunction compared with rate in case of adjusting the thawing time to achieve this stage at the transfer day.
naturally conceived mice. Notably, in vitro culture impaired blastocyst develop- Study design, size, duration: This randomized controlled trial was conducted
mental kinetics and differentiation, independently of ovarian stimulation. at two private ART centers in Egypt, and involved cycles of vitrified-warmed
Moreover, it has been shown that addition of melatonin to culture medium may blastocyst transfer after ICSI. Recruitment of first participant was on April 25,
improve embryo development and prevent cardiovascular alterations in IVF mice. 2017, and the last participant was on May 10, 2018. A total of 469 women
Study design, size, duration: A cross-sectional mouse model was utilized, undergoing frozen embryo transfer (FET) were randomized into two groups.
wherein blastocysts were generated by natural mating (control group) or IVF Participants/materials, setting, methods: Randomization was performed
with or without melatonin (10-6M) (mIVF and IVF group respectively) in clinical 2 days prior to a scheduled FET. Participants received FET either with hatching
grade fertilization and culture media. Blastocysts were transferred to pseu- or hatched blastocysts achieved by incubation of ≤ 3 grade of expansion blas-
do-pregnant ICR females. Cell lineage allocation was assessed for 30-40 blasto- tocysts after thawing 18-24 hours (study group, n=232) or with ≤ 3 grade of
cysts from each group. Body weight, glucose tolerance, energy expenditure, expansion blastocysts incubated only 2-4 hours (control group, n=237).
hepatic gene expression was measured in 6-10 mice from each group. Main results and the role of chance: Baseline and treatment characteristics
Participants/materials, setting, methods: C57BL/6 females and DBA2 of the study population were balanced for the two study groups.
males were used to generate blastocysts. Cell numbers of inner cell mass (SOX2) The implantation rate (45.6% vs. 38.2%; OR 1.35, 95% CI 1.03 to 1.78) and
or trophectoderm (Cdx2+) were determined by immunohistochemistry. Males ongoing implantation rate (38.2% vs. 30.5%; OR 1.4; 95% CI, 1.06 to 1.87) were
were weaned at 3 weeks of age onto a chow diet or a high-fat diet (60% fat) for statistically significantly improved in the study group compared with the control
8 weeks. Glucose tolerance was assessed by an intraperitoneal glucose tolerance group. There was no significant difference in the rates of livebirth (47.4% vs.
test (2g/Kg). Energy metabolism was examined in metabolic cages. Hepatic gene 38.8%; OR 1.42, 95% CI, 0.98 to 2.05), clinical pregnancy (56% vs. 50.2%;
expression was measured by RNA-Seq and validated by qPCR. OR1.26, 95% CI, 0.88 to 1.82), ongoing pregnancy (50% vs. 42.6%; OR 1.35,
Main results and the role of chance: Blastocyst rates were similar in the CI, 0.94 to 1.94), and miscarriage (5.6% vs. 5.9%; OR 0.95, 95% CI, 0.44 to 2.06)
two IVF groups. Reduced inner cell mass, trophectoderm and total cell number in the study group compared with the control group. The multiple pregnancy
were observed in IVF blastocysts, compared with the control group (P<0.05). rate was similar in the study group (62/232; 26.7%) compared with the control
IVF pups had significantly lower birth weight and body weight before weaning, group (48/237; 20.3%; OR 1.44; 95% CI, 0.93 to 2.21; P=0.1).
but exhibited increased body weight and liver weight at 11 weeks of age com- Limitations, reasons for caution: The age of the study population was
pared with controls (P<0.05), independently of diet. IVF mice were glucose relatively young, thus we should also be cautious to extend the results to older
intolerant shown as an increased glucose area under the curve, and had decreased patients. Further large scale multicenter randomized controlled studies are
energy expenditure as well as a large number of differentially expressed genes required to confirm our findings.
related to metabolic pathways in the liver tissue compared with the control Wider implications of the findings: Prolonged culture of blastocysts with
group (P<0.05), independently of diet. The mIVF group showed intermediate ≤3grade of expansion may not only provide better embryo–endometrium syn-
cell numbers in blastocysts, body weight, liver weight and differentially expressed chrony, but also induces self-selection of the most viable blastocysts which have
hepatic genes compared with the IVF group and control group, and normalized the capacity to resume its development.
glucose tolerance and energy expenditure compared with the IVF group Trial registration number: NCT03128970
(P<0.05), independently of diet. Importantly, there was no significant difference
in glucose tolerance and energy expenditure between the mIVF group and the
control group. P-203 Successful pregnancies following transfer of blastocysts
Limitations, reasons for caution: The findings in C57BL/6J x DBA2 F1 mice derived from delayed-intracytoplasmic sperm injection
may not represent human infertility which is complex and often multi-factorial. (delayed-ICSI)
Further studies are needed to confirm these findings in human settings and J.P. Sam1, A.Y.X. Lim1, Z.Q. Tee1, C.S.S. Lee2
investigate the underlying mechanisms by which melatonin improves embryonic 1
Alpha IVF & Women’s Specialists, IVF laboratory, Petaling Jaya, Malaysia ;
development and adult metabolic phenotypes. 2
Alpha IVF & Women’s Specialists, Medical Director, Petaling Jaya, Malaysia
Wider implications of the findings: The data shows that melatonin miti-
gated the detrimental effects of in vitro fertilization and culture on mouse embryo Study question: To evaluate the clinical outcome of delayed-ICSI on
development and catch-up growth after birth, and reversed glucose intolerant day1-matured oocytes.
in adult male offspring. Therefore supplementation of culture medium with mel- Summary answer: Blastocysts derived from day1-matured oocytes can result
atonin may be beneficial for human IVF. in successful pregnancies and thus can be considered for transfer when there is
Trial registration number: not applicable no blastocyst available from day0-ICSI.

What is known already: Delayed-ICSI is defined as the ICSI procedure per- What is known already: Female fertility is inversely correlated to maternal
formed on oocytes matured via extended culture (day1-matured oocytes). It is age due to a depletion of the oocyte pool and a reduction in oocyte develop-
a routine procedure in Alpha IVF & Women’s Specialists in order to improve mental competence. Few studies have addressed the effect of maternal age on
cycle outcomes of selected patients, especially poor responders. the mature oocyte (MII) transcriptome, which is established during oocyte mat-
Despite the lower fertilisation, blastulation and blastocyst utilization rates of uration. However, reduced sample numbers have been analysed, and the bias
delayed-ICSI compared to day0-ICSI, the pre-implantation genetic testing for represented by distinct genetic backgrounds was difficult to control for. Here,
aneuploidies (PGT-A) revealed that delayed-ICSI blastocysts can be chromo- we characterise and compare the transcriptional patterns of a large cohort of
somally normal (euploid) and can therefore be considered for embryo transfer fully grown GV and IVM-MII oocytes from women of varying reproductive age,
(Lee, C.S.S., 2018). age can be considered as a continuous variable.
This abstract reports the successful pregnancies following the transfer of Study design, size, duration: In this basic research study 38 women were
delayed-ICSI blastocysts. recruited from May 2018 to June 2019. The mean woman age was 28.8 years
Study design, size, duration: A total of 370 IVF treatments at Alpha IVF & (SD=7.6, range 18-43). A total of 82 immature GV oocytes were included in
Women’s Specialists between January 2018 and December 2019 had delayed- the study as: GV (n=40), IVM-MII (n=32) or GV unable to mature after in vitro
ICSI done on their day1-matured oocytes (mean maternal age: 37.6; range 23.0- maturation (n=10). Their transcriptome was compared by single-oocyte RNA-
47.0). Indications for a delayed-ICSI includes immature oocytes retrieved on seq analysis.
day-0 (n=50); fail or abnormal fertilization on day0-ICSI (n=36); poor fertilization Participants/materials, setting, methods: Oocytes were collected either
on day0-ICSI (<50%) (n=11); low oocyte maturation rates (<50%) (n=8); and as GV or after in vitro maturation for 30 hours in G2TM medium, and individually
poor responders (≤6 oocytes injected for day0-ICSI) (n=265). frozen in lysis buffer until processed according to Smart-Seq2 single-cell RNA-seq
Participants/materials, setting, methods: The immature oocytes were protocol. The RNA-seq data obtained was analysed using the Seurat R package,
cultured in incubators for 18-24 hours and delayed-ICSI (PIEZO, Japan) was which has been recently developed for the quality control, analysis and explo-
performed on oocytes that were mature on day-1. Injected oocytes were cul- ration of single-cell RNA-seq data.
tured up to 7 days and utilisable blastocysts were vitrified (Cryotec, Japan) on Main results and the role of chance: Data quality control was applied and
Day5, Day6 and/or Day7 with/without trophectoderm biopsy. Cells biopsied samples containing >30% of mitochondrial DNA and less than 1,000 genes were
were amplified and screened using Next Generation Sequencing (NGS) filtered out, leaving 75 oocytes for further analysis. Preliminary exploration of the
(IonTorrent, USA). Selected blastocysts were thawed for frozen single blastocyst RNA-seq data using an algorithm for dimensionality reduction identified two clus-
transfer (SBT) and the clinical outcome was analysed. ters of oocytes according to their maturation stage (GV and IVM-MII) with 10430
Main results and the role of chance: Delayed-ICSI was done on 980 and 8744 detected genes, respectively; and independently of maternal age. To
day1-matured oocytes, of which 644 were 2PN (65.7%), 122 were >2PN identify differentially expressed genes according to maternal age, we set a cut-off
(12.5%), 136 were <2PN (13.9%) and 78 were degenerated (8.0%). The blas- at 35 years old, which is the accepted clinical definition of advanced maternal age.
tulation and utilisation rates (per 2PN) were 36.5% (235/644) and 19.9% Gene expression was analysed in GV and IVM-MII comparing both age’s groups
(128/644) respectively. Of the 128 utilisable blastocysts, 108 blastocysts had (<35 vs. ≥35 years old); however, in our preliminary analysis, no candidate gene
PGT-A screening and 26.9% (29/108) were chromosomally normal (euploid). has been identified. Taking advantage of our dataset, which contains oocytes rep-
At the time of writing, a total of 12 delayed-ICSI blastocysts (10 euploid, 1 mosaic resenting a continuous age range, we looked for genes that would show gradual
and 1 untested) were thawed (post-thaw survival rate=100%) for frozen SBTs change with age. Here, we considered the set of genes that do not change their
as no suitable day0-derived blastocysts were available for transfer (mean age: expression due to maturation stage (i.e. those which would be necessary to provide
36.0; range:31.0-42.0). Nine (9) had positive β-HCG reading and gestational developmental competence to the oocyte) and we independently analysed both,
sacs were seen in all nine patients (clinical pregnancy rate = 75.0%; implantation GV and IVM-MII. Although we found some transcripts with a tendency to increase
rate = 75.0%). Seven (7) patients have ongoing pregnancies ranging from 11-36 or decrease according to age, a deeper analysis is required.
weeks whereas 2 patients were known to have miscarried at the 8th and 11th Limitations, reasons for caution: GVs were collected after ovarian stimu-
week of pregnancy. lation and MII were in vitro matured. Therefore, their transcriptome might not
Limitations, reasons for caution: The immature oocytes were further cul- represent the in vivo GVs or MIIs.
tured up to 18-24hours in-vitro. The long incubation duration may result in Non-polyadenylated transcripts, which regulate gene expression and could be
post-mature oocytes that could affect blastocyst development. Further studies responsible for impaired oocyte developmental potential with age, are not
could employ other maturity assessment tools such as spindle visualization prior detected with our sequencing method.
to delayed-ICSI, rather than having a fixed incubation duration to optimise Wider implications of the findings: Our analysis suggests that advanced
delayed-ICSI outcomes. maternal age does not strikingly affect oocyte gene expression at GV or IVM-MII
Wider implications of the findings: Good pregnancy and implantation rates stages. Nonetheless, some genes showed a tendency to change their expression
can be obtained with blastocysts derived from delayed-ICSI. Therefore, patients with age. A deeper analysis is required to discriminate whether those changes
who are unlikely to have a good number of normally fertilized oocytes on day-0 have clinically relevant effects on reproductive outcomes.
should have their immature oocytes cultured to day-1 for delayed-ICSI to gen- Trial registration number: not applicable
erate more utilisable blastocysts.
Trial registration number: Not Applicable P-205 Non-invasive metabolomics analysis of spent culture media
predicts embryo viability.
P-204 Impact of maternal age and in vitro maturation on the S. Cabello1, H. Abdulla2, M.L. Seth-Smith1, M. Escriba3, J. Crespo3,
transcriptome of denuded human oocytes. S. Munné1, J.A. Horcajadas1
S. Llonch1, M. Barragán2, P. Nieto3, A. Mallol1, M. Elosua3, S. Ruiz3, 1
Overture Life SL, Scientific department, Madrid, Spain ;
F. Zambelli2, H. Heyn3, B. Payer1, R. Vassena2 2
Texas A&M University Corpus Christi, Chemistry, Corpus Christi, U.S.A. ;
1 3
Center for Genomic Regulation- CRG, Gene Regulation Stem Cells and Cancer, Juana Crespo Clinic, Embryology, Valencia, Spain
Barcelona, Spain ;
2
Clínica EUGIN, Research and Development, Barcelona, Spain ; Study question: Are there metabolites in the spent media that can act as
3
National Center for Genomic Analysis- CNAG, Single Cell Genomics Team, biomarkers to predict embryo implantation?
Barcelona, Spain Summary answer: A set of culture-media specific biomarkers were identified
that highly predicted implantation potential in several fertility centers.
Study question: Does maternal age affect the transcriptome of human oocytes What is known already: A critical step in IVF cycles is the selection of the
at the germinal vesicle (GV) stage or at metaphase II (IVM-MII) after maturation best embryo to be transferred, since treatment success strongly depends on
in vitro? this choice. The standard of care tool for embryo selection has been morphoki-
Summary answer: Oocytes’ gene expression is predominantly affected by netics criteria, which has low predictability to ascertain implantation potential.
maturation stage and less by maternal age. The effectiveness of ART remains limited and only 10%-30% of embryos

replaced in the uterus implant. During the last 20 years, the alternative has been supernumerary thawed/warmed embryos that have not been transferred in the
PGT (Preimplantation Genetic Testing), an invasive method that requires embryo present cycle. However, there is almost no study about the effect of re-vitrifi-
biopsy. For this reason, novel non-invasive embryo screening methods are cation on genetic and epigenetics profile in human embryos.
required to optimize embryo selection using AI-morphokinetics, metabolomics Study design, size, duration: A total of 30 ICSI-derived human embryos were
and non-invasive PGT. used to evaluate the effect of re-vitrification on miRNAs expression with real-time
Study design, size, duration: This retrospective study was done on 197 sam- PCR and DNA fragmentation with TUNEL assay. Embryos were donated from
ples collected in three different clinics, using two different culture media (Sage fertile couples referring for family balancing program. Donated embryos were
n=129, and Vitrolife n=68), over a period of two years. Sixty-nine of the spent cultured to blastocyst stage and high quality blastocyst (AA-AB) assigned to three
media samples came from embryos that implanted (P) and 128 from non-implanted groups: fresh, vitrified and re-vitrified. Scoring of blastocysts were according to
embryos (NP). Embryos were obtained and cultured using routine IVF practices. Gardner et al. (2012) grading system. embryos were collected from 2017-2019.
20 - 40 µL of spent media were collected after incubation between days 3 and 5. Participants/materials, setting, methods: Collected Embryos were vitrified
Participants/materials, setting, methods: Patients undergoing infertility on Cryotech carriers, with the method described by Kuwayama. After warming
treatment in 3 clinics were included in this study. Spent media was frozen at -20ºC of blastocysts and 4 hours culturing of them, embryos were used individually for
after collection. Metabolites were extracted from the spent media samples Simultaneous RNA extraction and cDNA synthesis. The relative quantification of
(20 µL) using an ultrafiltering approach to remove molecules >3KDa and then miRna expression and their target genes expression was carried out by real-time
run in a UPLC-Fusion Orbitrap MS/MS system which determined the abundance PCR. TUNEL assessment was done using of an in situ cell death detection kit TMR
of metabolites in each sample. Different statistical techniques were applied to red (Roche, Mannheim, Germany) based on the company’s instructions
reduce the huge number of metabolites found to the most informative ones. Main results and the role of chance: The results of this research showed
Main results and the role of chance: More than 5,550 metabolites were that the re-vitrification of human blastocysts did not affect the ability of their
identified and measured. re-expand in culture. A significant decrease in miR-16 and miR-let7a expression
A first analysis was performed with 129 Sage media samples -30 from pregnant in re-vitrified and vitrified group was observed compared to the fresh (p<0.05)
(P) and 99 from non-pregnant cycles (NP)-. A Sage-specific MPI (Metabolite one. On the other hand, a significant upregulation of the target genes integrin
Pregnancy Index) was built with the most informative metabolites to determine β-3 and BCL2 in the re-vitrified and vitrified embryos was observed comparing
pregnancy potential. The non-supervised analysis (not telling the software which with the fresh group (p<0.05). The expression of BAX as a pro-apoptotic gene
sample is pregnant and which is not) identified 100% for P and 63% of for NP. Using gene showed a significant decrease in re-vitrification group comparing with the
supervised analysis, 100% of P samples were identified and 81% of NP samples. fresh one (P<0.05). In TUNEL assessment, there was no significance difference
A second analysis was performed with 68 Vitrolife media samples from two in the vitrification and re-vitrification groups compared to the fresh respect to
different clinics. With the data from the first clinic samples (n=37: 20 P and 17 the total number cells and also the apoptotic cell rate (P > 0.05)
NP), a Vitrolife-specific MPI (Metabolite Pregnancy Index) was built with the Limitations, reasons for caution: Clinical investigations are necessary to
most informative metabolites to determine pregnancy potential. For the first evaluate the responses of embryos belonging to sub-fertile/infertile couples
clinic, the analysis showed an ability to predict embryo viability of 100% for P which may have less resistance potential to stressful situations and show a dif-
and 88% for NP samples. A blind analysis with a third batch of Vitrolife samples ferent response to re-vitrification.
from a second center (n=31: 19P+12NP) showed 78% of P samples were iden- Wider implications of the findings: This study showed that the re-vitrification
tified and 61% of NP samples. of embryos changes the expression of genes involved in embryo survival and
Limitations, reasons for caution: The study was retrospective, but for one implantation. therefore, it seems the embryos with high-quality show adaptability
of the centers it was blinded and still yielded high implantation predictability. and resistance toward stress which this response can include increased expression
Nevertheless, a prospective clinical trial is underway. A small limitation of the of anti-apoptotic proteins and genes involved in apoptosis and implantation.
test is that each culture media has a different subset of informative biomarkers. Trial registration number: not applicable
Other media are being validated.
Wider implications of the findings: This metabolomics test is non-invasive,
inexpensive, does not required any change in embryology protocol, and can be P-207 Interaction of the hippo signaling pathway during bovine
combined with NI-PGT and or morphokinetics, having the potential in itself or follicle activation and growth in vitro
in combination with other methods to significantly improve embryo selection N. Donfack1, M. Devos1, P. Vidal1, I. Demeestere2
without causing embryo damage. 1
Research Laboratory on Human Reproduction, Université Libre de Bruxelles,
Trial registration number: NA Bruxelles, Belgium ;
2
Research Laboratory on Human Reproduction, Université Libre de Bruxelles and
Obstetrics and Gynaecology- Erasme Hospital, Bruxelles, Belgium
P-206 Differential expression of microRNAs associated with
apoptosis and implantation in human in vitro-produced blastocysts
Study question: How does biochemical inhibition of the Hippo pathway affect
following re-vitrification
the activation and the growth of primordial follicles in vitro?
M. Daneshvar1, M. Movahedin1, M. Salehi2, M. Noruzinia3 Summary answer: MAPK/ERK signaling pathway seems to promote follicles
1
Tarbiat Modares University, Department of Anatomical Sciences- Faculty of activation during in vitro culture independently of the hippo pathway.
Medical Sciences, Tehran, Iran ; What is known already: The Hippo pathway has been identified as the major
2
Shahid Beheshti University of Medical Sciences, Cellular and Molecular Biology suppressor of tissue overgrowth. It plays a crucial role in follicular activation in
Research Center, Tehran, Iran ; a non-physiological condition. Recent studies showed that hippo disturbance
3
Tarbiat Modares University, Department of Medical Genetics- Faculty of Medical takes place during the tissue preparation and that, fragmentation-induced follicle
Sciences, Tehran, Iran growth is partially blocked by verteporfin.
Study design, size, duration: Bovine ovarian tissue fragments were exposed
Study question: Does re-vitrification affect on the expression of microRNAs to a control medium containing 0.1% of DMSO or 3 µM of Verteporfin, a
involved in apoptosis and implantation of human in vitro-produced blastocysts? pharmacological inhibitor of YAP, in vitro for 3 or 24 h, and cultured for addi-
Summary answer: re-vitrification could change the expression of microRNAs tional 3 days.
involved in apoptosis and implantation of human in vitro-produced blastocysts Participants/materials, setting, methods: Bovine ovaries were obtained
What is known already: Numerous studies have been carried out about the from a slaughterhouse. The ovarian fragments were analyzed at Days 0, 3h or
effect of vitrification on genetic and epigenetic changes in human embryos, and 3 days of culture. Cortex was either directly processed for immunohistological
nowadays Embryo freezing is among useful and safe infertility treatment tech- or western blot analysis (AKT, p-AKT, p-ERK1/2, RPS6, p-RPS6) or subjected
niques. Also re-vitrification of the same embryos after thawing has both theo- to follicular isolation for assessment of gene expression (CCN2, BIRC1, Kit
retical and applied cryobiological implications. sometimes, due to the Ligand, Last 1 et 2). The follicle number and stage were evaluated to assess early
embryologist’s discretion, re-vitrification can be applied for good quality follicular development. Survival was evaluated using TUNEL.

Main results and the role of chance: At least, in vitro culture for three days Wider implications of the findings: Our finding suggested that the time
promoted follicular activation independently of treatment, triggered by the needed for embryos to develop from 2PB to PNf may be used as a predictor
upregulation of p-ERK. However, the tissue fragments exposed to verteporfin for aneuploid embryo.
for 3 h presented lower follicular activation than control. As observed in other Trial registration number: not applicable
species, p-AKT and p-RPS6 decreased throughout the culture period (these
results will be confirmed by the ongoing PCR and immunohistochemistry). P-209 Embryo morphokinetics and static morphology in the
Moreover, verteporfin seems to not affect follicle survival when compared to prediction of live birth: evidences that speed is more important
control. than beauty.
Limitations, reasons for caution: Transport and impact of in vitro culture A. Bartolacci1, C. Moutier1, D. Turchi1, M. Lain1, E. De Ponti2,
may cover up the potential benefit of the Verteporfin on follicle activation. M. Mignini Renzini1, J. Buratini1,3, M. Dal Canto1
Wider implications of the findings: Our results showed that exposure to 1
Biogenesi - Reproductive Medicine Centre, Istituti Clinici Zucchi, Monza, Italy ;
verteporfin during fragmentation partially blocked follicular activation and sug- 2
ASST Monza, Department of Medical Physics, Monza, Italy ;
gested the contribution of MAPK/ERK signaling pathway on follicular activation 3
Sao Paulo State University, Institute of Biosciences, Department of Physiology
during in vitro culture.
Trial registration number: not applicable Study question: Is embryo morphokinetics more accurate than conventional
embryo morphology in the prediction of live birth following day 3 fresh embryo
transfer?
P-208 Extended time from second polar body extrusion to two Summary answer: Embryo morphokinetics was more accurate than conven-
pronuclei formation is related to aneuploid embryo formation tional embryo morphology in ranking embryos in terms of live birth potential
T.B. Hsieh1, P. Chang2, J. Meriano2, R.F. Casper2 following a day 3 fresh embryo-transfer.
1
Wayne State University, Obstetrics & Gynaecology, Detroit, U.S.A. ; What is known already: IVF/ICSI success critically depends on the accuracy
2
TRIO Fertility- University of Toronto, Obstetrics & Gynaecology, Toronto, Canada of embryo selection, which has been traditionally performed with the utilization
of subjective and static morphological criteria. Time-lapse technology has
Study question: Can morphokinetic parameters be used to predict embryo recently allowed a dynamic morphological evaluation, and, through the utilization
aneuploidy? of several morphokinetic parameters, the comparison of embryos’developmen-
Summary answer: An extended time period from the second polar body tal performance aiming for more accurate embryo selection and higher chances
formation (2PB) to two pronuclei formation (PNa) is associated with an increased of a live birth. Indeed, this technology has been already reported to increase
chance of aneuploid embryo. accuracy of embryo selection, with positive impacts on pregnancy rates.
What is known already: Preimplantation genetic testing for aneuploid (PGT- Nevertheless, most IVF clinics still perform embryo selection based on static
a) is a useful technique to enhance the implantation rate and decreases the morphological criteria.
miscarriage rate, particularly in advanced female population. Without question, Study design, size, duration: This is a retrospective study including patients
PGT-a is an invasive procedure with potential harmful effects to the fetus. Time in their first ICSI cycle with embryo culture in a time-lapse system, performed
lapse microscopy (TLM) provides valuable morphokinetic parameters or mor- from January 2015 to June 2018. 828 embryos from 461 fresh embryo transfers
phological criteria for selecting better developed embryos to be transferred to (SET: n= 156; DET: n=305) were included in this study, in which each individual
improve implantation rate. However, current parameters still are not accurate embryo was evaluated for both morphological and morphokinetic quality.
enough to predict embryo ploidy or implantation potential. Patients giving birth to a single child after a double embryo-transfer were
Study design, size, duration: This is single center, retrospective cohort study excluded from the analysis.
conducted in a university affiliated fertility institute. A total of 1515 biopsied Participants/materials, setting, methods: The primary end-point was live
blastocysts were examined from 255 intracytosplasmic sperm injection (ICSI) birth. A generalized estimation equation was utilized to control for dependencies
cycles (female age, 23-48; average 37.64.1). We collected embryo records with within individuals, since some patients received two embryos at transfer. The
complete TLM recording from the time of ICSI until blastocyst stage and a day performance characteristics of the morphokinetics model and conventional
5/6 trophectoderm biopsy result for analysis. PGT-a results were obtained by embryo morphology was calculated using a ROC curve. Nine subgroups were
using either array comparative genomic hybridization or next generation created crossing three morphology degrees (A, B, C) with three morphokinetics
sequencing. degrees (A, B, C) attributed to each embryo, and live-birth rates were compared
Participants/materials, setting, methods: Morphokinetic parameters between these subgroups.
includes tPB2(from insemination to 2PB), tPNa (from insemination to 2PN, tPNf Main results and the role of chance: For each morphokinetic parameter, an
(from insemination to pronuclei faded), T2-8 (from insemination to 2 to 8 cells), optimal range was defined combining the two quartiles presenting the highest
tM (from insemination to morula), tSB (from insemination to blastulation), tB live-birth rate. The parameters to be inserted in the model were selected by logistic
(from insemination to full blastocyst), tEB (from insemination to expanded blas- regression, and were: t4 [OR = 2.04 (95% CI 0.87 – 4.77)] and t2 – tPNf [OR =
tocyst), CC2 (from 2 to 3 cells), and CC3 (from 3 to 5 cells). Morphokinetic 1.52 (95% CI 0.99 – 2.30)]. The resulting morphokinetic model was more accurate
parameters were analyzed with Mann-Whitney-U test. P-values less than 0.01 to predict live birth than conventional embryo morphology, as measured by AUC
were designated significant. on an external dataset not used for model development [AUC 0.658 (95% CI
Main results and the role of chance: Of the total 1515 embryos, 1153 0.606 – 0.681) and AUC 0.575 (95% CI 0.537 - 0.614), respectively]. Regression
embryos were reported euploid and 362 of them were aneuploid embryos with analysis showed a significant difference between the three subgroups in which
an embryo aneuploid rate of 31.4%. 456 of the embryos also underwent TLM morphokinetic quality was classified as poor (C; subgroups AC, BC, and CC)
examination. 317 of these embryos were reported euploid and 139 of them compared to the reference group AA (adjusted p < 0.001). Moreover, odds ratio
aneuploid by PGT-a with embryo aneuploid rate of 43.8%. From the original obtained in subgroups AC (good morphology/poor morphokinetics) and CA
Morphokinetic parameters, we further calculated time from 2PB to PNa (poor morphology/good morphokinetics) indicated that poor morphokinetics is
(2PB-PNa), PNa to PNf, and PNf to T2 as new parameters to compare with a better marker of low embryo competence than poor morphology (OR = 0.23;
the PGT-a results. The analysis of morphokinetic parameters and embryo ploidy 95% CI 0.09-0.60 and OR=0.55; 95% CI 0.17-1.87, respectively).
status showed that PB2-PNa were correlated with an increased rate of aneuploid Limitations, reasons for caution: We acknowledge that our study is limited
embryos. Aneuploid embryos have significantly longer time from 2PB to PNa by its retrospective nature and by the utilization of data generated in a single IVF
(euploid 2PB-PNa: 4.2 1.8 hours, aneuploid: 5.0 2.18 hours). The other param- center including only day 3 transfers.
eters didn’t show any statistically significant differences. Wider implications of the findings: Our results suggest that embryo mor-
Limitations, reasons for caution: The data variation is wide. These data phokinetics is a more reliable predictor of live birth potential after day 3 fresh
are single center, sample size of this study is small and it is a retrospective study. embryo transfers than embryo static morphology, thus providing valuable ref-
In addition, more scientific and strict criteria to define 2PB, PNa, and PNf may erences to define embryo selection strategies in IVF practice.
be able to be established. Trial registration number: Not applicable

P-210 The impact of synchronicity of blastomere division during 1

ProcreaTec, IVF Lab, Madrid, Spain ;
dynamic human embryo monitoring by a time-lapse on blastocyst 2
INIA, Animal Reproduction Department, Madrid, Spain ;
development and quality. 3
University College Dublin, School of Agriculture and Food Science-, Belfield-
N. Sayme1, M. Kljajic2, D.H.A. Maas1, T. Krebs1 Dublin, Ireland ;
4
1 Barcelona Institute of Science and Technology, Centre for Genomic Regulation,
Team Kinderwunsch Hannover, Dept. of Ob/Gyn, Hannover, Germany ;
2 Barcelona, Spain
Universitätsklinikum des Saarlandes, Reproduktionsmedizin, Homburg, Germany
Study question: Does cumulus cell transcript abundance correlate with oocyte
Study question: Does the synchronicity of blastomere division of embryo
developmental potential in the bovine model?
and type of media system have an impact on blastocyst development and quality.
Summary answer: The abundance of specific transcripts in cumulus cells from
Summary answer: The synchronicity of the cell cycle affects blastocyst devel-
oocytes developing to blastocysts following IVF differs from those arresting
opment and blastocyst quality, while the media system only affects blastocyst
before that stage.
quality.
What is known already: Cumulus cells serve essential functions during fol-
What is known already: The serial image capturing with time-lapse devices
liculogenesis, being connected to the oocyte through transzonal projections.
is a noninvasive method that offers the possibility for 24-hour continuous mon-
Following in vitro maturation, cumulus cells are often removed and discarded,
itoring of embryo development, increasing the quantity and quality of informa-
thereby constituting an interesting biological material on which to perform molec-
tion without disturbing the culture conditions. Some of the most recent
ular analysis aimed to predict oocyte developmental potential. Previous studies
publications show correlations between cleavage times until 8-cell stage and
performed to correlate the abundance of specific transcripts in cumulus cells
blastocyst rate, expansion, and implantation. These published data suggest that
with their developmental potential have yielded conflicting results. These studies
morphokinetic observations can yield valuable information to aid the selection
have been based on candidate gene qPCR or microarray analyses. RNAseq
of embryos for transfer. The impact of single versus sequential culture media
provides significant advantages over these methods as it’s more sensitive and
systems on time-lapse data also needs to be considered since the contradictory
specific and it is not limited to a subset of genes.
conclusions were reached recent studies.
Study design, size, duration: Bovine cumulus cells were allocated into three
Study design, size, duration: The synchrony of the second cell cycle (s2)
groups according to the developmental potential of the oocyte: 1) oocytes
was defined as the duration of the transition from a two-blastomere embryo to
developing to blastocysts following IVF (Bl+Cl+), 2) oocytes cleaving following
a four-blastomere embryo (s2 = t4–t3), while synchrony of the third cell cycle
IVF but arresting their development prior to the blastocyst stage (Bl-Cl+), and
(s3) as the duration from a five-blastomere embryo to an eight-blastomere
3) oocytes not cleaving following IVF (Bl-Cl-). RNAseq was performed in 4
embryo (s3 = t8–t5). The injection time of ICSI was designated as “time zero”
(Bl-Cl-) or 5 samples (Bl+Cl+ and Bl-Cl+) per group. Each sample contained
(t0), and computer software was used to calculate the time duration of the cycle.
cumulus cells from 10 cumulus-oocyte complexes (COCs).
Participants/materials, setting, methods: A total of 87 blastocysts from
Participants/materials, setting, methods: Bovine cumulus-oocyte com-
20 patients undergoing the antagonist cycle for ICSI treatment between
plexes were obtained from slaughtered cattle and individually matured in vitro
November 2019 and December 2019 were evaluated. All blastocysts were
(IVM). Following IVM, cumulus cells were removed by hyaluronidase treatment,
cultured in Embryoscope™ according to the manufacturer’s specifications
pelleted, snap frozen in liquid nitrogen and stored at -80 ºC until analysis.
(Vitrolife, Sweden). The Gardner and Schoolcraft scoring system was used to
Cumulus-free oocytes were fertilized and cultured in vitro individually and devel-
describe blastocyst quality. Correlations between the data were calculated using
opment was recorded for each oocyte. RNAseq was performed at Illumina
logistic regression analysis. Statistical significance was defined as p<0.05. All
platform with >30 M reads/sample. Differential expression was analysed by
statistical analyses were performed using SAS software.
DESeq2 software.
Main results and the role of chance: Morphokinetic data showed that
Main results and the role of chance: Although the abundance of most tran-
synchrony of the second cell cycle was significantly different (s2, p<0,05)
scripts detected in cumulus cells (19335) did not vary depending on the develop-
between embryos which reached the blastocyst stage and those embryos that
mental potential of their corresponding oocytes, RNAseq revealed 1609, 1466
did not. Although, in contrast with what was previously published, the more
and 1420 differentially expressed genes (DEGs) for the comparisons Bl+Cl+ vs.
careful analysis revealed that blastocyst quality was not affected by synchrony
of the second cell cycle as morphocinetic parameter. On the other hand, syn- Bl-Cl+, Bl+Cl+ vs. Bl-Cl- and Bl-Cl+ vs. Bl-Cl-, respectively, using a raw p value
<0.05. These DEGs were narrowed down to 77, 80 and 32 DEGs for the com-
chrony of the third cell cycle did not significantly correlate with blastocyst for-
parisons Bl+Cl+ vs. Bl-Cl+, Bl+Cl+ vs. Bl-Cl- and Bl-Cl+ vs. Bl-Cl-, respectively,
mation potential, but this parameter showed a significant correlation with
when an adjusted p value <0.05 was used. From these subsets of DEGs, 49, 50
blastocyst quality (s3, p<0,001).
and 18 DEGs, respectively, exhibited a fold change greater than 1.5. Focussing on
Besides Morphokinetic observations, the impact of single versus sequential
DEGs in cumulus cells obtained from oocytes developing to blastocysts, 10 DEGs
culture media systems was investigated as well. For a single media system
were common to both comparisons (10/49 from Bl+Cl+ vs. Bl-Cl+, 10/50 from
Gynemed 501CT (Lensahn, Germany) was used and for the sequential one
Bl+Cl+ vs. Bl-Cl-). These DEGs correspond to 6 genes upregulated (HBE1, ITGA1,
Sydney IVF Cleavage Medium and Sydney IVF Blastocyst Medium (Cook Medical,
PAPPA, AKAP12, ITGA5 and SLC1A4), and 4 genes downregulated (GSTA1,
Ireland) were used. Obtained results showed that there was not any difference
PSMB8, FMOD and SFRP4) in Bl+Cl+ compared to the other groups.
between the synchrony of the second or third cell cycle regardless on the media
Limitations, reasons for caution: Experiments were conducted in the
system. What is more, blastocyst formation potential, blastocyst quality or preg-
bovine model. Although bovine folliculogenesis, monoovulatory ovulation and
nancy rate did not differ between media system groups.
early embryo development exhibit considerable similarities with that of humans,
Limitations, reasons for caution: The disadvantage of the relatively small patient
caution should be taken when extrapolating these data to humans.
cohort was balanced with a wider range of patients’ age. Further research should
Wider implications of the findings: Selecting embryos with the highest
link morphokinetic parameters with pregnancy rate and live birth rate as well.
developmental potential is essential in order to achieve successful outcomes
Wider implications of the findings: The potential of the present findings
following single-embryo transfer. Our findings suggest that the abundance of
might be helpful for a better understanding of the association between embryo
specific transcripts in cumulus cells could be used as a predictor of the develop-
morphokinetic parameters and blastocyst development. Furthermore, the cul-
mental potential of their enclosed oocyte.
ture media system result indicates that morphokinetic benchmarks can be devel-
oped and used irrespective of a laboratory’s choice of culture media.
P-212 Epigenetic changes with maternal age in human eggs follow
P-211 RNAseq reveals cumulus cell transcripts associated with an inverse U-curve shape
developmental potential of bovine oocytes T. Imbar1, G. Karavani2, P. Pash3, N. Shecter-safrai2, M. Safrai2, T.
Á. Martínez Moro1,2, I. Lamas-Toranzo2, E. OĆallaghan3, Mordechai-daniel2, D. Bauman2, E. Reich3, R. Rothman3, T. Ajiro3,
A. Esteve-Codina4, P. Lonergan3, P. Bermejo-Álvarez2 M. Klutstein3

1
Hadassah-Hebrew University Medical Center, Fertility preservation center- IVF Summary answer: Traditional supervised and unsupervised ML methods can
unit, Jerusalem, Israel ; predict human embryo viability using a small number of time-lapse videos.
2
Hadassah-Hebrew University Medical Center, Fertility preservation center. IVF What is known already: There is insufficient evidence to conclude that the
unit, Jerusalem, Israel ; introduction of time-lapse technology has improved live birth outcomes after
3
Hebrew University, Institute of Dental Sciences, Jerusalem, Israel ART compared with traditional embryo assessment. ML methods have shown
to have much potential in analyzing images and may have predictive value in the
Study question: How do epigenetic modifications change in human MI oocytes assessment of embryos from time-lapse videos. All recent work uses some
collected from media of unstimulated ovarian pieces with aging? variation of deep neural networks that are known to require a lot of training
Summary answer: H3K9me2 repressive epigenetic mark in human oocytes data and are not easily interpretable. Thus, we propose to use interpretable and
may follow an inverse U-curve with age in a similar manner to the behavior of less data-demanding ML methods to predict embryo outcome.
aneuploidies. Study design, size, duration: We used eight time-lapse videos of embryo
What is known already: Natural fertility in humans follows an inverse development, up to day five. Two labels were assigned to the videos; positive
U-curve, where young females and women of advancing maternal age show for the embryos which resulted in a live birth and negative for the non-viable
reduced fertility rates. The mechanism or mechanisms that shape the fertility embryos. The dataset was evenly balanced with an equal number of positive
curve are unclear. A recent report shows that oocyte aneuploidies also follow and negative labels. 120 frames were extracted at evenly distributed intervals
such a U-curve, suggesting that chromosomal errors originating in oocytes deter- throughout the entire duration of each video in the dataset.
mine the behavior of natural fertility in humans. Epigenetic changes are consid- Participants/materials, setting, methods: Global image features (GF) were
ered as one of “hallmarks of aging”. These typically cause a decline of repressive extracted from each video frame. GF have the advantage of being manually
marks in repressed loci and a contaminant repression of active genes. This phe- engineered, which are more interpretable when compared to features extracted
nomenon, previously uninvestigated in human eggs, is termed “epigenetic aging”. through deep learning. We extracted 25 different features, where the best per-
Study design, size, duration: After IRB approval, all participants, undergoing forming features were Tamura and AutoColorCorrelogram. Due to the small
fertility preservation procedures, signed informed consent. From January 2019 number of real time-lapse videos, Synthetic Minority Over-sampling Technique
until October 2019, we searched and collected oocytes from media remains (SMOTE) was used to increase the sample size. Leave-one-embryo-out cross-val-
after ovarian cortex pieces cryopreservation. Eight women included in the study. idation was used to demonstrate generalizability and avoid overfitting.
Age ranged from nine to 30 years. Two women received chemotherapy prior Main results and the role of chance: The experiments can primarily be
to fertility preservation procedure. We used a cohort of immature oocytes split into two distinct groups; supervised and unsupervised ML methods. The
stained after retrieval from fertility age IVF patients as a control. supervised experiments used the algorithms naïve Bayes and random forests
Participants/materials, setting, methods: Medium solution obtained from and evaluated using leave-one-embryo-out cross-validation with and without
handling of ovarian cortical stripes, containing oocytes of different sizes and SMOTE (1000% oversampling for both classes). The unsupervised experiments
shapes together with cumulus cells. We separated different types of oocytes in were done using simple K-means clustering (using two clusters) and X-means
the solution using a modified Andreasen pipette technique based on Stokes law (number of clusters chosen automatically) with and without oversampling. The
of sedimentation. Oocytes found in the media were matured in vitro to MI and motivation behind choosing more traditional algorithms over modern ones, such
immuno-stained for the histone modification H3K9me2 . as deep learning, was due to their efficiency in terms of training and inference
Main results and the role of chance: Using our oocyte identification method time. Furthermore, the methods used are more interpretable and allow for a
we isolated oocytes that were not seen in the routine embryology lab medium higher understanding of why and how the algorithms make a prediction. The
search. After maturation in vitro we identified 3 GV , 23 MI and 6 MII oocytes. supervised methods using SMOTE performed well and classified all eight videos
MI oocytes staining for epigenetic histone modification H3K9me2 revealed a correctly (sensitivity and specificity of 1). Without SMOTE, specificity and sen-
low staining intensity in the younger patients (under 10 years) with a marked sitivity fell between 0.375 up to 0.750 depending on which feature was used.
increase in the more mature, post-menarche (over 14 years), patients. Staining The unsupervised methods also performed well. K-means with SMOTE correctly
of MI oocytes from women 30 years and older showed and even lower intensity clustered all videos into the correct clusters (live birth and not live birth).
of this marker. Staining of the few GV oocytes we retrieved corroborated this Without SMOTE, one video was wrongly clustered. X-means automatically
staining pattern. Thus, the intensity of the H3K9me2 repressive epigenetic generated two clusters and classified all instances correctly using SMOTE.
marker in human oocytes follows the shape of an inverse U-curve. Without SMOTE, X-means was able to cluster about 60% correctly.
Limitations, reasons for caution: Staining results are from human oocytes Limitations, reasons for caution: Since the size of the dataset was minimal,
during fertility preservation procedures, without gonadotropin induction. leave-one-embryo-out cross-validation and SMOTE were essential in the vali-
Moreover, the collection of oocytes was performed following a meticulous dation of this study. For future studies, larger datasets should be used with data
search and isolation of oocytes in the clinical lab. Thus, the epigenetic status may collected from different sources to increase generalizability and the robustness
not reflect only their age, but also additional factors such as cellular stress. of the validation.
Wider implications of the findings: Epigenetic changes involves in human Wider implications of the findings: Supervised ML methods have a potential
oocyte maturation and may be part of the U-curve natural fertility seen in for live birth prediction. Unsupervised learning may become an essential tool for
humans. Manipulating this effect, using epigenetic drugs, might enhance and the discovery of new embryo features for use in assessments. Our results suggest
improve our ability to preserve mature oocytes from young women with urgent that Tamura features might be visual biomarkers for embryo video analysis.
need of fertility preservation Trial registration number: not applicable
P-213 Big data is not always better – prediction of live birth using P-214 Ooplasmic DNA Repair Mechanisms of the Human Gamete
machine learning on time-lapse videos of human embryos Appear to Be Linked to Oocyte Maturity
S. Hicks1,2, T.B. Haugen3, M. Iliceto3,4, H.L. Hammer1,2, D. Keating1, A. Parrella1, Z. Rosenwaks1, G.D. Palermo1
1
J.M. Andersen3, O. Witczak3, M.A. Riegler1, M.H. Stensen4 Weill Cornell Medicine, The Ronald O. Perelman and Claudia Cohen Center for
1
Simula Metropolitan Center for Digital Engineering, Holistic Systems, Oslo, Reproductive Medicine, New York, U.S.A.
Norway ;
2
OsloMet – Oslo Metropolitan University, Faculty of Technology- Art and Design, Study question: Does ooplasmic maturity affect the ability of MII oocytes to
Oslo, Norway ; repair fragmented male genome DNA?
3
OsloMet – Oslo Metropolitan University, Faculty of Health Sciences, Oslo, Norway ; Summary answer: If the proportion of mature oocytes retrieved is not opti-
4
Fertilitetssenteret, IVF Laboratory, Oslo, Norway mal, the oocytes are unable to overcome abnormal sperm chromatin fragmen-
tation (SCF) and generate implantable embryos.
Study question: Can interpretable machine learning (ML) methods be used What is known already: It is known that following sperm penetration, the
on small amounts of data to predict human embryo viability? ooplasm replaces the protamines of the spermatozoon with histones and runs

through the male genome, repairing DNA for accuracy and eventual repair. This important for the neo-synthesis of another proteins important for embryo devel-
is particularly relevant in cases of high SCF, known to affect implantation. The opment, It was known that autophagy is highly activated immediately after fertil-
achieved nuclear maturation by extrusion of the first polar body, not concur- ization, and also found that fertilized embryos lack autophagy can not implant.
rently, is followed by ooplasmic readiness. We have shown previously that pro- Study design, size, duration: Our study is a prospective study, 60 embryos
portional nuclear maturity of the retrieved oocyte cohort has a crucial impact were derived from oocytes aspirated from twelve patients undergoing ICSI
on ICSI outcome in terms of fertilization, embryo implantation, and live birth. between September 2019 to November 2019 at Madina fertility center.Embryos
Study design, size, duration: This study included couples treated by ICSI were devided on day 5 in to two groups according to its developmental stage
using ejaculated spermatozoa between 2006 and 2019. Female partners were and its ability to form blastocyst:
limited to ≤37 years of age to control for female-related gamete aneuploidy.
Nuclear maturity of the retrieved oocyte cohort was assessed after the removal Group A ----> Developed embryos (blastocyst formation) n=31.
of cumulous cells. Cycles were allocated according to the proportion of nuclear Group B ----> Arrested embryos (No blastocyst formation) n=29.
maturity of the retrieved cohort: optimal (≥80%) and suboptimal (<80%).
Participants/materials, setting, methods: Ejaculate SCF was assessed by Participants/materials, setting, methods: Oocytes aspirated from twelve
terminal deoxynucleotidyl dUTP transferase nick-end labeling (TUNEL). Five- patients were injected with sperm by conventional ICSI , the injected oocytes
hundred spermatozoa were assessed with a normal threshold of ≤15%. Female were incubated till day 5.
patients were stimulated for ovarian superovulation by pituitary suppression Embryos on day 5 were scored and devided in to two groups according to its
utilizing either gonadotropin-releasing hormone antagonist or agonist, were developmental stage and its ability to form blastocyst:
treated with daily gonadotropins, and were triggered with human chorionic
gonadotropin when the lead follicle reached ≥17mm. Oocyte retrieval and ICSI Group A ----> Developed embryos (blastocyst formation)
were performed in the standard fashion. All cycles were performed at our center. Group B ----> Arrested embryos (No blastocyst formation)
Main results and the role of chance: A total of 126 couples underwent
220 ICSI cycles that had an optimal oocyte maturity, and 119 couples underwent then biopsied to evaluate expression of autophagic genes (ELISA) and proteins
202 cycles with suboptimal oocyte maturity. The average SCF for couples with (RT-PCR) : LC3, PI3K, E2F, mTOR.
normal and abnormal levels was 9.4±3% and 24.1±10%, respectively (P<0.0001). Main results and the role of chance: Developed embryos to blastocyst
When oocyte maturity was optimal, couples with normal and abnormal SCF stage on day 5 (Group A) shows significantly higher LC3 relative gene expression
had comparable fertilization (74.0% vs. 70.8%), embryo implantation (22.2% vs. (1.12±0.51), Beclin 1 relative gene expression (1.43±0.33) and Beclin protein
22.3%), and clinical pregnancy rates (CPR; 32.8% vs. 37.9%). expression (3.8±0.028) than their expression in embryos that failed to form
For the suboptimal maturity group, couples with normal SCF had a fertilization blastocyst on day 5 (Group B) (0.72±0.17,P=0.03), (0.35±0.12,P=0.0001),and
rate of 72.4%, an embryo implantation rate of 20.2%, and a CPR of 30.5%. (3.14±0.05, P=0.0001), respectively.
Couples with abnormal SCF had a comparable fertilization rate of 72.0%; how- While mTOR and PIK3C3 proteins expression were significantly higher in
ever, the implantation rate was compromised (9.8%; P<0.05). Similarly, in these Group B (arrested embryos) than their expression within developed embryos
couples, CPR trended lower (30.5% vs. 17.8%). (Group A) , P=0.007 and P= 0.0001 ,respectively.As well as the expression of
To confirm the impact that proportional oocyte maturity has on the alleviation E2F gene which is significantly lower within group A embryos (0.32±0.07) and
of male genomic defects, we assessed only cycles with abnormal SCF. In cycles remarkedly higher within group B embryos (4.38±1.16), P=0.0001.
where the proportion of MII oocytes was optimal, the fertilization rate was
70.8%, the implantation rate was 22.3%, and the CPR was 37.9%. Cycles with
suboptimal maturity had a comparable fertilization rate (72.0%); however,
Developed Arrested
embryo implantation (9.8%; P<0.05) and CPRs were impaired (17.8%; P<0.05). P
(Group A) (group B)
Limitations, reasons for caution: While we limited female age to ≤37 years
to control for female-related gamete aneuploidy, it is not possible to exclude all
mTOR (ng) 1.13±0.053 1.23±0.09** 0.007
confounding factors with certainty. This study demonstrated that not only nuclear
but also cytoplasmic maturity is required for a competent MII oocyte. Beclin (ng) 3.8±0.028 3.14±0.05*** 0.0001
Wider implications of the findings: These findings confirm the existence PIK3C3 (ng) 18.6±1.4 24.5±1.5*** 0.0001
of DNA repair mechanisms within the human oocyte and that this function is
LC3 relative gene expression fold 1.12±0.51 0.72±0.17* 0.03
impaired in oocytes with ooplasmic dysmaturity. This was evident in the retrieved
cohort with a suboptimal number of MII oocytes. Optimization of the super- Beclin 1 relative gene expression 1.43±0.33 0.35±0.12*** 0.0001
ovulation protocol can overcome male genomic impairment. fold
Trial registration number: not applicable E2F relative gene expression fold 0.32±0.07 4.38±1.16*** 0.0001
P-215 The role of autophagy on embryonic development at

blastocyst stage
N. Adel1, H. Elmaghraby2, D. Ghareeb1, M. Elmahdy2, M. Nabil3 Limitations, reasons for caution: larger sample size is needed to support
1
our results.
Alexandria university- Faculty of science, Biochemistry, Alexandria, Egypt ;
2
Wider implications of the findings: Investigation of autophagy function (spe-
Alexandria university- Faculty of medicine, Obstetrics and Gynecology, Alexandria,
cially evaluation of expression of Beclin 1 and LC3 genes and proteins) in embryos
Egypt ;
3
at late developmental stage introduce a new diagnostic tools for embryos with
City of scientific researches and technological applications., Pharmaceutical and
impaired development and those with impaired implantation potential.
fermentation industries development center, Alexandria, Egypt
Study question: Does autophagy genes and proteins expression is deifferent
between preimplantation embryos that developed to blastocyst and those which P-216 The influence of post-thaw culture period on the pregnancy
are arrested on day 5? outcomes of poor-quality blastocyst transfer
Summary answer: From all autophagy genes and proteins, Beclin 1 and LC3 S.H. Shin1, J.H. Cha1, Y.J. Kim1, S.K. Lee1, J.H. Kim1, D.H. Kim1,
expression is important for embryonic development to blastocyst on day 5. H.Y. Kim1, S.H. Back1, M.J. Choo1, J.H. Ahn1, H.Y. Kim1, K.A. Pak1,
What is known already: Autophagy is one of the molecular mechanisms that J.S. Yoon1, S.Y. Park1
is important for the early embryonic development. It is a massive cytoplasmic 1
Agaon Fertility Clinic, IVF center, Seoul, Korea- South
degradation pathway mediated by the lysosome.
Autophagy provides a“recycling system”, As it engulfs unnecessary macromolecules Study question: Does post-thaw culture period affect the pregnancy outcomes
proteins and organelles, degraded them to amino acids afree fatty acids which are of poor-quality blastocyst in single vitrified-thawed embryo transfer?

Summary answer: In poor-quality blastocyst transfer, short post-thaw culture Main results and the role of chance: ICM Grade A embryos (N=2214)
period was higher the pregnancy outcomes compared to long post-thaw cul- were euploid in 57.1% of cases versus 45.5% and 31.2%, respectively, for ICM
ture period. Grade B (N=1090) and Grade C (N=404), (P<0.0001). TE grading resulted
What is known already: During vitrification and thawing, embryo can be similarly for euploidy (Grade A [N=1551] 62.0% vs. Grade B [N=1627] 48.3 %
damaged and stressed. Stressed embryo can be particularly sensitive to envi- vs. Grade C [N=530] 26.2%, P<0.0001). Neither ICM (P=0.257) nor TE
ronmental influences during post-thaw culture. Therefore, it raised hypothesis (P=0.079) correlated with gender.
that the post-thaw culture period may be affect the pregnancy outcomes. In transferred, euploid embryos, the distribution included ICM Grade A (N=392),
Study design, size, duration: A retrospective cohort study of 428 vitri- Grade B (N=116), Grade C (N=31) and TE Grade A (N=302), Grade B
fied-thawed single embryo transfer cycles was performed from January 2014 to (N=209), Grade C (N=28).
October 2019. Cycles with surgical retrieved sperm, genetic diagnosis, oocyte Implantation rate (IR) (ICM Grade A 71.7% vs. Grade B 59.5% vs. Grade C
donation, and advanced maternal age (≥38 years) were excluded. We compared 48.4%, P<0.0001), clinical pregnancy rate (CPR) (ICM Grade A 69.4% vs. Grade
rates of survival, biochemical pregnancy, clinical pregnancy, ongoing pregnancy, B 57.8% vs. Grade C 45.2%, P=0.0034), and ongoing pregnancy rate (OPR)
and implantation according to post-thaw culture period. (ICM Grade A 59.7% vs. Grade B 48.3% vs. Grade C 38.7%, P=0.013) are higher
Participants/materials, setting, methods: Cycles, that transferred single with better ICM grades.
embryo of poor-quality, were divided into two groups according to post-thaw TE grading resulted: IR (TE Grade A 73.8% vs. Grade B 63.2% vs. Grade C
culture period; cultured overnight (21-23 h) in group A (n = 281) and short 35.7%, P<0.0001), CPR (TE Grade A 71.5% vs. Grade B 60.8% vs. Grade C
culture (3-5 h) in group B (n = 147). Blastocyst was graded according to the 35.7%, P=0.0001), and OPR (TE Grade A 61.6% vs. Grade B 51.7% vs. Grade
Gardner and Schoolcraft’s classification system. Poor-quality was defined as C 28.6%, P=0.0009). ICM and TE do not correlate with miscarriage, chemical,
excluding expansion grade 4-6 and ICM/trophectoderm grade A or B. or ectopic pregnancy.
Main results and the role of chance: Cycle characteristics such as female Limitations, reasons for caution: Limitations of this study include a patient
age (33.4 ± 2.5 vs. 33.4 ± 2.4, p = 0.959), survival rate (96.6 % vs. 97.4 %, p = population derived from a single facility. Additionally, the retrospective nature
0.651) were similar in the two groups. Rates of biochemical pregnancy (25.6 % of this study leaves the possibility for introduction of unaccounted for confound-
vs. 44.9 %, p < 0.001), clinical pregnancy (18.1 % vs. 31.3 %, p = 0.002), and ing factors.
implantation (18.5 % vs. 32.0 %, p = 0.002) in Group B were higher than those Wider implications of the findings: This large-scale study elaborates on
of Group A. Group B tends to have a higher ongoing pregnancy rate (14.9 % the use of ICM and TE grading in predicting embryo gender, ploidy, and successful
vs. 22.4 %, p = 0.053) than Group A, but no significant difference. IVF outcomes. Our data suggest that both ICM and TE grading are important
Limitations, reasons for caution: This is a retrospective study. The quality tools and should both be considered when selecting embryos for successful IVF
at the time of blastocyst vitrification was not considered. Therefore, further outcomes.
studies are needed in order to support our results. Trial registration number: None
Wider implications of the findings: We analyzed poor-quality blastocyst, which
can be assumed to be more stressed. This study suggests that poor-quality blastocyst P-218 Human sperm selected by a direct swim-up without
may be sensitive to the in vitro environment. Therefore, the short post-thaw culture centrifugation is positively associated with high blastocyst
period of poor-quality blastocyst can improve the pregnancy outcomes. formation rate in normal responders after intracytoplasmic sperm
Trial registration number: Not Applicable injection (ICSI)
J. Lee1, S. Shim1, M. Kim1, S. Choi1, I. Park1, H. Sun1, J. Kim1
P-217 The effect of inner cell mass and trophectoderm 1
Liora Fertility Center, IVF center, Busan, Korea- South
morphology on embryo ploidy, gender, and in vitro fertilization
outcomes Study question: Does human sperm selected by the direct swim-up (DU)
without centrifugation affect development and quality grade of embryo and
C. Moutos1, W. Kearns2,3, S. Farmer4, J. Richards4, A. Saad1, J.R.
blastocyst formation rate after ICSI?
Crochet1,4
1
Summary answer: The human sperm selected by DU without centrifugation
University of Texas Medical Branch -- Galveston, Obstetrics and Gynecology, was significantly associated with high blastocyst formation rate, but unrelated to
Galveston, U.S.A. ;
2
development and quality grade of embryo.
AdvaGenix, AdvaGenix, Rockville, U.S.A. ;
3
What is known already: Centrifugation could produce reactive oxygen spe-
Johns Hopkins University School of Medicine, Gynecology and Obstetrics, cies and impair sperm functions, and also induce sub-lethal damage. Therefore,
Baltimore, U.S.A. ;
4
we are still concerned about using centrifugation in the procedure of sperm
Center of Reproductive Medicine, Center of Reproductive Medicine, Houston, U.S.A. preparation. Previous study suggested that the DU without centrifugation can
minimize additional sperm preparation and should be used in assisted reproduc-
Study question: Is there a relationship between inner cell mass (ICM) or tive technology (ART) treatment as a reasonable alternative to improve the
trophectoderm (TE) grading and embryo gender, ploidy and in vitro fertilization embryo development and clinical outcomes. However, there is insufficient evi-
(IVF) outcomes? dence from previous studies to support this suggestion.
Summary answer: Higher ICM /TE grade embryos were more likely to be Study design, size, duration: A retrospective study of 76 (n = 1,365
euploid and have better IVF outcomes. while no correlation exists between retrieved oocytes) fresh ICSI cycles with normal responders was conducted
ICM/TE and embryo gender. from May 2018 to December 2019 at Liora Fertility Center. All cycles with severe
What is known already: Along with preimplantation genetic testing for aneu- oligoasthenoteratozoospermia, frozen sperm and surgically retrieved sperm
ploidy (PGT-A), embryo morphology has been used in selecting which embryo were excluded. All women had undergone a GnRH antagonist protocol. Cycles
to transfer during IVF/embryo transfer (ET) procedures. The relationship were divided into two groups; DU without centrifugation (Group A, n = 49)
between embryo morphology and euploidy has been well established; the effects and DU with centrifugation (Group B, n = 27).
of ICM and TE grading separately on PGT-A and subsequent IVF outcomes Participants/materials, setting, methods: Sperm medium (1㎖) was placed
remain uncertain. on the top of raw semen (1㎖) in 5 ㎖ tube. The supernatant with motile sperm
Study design, size, duration: A retrospective chart review was conducted of was gently collected after the DU for 20 minutes, and then placed into 15㎖ tube
patients age 21 to 47. Embryos that underwent PGT-A (N=3708) and subsequent without the washing step using centrifugation. Then the 15㎖ tube was stored in
elective single ET (N=539) from June 2007 to December 2018 were evaluated. 6% CO2 incubator at 35°C before ICSI. We compared the fertilization rate,
Participants/materials, setting, methods: Embryo data originated from embryo development, embryo quality grade and blastocyst formation rate
patients receiving care at a single reproductive endocrinology practice. Primary between two groups.
outcomes of this study consisted of IVF outcomes among different embryo ICM Main results and the role of chance: There were no significant differences
and TE morphological grades. Secondary outcomes included embryo gender between Group A and B regarding mean female age (33.5 ± 3.9 vs. 34.6 ± 4.4,
and ploidy. Statistical analyses were performed via Pearson’s chi-squared tests. p = 0.257), mean male age (35.4 ± 4.5 vs. 37.1 ± 5.1, p = 0.137), mean number

of in in-vitro fertilization (IVF) attempts (0.3 ± 0.6 vs. 0.3 ± 0.8, p = 0.864), mean Wider implications of the findings: These results demonstrate while net-
number of retrieved oocytes (18.8 ± 7.2 vs. 16.4 ± 7.2, p = 0.156), mean works can identify non-invasive markers for genetically-abnormal embryos, the
number of Metaphase II oocytes (13.2 ± 4.7 vs. 11.5 ± 5.0, p = 0.140), matu- need for markers to selecting genetically-normal embryos for transfer remains.
ration rate (87.0% vs. 90.0%, p = 0.272), fertilization rate (76.4% vs. 75.4%, This was the first report to our knowledge that’s able to objectively determine
p = 0.707), arrested embryos rate (1.0% vs. 0.7%, p = 0.309). We also observed without technician-bias or input that morphology-alone can be used to identify
similar rates of high-quality 4-cell stage embryos on day 2 (22.1% vs. 26.9%, genetically abnormal embryos.
p = 0.256), high-quality cleavage stage (5- to 9-cell stages) embryos on day 3 Trial registration number: not applicable
(26.3% vs. 30.0%, p = 0.434), and high-quality blastocysts stage (≥BB grade,
32.8% vs. 33.5%, p = 0.898) in Group A and B, respectively. However, the Group P-220 One step further: Prospective randomized controlled trial
A had significantly higher blastocyst formation rate (58.4% vs. 50.1%, p = 0.036) comparing one and two steps technique of embryo transfer.
than Group B. Lastly, any microbial contamination during embryo culture was P.E. Levi Setti1, F. Cirillo1, E. Morenghi2, V. Immediata1, V.
not discovered in two groups. Caccavari3, A. Baggiani1, E. Albani1, P. Patrizio4
Limitations, reasons for caution: This retrospective study is based on a 1
Humanitas Research Hospital, Department og Gynecology- Division of
small sample size so that further study in a large sample is needed. We also need Gynecology and Reproductive Medicine- Humanitas Fertility Center, Milano, Italy ;
to investigate whether the human sperm selected by DU without centrifugation 2
Humanitas Research Hospital, Department of Biostatistics, Milano, Italy ;
positively affect pregnancy rate in IVF cycles. 3
Istituto Clinico Città Studi, Department of Gynecology- Division of Assisted
Wider implications of the findings: This method is possible to reduce the Reproduction, Milano, Italy ;
influence of impaired sperm motility and DNA damage by centrifugation. 4
Yale University- School of Medicine, Department of Obstetrics- Gynecology and
Additionally, it can help to reduce an embryologist workload as a simple sperm Reproductive Sciences, New Haven, U.S.A.
preparation and provide the cost effective in ART. We suggested that this method
will improve safe and useful clinical application. Study question: To determine if there are differences in ease of use between
Trial registration number: not applicable two different embryo transfer’s techniques: preload single step and post load
double step approach.
P-219 Can deep convolutional neural network (CNN) be used as a Summary answer: Two step technique seems to reduce the difficult embryo
non-invasive method to replace Preimplantation Genetic Testing transfer rate.
for Aneuploidy (PGT-A)? What is known already: Numerous published trials now document that the
A. Meyer1, J. Dickinson1, N. Kelly1, H. Kandula2, M. embryo transfer (ET) procedure has a huge impact on pregnancy and delivery
Kanakasabapathy2, P. Thirumalaraju2, C. Bormann1, H. Shafiee3 rates after IVF. Difficult transfers should be avoided, as they are likely to reduce
1
Massachusetts General Hospital, Obstetrics and Gynecology, Boston, U.S.A. ; the chances of implantation and subsequent pregnancy rates. Free hand embryo
2
Brigham and Women’s Hospital, Division of Engineering in Medicine- Department transfers with soft catheters under ultrasound guidance is currently considered
of Medicine, Boston, U.S.A. ; the best option in transferring embryos. However, in technically difficult ET the
3
Brigham and Women’s Hospital, Department of Medicine- Harvard Medical use of harder catheters is needed and this could impact in uterine contraction.
School, Boston, U.S.A. Among soft catheters, it is not known which technique is preferable.
Study design, size, duration: prospective randomized unblinded controlled
Study question: Can deep convolutional neural network (CNN) be used as clinical trial, including 352 ecoguided ETs assigned to single step ET with Cook
a non-invasive method to accurately classify embryos based on their karyotype? k-soft-5000 soft catheter or double step ET with Cook K-JETS-551910-S bulb
Summary answer: Trained CNN algorithm of cleavage and blastocyst-stage tip catheter, from September 2017 to September 2019. The sample size was
embryo images performed very well in identifying aneuploid embryos. calculated on the basis of historical difficult ET rate encountered between 2014
What is known already: Preimplantation genetic testing for aneuploidy (PGT- and 2015. The randomization to one of the study branches was done by the
A) has become a widely utilized tool for screening and selecting embryos for operator just before the procedure, with a closed envelope.
transfer. There are limited studies showing the efficacy of this procedure in Participants/materials, setting, methods: Inclusion criteria: 18 - 38 years
women under 35 years of age. Conversely, there are some reports demonstrat- old, BMI between 18 and 28, one thawed blastocyst transfers. Exclusion criteria:
ing that younger patients have reduced cumulative pregnancy rates when utilizing ICSI-TESE and pregestational test cycles. The primary outcome was difficult
PGT. These lower outcomes are likely due to inaccurate PGT-A results or due transfer rate, defined as presence of blood, required manipulation, need for
to the invasive procedures involved with embryo biopsy and cryopreservation. instrumentation, stylet or tenaculum, multiple attempts, force and dilatation, for
The goal of the study was to determine if deep-learning CNNs could be used both groups. For the single step approach, the need for the outer sheath was
as a non-invasive method to select chromosomally normal embryos for transfer. considered as a difficult ET. The secondary outcome was clinical pregnancy rate.
Study design, size, duration: Using a retrospective dataset of embryos Main results and the role of chance: A total of 352 ET’s were performed.
analyzed using a modified FAST-SeqS next generation sequencing method The two arms were homogeneous for age at ET and at freezing, endometrial
(Invitae, San Francisco, Ca) a deep neural network model was trained and tested preparation protocols, BMI, and duration of infertility. Among the entire popula-
using 3112 images to classify embryos as aneuploid or euploid. Embryo images tion, 24.15% of the ET was defined as difficult. In the two branches, difficult transfer
captured on day 3 (D3) and day 5 (D5) were used to classify embryos based on rate was significantly higher in the single step group than in the double step’s one:
karyotype. 38.76% versus 9.20% respectively (p<0.001); the OR of difficult transfer with the
Participants/materials, setting, methods: For D3 karyotype classification double step was 0.16 (IC 95% 0.09 – 0.29). Clinical pregnancy rate was lower,
we developed a CNN with 2910 annotated images of cleavage embryos. We however not significantly, in the single step group (42.13% versus 48.28%).
tested the performance using 202 D3 embryo images. For D5 karyotype classi- Limitations, reasons for caution: 18 different experienced operators par-
fication we developed a CNN using 2906 images of annotated blastocysts. We ticipated in the trial, using a fixed distance transfer protocol. The differences in
tested the algorithm’s performance using 201 blastocyst images. difficult transfer rate between operators were not analyzed. Conclusions about
Main results and the role of chance: The deep learning CNN trained to the pregnancy rate should not be generalized, since the sample analysis was not
classify D3 embryos as aneuploid or euploid performed with a high specificity performed on this outcome.
and thus was able to sufficiently identify 85.45% (CI: 77.46% to 91.45%) of Wider implications of the findings: Difficult transfer rate was significantly
aneuploid embryos. Similarly, the CNN used to classify embryos based on karyo- higher in the single step group. Further study about the correlation with ART
type on D5 of development had a specificity of 75.23% (CI: 66.04% to 83%). outcomes are required.
These results objectively demonstrate that morphology-alone may be useful in Trial registration number: NCT03161119
identifying genetically abnormal embryos.
Limitations, reasons for caution: Images were obtained using a single P-221 Euploidy rates between either oral dydrogesterone (DYG)
imaging platform (EmbryoScope) at only 2 timepoints for karyotype primed ovarian stimulation protocol or the GnRH antagonist
classification. (GnRH-ant) protocol in 780 patients in the first PGT-A cycle

L. Yang1, K. Luo1, G. Lu1,2,3, G. Lin1,2,3, F. Gong1,3 5

Monteleone - Human Reproduction Center, Clinical Director, Sao Paulo / SP, Brazil
1
Reproductive and Genetic Hospital of CITIC-XIANGYA, Reproductive and Genetic
Hospital of CITIC-XIANGYA, Changsha, China ; Study question: Is very early-cleavage embryos parameters, as soon as the
2
Key Laboratory of Reproductive and Stem Cell Engineering- National Health and second cell cycle (CC2), based on time-lapse monitoring (TLM) useful to predict
Family Planning Commission, Key Laboratory of Reproductive and Stem Cell blastocyst formation?
Engineering, Changsha, China ; Summary answer: The time to embryos progress from two to three-cell
3
Institute of Reproducitve and Stem Cell Engineering- Basic Medicine College- (CC2) predict blastocyst formation. Optimal CC2 time-range is 8 to 13 hours,
Central South University, Institute of Reproducitve and Stem Cell Engineering-, leading to 78.2% blastocysts
Changsha, China What is known already: Assisted reproductive technologies success is depen-
dent of a number of factors, but embryo selection is a crucial step. The cell number
Study question: whether differences in euploidy rates between oral dydro- during cleavage has been described as a key factor for selecting embryos, as slow
gesterone (DYG) primed ovarian stimulation protocol and the GnRH-ant pro- or fast cleavage embryos shows reduced implantation potential. The introduction
tocol in the first PGT-A cycle. of TLM allowed additional noninvasive criteria to best embryo selection and mark-
Summary answer: DYG is comparable to GnRH-ant protocol in terms of ers of embryonic kinetic from the first-cleavage onwards has been studied and
euploidy rates at OPU in the first PGT-A cycle. applied in algorithms for best embryo choice. Although early cleavage parameters
What is known already: It is well known that successful meiosis in the IVF may be predictive of blastocyst development and even implantation, this approach
laboratory is inversely correlated with maternal age. However, it is unclear has gathered little attention and a consensus is not stablished.
whether other factors, particularly the IVF process, can interfere with normal Study design, size, duration: Retrospective cohort study used prospectively
oocyte physiology. Oral treatment with DYG inhibits the pituitary LH surge collected data from 671 ICSI cycles from 486 patients (37.7±4.2 years) between
during ovarian stimulation. Previous reports indicated that both the number of April/2018 and September/2019 in a private reproductive medicine center. A
MII retrieved and pregnancy rates from these oocytes are comparable to short total of 4724 mature (MII) oocytes were fertilized by ICSI and cultured in a
protocol of GnRH agonists during IVF cycles. It is worth investigating whether time-lapse monitoring system (Embryoscope®). Embryos were recorded since
ovarian stimulation protocol in the first PGT-A cycle affects euploidy status in fertilization until the blastocyst formation. The time for pronuclei fading (tPNf ),
DYG protocol and the GnRH-ant protocol. two-cell (t2), three-cell (t3), four-cell (t4), five-cell (t5), eight-cell (t8), nine-cell
Study design, size, duration: This single-center, retrospective cohort study (t9) were evaluated.
included patient who underwent the dydrogesterone (DYG, study group) or the Participants/materials, setting, methods: The early-cleavage characteristics
GnRH-ant protocol (control group) in the first PGT-A cycle from November were analyzed and correlated to blastocyst formation. The periods between t2-t3
2017 to May 2019 in Reproductive and Genetic Hospital of CITIC-XIANGYA. (cell cycle two – CC2), between t3-t5 (CC3) and between t5-t9 (CC4) were
The study group and control group both included 390 cycles, obtained from 408 calculated. The blastocysts were also classified according to morphology by
and 534 cycles respectively by propensity matching. Gardner’s classification and those with trophoectoderm grade ≥3 and inner cell
Participants/materials, setting, methods: We retrospectively evaluated mass A or B were considered top-quality. The blastocyst rate (BlastR) was calcu-
in the first PGT-A cycle patients who underwent the dydrogesterone (DYG, lated by the number of blastocysts formed per number of 2PN in each category.
study group) or the GnRH-ant protocol (control group). Propensity matching Main results and the role of chance: From the 4724 MII oocytes injected,
was used to identify a propensity matched control group from a cohort of 534 3622 were normally fertilized (2PN). From those, 3552 cleaved into two-cells
cycles, based on age, BMI, and AMH with a 1:1 ratio. Patient’s demographic, (t2=28.5±6.7 hours), 3500 attained three-cell (t3=38.6±7.3 hours), 3399 got
baseline, and cycle characteristics were obtained for each cycle from our elec- five-cell (t5=51.0±10.0 hours) and 3077 attained nine-cell (t9=73.7±12.2 hours).
tronic medical record. The main outcome was euploidy rate. The mean time of CC2 was 10.2±4.7, CC3 was 12.6±6.7 and CC4 was 23.3±9.1
Main results and the role of chance: 780 cycles were included in the study. hours. A total of 2341 embryos developed to blastocyst stage (64.6%) on days
The study group and control group both included 390 cycles. There was no 5 (n=2108, 90.0%) or 6 (n=233, 10.0%). We evaluated the blastocyst rate
difference in patient baseline and cycle characteristics between the two groups. (BlastR) according to CC2 varying from zero (for embryos who had tripolar
The number of oocytes retrieved (6.08±5.02 vs. 6.41±4.68, P =0.34), MII mitosis and did not had two cells observed) to ≥20 hours. For CC2<1.0 hour
(4.97±4.09 vs. 5.15±3.78, P = 0.42), embryos biopsied (1.88±2.57 vs. the mean BlastR was 20.4%, for CC2 between 1 and 7 hours the mean BlastR
1.79±2.01, P = 0.59), and embryos testing (1.88±2.57 vs. 1.79±2.01, P = 0.59) was 46.2%. The highest BlastR (78.2%) occurred for embryos presenting CC2
were no statistic difference between study and control groups. Due to some between 8 and 13 hours. After that, the BlastR progressively decreased for
cycles did not form biopsy blastocysts. Therefore, cycles using NGS testing were embryos with CC2≥14 hours (38.9%), attaining a mean of 12.2% for CC2≥20
262 in study group and 263 in the control group respectively. The main outcome hours. Among the blastocysts developed, we assessed the top-quality blastocyst
was that the euploid rate per embryo biopsied was not significantly different rate (TQ-BlastR) and the same pattern was observed: CC2<1.0 hour:
between study and control groups (DYG vs. GnRH-ant: 34.9%±2.4% TQ-BlastR=17.0%; CC2 between 1 and 7 hours: TQ-BlastR=20.0%; CC2
vs.40.2%±3.97%, P = 0.116). between 8 and 13 hours: TQ-BlastR=55.5%; CC2≥14 hours: 42.7%. There was
Limitations, reasons for caution: It is a retrospective study and subject to no TQ-Blast formed when the CC2 was ≥18 hours.
bias from confounding factors. Limitations, reasons for caution: This is a retrospective study evaluating
Wider implications of the findings: DYG and GnRH-ant protocol result in the association of CC2 and blastocyst formation. The choice of blastocysts for
comparable euploid rates. Therefore, ovarian stimulation protocol with DYG transfer was based on blastocyst morphology on day 5 or 6 and did not take
should be considered a valid option in freeze-all PGT-A cycles, in view of its into account the CC2. Then, we could not evaluate the impact of CC2 on
demonstrated effectiveness and known safety enhancement. pregnancy success.
Trial registration number: not applicable Wider implications of the findings: Based on our findings, we can underline
that time of CC2 predict blastocyst formation and can be used for selecting
day-3 embryos for transfer and contribute to embryo development prognostic.
P-222 Prediction of blastocyst formation based on very early- The CC2 is an auspicious candidate to be included in the time-lapse algorithm
cleavage embryos parameters by time-lapse monitoring and should be further validated for that.
H. De Martin1,2, G. Alecsandra1, M. Fujii1, M. Conatti1, M. Nakano2, Trial registration number: not applicable
T. Bonetti3,4, P. Monteleone2,5
1
Monteleone - Human Reproduction Center, Embriology, Sao Paulo / SP, Brazil ;
2 P-223 Comparison between two oil layer thickness covering on
Universidade de São Paulo USP, Gynecology Discipline - Obstetrics and Gynecology
human embryo culture: a prospective study
Department, São Paulo, Brazil ;
3
Monteleone - Human Reproduction Center, Scientific Department, Sao Paulo / SP, H. Bahri1, M. Benkhalifa2, W. Zidi3, Z. Mhamdi4, H. Garai4, O.
Brazil ; Triki4, K. Ben Rhouma4, G. Merdassi4
4 1
Universidade Federal de São Paulo, Gynecology Department, São Paulo, Brazil ; Alyssa Fertility Group, Alyssa Clinic / HB laboratory, tunis, Tunisia ;

2
Alyssa Fertility Group / Faculty of sciences of Bizert, Alyssa Clinic / Biochemistry What is known already: It is well established that culturing embryos in an
department of La Rabta Hospital, tunis, Tunisia ; oxygen concentration lower than the atmospheric one will increase clinical and
3
Faculty of sciences of Bizert, Biochemistry department of La Rabta Hospital, live birth rates. The oxygen tension most widely employed is 5%. Based on the
tunis, Tunisia ; observation that the levels of oxygen in the uterus are lower than in the oviduct,
4
Alyssa Fertility Group, Alyssa Clinic, tunis, Tunisia it has been postulated that a further reduction in oxygen tension after embryo
compaction could more closely mimics the natural environment, better support-
Study question: Does the light paraffin oil layer thickness used for in vitro ing the embryo’s physiology. Few studies comparing different oxygen values have
embryo culture improve in vitro development of human embryos and preg- been published with conflicting results and the optimal range of oxygen concen-
nancy rate? tration has still to be determined.
Summary answer: The success rate of embryo implantation and biochemical Study design, size, duration: From September to December 2018, 12
pregnancy (BP) was significantly higher with in vitro embryo culture (EC) using a patients undergoing to preimplantation genetic testing for aneuploidy cycles were
thicker cover of light-paraffin-oil. enrolled in the study. The mean female age was 36.7±4.1 years old. Inclusion
What is known already: The oil overlay in embryo culture systems prevents criteria were at least 6 normal fertilized oocytes. Exclusion criteria were: genetic
medium evaporation helps to maintain appropriate pH and osmotic conditions and diseases, severe male factors and egg donation cycles.
protects from volatile organic compounds and microbial contamination. In classic Participants/materials, setting, methods: Sibling zygotes were divided in
procedures, once microinjected, the oocytes are transferred into a culture dish of 1:1 ratio in two groups. All cultures were performed in sequential media with a
separated embryo culture medium drops (1 – 3 oocytes per drop of 25µl) covered change-over on day-3. In the standard oxygen (SO) group, zygotes were main-
with approximately 1 mm of paraffin oil (from the top of the drops). The risk of tained at 5% of oxygen tension for the whole culture. In the low oxygen (LO)
the infiltration of contaminating agent becomes higher after 3 days of culture. group, zygotes were maintained at 5% until day-3 of culture and then moved to
Study design, size, duration: 168 patients (20-40years) undergoing in-vitro 3% of oxygen tension for the remaining days.
fertilization (IVF) treatment by intracytoplasmic-sperm-injection (ICSI) were Main results and the role of chance: A total of 119 normal fertilized zygotes
enrolled in our prospective study between October 14 – December 23, 2019. were obtained, 62 and 57 of them were cultured in SO and LO groups, respectively.
After ICSI, 833 oocytes were allocated into 2 groups according to the thickness The good/excellent day-3 embryo quality was comparable in the two groups:
of oil covering the embryo culture medium (ECM): 1mm or 4mm. Statistical- 80.6% (N=50) and 87.7% (N=50) in SO and LO groups, respectively (NS). The
analyses by SPSS 20.0 (SPSS Inc., Chicago-IL-USA) with Kolmogorov-Smirnov blastocyst formation rate was similar in the two groups: 56.5% (N=35) and 52.6%
test for normality, Student-t test for parametric comparisons and Mann-Whitney (N=30) in SO and LO groups, respectively (NS). In SO group, 15 blastocysts were
test for non-parametric comparisons with p<0.05 as statistically-significant. obtained on day-5, 15 on day-6 and 5 on day-7. In LO group, 14 blastocysts were
Participants/materials, setting, methods: Group-1: Protected culture obtained on day-5, 16 on day-6 and none on day-7. A total of 30 and 27 blastocysts
(PC+): 520 matured oocytes with EC in dishes containing single layer of 400µl were biopsied in SO and LO groups, respectively. The percentage of blastocyst
ECM covered by ~200µl of light-paraffin-oil (Ovoil, Vitrolife-Sweden) (~4mm available for the transfer was statistically higher in LO group (81.5%, N=22) com-
from the ECM surface). Group-2 (PC-) 313 matured oocytes with EC in dishes pared to SO group (46.7%, N=14; p=0.0124). In 3 and 1 blastocysts it was not
covered by light-paraffin-oil (1mm of thickness from the top of the drops). possible to obtain the genetic result, in the SO and LO groups, respectively.
Sage1-step (Origio-Denmark) was used as ECM. EC was into trigaz incubators. Limitations, reasons for caution: This is a pilot study enrolling few patients
BP was defined as a positive βHCG-test after 2 weeks from the embryo transfer. with a high oocyte’s number. Data on clinical outcomes, such as implantation,
Main results and the role of chance: Embryo morphology was recorded clinical pregnancy, live birth and miscarriage rates as well as follow-up data on
on days 3 and 5. A total of 139 PC+ and 97 PC- day3 embryos and 59 PC+ and babies born, are still missing. These preliminary results need to be confirmed on
46 PC- day5 embryos were analyzed. an increased sample size.
There were no differences between the two groups in terms of mean of age, Wider implications of the findings: It has been reported that euploidy rate
number of collected oocytes, and number of matured oocytes, cleavage rate, can greatly vary among different infertility centers demonstrating that chromo-
top quality day3 embryo rate and the number of transferred embryos. The somal abnormalities can be partially iatrogenic. To set-up the optimal culture
proportion of top quality day3 embryos was 66.66% [0% – 100%] vs 75.00% conditions in order to obtain the highest number of transferable blastocysts
[0% – 100%] in PC+ and PC- groups respectively. However, Blastulation rate could enhance the efficiency of infertility treatments.
was significantly higher in PC+ group than in PC- with 61.55% ±2.77% vs 57.73% Trial registration number: not applicable
±2.54%; (p=0.001) respectively. Pregnancy rate per transfer was evaluated in
69% of the total patients. When the unknown βHCG test results were elimi-
nated, the biochemical pregnancy rate was higher in the PC+ group compared P-225 The effect of women’s age on timing, morphology and
to PC- group with 41.2% vs 35.4%; (p< 0.001) respectively. implantation of the competent blastocyst – a multicenter cohort
Limitations, reasons for caution: The design in this study is not a random- study
ized controlled trial, although it is prospective. Moreover, different origin of M. Buhl Borgstrøm1, M.L. Grøndahl1, T. Wirenfeldt Klausen2,
sperm (Ejaculated and testicular) were used to fertilize corresponding oocytes. A. Kjærgard Danielsen3, T. Thomsen4, U. Schiøler Kesmodel5,
Wider implications of the findings: These results should interest embry- I.T.C.B.P.G. Inclusive: The Competent Blastocyst Project Group1
ologists who want to improve embryo culture conditions. More studies must be 1
Herlev University Hospital, The fertility clinic, Herlev, Denmark ;
done with a larger cohort of oocytes. 2
Herlev University Hospital, Department of hematology, Herlev, Denmark ;
Herlev University Hospital, Department of gastroenterology, Herlev, Denmark ;
4
Herlev University Hospital, Department of Anaesthesiology, Herlev, Denmark ;
P-224 Reducing oxygen concentration from 5% to 3% after 5
Aalborg University Hospital, Department og Obstetrics and Gynecology, Aalborg,
embryo compaction decreases the aneuploidy rate of the derived
Denmark
blastocysts. Analysis of 119 sibling zygotes
A. Ruberti1, M.G. Minasi1, E. Cursio1, A. Colasante1, S. Muzzì1, G. Study question: Is the age of women undergoing assisted reproductive tech-
Pirastu1, D. Paccagnini1, S. Gatti1, A. Caragia1, P. Greco1, E. Iovine1, nology (ART) associated with timing, morphology and implantation of the com-
A. Greco1, M.T. Varricchio1, E. Greco1 petent blastocyst?
1
European Hospital, Medicina della Riproduzione, Rome, Italy Summary answer: The effect of age on timing and morphology was modest,
while initial hCG rise was lower for the youngest women.
Study question: Does a reduction in incubation oxygen concentration from What is known already: Within ART, the age of a woman is the most prog-
5% to 3%, from day-3 of culture in advance, affect blastocyst’s quality? nostic parameter for her chance to conceive. Both women’s age and ploidy fail
Summary answer: Reducing oxygen concentration from day-3 of culture to be reflected in routine embryo assessment. We have previously shown that
allows to obtain a higher blastocyst number available for the transfer compared initial hCG rise in singleton pregnancies after single cleavage stage embryo trans-
to fixed 5% oxygen concentration. fer is higher with increasing age in both in vitro fertilization (IVF) and

intracytoplasmic sperm injection (ICSI) suggesting that age may influence tro- extent, the sperm genome. In this study, we hypothesize that poor sperm quality
phoblast differentiation and placentation. Hence, the question remains whether may modify the oocyte’s mtDNA levels as a part of the repair mechanism.
the competent blastocyst develops in the same way regardless of women’s age Study design, size, duration: A retrospective pilot study was conducted at
or if age imposes an alteration in timing, morphology and implantation? AL Hadi Laboratory and Medical Center, between January 2019 and January
Study design, size, duration: This is a multicenter historical cohort study 2020.The study included the data analysis of 170 embryos obtained from 28
based on exposure and outcome data from 7246 women, who between 2014 infertile couples undergoing pre-implantation genetic screening (PGS) for aneu-
and 2018 underwent controlled ovarian stimulation (COS) or Frozen Embryo ploidy. Couples were categorized based on sperm indices into: normozoosper-
Transfer (FET) followed by single blastocyst transfer and hCG measurement, mic (n= 13) and non-normozoospermic (n= 15) groups. Non-normozoospermia
resulting in singleton pregnancy. These data were linked to the Danish Medical was defined by the presence of at least one abnormal sperm parameter according
Birth Registry using the women’s unique personal identification number (CPR). to the World Health Organization 2010.
Resulting in a total of 5466 women with a live birth being included. Participants/materials, setting, methods: PGS data collected from couples
Participants/materials, setting, methods: Exposure (age) and outcome with maternal age <38 years, a history of unsuccessful IVF treatments, and/or
(Gardner’s score (1-6), inner cell mass (ICM)(A,B,C), trophectoderm (TE) previous spontaneous miscarriages were included. Evaluation of embryo quality
(A,B,C) and hCG) data were collected from the fertility database, Danish Medical was carried out in accordance with Alpha-ESHRE consensus. Fresh embryos
Data Center, used by the 16 participating private and public fertility clinics. All derived from fresh gametes were biopsied on day 3. Genomic and mitochondrial
COS cycles (IVF and ICSI) and FET cycles (natural and substituted), were DNA levels were quantified using next generation sequencing (Ion Torrent,
included. Exclusion criteria were cycles with pre-implantation genetic testing and ThermoFisher Scientific, Lebanon). Implantation rates per transferred fresh
donated oocytes. The analysis was adjusted for male age, female smoking, female embryo were analyzed.
BMI, diagnosis and clinic. Main results and the role of chance: From the total of 170 assessed
Main results and the role of chance: The women’s mean age for COS/ embryos, 73 were identified as euploid. No statistical significant trend was
FET was 32.4/32.6 years. Adjusted analysis of age and stage in COS treatments observed in the percentage of euploid embryos between the normozoospermic
showed that for every one-year increase in age there was a 5% reduced prob- and non-normozoospermic groups (49% and 38% respectively). In contrast, a
ability of the competent blastocyst assessed as being in a high stage at transfer higher mtDNA levels were detected in the non- normozoospermic group com-
(OR 0.95, 95% CI (0.93;0.98)). Adjusted analysis of age and TE (A,B,C; A being pared to the normozoospermic (p = 0.0232). Implantation and fertilization rates
the highest category) in FET treatments showed that for every one-year there showed no significant difference between the two comparable groups (p >0.05).
was a 3% increased probability of the competent blastocyst assessed as being Limitations, reasons for caution: This study provided a possible effect
in a high TE category. Other comparisons related to the effect of age on timing caused by paternal factors on embryonic mtDNA levels. However, to determine
and morphology did not reach statistical significance. the true extent of this clinical outcome, a prospective design with a larger sample
HCG values followed an identical pattern in COS and FET, with the lowest hCG size is necessary to corroborate the current findings.
values in the youngest age group for both treatments, 95.3 (IU/L) for COS and Wider implications of the findings: The biological mechanism(s) underlying
130.9 (IU/L) for FET. Comparison between hCG values in women 18-24 years the association between paternal factors, oocyte repair system, and mitochon-
and 25-29 years in those receiving respectively COS and FET treatment, showed drial DNA are unknown. Hence, this topic is of considerable importance in
a significantly lower levels in the youngest women, with a significant mean difference future assessment of the role of mitochondria in oocyte ability to repair any
in the adjusted analyses COS: -20.9, 95% CI (-35.7;-6.1)); FET: -21.8, 95% CI sperm defect that could alter embryo development.
(-42.1;-1.5). Other comparisons did not reach statistical significance. Trial registration number: Not applicable
Limitations, reasons for caution: The blastocyst morphology was subjec-
tively assessed and inter observer differences may have influenced the results.
However, adjusting for clinic takes the potential inter clinic variation in embryo P-227 Clinical factors affecting meiotic spindle imaging of oocytes
scoring into consideration. by PolScope before ICSI
Wider implications of the findings: Our study supports an association S.J. Kim1, J. Jeong1, S. Kim1, T.H. Kim1, J.H. Eum1, W.S. Lee1,
between young age and lower initial hCG-values after ART-treatment irrespective S.W. Lyu1
of mode of treatment, which should be born in mind when evaluating 1
Fertility Center of CHA Gangnam Medical Center- CHA University School of
hCG-measurements. Medicine, Department of Obstetrics and Gynecology, Seoul, Korea- South
Whether these findings correlate to the embryo or the endometrium need
further clarification. Study question: Are there clinical factors associated with the visibility of mei-
Trial registration number: not relevant otic spindle of oocytes when performing intracytoplasmic sperm injection (ICSI)?
Summary answer: Longer gonadotropin stimulation was associated with
P-226 Paternal influence on mitochondrial DNA levels in day 3 oocytes of no visible spindle, but not the ovarian reserve and female age.
euploid embryos: a retrospective pilot study. What is known already: In the laboratory, Polarized light microscopy
Y. Mourad1, M. Bazzi1, V. Massaad2, C. Afeich2, G. Raad1, J. Mortada3, (PolScope) has been used to locate the meiotic spindle and inject oocytes without
F. Fakih4, R. Yazbeck5, M. Chartouny5, R. Zahwe1, C. Fakih1 damaging spindle. Meiotic spindle imaging by PolScope has been studied to pre-
1
Al Hadi Laboratory and Medical Center, Ivf, Beirut, Lebanon ; dict oocyte quality. Moreover, some studies analyzed various environmental
2
Lebanese University, Master in Reproduction and Development, Fanar, Lebanon ; conditions affecting meiotic spindle. However, there are few papers on the rela-
3
Al Hadi Laboratory and Medical Center, Molecular Biology, Beirut, Lebanon ; tionship between clinical factors and oocyte meiotic spindle.
4
Al Hadi aboratory and Medical Center, Ivf, Beirut, Lebanon ; Study design, size, duration: This retrospective study was conducted at a
5
Lebanese University, Faculty of Medicine, Hadath, Lebanon single fertility center. A total of 717 mature oocytes of 140 patients were selected
for spindle analysis. Fertilization and embryo development of each oocyte were
Study question: Is there any paternal influence on the mitochondrial DNA analyzed with respect to the observed spindle pattern. We evaluated multiple
levels in day 3 euploid embryos? clinical factors as a predictor of oocytes with no visible spindle.
Summary answer: Higher mitochondrial DNA (mtDNA) levels were detected Participants/materials, setting, methods: From October 2017 to April
in day 3 euploid embryos of non-normozoospermic men compared to 2019, patients who underwent ICSI in previous in vitro fertilization treatment
normozoospermic. but have failed pregnancy more than 2 times were recruited. The meiotic spindle
What is known already: Early paternal influence on pre-implantation embryo angle of oocytes was defined as the angle of spindle deviation to the first polar
development was considered to be related to sperm cytoplasmic factors such as body and graded as the follows: a, 0-5°; b, 6°-15°; c, 16°-30°; d, 31°-45°;
centrioles. Recently, double-stranded sperm DNA damage was shown to be asso- e, >46°; non-visible spindle. Statistical analysis was carried out using the Chi-
ciated with a delay in cleavage embryo kinetics. Furthermore, altered cleavage square test and multivariate regression analysis.
divisions may lead to an abnormal distribution of mitochondria between blasto- Main results and the role of chance: The mean age, anti-Mullerian hormone
meres. In parallel, oocyte may have the capacity to modulate and repair, to certain (AMH), and duration of stimulation were 39.6 ± 3.6 years, 2.25 ± 3.04 ng/ml,

and 9.0 ± 2.5 days, respectively. The number of oocytes retrieved and the 363 of them it was obtained the genetic result (95.5% and 97.1%, NS) in LAH
number of mature oocytes retrieved were 7.8 ± 6.6 and 5.3 ± 4.8, respectively. and NO-LAH groups, respectively. The percentages of transfereable blastocysts
The distribution of the oocytes according to grade were as follows: a (32.6%), were comparable between the two groups: 40.2% (N=452) and 41.2% (N=154)
b (31.7%), c (18.0%), d (7.1%), e (6.0%), and non-visible (4.6%). Fertilization in LAH and NO-LAH groups (NS), respectively. In total, 275 and 47 frozen
rates were similar except non-visible grade: a (73.9%), b (74.0%), c (72.9%), d embryo transfer were performed in LAH and NO-LAH groups, respectively. In
(72.5%), e (76.7%), and non-visible (51.5%). However, the cleavage rate and LAH group, 281 blastocysts were transferred and 139 of them implanted
good quality of cleavage embryo formation rate were not different among the (49.5%). In NO-LAH group, 48 blastocysts were transferred and 22 of them
grades. When evaluating for factors predicting oocyte of non-visible spindle, the implanted (45.8%, NS). The miscarriage rate was the same in two groups
duration of gonadotropin stimulation was the only relevant factor (adjusted (19.0%). Statistical analysis was performed using Sudent’s t-test and Chi-squared
Odds Ratio (aOR) 1.218, 95% CI (1.005-1.477)). However, Female age (aOR test at the level of p<0.05
1.064 (0.929-1.220)), body mass index (aOR 1.092 (0.901-1.323)), AMH (aOR Limitations, reasons for caution: Although the incubator employed for the
0.976 (0.806-1.182)), and the number of oocytes retrieved (aOR 1.124 (0.948- culture was the same in all analyzed embryos, the use of different culture media
1.333) were not associated with spindle nondetection. in the two groups could have introduced a bias in the study. The study has a
Limitations, reasons for caution: The retrospective study design and the retrospective design
small number of study subjects are limitations. We only focused on clinical Wider implications of the findings: Despite NO-LAH group includes
factors; therefore, further studies are needed to evaluate the laboratory factors younger women and higher fertilization and blastocyst formation rates compared
such as temperature and pH. to LAH group, the percentage of blastocyst available for transfer and the implan-
Wider implications of the findings: Oocytes with non-visible spindle tation rate were comparable. This outcome should reassure the operators about
showed significantly lower fertilization rates. Meiotic spindle angle does not affect the LAH day-3 ZP opening in order to simplify the blastocyst biopsy procedure
the oocyte quality represented by fertilization rate, cleavage rate, and good Trial registration number: Not applicable
quality of cleavage embryo formation rate. The duration of gonadotropin stim-
ulation was the only clinical factor related to oocytes of no visible spindle. P-229 Can we predict aneuploidy or mosaicism considering
Trial registration number: Not Applicable blastocyst morphology?
A. Rodríguez-Arnedo1, L. Herrero2, L. Cascales2, J.A. Ortiz3,
P-228 Laser assisted hatching at cleavage stage before the A. Bernabeu4, J. Llácer4, A. Llaneza5, J. Ten1, R. Bernabeu4
subsequent trophectoderm biopsy: to perform or not to perform? 1
Instituto Bernabeu, Embryology Dept., Alicante, Spain ;
M.G. Minasi1, E. Cursio1, A. Colasante1, S. Muzzì1, G. Pirastu1, 2
Instituto Bernabeu, Embryology Dept., Madrid, Spain ;
D. Paccagnini1, S. Gatti1, C. Arrivi1, A. Greco1, P. Greco1, 3
Instituto Bernabeu, Molecular Biology, Alicante, Spain ;
M.C. Greco1, M.T. Varricchio1, A. Ruberti1, E. Greco1 4
Instituto Bernabeu, Reproductive Medicine, Alicante, Spain ;
1 5
European Hospital, Center for Reproductive Medicine, Rome, Italy Instituto Bernabeu, Reproductive Medicine, Madrid, Spain
Study question: Is the laser assisted hatching (LAH) day-3 zona pellucida (ZP) Study question: Can we correlate blastocyst morphology with aneuploidy or
opening, performed in order to facilitate the subsequent biopsy at blastocyst mosaicism rates?
level, safe? Summary answer: Blastocyst morphology: quality and expansion (percentage
Summary answer: To perform LAH at cleavage stage is safe and does not of trophectoderm (TE) hatching out of the zona) as well as maternal age are
impair the ploidy nor the implantation rate in the deriving blastocysts associated with aneuploidy and mosaicism.
What is known already: Preimplantation genetic testing (PGT) is widespread What is known already: Preimplantation genetic testing of aneuploidies
used to select the embryo with the highest implantation potential. Currently, (PGT-A) has significantly grown in assisted reproduction treatments in the last
the blastocyst is the preferred developmental stage to be biopsied. A strategy decade. PGT-A aims to find the best euploid embryo, improving birth rates and
is open the ZP with laser at embryo level in order to favorite the subsequent allowing single embryo transfer. Embryo morphology is considered a predictor
blastocyst hatching, simplifying the biopsy procedure. This method, however, of implantation potential and several publications support a correlation between
has been suggested to negatively interfere with embryo development. Another embryo development parameters with the chromosomic status of the embryo.
possibility, hypothesized to be safer, consist of let the embryos undisturbed Nonetheless, many “top-quality” embryos still turn out to be aneuploid after
during the whole culture performing the ZP opening directly during the biopsy. PGT-A. The aim of this study is to identify which embryo features are correlated
A complete evaluation of the risks associated with these methods is still missing. with higher aneuploidy or mosaicism rates, and if it is possible to predict them.
Study design, size, duration: Retrospective analysis of 491 PGT cycles. Study design, size, duration: Retrospective database study performed
According to our laboratory procedure, when a cycle was allocated in sequential between August 2017 and December 2018 including 162 PGT-A cycles and 499
culture media all the developing day-3 embryos underwent to LAH during the biopsied embryos. Cycles with severe male factor (sperm samples concentrations
media change-over (LAH group). When single-step media was employed, all under one million/mL) were excluded.
embryos were maintained undisturbed in culture and LAH was performed Participants/materials, setting, methods: Blastocyst quality was assessed
directly during the biopsy at blastocyst stage (NO-LAH group). The allocation according to Gardner and Schoolcraft score. Expansion was classified according
in the two culture media is randomly based on weekly rotation to the percentage of Trophectoderm (TE) hatching out of the zona. Mosaic
Participants/materials, setting, methods: All cycles were performed from embryos with ≤25% aneuploid cells were considered euploid, and those
June 2015 to December 2019: 393 were allocated in LAH and 98 in NO-LAH with >50% were considered aneuploid.
groups, respectively. All culture were carried out in the same time-lapse incu- Student’s t, Pearson’s chi-square and multivariate analysis (linear and binary
bator at 37°C, 5%O2 and 6%CO2. Mean female ages were 38.8±4.1 and logistic regressions) were used. Maternal age, embryo quality and expansion,
37.4±3.9 year old in LAH and NO-LAH groups, respectively (p=0.002). day of biopsy and oocyte origin were introduced as confounding variables.
Monogenic diseases, structural rearrangements and egg donation cycles were Main results and the role of chance: A total number of 310 blastocysts
excluded from the analysis. All embryo transfers were performed on a subse- were biopsied on day-5 (62.1% out of 499). Three-hundred and twenty-eight
quent cryopreserved cycle blastocysts came from donor eggs cycles (65.7%) and 183 from frozen/thawed
Main results and the role of chance: In LAH group, 3706 oocytes were oocytes (36.7%). The diagnosed blastocysts (4.6% amplification failure) were
collected, 2747 of them were mature and were injected. In NO-LAH group 720 classified in: 57.7% euploid, 13.7% euploid mosaic and 42.4% aneuploidy blas-
out of 1013 retrieved oocytes were mature and injected. The fertilization rates tocysts. A tendency of lower aneuploidy rates as the blastocyst expansion
were 76.2% (N=2093) and 80.0% (N=576) in LAH and NO-LAH groups, increases was observed: 5-25% extruded TE: 47.7% aneuploidy, 25-50% extruded
respectively (p=0.0324). A total of 2081 and 576 day-3 embryos were obtained TE: 44.0% aneuploidy, >50% extruded TE: 32.6% aneuploidy, 100% extruded
in LAH and NO-LAH groups, respectively. The blastocyst formation rates were TE: 28.6% aneuploidy (p=0.052). Maternal average age also decreased as the
54.2% (N=1127) and 65.3% (N=376) in LAH and NO-LAH groups, respectively blastocyst expansion increased (p=0.089). TE quality was found to be significative
(p<0.0001). Overall, 1123 and 374 blastocysts were biopsied and for 1072 and in predicting aneuploidy rate: 40.8% aneuploidy (TE A/B), 93.3% (TE C/D;

p=0.004). Maternal average age of blastocyst with A/B TE was lower than those ±28.45%; p=0.002). No difference was observed in the rate of mosaicism. In
blastocysts with C/D TE (29.8±9.8 vs 31.8±6.8 years), and also predictive for the grouped culture cycles, the rate of day 5 biopsy was higher (38.73 ±18.89%
aneuploidy: p<0.001. TE quality (p=0.016) and maternal age (p=0.012) were vs 24.76 ±19.28%; p<0.0001) and of day 6 lower (52.39±29.07% vs 63.69
found to be the most important variables predicting mosaicism. Logistic regres- ±33.75%; p=0.0012) compared to single culture cycles. A generalised linear
sion and multivariate aneuploidy risk score were performed. AUC was used to estimation corrected for variables known to be associated with ploidy according
assess the predictive performance of the aneuploidy risk score: AUC=0.719 to the literature. Single embryo culture had 7.1% (13.2-1.1 %CI) lower euploidy
(95% CI 0.672-0.767). The same multivariate analysis was performed to predict rate compared to grouped embryo culture (p=0.02).
euploid mosaic presence, and the AUC was 0.672 (95% CI 0.595-0.748). Limitations, reasons for caution: The study is limited by its retrospective
Limitations, reasons for caution: Expansion and quality assessment is oper- design. The grouped embryo culture contained embryos cultured individually
ator dependent and a potential source of subjective bias. Further studies focused when the cycle had 1 embryo in excess. Cycles including at least 1 blastocyst
on non-invasive embryo biomarkers for implantation potential to improve selec- with inconclusive diagnosis were included. The methodology used for assessment
tion are warranted. PGT-A cannot yet be replaced by a genetic status statistical of chromosomal constitution may have introduced a bias.
estimation, but it will be helpful in patient counselling. Wider implications of the findings: Our findings suggest that grouped
Wider implications of the findings: Our results suggest a correlation embryo culture may benefit PGT-A cycles by increasing the number of blasto-
between aneuploidy and blastocyst expansion/TE quality. This information might cysts diagnosed as euploid. This phenomenon could be attributed to the para-
be helpful for patient counselling especially when PGT-A is not considered. Based crine communication that may aid cell correction and free the blastocyst from
on our findings, poor morphology and slow blastocysts from older patients carry aneuploid cells, which if persist may result in a mosaic diagnosis.
a high risk for aneuploidies. Trial registration number: non applicable
Trial registration number: no trial rnumber
P-231 Suitability of 3D culture system with GrowDex for long-
P-230 Comparison of ploidy outcomes between grouped and term culture of granulosa cells
single embryo culture in preimplantation genetic testing for E. Chmelíková1, M. Sedmíková1, M. Ješeta2, P. Postlerová1,
aneuploidy (PGT-A) cycles. R. Krejčířová1, T. Krejčová1, K. Havlíková1, J. Žáková2, P. Ventruba2
S. Makieva1, L. Pagliardini1, L. Corti1, A. Alteri1, E. Papaleo1, 1
Czech University of Life Sciences Prague, Department of Veterinary Sciences,
G. Cermisoni1, A. Biricik2, F. Spinella2, F. Fiorentino2, P. Viganò1 Prague, Czech Republic ;
1 2
IRCCS San Raffaele Scientific Institute, Obstetrics and Gynecology Unit, Faculty of Medicine- Masaryk University Brno and University Hospital Brno,
Milan, Italy ; Department of Obstetrics and Gynecology, Brno, Czech Republic
2
Genoma, Molecular Genetics Laboratory, Rome, Italy
Study question: Is it possible to increase the suitability of in vitro culture system
Study question: Does grouped embryo culture influence ploidy outcomes in for long-term granulosa cells cultivation using three dimensional (3D) culture
PGT-A cycles? with hydrocellulose GrowDex?
Summary answer: PGT-A cycles performed using grouped embryo culture Summary answer: Using of 3D long-term culture system with GrowDex
had 7.1% (p=0.02) higher euploidy rate compared to single embryo culture. resulted in increasing of anti-apoptotic Bcl-2 and decreasing of pro-apoptotic
What is known already: Animal and human studies have shown that single Bax factors in porcine granulosa cells.
embryos in culture lack the benefits of embryotrophic factors of group culture. What is known already: Granulosa cells (GC) play important role in meiotic
In mice, single culture is associated with suboptimal neonatal outcomes while in maturation of oocytes: by steroidogenesis, hormone activity regulation and
humans grouped culture reduces embryo development rates, blastocyst quality prevention apoptosis. Apoptosis is regulated by pro- and anti-apoptotic factors;
and pregnancy rates. Thus, single culture may be detrimental to human embryos well-known of them are Bcl-2 and Bax. GC represent suitable model for in vitro
compared to grouped culture plausibly due to deprivation of paracrine signaling culture, however when GC are cultured in monolayers they gradually lose their
occurring between neighboring embryos. To date no study has assessed whether functions. 3D culture systems more closely mimic cell physiology environment
grouping embryos can influence their genetic composition and, hence, PGT-A and preserve interactions between cells. GrowDex hydrogel could be used as
outcomes. a matrix for 3D cultivation. GrowDex is a nanofibrillar cellulose hydrogel that
Study design, size, duration: A retrospective analysis of 342 PGT-A cycles forms fibres with length of several micrometres while their diameter is only
involving 1265 biopsied blastocysts performed between December 2014 and 4-100 nanometres.
July 2019. The blastocysts were cultured either in groups of 1-6 (grouped culture; Study design, size, duration: In this study, porcine cumulus oocyte com-
n=616) between December 2014 and March 2017 or individually (single culture; plexes (COCs) were matured in culture medium with GrowDex hydrogel as a
n=649) between May 2017 and July 2019, under similar conditions. The biopsy matrix for 3D culture system (20 COCs; 4-well NUNC dish; 38°C; 5% CO2).
was performed between day 5 and 7 of development. The PGT-A diagnosis and Conventional (2D) culture system was used as a control. After 48-hour culture
other IVF outcomes were compared between the two groups. matured oocytes were removed from COCs and GC were further cultured for
Participants/materials, setting, methods: PGT-A cycles were performed 16 days. After pro-long culture, apoptotic factors (Bcl-2 and Bax) were deter-
at the IVF center of San Raffaele Hospital and included patients with an indication mined in GC samples. Experiment was four times repeated.
for advanced maternal age, recurrent implantation failure or a risk for a genetic Participants/materials, setting, methods: BCl-2 and Bax proteins were
disease. Grouped and single embryo culture involved a sequential system using detected by blot immunodetection. The antigen-response signal to antibody was
culture media supplemented with SSS. All procedures were performed according expressed on the basis of integrated optical density (IOD), which is equal to the
to standard clinical practice. The PGT-A diagnosis was based on WGA+aCGH average optical density and the area at the antigen-antibody response. In order
or NGS, which was predominantly performed at the genetics laboratory to assess the amount of detected proteins, the IOD values were compared to
GENOMA. the integrated optical density of non-cultured GC (RIOD=1). Statistical evalu-
Main results and the role of chance: The analysis of 1265 blastocysts ation was performed using one-way ANOVA.
revealed that grouped culture had a higher number of euploid (176/616; 28.5%; Main results and the role of chance: 3D culture system with GrowDex
p=0.027) and lower number of mosaic (72/616;11.6%; p=0.025) blastocysts was shown to be more suitable for long-term culture of porcine granulosa cells
when compared to single culture (150/649;23.1% and 104/649;16% respec- as compared with conventional 2D culture system. Granulosa cells cultured in
tively). The number of grouped blastocysts biopsied on day 5 (261/616; 42.3%) 3D culture system expressed pro-apoptotic factor Bax in significantly lower
was higher (p<0.0001) when compared to single blastocysts (166/649;26.9%). manner than those in 2D conventional system (RIOD = 0.3 vs. 0.73). On the
Moreover, single cultured blastocysts were more likely to be biopsied on day 6 contrary, the expression of the anti-apoptotic factor Bcl-2 was significantly higher
(414/649;67.2%; p<0.0001) and day 7 (69/649;11.2%; p=0.0015) compared in GrowDex culture system (RIOD = 2.31 vs. 1.61).
to grouped (366/616;49.6% and 49/616;7.9% respectively). Overall, the rate Limitations, reasons for caution: A limitation is the number of samples
of euploidy in the 159 cycles in the grouped culture group was significantly higher included and analysed in this study, which slightly reduced the power of statistical
compared to that in the 183 cycles of single group (28.66 ±29.19% vs 20.95 analysis.

Wider implications of the findings: 3D culture system with GrowDex could 1

A.G.I. Medica, Reproductive Medicine, Siena, Italy ;
2
be tested for human granulosa cells when the long-term culture is required. Università degli studi di Siena, Dipartimento di Scienze della Vita, Siena, Italy
Study question: is there a difference in the expression of some genes involved
P-232 Blastocyst re-expansion after warming is not predictive of in lipid metabolism, between different categories of patients?
implantation after euploid blastocyst transfer. Summary answer: Some genes involved in lipid metabolism are differently
expressed overall in patients with reduced ovarian reserve.
S. Mateo cuadros1, M. Parriego1, M. Solé1, L. Coll1, M. Tresanchez1,
What is known already: Successful reproduction is dependent on the ovu-
F. Martínez1, M. Boada1, A. Veiga1
lation of competent oocyte, capable of undergoing fertilization and subsequent
1
Dexeus Mujer, Reproductive Medicine Service- Department of Obstetrics- Gynecology embryo and fetal development, and requires the co-ordinated growth and mat-
and Reproduction, Barcelona, Spain uration of the oocyte and its somatic cells. Oocytes, but also cumulus cells are
well known to contain lipid droplets, but not yet much is known about their role
Study question: Is blastocoel re-expansion a predictor of implantation in play during the maturation of the oocyte. Further studies are needed to under-
euploid blastocyst transfer? stand the expression profile of those genes that regulate folliculogenesis espe-
Summary answer: Blastocoel re-expansion degree before embryo transfer cially during the last stages of follicular maturation that could be useful in
does not correlate with implantation potential in warmed euploid blastocysts. identifying the lipids most involved during oocyte development.
What is known already: Vitrification has improved the efficiency of embryo Study design, size, duration: Present study included 16 IVF patients referred
cryopreservation allowing the achievement of similar results after frozen/ to our fertility Center A.G.I. Medica. Patients were subdivided into 3 groups
warmed embryo transfer compared to the fresh ones. In PGT cycles where a according to the type of infertility: male infertility (control Group, n=7), Group
freeze-all strategy is needed, vitrification results have enabled the transition to 1 (G1, n=5) idiophatic infertility and Group 2 (G2, n=4) reduced ovarian reserve.
blastocyst biopsy. Nonetheless, little is known about embryo characteristics after The study was performed in cumulus cells obtained from cycles of intra-cyto-
warming that can impact on implantation after euploid embryo transfer. Time plasmatic injection between March and October 2019.
between biopsy and vitrification, as well as time between warming and transfer, Participants/materials, setting, methods: Following oocytes denuda-
have been matter of study without conclusive results. Similarly, blastocyst re-ex- tion, cumulus cells (CC) were collected and cumulated for each patient. CC
pansion after warming has been proposed as a good prognosis parameter but were then washed and centrifuged and their number and viability was evalu-
without consensus in the literature. ated with Trypan Blue dye. In addition, mRNA extraction and retrotranscrip-
Study design, size, duration: Prospective observational study of 113 euploid tion was performed and through the Real Time Quantitative PCR were
blastocysts replaced in single frozen-thawed embryo transfers performed from analyzed and quantified following genes: 11β-HSD 1, 11β-HSD 2, NR3C1
June to December 2019. Characteristics of the implanted and non-implanted FASN, PNPLA2.
blastocysts were compared. Blastocyst re-expansion index was the main variable Main results and the role of chance: our data reported low viability of
analysed. Secondary variables were blastocyst morphology before biopsy, CC in the G2, which is also the group with the highest average age (39.5-2).
elapsed time between biopsy and vitrification and between warming and transfer. These results suggest that the incidence of CC viability could predict the age-re-
Participants/materials, setting, methods: Blastocyst morphology included: lated decline in fertility. Molecular studies indicate that the expression of 11β-
inner cell mass (ICM) and trophectoderm (TE) grade (A, B or C, according to HSD-1 or 2 in the ovary depends on the prevailing hormonal environment
ASEBIR classification). Biopsied blastocysts were vitrified/warmed as described and/or functional phenotype of cells. Compared to the control group our
by manufacturer (Kitazato®). In order to assess the degree of blastocoel re-ex- results indicate reduced levels of 11β-HSD-1 in G1 and G2 whereas 11β-
pansion, two pictures of the blastocysts were obtained: one taken immediately HSD-2 is down regulated in G2 only. Considering the 11β-HSD-1/11β-HSD-2
after warming (AW) and the second before transfer (BT). Re-expansion index ratio, which should increase physiologically during the ovulatory phase, this
was calculated as follows: diameter BT/diameter AW. Presence or absence of appears to have a clearly decrease especially in G2: this could mean that in
degenerated cells was also evaluated. this group an adequate conversion to cortisol does not take place. In addition
Main results and the role of chance: The mean maternal age was 35.5±6.1. in G2 the NR3C1 (glucocorticoid receptor) expression increase significantly
Sixty-three of 113 blastocysts implanted (IR=55.8%). Implanted blastocysts compared to control; this could indicate a greater presence of receptor for
exhibited significantly better ICM and TE quality than those that failed to implant low hormone levels as physiological feedback. PNPLA2 (involved in lipolysis)
(ICM A: 78.6% vs 21.4%, p=0.018; TE A: 80% vs 20%, p=0.012). Blastocoel is significantly increased in G2, this could indicate a greater presence of free
re-expansion index ranged from 0.2 to 2.4 without differences between fatty acids. The FASN expression (involved in the synthesis of fatty acids)
implanted and non-implanted blastocysts (1.3±0.3 vs 1.2±0.3 respectively; significantly increases in G1 and G2, compared to the control group and this
p=0.096). Degenerated cells after warming were detected in 39 blastocysts could indicate an increased cellular metabolism.
(34.5%). This feature was associated with an impaired implantation ability after Limitations, reasons for caution: Additional studies with an increased sam-
embryo transfer (64.9% vs 38.5%; p=0.007). Elapsed time between biopsy and ple size in different clinical group will help to validate present results. In addition,
vitrification ranged from 0.2h to 7.2h with no differences between implanted it would be appropriate to evaluate the viability and expression of the lipid
and non-implanted blastocysts (1.9h vs 2.0h; p=0.938). Elapsed time between metabolism genes in CC of individual oocytes and not by cumulating them as
warming and embryo transfer ranged between 0.2h and 5.2h with no differences we did in this study.
between implanted and non-implanted blastocysts (3h vs 3.2h; p=0.412). Wider implications of the findings: none
Limitations, reasons for caution: These are preliminary results. For a deeper Trial registration number: None
analysis controlling for potential confounding factors more data are needed. As unique
SOPs were applied in a single IVF setting the time ranges included were limited.
Wider implications of the findings: Time to vitrification after biopsy, as P-234 Intracytoplasmic sperm injection versus conventional in
well as time to transfer after warming can be adjusted according laboratory vitro fertilization in non-male factor infertility. Interim analysis of
needs. When dealing with poor morphology blastocysts before vitrification and a prospective randomized study
degenerated cells arise after warming, double embryo transfer might be pro- P. Fancsovits1, Z. Kaszas1, A. Nemes1, K. Szedlak1, A. Murber1,
posed to selected couples. D. Pocsi1, A. Arokszallasi1, J. Urbancsek1
Semmelweis University, Division of Assisited Reproduction- Department of Obstetrics
and Gynecology, Budapest, Hungary
P-233 Expression of genes related to lipid metabolism in cells of Study question: Does intracytoplasmic sperm injection (ICSI) superior to
the cumulus oophorus retrieved from patients undergoing assisted conventional in vitro fertilization (IVF) as a method of fertilization in non-male
reproductive treatments. factor infertility?
S. Pizzasegale1, A. Stendardi1, T. Ruocco2, F.L. Vellucci1, F. Ietta2, Summary answer: Conventional IVF resulted in higher fertilization rate while
B. Fineschi1, L. Gambera1 pregnancy and implantation rate was similar in IVF and ICSI cycles.

What is known already: ICSI is the most effective assisted reproductive one using NT, will improve oocyte competence, resulting in better IVF outcomes.
technique to treat male infertility. However, it is also used for non-male factor Nevertheless, there are limited data on applying NT in human for overcoming
indications, such as low number of oocytes or advanced maternal age. Thus, the mitochondrial diseases and female infertility.
incidence of ICSI fertilization method in fresh IVF cycles exceeds 70 % worldwide. Study design, size, duration: ST and recently proposed early (e)PNT were
However, data on the superiority of ICSI over conventional IVF in case of non- performed in the first patient using in vivo matured (n=5) and an in vitro matured
male factor infertility are still lacking. (IVM) oocyte, respectively. Donated enucleated frozen-thawed in vivo matured
Study design, size, duration: Data of interim analysis from an ongoing pro- oocytes with smooth endoplasmic reticulum aggregates (SERa) and a healthy
spective randomized study performed in a university setting are presented here. fresh IVM oocyte (after ICSI and enucleation) served as cytoplast recipients,
Two hundred thirty two IVF cycles were randomized into ICSI or conventional respectively. The second patient’s spindles were transferred from fresh IVM
IVF group between January 2018 and November 2019. oocytes (n=5) into enucleated frozen-thawed SERa oocytes from other patients,
Participants/materials, setting, methods: Inclusion criteria: semen param- followed by ICSI.
eters suitable for conventional IVF fertilization, number of oocytes ≤4 and/or Participants/materials, setting, methods: The first patient was a 30-year-
female age ≥40 years. Primary outcome was fertilization rate, secondary out- old woman carrying a homoplasmic mtDNA mutation m.11778G>A, which is
come was clinical pregnancy and implantation rates. Embryo development and known to cause Leber’s hereditary optic neuropathy (LHON) syndrome. The
quality (cell number, fragmentation, morphology score, rate of good quality second patient was a 26-year-old woman experiencing two failed ICSI cycles
embryos) on day 2 were also compared. Cycles with low oocyte number (A), characterized by almost all oocytes having fertilization failure. Finally, using
advanced maternal age (B) and both factor together (C) were also investigated B6D2F1 mice, we conducted RNA sequencing (RNA-seq) on PNT-blastocysts
in a subgroup analysis. (n=3). Blastocysts from ICSI-oocytes (n=3) and unmanipulated oocyte pools
Main results and the role of chance: Oocytes were fertilized by conven- (n=2) served as controls to assess safety concerns of NT technology.
tional IVF in 118 and by ICSI in 114 cycles. Fertilization rate was significantly Main results and the role of chance: For the patient harboring m.11778G>A
higher in IVF group (IVF:62,7%, ICSI:50,8%; P<0,001). Clinical pregnancy rate mutation, PGD confirmed almost 100% homoplasmic mutation load in all
was 24,5% (25/102) in IVF group and 18,8% (18/96) in ICSI group. The differ- embryos analyzed, which could not be used for embryo transfer. After ICSI, 5/5
ence was not significant (P=0,326). Implantation rate was also similar in the two of ST-oocytes were normally fertilized, 5/5 cleaved, and 1/5 progressed to the
group (IVF:13,0%, ICSI:9,6%; P=0,270). Number of blastomeres (IVF:3,79±1,34, blastocyst stage (scored as 4AA); while 1/1 reconstructed ePNT-zygote reached
ICSI:3,85±1,14; P=0,596), embryo score (IVF:2,17±0,87, ICSI:2,21±0,78; morula stage. NGS data revealed that mtDNA carry-over in two trophectoderm
P=0,495) and frequency of good quality embryos (IVF:29,4%, ICSI:26,9%; biopsy samples from the ST-blastocyst and the ePNT-morula was 3.2%, 2.9%
P=0,510) was comparable in the two group. and 3.1%, respectively, Carry-over of remaining arrested ST-embryos was 2.9%.
In cycles with ≤4 oocytes (Subgroup A) the fertilization rate, clinical pregnancy For the patient displaying fertilization failure, we first applied the mouse oocyte
rate and implantation rate was similar in the two group. In case of advanced activation test to evaluate sperm activation potential of patient’s partner. After
maternal age (≥40 years; Subgroup B) the fertilization rate was significantly higher injecting human sperm into mouse oocytes, more than 85% of mouse oocytes
in IVF group (IVF:64,1%, ICSI: 49,5%; P<0,001), but clinical pregnancy rate and were activated (suspected oocyte-related problem). The application of assisted
implantation rate did not differ between the two fertilization method. In cycles oocyte activation (AOA) resulted in only one oocyte fertilized (1/12), showing
where advanced maternal age was associated with low number of oocytes vague 2PN and no division further. Following ST and routine ICSI without AOA,
(Subgroup C), the fertilization rate (IVF:60,9%, ICSI:47,1%; P=0,043), clinical 2/4 reconstituted oocytes were normally fertilized with 2PN, one showed 1PN,
pregnancy rate (IVF:23,7%, ICSI:3,0%; P=0,013) and implantation rate (IVF:14,3%, and the last one underwent immediate cleavage, and 3/4 subsequently cleaved
ICSI:2,0%; P=0,018) was significantly higher in case of conventional IVF com- further but did not progress to blastocysts. RNA-seq data showed that mouse
pared to ICSI. PNT-blastocysts clustered very closely with ICSI controls, indicating that NT did
Limitations, reasons for caution: This is an interim analysis of a prospective not significantly change global gene expression.
randomized study. Number of cases are low to draw a powerful conclusion on Limitations, reasons for caution: The scarce availability of human oocytes
the effect of fertilization method on pregnancy an implantation rates. donated for research purposes is a limiting factor to obtain more pre-clinical
Wider implications of the findings: Our data show a clear evidence that evidence of this new NT technology. In addition, molecular analysis of recon-
ICSI is not superior over conventional IVF in non-male factor infertility as constructed embryos following NT is still required to verify the safety concerns.
ventional IVF resulted in higher fertilization rate. Furthermore, in case of Wider implications of the findings: Both ST and ePNT show the capacity to
advanced maternal age combined with low oocyte number, conventional IVF prevent the transmission of pathogenic mtDNA mutations of human, with minimal
could be the most effective method of fertilization. carry-over in the reconstructed embryos. Moreover, as a proof-of-concept, ST
Trial registration number: ClinicalTrial: NCT03513913 might be able to overcome fertilization failure in case of an oocyte-related activa-
tion deficiency causing failed fertilization after ICSI and AOA.
P-235 Germline nuclear transfer to overcome mitochondrial Trial registration number: Not applicable
disease and female infertility
M. Tang1, A. Boel1, R.R. Guggilla1, B. Bekaert1, P. De Sutter1, P-236 Performance variability among Spanish oocyte cryobanks
B. Menten2, F. Vanden Meerschaut1, D. Stoop1, P. Coucke2, G. Terzuoli1, M.C. Magli1, L. Maresca1, D. Visone1, A.P. Ferraretti1,
B. Heindryckx1 L. Gianaroli1
1
Ghent University Hospital Ghent Fertility and Stem cell Team, Department for 1
S.I.S.Me.R., Reproductive Medicine Unit, Bologna, Italy
Reproductive Medicine, Gent, Belgium ;
2
Ghent University Hospital, Center for Medical Genetics, Ghent, Belgium Study question: Do vitrified oocytes coming from different cryobanks have a
different performance in the same oocyte donation program?
Study question: Can germline nuclear transfer (NT) technique serve as a novel Summary answer: In our oocyte donation program using vitrified oocytes,
reproductive approach to overcome mitochondrial DNA (mtDNA) diseases as there are significant, cryobank-dependent differences in the clinical performance
well as female infertility? of the thawed oocytes.
Summary answer: The NT technique has a high potential to overcome mito- What is known already: Several studies show that the use of vitrified oocytes
chondrial disease transmission and certain forms of female infertility in human. does not negatively affect the clinical pregnancy rate compared to fresh oocytes.
What is known already: Mitochondrial disease prevention may currently be Therefore, the import of vitrified oocytes from cryobanks has been adopted in
possible using NT, such as spindle transfer (ST) and pronuclear transfer (PNT). many oocyte donation programs as a practical solution to the major problems
These techniques involve transferring nuclear genome from an oocyte/zygote related to the realization of this program. These include the difficulty of recruiting
carrying mtDNA mutations to an enucleated donor counterpart with unaffected local donors, and the need to perform the screen tests requested by the
mtDNA. Besides, NT has also been considered for certain infertility indications, European Directives on Tissues and Cells.
such as infertile women experiencing poor embryo development. This is mainly Study design, size, duration: This retrospective study includes 275 oocyte
based on the rationale that replacing a low-quality cytoplasm with a competent donation cycles from January 2016 to December 2019. We used vitrified oocytes

from four different cryobanks that were selected on the basis of their oocytes’ and KID+/-, Chi-Squared test was performed. To determine correlation
availability. Laboratory data and clinical outcome were analyzed and compared between type and when strings were expressed with KID+/-, ANOVA test
among the four cryobanks, including oocyte survival rate, clinical pregnancy rate was used. Correlation between presence/absence and embryo quality was
at the first transfer, and ongoing cumulative pregnancy rate. We also calculated studied using Kruskal-Wallis test. All statistics were performed using SPSS©
the number of oocytes needed to have a baby born. v22.0 (IBM, EEUU) software (p<0,05).
Participants/materials, setting, methods: The oocytes from the four Main results and the role of chance: Out of 154 embryos, 123 showed
cryobanks were used as follows. Cryobank bank 1, 67 cycles; cryobank 2, 22 cytoplasmic strings. 66,3% of them were KID+ and 56,7% were KID-. 31
cycles; cryobank 3, 116 cycles; and cryobank 4, 70 cycles. A minimum of 6 embryos did not show any cytoplasmic strings; in this case 16,7% were KID+
oocytes were allotted to each cycle and the same transfer policy was applied to and 14,3% were KID-. Significant differences were found (p=0,021) between
all cycles. All thawing procedures were performed by experienced embryologists presence or absence of cytoplasmic strings groups. Regarding type and moment
that strictly followed the given protocol and after a specific training provided in when strings were expressed, no correlation was found between those two
loco by each cryobank. variables and implantation outcomes (p=0 ,742 and p=0,585, respectively).
Main results and the role of chance: The characteristics of the recipients According to embryo quality and presence of cytoplasmic strings: 62,3% of
in the four groups were similar. No relevant differences among the four cryo- embryos were A quality, 25,5% B quality, 10,2% C quality and 2% D embryos.
banks were found in the survival rate (mean value 82.7%, range 79.08-89.4%). However, in the absence of cytoplasmic strings: 38,5% were A quality, 53,8% B
Conversely, the clinical outcome showed significant disparities with cryobank 1 quality and 7,7% C quality. Significant differences were registered between these
having the lowest clinical pregnancy rate per thawing cycle at the first transfer two groups (p=0,049).
(24.1%) in comparison with cryobank 3 (40.5%, P<0.025) and 4 (40%, P<0.05). Limitations, reasons for caution: To confirm if presence of strings in the
The same trend was registered in the cumulative pregnancy rate (29.8% vs. expanded blastocyst is not related with a poor embryo quality (C and D classi-
55.1%, P<0.005; and 51.4%, P<0.025 respectively). Clinical and cumulative fication in ASEBIR’s embryo selection criteria) and that could be a good embryo
pregnancy rates in oocytes from cryobank 2 were 22.7% and 36.3% respectively, implantation predictor, further studies should be carried out.
but due to the low number of treated cycles, these data were not used in the Wider implications of the findings: According to these findings and con-
statistical examination. To calculate the number of oocytes needed to have a versely to previous bibliography, the presence of cytoplasmic strings could have
baby born, the analysis of the data was restricted until December 2018. In all, positive implication on embryo development and implantation outcomes. This
the number of oocytes to be thawed to have a birth were 23.1 in cryobank 1, filopodia could provide cells from ICM, which is mitotically active, to TE, which
21.2 in cryobank 2, 15.7 in cryobank 3 and 14.6 in cryobank 4. is mitotically less active and more differentiated.
Limitations, reasons for caution: The number of cycles in the four cryo- Trial registration number: Not applicable
banks was not homogenous.
Wider implications of the findings: The different quality of thawed oocytes P-238 Clinical pregnancy in euploid blastocyst transfer is
from different cryobanks is not necessarily reflected by their survival, but by their correlated to early cleavage and blastulation morphokinetic
developmental potential. Considering a tight control of thawing conditions, parameters
possible causes could be the donors’ ovarian stimulation and/or the C. Jacobs1, M. Nicolielo2, L. Cremonesi2, E.L.A. Motta3,
operator of the vitrification procedure, suggesting that oocyte viability is J.R. Alegretti2, A. Lorenzon4
cryobank-dependent. 1
Huntington Medicina Reprodutiva, Embryology, Sao Paulo, Brazil ;
Huntington Medicina Reprodutiva, Embryology, São Paulo, Brazil ;
3
Huntington Medicina Reprodutiva / Federal University of São Paulo, Clinical
P-237 Cytoplasmic strings, who would say that they are not as bad Director / Associate Professor, São Paulo, Brazil ;
as we thought? 4
Huntington Medicina Reprodutiva, Research & Development, São Paulo, Brazil
M. Palma Rodríguez1, C. Olmedo-Illueca1, V.M. Chapero-Ciurana1,
D. Domingo-Fernandez-Vermejo1, M. Barea-Gomez1, L. Abad-De- Study question: What are the differences to achieve clinical pregnancy cor-
Velasco1, S. Royo-Bolea1, I. Cuevas-Saiz1 related to morphokinetic parameters with euploid embryos?
1
Hospital General de Valencia, Unidad de Medicina Reproductiva, Valenca, Spain Summary answer: Euploid blastocysts that results in clinical pregnancy
achieved time to 5 (t5), to 8 cells (t8) and blastulation (tB) at an earlier point
Study question: Is there any relation between cytoplasmic strings and embryo in time.
quality as well as implantation outcomes? What is known already: Embryo aneuploidy is known as the main cause of
Summary answer: In this study correlation was found between presence or embryo implantation failure. The next-generation sequencing analysis (NGS) of
absence of cytoplasmic strings and implantation outcomes in addition to blas- blastocysts biopsied cells is the current most advanced technique to diagnose
tocyst quality. embryo genetic constitution allowing the selection of an euploid embryo for
What is known already: It has been described that, during in-vitro embryo transfer. Beside the ploidy status, the morphokinetic evaluation for embryo
development, trophectoderm (TE) cells project cytoplasmic extensions to the selection can enhance the chance to achieve an ongoing pregnancy. The rapid
inner cell mass (ICM), known as cytoplasmic strings or filopodia. Cytoplasmic spread of the time-lapse technology increased the demand of algorithms to
strings are mainly classified into two groups: Short strings, which are extended predict embryo potential. However, intrinsic differences in each laboratory
from the ICM to the mural TE and long strings, which are extended from the should be taken into consideration before applying global algorithms in routine
ICM, trough the blastocoelic cavity, to the polar ICM. According to bibliography, before validation.
cytoplasmic strings are still present during expansion in poor quality blastocysts Study design, size, duration: Retrospective cohort study analyzing the mor-
and related to poor media conditions. phokinetic parameters of 227 euploid biopsied embryos (NGS) from autologous
Study design, size, duration: Presence/absence of cytoplasmic strings, type cycles (n= 132 patients) with known implantation outcome (single embryo
and the moment of development when were expressed (before/after or during transfer: positive or negative gestational sac (GS) and fetal heartbeat (FHB),
blastocyst expansion) were retrospectively analyzed in 154 blastocyst that under- double embryo transfer: negative or two GS and FHB). Data was collected
went fresh or frozen embryo transfer with known implantation data (KID). All between December 2017 and December 2019 in a private IVF center.
data were collected from 2015 until 2019. Two groups were defined regarding Participants/materials, setting, methods: Blastocyst biopsied embryos
KID+ or -. Furthermore, presence or absence of cytoplasmic string was also from patients undergoing IVF with clinical indication for PGT-A were included.
related to embryo quality. ASEBIR’s embryo selection criteria was used to assess All oocytes were fertilized by ICSI and cultured in time-lapse system incubator
embryo quality. (Embryoscope Plus, Vitrolife). Embryos were analyzed for the following mor-
Participants/materials, setting, methods: 131 patients underwent elec- phokinetic parameters: time of pronucleous fading (tPNf ), time to 2-cell(t2),
tive embryo transfer. 154 embryos were cultured at MIRI-TL® (Esco, time to 3-cell (t3), time to 4-cell (t4), time to 5-cell (t5), time to 8-cell (t8) and
Denmark) incubator using Continuous Single Culture Media (Irvine Scientific, time to blastulation (tB). Embryos were graded according to Gardner’s mor-
EEUU). To set correlation between presence/absence of cytoplasmic strings phology parameters.

Main results and the role of chance: Two-hundred twenty-seven blasto- in the DET group, without significant difference (OR of 0.74, 95%CI: 0.48-1.13;
cysts were analyzed: 165 single embryo transfers (sET) and 62 embryos from p=0.16). However, multiple pregnancy rate was significantly higher in the DET
31 double embryo transfers (dET). From those, 119 blastocysts resulted in group compared to the SET group (37.5% Vs 0%; p= 0.006), Moreover, preterm
positive clinical pregnancy (49%, 81 from sET and 38 from dET) and 108 in birth rate was 32.3% in the DET group compare to 13.9% in the SET group with
negative (51%, 84 from sET and 24 from dET). Maternal age was similar between OR of 2.94 (95%CI: 1.12-7.7; P value=0.03). Other obstetrical parameters were
positive and negative clinical pregnancy (38,43 ± 3,15 versus 38,31 ± 4,07, without significant difference between the two groups.
p=0.9090). Morphokinetics parameters were earlier in euploid embryos that The similar implantation rate following two cycles of SET compared to DET
achieved clinical pregnancy in three time-points: t5, t8 and tB (t5: 48,55 ± 6,42 in egg donation treatment means that the impaired endometrial receptivity fol-
versus 50,46 ± 6,63, p=0.028; t8: 56,93 ± 8,43 versus 60,81 ± 9,57, p=0.001 lowing the ovarian hyperstimulation in autologous IVF treatment is not the main
and tB: 106,58 ± 9,71 versus 109,64 ± 11,05, p=0.023). Regarding other param- factor to explain this finding. It may support the existing information that the
eters analyzed, all earlier time-points were similar between positive and negative transfer of frozen-thawed embryo has similar success rates as the transfer of
clinical pregnancy (tPNf: 23,11±3,76 versus 23,43±3,62, p=0.278, t2: 26,32±3,56 fresh embryo.
versus 26,66±2,99, p=0.239; t3: 36,84±4,29 versus 37,16±3,73, p=0.180; t4: Limitations, reasons for caution: The retrospective design of our study
38,11±4,92 versus 38,56±3,50, p=0.162). Morphology grades for positive and makes it more prone to bias.
negative clinical pregnancy were different between good quality embryos (grades Wider implications of the findings: This study can give us an insight on the
A and B – 86% versus 73%) and poor quality embryos (at least one grade factors that contribute SET, DET outcomes in autologous and egg donation
C – 14% versus 27%, p=0.021). Statistical significances were calculated using chi treatments. In addition, the study results support the use of elective SET in egg
square or t-test as appropriate. donation program in order to decrease the risk of multiple pregnancy without
Limitations, reasons for caution: This is an observational study based on decreasing implantation rate.
a retrospective database analysis, limited to biopsied embryos, generated by a Trial registration number: Not applicable
population with PGT-A indication.
Wider implications of the findings: Our study highlights differences in P-240 A novel microfluidic-based organoid mimicking the oviduct
morphokinetic parameters in euploid blastocysts that achieved clinical pregnancy. environment in vivo promote the embryo development via
Since the differences in these parameters were independent from aneuploidy reducing cellular reactive oxygen species (ROS) level
status, they represent potential time-points to be better analyzed in laboratory M. Wang1, B. Zhang1, L. Jin1
algorithms, reinforcing the potential use of morphokinetics as a tool for non-in- 1
vasive embryo selection. Tongji Hospital- Tongji Medical College- Huazhong University of Science and
Trial registration number: Not applicable Technology, Reproductive Medicine Center, Wuhan, China
Study question: Could a microfluidic-based oviduct organoid promote the

P-239 The effectiveness of two cycles with single embryo transfer embryo development?
(fresh + frozen) versus one fresh double embryo transfer in Summary answer: A microfluidic-based organoid mimicking both anatomical
women treated in egg donation program and biochemical environment of oviduct in vivo could promote the embryo
S. Herzberg1, S. Kraus1, N. Schachter-Safarai1, G. Karavani1, A. development via reducing cellular ROS level.
Simon1, A. Ben-Meir1 What is known already: In current methods, embryos were cultured in
1
Hadassah Ein Cerem- Jerusalem, Obstetrics and Gynecology, Jerusalem, Israel petri-dish under substantially static conditions, conversely during the most of
time, in vivo pre-implantation embryos experience dynamic micro-environments.
Study question: What is the effectiveness of two cycles of Single Embryo In addition, the processes of fertilization and development from zygote to cleav-
Transfer (SET) versus Double Embryo Transfer (DET) in egg donation IVF age embryo were completed in the oviduct. Consequently, oviduct is extremely
treatment. significant for pre-implantation embryo. However, due to the intra-abdominal
Summary answer: Implantation rate in two cycles of SET is similar to DET anatomical location, it is difficult to access an available oviduct model for embryo
in egg donation IVF treatment. Multiple pregnancy rate is significantly development at present. Microfluidic chips are widely applied in various walks
higher in DET. of life due to the extraordinary characteristics. Therefore, a microfluidic-based
What is known already: The practice of transferring multiple embryos results oviduct organoid was designed for embryo culturing.
in a high incidence of multiple births. Several studies on autologous IVF treatment Study design, size, duration: The microfluidic devices were fabricated fol-
have shown that two cycles of SET compared to DET result in a significant lowing standard rapid prototyping methods. Mouse primary oviduct epithelial
decrease of multiple pregnancy with similar pregnancy rates. Several factors can cells and embryos were obtained from ICR, and then identified by anti-Keratin
influence the results, including: the freezing and thawing process and an endo- immunofluorescence staining. Mouse embryos were cultured in chip and in dish
metrial receptivity impairment secondary to the ovarian hyperstimulation. In egg separately and cellular ROS level were measured.
donation we effectively neutralize the possible effect of exposing the endome- Participants/materials, setting, methods: The PDMS-PDMS devices were
trium to high levels of estrogen. This allows us to examine more clearly whether fabricated by multi-layer soft lithography technology. Oviduct epithelial cells and
differences between fresh embryo and frozen-thawed transfers are present. embryos were obtained from ICR. Mouse embryos were separated into two
Study design, size, duration: A retrospective cohort study conducted at groups for further development. In microfluidic group, embryos were co-cul-
two IVF units in the years 2010-2019. tured with oviduct epithelial cells in the microfluidic-based organoid, and in
The patients were divided into 2 groups: Group A - DET - transferring of two control groups were cultured in traditional dish. The numbers of embryos in
fresh embryos in one cycle, Group B - SET - transferring of a single fresh embryo different stages were recorded, and then retrieved and exposed in H2DCFDA
and later a second single frozen thawed embryo transfer in another cycle. to measure cellular ROS level.
Implantation rate, multiple pregnancy rate and obstetrical outcomes were com- Main results and the role of chance: The novel microfluidic-based organoid
pared between the two groups. in this study was able to mimic oviduct environment in terms of both anatomy
Participants/materials, setting, methods: Women treated with oocyte and biochemistry of fallopian tubes. The oviduct-on-chip models were with a
donation and who had at least two A-grade embryos were included in the study. length of 2 cm and a diameter of 1 cm, which were in accordance with the
The donors were healthy women 25-32 years. The endometrial preparation of anatomical characteristics of the oviduct in vivo. When the microfluidic chips
the recipient women included estradiol for two weeks. On the oocyte retrieval were connected to the computer control system, the liquid can flow into the
day, the recipient began treatment with progesterone. The recipients were micro-channels through the inlets to outlets at a slow speed. Mouse primary
treated with Long-acting GnRH agonist, to prevent unplanned ovulation. oviduct epithelial cells were obtained and identified by immunofluorescence
Main results and the role of chance: 251 women met the criteria set for staining, and then loaded into the chip to provide a suitable biochemical envi-
the study. 168 women were included in the DET group and 81 women were ronment. The levels of embryo ROS in microfluidic groups were statistically
included in the SET group. The two groups were similar in the demographic and significant lower than that in control group (P < 0.001). In order to explore the
clinical characteristics. Implantation rate was 32.3% in the SET group and 39.3% effect of different conditions on the development of embryos, fertilization rate,

4-cell embryo rate, 8-cell embryo rate and high quality embryo rate were cal- D. Freitas1, C. Teixeira1, A. Almeida1, L. Queiroz1, R. Carvalho2,
culated. Compared with control group, the rates of different stages embryos in T. Oliveira2, C. Santos2, T. Abreu2, M. Niag2, G. Coelho3
microfluidic group were higher, although there were no significant differences 1
IVIRMA Brazil- Clinic IVI Salvador, IVF LAB, Salvador, Brazil ;
(P=0.159, 0.246, 0.823, 0.258 respectively). 2
Universidade Federal do Recôncavo da Bahia., Medicine, Santo Antonio de Jesus,
Limitations, reasons for caution: The reduction of ROS concentration can Brazil ;
definitely benefit to the embryo quality. However, the gold standard of embryo 3
IVIRMA Brazil- Clinic IVI Salvador, Clinical director, Salvador, Brazil
quality is DNA integrity of euploid embryo. In addition, mouse oocytes and
sperms were used instead of human due to ethical requirements. More exper- Study question: How is the embryonic development, cytogenetic constitution
iments are needed using human tissue. and blastocyst rates of apronuclear zygotes presenting two polar bodies (“0PN-
Wider implications of the findings: This organ-on-chip device allows the prob- pronuclei / 2CP- polar body”)?
ability of mammalian embryo culture in a microfluidic-based manner. Further devel- Summary answer: Apronuclear zygotes with two polar bodies (“0PN / 2CP”)
opment might promote the combination of this novel embryo development methods can be euploid and have appropriate embryonic development.
and clinical IVF practice, and enhance the cumulative clinical pregnancy rate. What is known already: The fertilization process involves ordered morpho-
Trial registration number: not applicable logical changes that affect the gametes, cellularity, allowing to identify failures or
changes in the zygote. A fertilization assessment comprises complex and dynamic
classification systems for embryo morphology at various stages of development,
P-241 Universal warming protocol for vitrified blastocysts: The which impact on embryo selection. The average observation time for fertilization
“coming-out” of “off-label” use of warming kit brands is 16 to 22 hours. The interpretation of the absence of pro-nuclei (0PN) in
L. Parmegiani1, C. Garello2, S. Canosa2, F. Granella2, F. zygotes with 2 polar body and the use of these zygotes in IVF has been a topic
Evangelista2, G. Monelli2, D. Guidetti2, F. Bongioanni2, M. Filicori3 of debate. Cytogenetic analyzes reveal that a significant subset of these zygotes
1
GynePro Medical, Reproductive Medicine Unit, Bologna, Italy ; is diploid and with competent development.
2
LIVET Srl, LIVET Srl, Torino, Italy ; Study design, size, duration: This is a descriptive retrospective, cross-sec-
3
GynePro Medical NextClinics International, Reproductive Medicine Unit, Bologna, tional study. A convenience sample was used, covering the analysis of medical
Italy records of couples diagnosed with infertility, who underwent treatment at an
assisted reproduction clinic in Salvador, Bahia, Brazil. This work was developed
Study question: Is it possible to use warming kit brands “off-label” to efficiently during the period from January to December 2019.
warm vitrified blastocysts? Participants/materials, setting, methods: This study was carried out
Summary answer: Different vitrification and warming kits can be combined through the analysis of medical records of couples submitted to assisted repro-
without affecting survival and implantation rates. duction techniques for conjugal infertility. The analysis included 309 cycles of
What is known already: Embryos are routinely vitirified using different com- couples submitted to in-vitro fertilization and who had at least one apronuclear
mercial brands of ready-to-use vitrification solutions which differ only slightly in zygote. These analyzed embryos were cultured using sequential media until
their composition. Every brand of vitrification kit requires the use of its own blastocyst (D5 / D6). Fertilization was evaluated between 18 and 20 hours after
warming kit. However, it is useful for Assisted Reproduction centres to have ICSI (Intracytoplasmic Sperm Injection) of 2642 MII oocytes (in metaphase II).
alternative options for carrying out any procedure. The application of “Universal Main results and the role of chance: 2129 oocytes fertilized from 2642
Warming” through the combination of different vitrification/warming kits can oocytes MII injected, performing a fertilization rate of 80.58%. From these fer-
offer this flexibility, but this action may have potential legal consequences for AR tilized eggs, 295 were apronuclear, with the presence of 2 polar bodies (0PN /
centers. This study aims to give scientific validation to this common but “off-label” 2CP), representing 13.85% of the total number of zygotes. Among the 295 0PN
and thus often covert usage. / 2CP zygotes, a group of 36 embryos, belonging to 15 cycles of patients with
Study design, size, duration: Prospective randomized study. Vitrified blas- an average age of 36.72 years, had their development monitored, since they
tocysts were randomized at warming. Group randomization 1:1:1 via www. would be submitted to cytogenetic study. 24 blastocysts were generated, with
random.org. Total number of vitrified/warmed blastocysts: 231. Duration: a blastocyst rate of 66.67%, being submitted to PGT (Preimplantation Genetic
January – October 2018. Testing) by NGS (New Generation Sequencing). Among the 24 embryos ana-
Participants/materials, setting, methods: 107 blastocysts from patients’ lyzed, 13 embryos (54.17%) had a normal karyotype, and 11 embryos (45.83%)
own oocytes, 124 blastocysts form donors. Each patient’s/ donor’s embryos had an abnormal result. Of the 13 euploid blastocysts, only 9 embryos have
were vitrified with Kitazato Vitrification Kit (Japan) and warmed with three dif- already been transferred and 3 of these embryos have implanted, which gives
ferent warming kits: i) Kitazato, ii) Sage - Coopersurgical, Denmark, iii) an implantation rate of 33.33%.
Irvine -Fujifilm USA . Group names: KK, KS, KI, (patient’s blastocysts); eKK, eKS, Limitations, reasons for caution: The reduced number of embryos analyzed
eKI (donor’s blastocysts). Primary outcome measure: cryo-survival rate (embryos in this study does not allow the establishment of more precise correlations
surviving/embryos warmed). Secondary outcome measure: embryo implanta- between analyzed variables. This study will be complemented with new cycles
tion rate (gestational sacs/embryos transferred). to increase the investigation of the apronuclear zygotes (0PN / 2CP).
Main results and the role of chance: Cryo-survival rate was statistically Wider implications of the findings: Embryos from apronuclear zygotes
comparable between the study groups: KK 97.9% (47/48), KS 96.5% (53/55), tend to be discarded due to erroneous attribution of ploidy based on numbers
KI 100% (47/47), eKK 97.5% (39/40), eKS 100% (37/37), eKI 95.7% (45/47). of pronuclei. Studies that evaluate the development in blastocysts and the genetic
Embryo implantation rate was comparable between groups: KK 40.4% (19/47), evaluation, demonstrate that many apronuclear zygotes present diploid, resulting
KS 43.4% (23/53), KI 48.9% (23/47), eKK 46.1% (18/39), eKS 35.1% (11/37), in pregnancies with the birth of healthy babies.
eKI 46.7% (21/45). Trial registration number: not applicable
Limitations, reasons for caution: These results need to be confirmed with
larger studies investigating different “off-label” combinations of vitrification and
warming kits. P-243 Is the embryo utilization rate different after conventional
Wider implications of the findings: This study confirms that the combination IVF or ICSI in sibling oocytes?
of different kits for vitrification/warming permits efficient survival and implantation, I. Mateizel1, P. Drakopoulos1, H. Tournaye1, G. Verheyen1
irrespective of brand, cryoprotectants and basic medium in the kits. This result 1
UZ Brussel, Centre for Reproductive Medicine, Jette- Brussels, Belgium
supports the “off-label” use of different combinations of vitrification and warming
kits, allowing practitioners worldwide to “come-out” of the closet. Study question: Is the embryo utilization rate affected by the insemination
Trial registration number: Not Applicable procedure (conventional IVF or ICSI) in patients with split insemination of sibling
oocytes?
P-242 Embryonic development and cytogenetic evaluation of Summary answer: Embryo utilization rates are similar after IVF and ICSI in
apronuclear zygotes after 18 to 20 hours of in vitro fertilization sibling oocytes cycles.

What is known already: The use of ICSI in assisted reproduction has live-birth rates after transferring one or two Gardner’s grade BB or lower quality
increased exponentially since its clinical introduction in 1991. Initially developed frozen blastocysts.
for severe male factor infertility, it is now offered for all causes of infertility, Study design, size, duration: A retrospective cohort study including 1104
despite lack of data to justify this approach. Few studies have addressed the issue FET cycles was performed. All frozen Day 5 or 6 blastocyst of Gardner’s grade
of the optimal insemination technique, with conflicting results and possible biased BB or lower quality blastocysts transferred between January-2008 to
by the inclusion of women with poorer prognosis in the ICSI group. Hence, the December-2019 at an Academic Fertility center were included. 189 cycles were
ideal population to study the effectiveness of each insemination method, con- egg donor. Data was stratified for age, autologous or donor embryo and embryo
sidering also the ovarian response, would be patients undergoing IVF and ICSI quality. In the case of a double embryo transfer quality was based on the best
within the same cycle. grade of the two embryos.
Study design, size, duration: The aim of our retrospective study was to Participants/materials, setting, methods: 96 cycles were in patients >40-
compare the embryological outcome between IVF or ICSI insemination. All years of age and 819 cycles in women <40 years of age, at collection. Clinical
consecutive patients undergoing IVF versus ICSI procedure in their first ovarian pregnancy (CP), multiple pregnancy (MP) and live birth rates (LBR) per cycle
stimulation cycle (only antagonist protocol) between 2009 and 2014 were were compared between SET (n=969) and DET (n=135) in each group. Clinical
included. Only patients with fresh semen samples fulfilling the minimum criteria pregnancy and multiple pregnancy rates were compared between SET and DET
for IVF were considered. per cycle in grade BB (n=894), BC (n=86), CB (n=52) and CC (n=72) groups
Participants/materials, setting, methods: Following ovarian puncture, the individually.
sibling cumulus–oocyte-complexes (COCs) were randomly allocated to either Main results and the role of chance: In SET group, Gardner’s grade BB
IVF or ICSI procedure. In cases of an uneven COC number, the extra COC was blastocysts had higher pregnancy 48% (P=0.000), CP 34% (P=0.000) and LBR
included in the ICSI arm. The embryos were cultured in sequential media until 19% (P=0.059) in comparison to Gardner’s grade BC, CB and CC single embryo
day 3 or day 5/6. The main outcome was the embryo utilization rate per fertilized transfers. Within BB FET cycles, MP rate was significantly higher when transfer-
oocyte (total number of embryos transferred and cryopreserved divided by the ring two vs. one (5.9 vs. 1.9, P =0.009). Transferring two Gardner’s grade CB,
number of 2PN). BC or CC embryos did not increase the multiple pregnancy rate (P=0.78). If
Main results and the role of chance: A number of 418 patients were age at egg collection was ≥40 no difference was found in CP (11.1 vs. 11.7,
included and divided according to their ovarian response in: poor (≤ 3 COC P =0.9), MP (0 vs. 0) and LBR (6.3% vs. 0%, P=0.13) when SET or DET was
retrieved, suboptimal (4-9 COC), normal (10-15 COC) and high (>15 COC) performed. If age at egg collection was <40(N=819) the MP rate was significantly
responders.The median age [interquartile range (IQR)] was 32 (29-36), while higher in DET than SET (6.75% vs 1.63%, P=0.004) even though, age at collection
the number of oocytes retrieved per insemination method was 5 (3-7). (37 vs. 33years, P=0.001) and transfer (39 vs. 35years, P=0.001) were signifi-
Fertilization rates defined as the ratio of 2PN per number of COCs were sig- cantly older, respectively. However, no difference was found in CP and LBR,
nificantly higher in ICSI (67.5 vs. 54.1, p<0.001). However, embryo utilization overall. Among egg donors (N=189), there were no difference in CP (25.9% vs.
rates per 2PN were similar between oocytes inseminated with IVF and those 29.62%, P=0.68), MP (3.12% vs.7.4%, P=0.28) and LBR (11.72% vs. 22.2%,
inseminated with ICSI (53.8% vs. 57.4%, p value =0.3). In order to assess the P=0.137) between SET and DET. Among egg donors the LBR for Gardner’s
effect of the insemination technique on embryo utilization rates after accounting grade BB, BC, CB and CC were (8.2%, 6.3% 9% and 0%), respectively (P=0.74).
for several confounders (age, BMI, cause of infertility, initial stimulation dose, Limitations, reasons for caution: This is a retrospective cohort study with
number of COCs), a regression model with estimation by generalized estimating the inherent risk of undetected bias. The small number of cycles in women ≥40
equations (GEE) was used. According to GEE, the insemination method was not years of age at collection should be considered.
significantly associated with embryo utilization rate (coefficient 3.8, 95% CI -1.3 Wider implications of the findings: Among women with grade BB or lower
to 9, p=0.14). Also when considering subpopulations according to ovarian blastocyst at ≥40 years-of-age transferring 2 did not increase the MP rates. In
response, embryo utilization rate after IVF and ICSI were comparable (coefficient women <40 years-of-age transferring 2 BB blastocysts increased the MP rate,
3.4, 95% CI -1.8 to 8.5, p=0.2). but may fall into acceptable limits. MP rates did not increase when transferring
Limitations, reasons for caution: The retrospective nature of the present two CB, BC or CC blastocysts.
study can be considered as a limitation. Trial registration number: MUHC REB arsu6002/2020-6228
Wider implications of the findings: The robust design of our study, allowing
each patient to serve as its own control, provides reassuring evidence that
P-245 Assessment of molecular determinants underlying
embryo quality is not impaired by the insemination method used.
recurrent embryo developmental arrest and value of assisted
Trial registration number: B.U.N.143201939166
oocyte activation as a treatment option
A. Boel1, D. Bonte1, A. Cardona Barberan1, V. Thys1, D. Stoop1,
P-244 A comparison of live birth and multiple pregnancy rates P. De Sutter1, P. Coucke2, F. Vanden Meerschaut1, B. Heindryckx1
when 1 or 2 low quality blastocysts are transferred in a frozen 1
Ghent University Hospital, Ghent-Fertility And Stem cell Team- Department for
cycle, stratified for age Reproductive Medicine, Ghent, Belgium ;
S. Aldhaheri1, S. Arab2, A. Baeghiesh1, M.H. Dahan1 2
Ghent University, Center for Medical Genetics Ghent- Department of
1
McGill University, Obstetrics and Gynecology, Montreal, Canada ; Biomolecular Medicine, Ghent, Belgium
2
McGill University, Gyncecologic and Reproductive Endocrinology and Infertility -
Obstetrics and Gynecology Department, Montreal, Canada Study question: Can recurrent embryo developmental arrest be linked to
specific molecular defects and be overcome by assisted oocyte activation (AOA)?
Study question: What are the in-vitro fertilization (IVF) outcomes in patient Summary answer: Applying stringent inclusion criteria, AOA did not show
with Gardner’s grade BB or lower frozen blastocysts transfers and the effect on beneficial effects in our patient cohort, for which the molecular cause of embryo
multiple pregnancy rates? arrest remains mainly unexplained.
Summary answer: If age was <40 but not ≥40, the multiple pregnancy rates What is known already: Recurrent embryo developmental arrest has recently
were significantly higher in double embryo transfer (DET) than single embryo been linked to genetic defects in maternal effect genes. Treatment options for
transfer (SET) cycles. these patients are limited, however, assisted oocyte activation (AOA) has been
What is known already: Most studies have investigated outcomes based on proposed. AOA is routinely used to overcome fertilization failure after ICSI and
high quality blastocyst transfer or a combination of good and poor quality blas- involves the artificial induction of Ca2+ rises in the oocyte. The lack of Ca2+
tocysts. Outcomes among women with only Gardner’s grade BB, BC, CB and oscillations, normally evoked by the sperm factor phospholipase C zeta (PLCζ),
CC frozen embryos transferred (FET) are not well known. The outcomes strat- has been suggested to both contribute to failed fertilization, as well as to post-fer-
ified for age with low quality frozen blastocysts has also not been investigated, tilization processes, such as embryo arrest or inferior blastocyst formation.
nor has the role of transferring two poor grade blastocysts on multiple pregnancy Study design, size, duration: A prospective study was performed between
rates. Our study will be the first one to look at the multiple pregnancy and January 2018 and August 2019, including 11 couples, presenting with embryo

arrest or inferior blastocyst formation. Patients were only included when they The main objective of this study is to find out how the biopsy technique influ-
underwent at least two fresh ICSI cycles. Furthermore, high fertilization rates ences the subsequent genetic results and clinical outcomes.
(≥60% 2PN or zygote) with a minimum of 10 zygotes but low blastocyst rates Study design, size, duration: Retrospective database study including 499
(≤15% of at least early blastocyst quality on day 5) were required. embryos belonging to 162 PGT-A cycles performed between August 2017 and
Participants/materials, setting, methods: All couples underwent a fresh December 2018. Cycles with semen sample concentrations under 1 million/mL
AOA cycle (CaCl2 injection and ionomycin exposure (2x)). Both partners donated were excluded. Blastocyst were biopsied by two expert embryologists using the
a saliva sample for genetic screening of six MEGs (female) and PLCZ1 (male). same micromanipulation equipment. They performed two different biopsy tech-
Except for one lesbian couple, the Ca2+ oscillatory patterns induced by the male niques: pulling (292 embryos) and flicking (207 embryos).
patient’s spermatozoa were examined through mouse oocyte Ca2+ analysis Participants/materials, setting, methods: In pulling, cells were suctioned,
(MOCA). The mean maternal age was 33.4 years ± 3.4 years. Most couples suf- while laser pulses were applied (4-9). In flicking, cells were aspirated and after
fered from male factor infertility (n=3) or unexplained female infertility (n=3). 3-4 laser pulses they were removed by a quick movement of the biopsy needle
Main results and the role of chance: In total, 11 AOA cycles were per- against the holding.
formed and compared to 31 previous routine ICSI cycles. The overall fertilization Mosaic embryos with ≤25% aneuploidy were considered euploid. Euploid
rate after AOA application was 70.1% (96/137) which was similar to the fertil- embryo fragments with 25-40% of aneuploidy were classified as low-grade mosa-
ization rate of 74.8% (237/317) after conventional ICSI. Blastocyst rates did not ics, and those with 40-50%, were sorted as high-grade mosaics.
significantly increase after AOA (9.4%, 9/96) compared to the rates obtained Student’s t and Pearson’s chi-square were used.
in previous ICSI attempts (9.3%, 22/237). Furthermore, pregnancy rates were Main results and the role of chance: Pulling and flicking techniques achieved
not significantly different between AOA (28.6%, 2/7) and conventional ICSI significant differences in average number of laser shots needed (6.7±2.7 vs.
(36.8%, 7/19). Previous conventional ICSI cycles resulted in one miscarriage, 3.2±0.8, respectively; p<0.0001) to obtain similar number of trophectoderm
two biochemical pregnancies, and four healthy singletons. Regarding the 11 AOA cells biopsied (5.4±1.4 vs. 5.6±1.6, respectively). No differences in amplification
cycles, one embryo transfer resulted in an ectopic pregnancy, while the other failure between techniques were reported (4.8% vs. 4.3%, respectively).
pregnancy is still ongoing. Similar aneuploidy rate was observed between pulling and flicking (43.0% vs.
For all male patients, MOCA displayed Ca2+ oscillatory patterns with high 41.3%, respectively; p=0.812) but a tendency of higher euploid mosaicism rate
Ca2+ spike frequencies, comparable to controls, confirming a normal oocyte in the pulling group was noticed (16.1% vs. 10.2%; p=0.061). Furthermore,
activating capacity. Sequencing of the selected MEGs (NLRP2, NLRP5, NLRP7, flicking mosaics resulted to be more complex than pulling ones: more high-grade
TLE6, PADI6, KHDC3L) in female and PLCZ1 in male patients revealed several mosaics (35.6% vs. 60.0%; p=0.066), higher number of chromosomes involved
variants, most of which were categorized as benign and likely benign. No PLCZ1 in the mosaicism (1.6±1.0 vs. 2.5±2.1; p=0.025), and higher number of seg-
pathogenic mutations were identified in the ten assessed male individuals. mentary alterations (1.0±1.0 vs. 1.9±2.1; p=0.029).
However, amongst the eleven tested female individuals who underwent AOA, Until December 2018, 139 euploid single embryo transfers (SET) were per-
we identified three unique heterozygous variants of uncertain significance. formed (78 from pulling group and 61 from flicking group) and no differences in
Limitations, reasons for caution: Our study, showing that AOA is not able clinical pregnancy rate (34.7% vs. 32.8%) or live birth rate (32.1% vs. 27.8%)
to overcome embryo developmental problems, which is in contrast with earlier were observed, respectively. Nevertheless, it seems to be a tendency of lower
findings, is limited by its small sample size and displays therefore only preliminary implantation rate and delivery rates when flicking euploid mosaics were trans-
results. Genetic screening targets are too restricted to identify a genetic cause ferred: 45.5% implantation and live birth rate in the pulling group (11 SET) vs.
underlying the phenotype for all patients. 20% in the flicking group (5 SET).
Wider implications of the findings: Strong evidence for AOA to overcome Limitations, reasons for caution: This is a retrospective study, based on
impaired embryonic development is still lacking, therefore, embryo arrest should observation of current practice and its impact on subsequent results. Biopsy
not be an indication for AOA application. Genetic screening on a routine basis techniques were carried out by different embryologists, with the implicit bias on
might be valuable to improve clinical management. it. The sample size exploring reproductive outcome is small. More data must be
Trial registration number: NA compiled, and prospective studies carried out.
Wider implications of the findings: Even though laser used had been proved
safe, more than 4 shots employed in the pulling method could modify the genetic
P-246 Is mosaicism and its characteristics influenced by the result of the biopsy, increasing the euploid mosaicism rate.
method of trophectoderm-biopsy technique employed in PGT-A Mosaics derived from pulling are less-complex and may implant as euploid
cycles? Genetic and reproductive outcomes after pulling versus embryos, suggesting a plausible over-diagnosis of mosaics.
flicking. Trial registration number: None
L. Herrero Grassa1, M. Aparicio Gonzalez1, L. Cascales Romero1,
J.A. Ortiz2, J.C. Castillo3, C. García-Ajofrin4, J. Ten5, R. Bernabéu3
1
Instituto Bernabéu Madrid, IVF Laboratory, Madrid, Spain ; P-247 Association between IL-17A levels, oocyte fertilisation and
2
Instituto Bernabéu Biotech, Molecular Genetics Laboratory, Alicante, Spain ; embryo development in unexplained infertility
3
Instituto Bernabéu Alicante, Ginaecology, Alicante, Spain ; F. Reidy1, R. Kavanagh2, L. Glover1, C. O’Farrelly2, M. Wingfield1
4
Instituto Bernabéu Madrid, Gynaecology, Madrid, Spain ; 1
5
Merrion Fertility Clinic- National Maternity Hospital, Obstetrics and Gynaecology,
Instituto Bernabéu Alicante, IVF Laboratory, Alicante, Spain Dublin, Ireland ;
2
Trinity College Dublin, Trinity Biomedical Sciences Institute, Dublin, Ireland
Study question: Are pulling and laser (over)exposure influencing a (over)
diagnose of euploid mosaic embryos in PGT-A cycles?
Summary answer: Pulling and number of laser shots employed might influence Study question: Does dysregulation of the IL-17 pathway impact on fertilisa-
the genetic condition of the biopsied fragment leading to an over-diagnosis of tion and embryo development?
normal embryos as mosaic. Summary answer: Raised IL-17A levels prior to an ART cycle are associated
What is known already: Preimplantation genetic testing for aneuploidies with a reduced fertilisation rate.
(PGT-A) and blastocyst biopsy is a fashion trend in assisted reproduction, but it What is known already: The inflammatory cytokine IL-17A plays a multifac-
must be performed without damaging the embryo or disrupting its genetic con- eted role in modulating the immune response, and is thought to contribute to
dition. Laser technology is commonly used to manipulate embryos in the IVF successful embryo implantation. We recently showed that dysregulation of the
laboratory, optimizing the efficacy and reducing the exposure to suboptimal IL-17 pathway influences the outcome of ART cycles, such that elevated levels
conditions. of endometrial and serum IL-17 were associated with a reduced live birth rate.
Blastocyst stage biopsy is associated to high resolution and sensitive genetics In the current study, we analysed the impact of IL17 on ovarian innate immune
technologies that enable mosaicism detection, increasing the reproductive responses, and investigated whether elevated circulating IL-17A is associated
chances per PGT cycle. with impaired oocyte fertilisation and subsequent embryo development.

Study design, size, duration: Our aim was to investigate if serum IL-17A (E-SMs) obtained from 539 transferred embryos from 300 sub/infertile women.
levels impact on embryo fertilisation and embryo development in women under- Following ICSI and embryo cultures, E-SMs were stored frozen (-80º C) until
going ART cycles. Peripheral blood samples were taken at mid-luteal cycle phase, subjecting them to sandwich ELISA for sHLA-G (Mybiosource, USA). Measured
timed with luteinising hormone testing. Serum IL-17 levels were correlated with sHLA-G in E-SMs was correlated with embryo stages and their morphology and
oocyte fertilisation rates, embryo quality and pregnancy outcomes. This study clinical outcomes, in terms live-births.
also examined the effect of IL-17A, a potent proinflammatory cytokine, on the Participants/materials, setting, methods: We analysed E-SMs, for sHLA-G
regulation of three antimicrobial peptides, Human b Defensin 1, Psoriasin (ng/ml), from three embryonic stages i.e., 4-8-cells (cleavage-stage; n = 247),
and Elafin. morulae (n = 112) and blastocysts (n = 180). Obtained values for statistical
Participants/materials, setting, methods: 20 patients undergoing IVF/ differences were analysed between/among embryo-groups using the Student’s
ICSI were recruited. Patients were ≤38 years, with no previous pregnancy and t-test/ANOVA (P ≤0.05), followed by Tukey posthoc test. Levels of sHLA-G and
BMI<30kg/m². Serum IL-17A levels were measured using ELISA. An in vitro embryo morphology were correlated using Pearson correlation analysis. We
study was also performed utilizing a commercially available model of ovarian compared sHLA-G positive embryos with their pregnancy outcomes in terms
epithelia. Ovcar-3 epithelial cells were treated with recombinant human IL-17A of live-births.
and expression levels of the antimicrobial peptide (AMP) genes beta-defensin-1, Main results and the role of chance: Concentrations of sHLA-G were
PI3 and S100A7 were assessed by qPCR. significantly different (P<0.05) in E-SMs of cleavage-stage (0.89 ± 0.12; n = 23)
Main results and the role of chance: Mean fertilisation rate for oocytes embryos vs blastocysts (1.81 ± 0.21; n = 11; fresh-ETs). Similarly, sHLA-G con-
retrieved from women with a serum IL-17A level of <50pg/ml (0.7283; n=9) is centrations between E-SMs of cleavage-stage embryos (1.58 ± 0.17, n = 63) vs
significantly increased compared to that of women with a serum IL17A level blastocysts (3.06 ± 0.28; n = 46; frozen-ETs) were different (P<0.05). Within
of >50pg/ml (0.5857; n=11); p= 0.0193. With lower circulating IL-17A there and across clinic comparisons showed that concentrations of sHLA-G in E-SMs
is also a trend towards improved embryo quality, but does not reach statistical of cleavage-stage embryos, morulae and blastocysts were variable. A significant
significance. difference (P<0.001) was observed in sHLA-G concentrations only with cleav-
IL-17A stimulation of female reproductive tract epithelial cells in vitro increased age-stage embryos but not with morulae or blastocysts, indicating embryo devel-
antimicrobial peptide expression. This further connects this cytokine to the opment stage-wise correlations, albeit, with variability within and across clinics.
mediation of the innate immune response in the ovary. Furthermore, there was a moderate relationship between cleavage-stage embryo
Limitations, reasons for caution: This study suggests that IL-17A can mod- morphology and sHLA-G levels and, a positive correlation of the blastocyst
ulate innate immune responses in the female reproductive tract, including the grades with sHLA-G concentrations. Interestingly, sHLA-G levels were higher
ovarian microenvironment, in successful pregnancy. Our study is limited by the in live-births cases vis-à-vis no-birth cases; the sHLA-G concentration was higher
small sample size, and further study is ongoing to elaborate on these findings. in live-births out of blastocyst-ETs (1.79 ± 0.28, n = 22) vs those out of cleav-
Wider implications of the findings: Our previous work has found that age-ETs (1.44 ± 0.24; n = 42). Live-births were observed only when at least one
dysregulation of IL-17A impacts on success of ART. This further analysis suggests sHLA-G positive embryo was a part in twin- or triple- ETs. These results show
an effect on egg quality and subsequent embryo development. We have also a clear association of embryo-secreted sHLA-G levels with pregnancy outcome
shown that IL-17A induces expression of antimicrobial peptides,highlighting the with live-births.
role it may have in activation of the innate immune response. Limitations, reasons for caution: Variations of sHLA-G contents in E-SMs
Trial registration number: n/a between different IVF clinics could be attributed to protocol-practices differences
of clinics. More multi-centric larger-cohort studies and a large scale meta-analysis
P-248 Soluble human leucocyte antigen-G (sHLA-G) in embryo- are required to establish the diagnostic accuracy power of the sHLA-G for
spent medium is a potential embryo viability biomarker and a predicting the IVF outcomes in term of child-birth.
positive predictor of live-births: a retrospective Indian study Wider implications of the findings: For the first time, our multi-centric
Indian study, show that the sHLA-G could be an embryo viability biomarker to
V. Venkatappa1, S.S. Vasan2, S.K. Adiga3, V. Samson Roy4,
predict child-birth. The sHLA-G could potentially be an immunodiagnostic point-
G. Sachdeva5, P.B. Seshagiri6
1
of-care diagnostic test, a valuable adjunct to other criteria practiced for selection
Indian Institute of Science, Molecular Reproduction and Developmental Genetics, of potentially ‘top-quality’ embryo(s) for ET.
Bangalore, India ;
2
Grant: MHRD-ICMR, India.
Manipal Ankur Andrology & Reproductive Services-, IVF Clinic, Bangalore, India ;
3
Kasturba Medical College-, Department of Clinical Embryology-, Manipal, India ;
4
Advanced Fertility Clinic, IVF Clinic, Bangalore, India ;
5 P-249 The removal of a quarter of the Zona Pellucida improves
National Institute for Research in Reproductive Health-, Department of Primate
pregnancy and implantation rates in vitrified blastocysts that
Biology, Mumbai, India ;
6 underwent Artificial Shrinkage
Indian Institute of Science, Department of Molecular Reproduction- Development
and Genetics-, Bangalore, India A. Volpes1, F. Sammartano1, M. Modica1, S. Gullo2, A. Marino1,
A. Allegra1
Study question: Can embryo-secreted soluble human leucocyte antigen-G 1
ANDROS Day Surgery Clinic, Reproductive Medicine Unit, Palermo, Italy ;
(sHLA-G) serve as a predictive biomarker for embryo viability with successful 2
University of Palermo, Department of Psychology- Educational Science and
pregnancy outcome? Human Movement- Statistics Unit-, Palermo, Italy
Summary answer: Embryo-secreted sHLA-G concentrations showed positive
correlations with (i) developing embryo stages and (ii) live-births. Their altered Study question: Is the partial removal of the Zona Pellucida (ZP) effective in
concentrations were associated with pregnancy loss or miscarriages. improving pregnancy and implantation rates in vitrified blastocysts undergoing
What is known already: Human infertility is treated in IVF clinics with the Artificial Shrinkage (AS)?
live-birth rate being only <15%. Poor embryo viability contributes to pregnancy Summary answer: Partial removal of the ZP significantly improves pregnancy
loss. Current embryo-grading approach has limitations in terms of identifying and implantation rates in blastocysts that underwent AS either before vitrification
biologically-viable embryos that are capable of producing live-births. Several or after thawing.
reports have indicated that sHLA-G could act as a biomarker for elective-embryo What is known already: The hatching process is an essential mechanism
transfer, with positive IVF outcomes. But, there were controversies in terms of enabling blastocyst implantation. It is well known that vitrification may cause
correlative analyses, lack of cross-clinics comparisons and to predict live-births. hardening of the ZP and this event could explain implantation failure after blas-
Hence, there is a need for a cross-clinics comparison on the embryo viability tocyst thawing and transfer.
biomarker i.e., sHLA-G with embryo quality assessment and correlative analysis However, due to the thinness of the ZP in the expanded blastocysts, Laser
with live-births. Assisted Hatching (LAH) is almost impossible to perform.
Study design, size, duration: This was a three-IVF clinics retrospective study Moreover, published articles state that the blastocoelic fluid may cause inad-
performed between Oct 2017 and May 2019, using 539 embryo-spent media equate vitrification resulting in lower pregnancy and implantation rates.

To overcome this problem, a reduction in the volume of the blastocyst has far has assessed the impact of maternal age on the morphokinetics of embryos
been proposed, by using pre-vitrification AS to allow the blastocoelic fluid leak. competent to provide a LB.
Study design, size, duration: Retrospective observational study. All proce- Study design, size, duration: Retrospective study including 816 ICSI first
dures were performed at Andros Day Surgery Clinic, Reproductive Medicine cycles, from 2014 to 2018. 4915 embryos were produced, from which 2093
Unit, Palermo, Italy. were morphologically selected to be transferred in SET or DET 3 days after ICSI,
We included 409 warming cycles ( January 2017-October 2019) (625 blasto- resulting in 168 LB and 1176 non-implanted (NI) embryos with trackable mor-
cysts; 397 embryo transfer). phokinetics. Morphokinetic parameters were compared between LB and NI
Participants/materials, setting, methods: We compared outcomes in embryos, and within each embryo type, between patients <37 or ≥37 years.
cycles with blastocysts cryopreserved performing AS and LAH (group A) and The correlation between morphokinetic parameters and LB rates was tested.
blastocysts transferred without performing AS nor LAH (group B). Participants/materials, setting, methods: After ICSI, embryo culture was
Moreover, group A was divided in A1 (where AS was performed before performed in a TLM incubator. The morphokinetic parameters assessed were
vitrification; after thawing, LAH was applied to partially remove ZP) and A2 tPNf, t2, t3, t4, t5 and t8, which were compared between groups with different
(where blastocysts were vitrified without AS; after thawing, in the expanded embryo performance (LB vs. NI), and between LB embryos from patients < 37
blastocysts, AS and LAH were performed; if blastocysts were still spontaneously and ≥ 37 years by Wilcoxon test. The association between morphokinetic param-
collapsed, only LAH was conducted). eters and LB rates was assessed by multivariate regression analysis including
Main results and the role of chance: Significant differences were deter- maternal age and BMI as continuous variables in the model.
mined using chi-square with Yates’ correction and Fisher’s exact tests. Main results and the role of chance: All morphokinetic parameters were
Independent t-test was computed for continuous variable (age). A P-value < reached significantly earlier by LB embryos compared to NI embryos
0.05 was considered significant. [means(hours)±SD; tPNf: 22.5±2.7 vs. 24±3.4; t2: 25±2.7 vs. 26.8±3.7; t3:
Group A included 248 blastocysts transferred in 162 cycles. Group B included 36.1±3.3 vs. 37.7±4.5; t4: 36.8±3.6 vs. 39.1±5.1; t5: 48.8±4.9 vs. 50.1±6.4; t8:
345 blastocysts transferred in 235 cycles. 52.7±6.1 vs. 55.1±6.6; (p<0.0001)]. LB embryos from patients < 37 years
There is no significant difference in maternal age between two groups reached all morphokinetic endpoints faster than LB embryos from patients ≥ 37
(33.1±5.4 vs 32.05±5.5 p=0.06). years [tPNf: 22.3±2.7 vs. 23.2±2.4; t2: 24.8±2.8 vs. 25.7±2.5; t3: 35.7±3.1 vs.
Survival rate is the same in both groups (group A: 96.9% and group B: 93.5% 37.4±3.4; t4: 36.4±3.5 vs. 37.9±3.6; t5: 48.1±4.7 vs. 50.8±5; t8: 52.2±6 vs.
p=0.07). 54.2±6.2; (p<0.05)]. There were no differences between morphokinetic param-
Both pregnancy and implantation rates are significantly higher in group A than eters from NI embryos obtained from patients < 37 and ≥ 37 years. Consequently,
group B (pregnancy rate=47.5% vs 35.3% p=0.019; implantation rate=38.3% vs differences between LB and NI morphokinetics were more evident in patients
28.7% p=0.018). < 37 years, compared to those ≥ 37 years. Multivariate regression analysis
No differences are shown regarding multiple pregnancy rates (22.1% vs 19.3% revealed that, independently of maternal age and BMI, tPNf, t2, t5 and t8 are
p=0.81) or miscarriage rates (26.0% vs 25.3% p=0.93). significantly associated with LB rate in the overall patient population [tPNf: 0.7
We also compared group A1 and group A2. OR, (0.4-1.0) 95% IC, p=0.048; t2: 0.7 OR, (0.4-1.0) 95% IC, p=0.048; t5: 1.1
Group A1 included 200 blastocysts transferred in 127 cycles. (0.6-1.0), p=0.01; t8: 0.9 (0.9-1.0, p=0.047)].
Group A2 included 48 blastocysts transferred in 35 cycles. Limitations, reasons for caution: Our study is subjected to the intrinsic
Survival (97.6% vs 94.1% p=0.20), pregnancy (48.8% vs 42.9% p=0.53), limitations of a retrospective analysis, the results presented could have been
implantation (38,0% vs 39.6% p=0.84), multiple pregnancies (26.7% vs 21,0% affected by variables that are uncontrolled for.
p=0.89) and miscarriage (24.2% vs 33.3% p=0.69) rates are not statistically Wider implications of the findings: The present data indicate that early
different. morphokinetics can be used to predict embryo developmental competence, but
Limitations, reasons for caution: The study, being retrospective, limits its its predictive power decreases with maternal age. Therefore, our study provides
value. Furthermore, group A2 is less numerous than the others. valuable references for the design and application of embryo selection strategies
Wider implications of the findings: Our results seem to indicate that the utilizing TLM-derived information.
increased outcomes are not correlated to the AS. The removal of a quarter of Trial registration number: Not Applicable.
the ZP before embryo transfer (as in blastocysts collapsed before vitrification and
blastocysts collapsed after thawing) could be the key to the improved outcomes.
Trial registration number: Non applicable P-251 How many donor eggs are needed in an IVF-DO cycle?
B. Marqués López-Teijón1, A. Garcia-Faura1, S. Novo1, F. Garcia1,
C. Castello1, M. Lopez-Teijon1
P-250 Maternal age alters morphokinetics of embryos competent 1
Institut Marques, Reproductive Medicine Service, Barcelona, Spain
to provide a live birth
M.C. Guglielmo1, A. Bartolacci1, E. De Ponti2, F. Brambillasca1, Study question: Is the largest number of donor eggs always associated with
M. Mignini Renzini1, J. Buratini1,3, D.C. Mariabeatrice1 better results?
1
Biogenesi - Reproductive Medicine Centre, Istituti Clinici Zucchi, Monza, Italy ; Summary answer: Good results can be obtained with an adjusted number
2
ASST Monza, Dept Medical Physics, Monza, Italy ; of mature oocytes and a greatly increasing the number of ovules provides a
3
Sao Paulo University - Institute of Biosciences, Department of Physiology, limited value.
Botucatu, Brazil What is known already: There is much literature on the number of eggs
necessary to obtain success in an IVF of their own, but scarce about how many
Study question: Do morphokinetic parameters until the eight-cells stage differ mature eggs are necessary for an optimal result in case of IVF with donor eggs.
between embryos competent and noncompetent to provide a live birth, and Fertility clinics suffer the pressure of patients who need donor eggs because they
are they influenced by maternal age? believe that the greater the number, the more likely they are to succeed.
Summary answer: Embryos competent to provide a live birth display faster Study design, size, duration: We have conducted a single center retrospec-
early developmental kinetics than those noncompetent. Maternal age reduces tive observational study. The result of IVF cycles with egg donation is compared
kinetic differences between competent and noncompetent embryos. according to the number of mature oocytes assigned. 1,648 consecutive IVF
What is known already: Time-lapse microscopy (TLM) has been increasingly with donor eggs cycles were analyzed over a period of two years (2017-2019).
applied to improve accuracy in embryo selection. However, the lack of studies Three groups were created according to the number of mature oocytes assigned:
assessing the association of morphokinetic markers in embryos with their com- A. 4 mature oocytes in metaphase II (396 cycles), B. 5-7 oocytes (620 cycles)
petence to provide a live birth (LB) represents a significant knowledge gap. The and C. 8-10 oocytes (632 cycles).
performance of TLM-based algorithms has varied among different clinical set- Participants/materials, setting, methods: The groups are comparable in
tings, and patient profile, particularly maternal age, has been suggested as a mean age of donors (26.2 years) and recipients (43.0 years), in percentage of
significant driver of this variation. Nevertheless, to our knowledge, no study so fresh (91-94%) and vitrified (6-9%) oocytes, and in percentage of fresh embryo

transfers (35-37%) and frozen (63-65%). All embryo transfers were of single P-253 Oocyte incubation time prior to ICSI and its impact on
blastocyst of superior or equal quality to 3BB (Gardner). assisted reproduction results. A randomized controlled trial
Main results and the role of chance: Statistically significant differences (p R. Pesce1, A. Piazza1, R. Rolando1, C. Elizondo2, G. Perman3
<0.05) have been found in the average of good quality blastocysts obtained per 1
Hospital Italiano de Buenos Aires, Gyneacologucal Department, Buenos Aires,
cycle: 2.1 in group A, compared to 3.1 and 3.2 in groups B and C. There are Argentina ;
also statistically significant differences in the pregnancy rate per cycle: 85% in 2
Hospital Italiano de Buenos Aires, Epidemiology Department, Buenos Aires,
group A, compared to 91% and 90% in groups B and C. No statistically significant Argentina ;
differences were found between the groups in the pregnancy rate per transfer 3
Hospital Italiano de Buenos Aires, Public Health Department, Buenos Aires,
(61,5% to 65,2%) or in the newborn rate per transfer (44,2% to 47,2%). Argentina
Limitations, reasons for caution: It’s a retrospective single center study,
these results will vary depending on the success rates of each laboratory. Study question: Are there differences in oocyte maturity, fertilization rate and
Wider implications of the findings: Increasing the number of eggs up to a early embryo evolution (cleaving embryo) between early and late oocyte denu-
250%, the pregnancy rate per cycle rises just a 5%. These data suggest that good dation immediately prior to ICSI?
results can be obtained with an adjusted number of mature oocytes and that Summary answer: Early oocyte denudation is better than late denudation in per-
greatly increasing the number of eggs provides limited value. centage of: MII oocytes (OR 2.38; p 0.013) and cleaving embryos (OR 2.30; p 0.028).
Trial registration number: not applicable What is known already: The time of contact between cumulus cells and the
oocyte play an important role in the achievement of oocyte competence, oocyte
P-252 Reproductive potential of conventional IVF and ICSI on aging process and early embryo development. In addition, the time from trigger
sibling oocytes in the case of isolated teratozoospermia. administration to retrieval, and the time in culture before insemination affect
N. Thondehal Math1, G. Devika1, G. Deepanjali1, R. Sajana1, oocyte maturation and ovarian ageing in vitro.
R. Anjana1, B. Vidhya1, W.Y. Son2 There is controversy in international scientific literature regarding oocyte
1
Gunasheela Surgical & Maternity Hospital, embryology, Bangalore, India ; incubation time. Studies thus far were done in women with fertility issues, or in
2
McGill University Health Centre, Embryology, Montreal, Canada donor women but with retrospective designs.
The best timing of incubation prior to ICSI is yet to be determined.
Study question: Does ICSI have any embryological and clinical benefits for Study design, size, duration: Design: randomized controlled trial. Oocytes
the couple who have isolated teratozoospermia compared to conventional IVF were randomized to early or late denudation time (two arms). They were the unit
insemination? of analysis. Since oocytes coming from each donor woman were considered to
Summary answer: Reproductive potential is similar between both insemina- have a cluster effect, we adjusted for this with a random-effects multilevel model.
tion methods and so, couples with isolated teratozoospermia need not be sub- Sample size: 184 oocytes per treatment arm (accounting for the design effect)
jected to ICSI. to have 80% power and 5% alpha error to detect a 20% difference in results.
What is known already: In many studies, semen samples with teratozoosper- Duration: January 2016-December 2018.
mia produced lower fertilization rates when conventional IVF was used. On the Participants/materials, setting, methods: Setting: Oocyte donation pro-
other hand, in some studies, patients with teratozoospermia achieved good gramme in a university hospital.
fertilization in conventional IVF as long as the sperm concentration and motility Participants: Randomized oocytes came from healthy women (donors).
were within the normal range (isolated teratozoospermia) according to WHO Methods: Treatment arms: Early denudation: thirty minutes after oocyte pick-up
standards. Few studies observed the difference in embryo quality and further (OPU), oocytes were denudated from its cumulus. Late denudation: four hours
clinical outcomes of patients with isolated teratozoospermia between conven- later than OPU. All oocytes were subject to ICSI immediately after denudation.
tional IVF and ICSI. This study compares reproductive potential between con- All participants, professionals and researchers were blinded to treatment
ventional IVF and ICSI from the same patients with isolated teratozoospermia. allocation except for the biologist that performed the process (who did not
Study design, size, duration: These prospective studies were conducted in participate in other activities).
a private hospital between April 2015 and December 2019. In each case, sibling Main results and the role of chance: Out of a total of 375 included oocytes,
oocyte cumulus complexes were randomized to be inseminated either by ICSI 191 were randomly assigned to early denudation (ED) arm, and 184 to late
(n=1555) or IVF (n=471) (4: 1 ratio) to prevent completely failed fertilization. denudation (LD). The mean time to denudation was: ED, 0.5 hours (95% CI
Fertilization, good blastocyst rates (≥ 4BB) and clinical outcomes in ICSI group 0.42 – 0.58); LD, 4 hours (4 – 4.5); p <0.001.
were compared to IVF group. The crude percentage of MII oocytes was 91.6% and 83.7% (ED vs LD, respec-
Participants/materials, setting, methods: This study was conducted in tively), p 0.033. The crude percentage of fertilized oocytes was 55.3% vs 44.7%,
patients (n= 102, age= 34.7 ± 4.6 years old) for whom at least 10 oocytes were p 0.642. The crude percentage of cleaving embryos was 56.9% vs 43.1%, p 0.012.
retrieved and partner had isolated teratozoospermia. In the ICSI group, 1159 The adjusted odds ratio (OR) for MII oocytes (comparing early vs late denu-
oocytes out of total 1555 oocytes were mature and underwent ICSI, and the rest dation) was 2.38 (95% CI 1.20 – 4.72); p 0.013. The adjusted OR for fertilization
underwent conventional IVF. All fertilized embryos were cultured to blastocyst stage. was 1.30 (0.68 – 2.48); 0.423. The adjusted OR for cleaving embryo was 2.30
The best blastocysts produced were transferred in either fresh or frozen cycles. (1.10 – 4.85); p 0.028Limitations, reasons for caution: This is the first
Main results and the role of chance: Fertilization and blastocyst utilization randomized controlled trial that evaluates early vs late denudation in oocytes
rates were similar between ICSI group (75.8%, 878/1159; 67.8%, 595/878) and from healthy women. Its results can encourage the production of new studies
IVF group (74.5%, 351/471; 71.5%, 251/351) (P > 0.05). Failure of fertilization that confirm findings and assess its potential impact in clinical outcomes (preg-
after IVF insemination occurred in 5 of the 102 couples (4.9%) and in none with nancy rate and live birth rate).
ICSI. After transferring the embryos either in fresh or frozen cycles (n=94) there Wider implications of the findings: These findings suggest that early denu-
was no statistically significant difference in clinical pregnancy and ongoing/live dation could improve the odds of having cleaving embryos at third day after ICSI.
birth rates among cycles transferred from ICSI (58.13%, 25/43; 53.4%, 23/43), This could mean a need for change in usual embryology laboratory processes.
IVF (64.7%, 22/34; 47.1%, 16/34) and combined groups (61.7%, 11/17; 58.8%). Trial registration number: ClinicalTrials.gov Identifier: NCT03121924
Limitations, reasons for caution: The sample size is low. Although number
of sibling oocytes should have been put into IVF group, the design was intended
P-254 Make hey while sun shines! Hormone 25 OH D (vitamin D3)
to prevent total fertilization failure.
in follicular fluid: a determinant factor for top grade blastocyst
Wider implications of the findings: Since failed fertilization rate in IVF
formation?
insemination on normospermic infertility factor patients is also over 5%, our
study implies that IVF insemination is an option to avoid unnecessary use of ICSI N.M. Chimote1, B. Chimote2
1
which is time-consuming, costly and has potential risks for the couples with Vaunshdhara Fertility Centre, Embryology & Reproductive Endocrinology,
isolated teratozoospermia. Nagpur, India ;
Vaunshdhara Fertility Centre, Embryology- Endocrinology, Nagpur, India

Study question: To investigate relationship of 25(OH) D (vitamin D3) in What is known already: Theoretically, freeze-thawed embryo-transfer (ET)
follicular fluid (FF) with oocyte competence to fertilize, cleave and for top quality may have better ongoing pregnancy rate than fresh ET due to a more favorable
blastocysts. intrauterine environment by avoiding the condition after ovarian stimulation. It
Summary answer: Follicular-fluid level of 25(OH)D (Vitamin D3) is a poten- is although thought that the pregnancy potential of blastocyst frozen on day 5
tially predictive marker for oocyte competence to fertilize, cleave and form and slow developing day 6 blastocysts, after a freeze-thawed strategy, are com-
top-grade blastocysts in women undergoing IVF. parable. Synchronization of the donor-recipient cycle may have some difficulties
What is known already: There is growing evidence that [25(OH)D has very such as cancellation of the treatment, either of the donor stimulation or the
important role in human reproduction. Vitamin D3 contributes to restoration recipient endometrial preparation or the possibility of the absence of blastocyst.
of the menstrual cycle and endometrial proliferation, growth of follicles, improves These challenges may disappear with a freeze-all strategy.
primary dysmenorrhea, and reduce occurrence of uterine fibroids. However, Study design, size, duration: This was a retrospective cohort study con-
owing to conflicting results, the relationship of serum levels of Vitamin D3 with ducted at CEGyR, Buenos Aires, Argentina. A total of 650 women who were
ovarian stimulation characteristics or with embryo quality has been rather recipients having her first single embryo transfer from March 2016 and November
obscure. IVF provides a unique opportunity to explore such a relationship as 2019 were included.
measurement of vitamin D3 in follicular-fluid can help trace the fate of individual Participants/materials, setting, methods: A total of 650 recipients
oocytes, their fertilization and embryonic development. between 28-51 years who were having her first single ET were included. All ET
Study design, size, duration: This non-randomized prospective study were performed at blastocyst stage. The patients were separated in two groups:
included women (n=300, 22-42 years) undergoing IVF during January2017- (1) ET was made with a synchronization of donor-recipient cycle (2) ET was
December2019. None of the patients received vitamin D3 supplementation made after a freeze-all strategy. The analyzed variables were ongoing pregnancy
before Controlled Ovarian Hyperstimulation (COH). Follicular-fluid collected rate and miscarriage rate by chi square test. (The ongoing pregnancy rate deter-
from first aspirate of individual follicle was pooled, for each patient, to measure mined by transvaginal ultrasonography at gestational weeks 8-10).
Vitamin D3 levels using RIA kits. Embryonic development from fertilization to Main results and the role of chance: A total of 392 patients were included
blastocyst formation was recorded. Embryo gradation was done as per conven- in group 1 and 258 patients in group 2. The average age in group one was 41.5
tional criteria. All blastocysts were vitrified for next natural cycle embryo transfer. years old and in group two 42.1 years old. In the former group the ongoing
Participants/materials, setting, methods: Women with endometriosis, pregnancy rate was 47,70% (n=187) vs 43,41% (n=112) in the latter group (IC95
tuberculosis and hydrosalpynx and their male partners with severe or moderate 0,76-1,08 p=0.14). The miscarriage rate in the first group was 11,48% (n=45)
male factor were excluded from this study. Women with Poor ovarian response vs 14,34% (n=37) in the second group (IC95 0,86-1,86 p=0.11). Both groups
(≤ 3 retrieved oocytes) to recombinat FSH / gonadotropin stimulation were have similar results and they were not statistical significance.
also excluded. FF Vitamin D3 levels were divided into Low and High groups as Limitations, reasons for caution: The main limitation of this study was its
per their median value. Fertilization, cleavage and blastocyst formation rates retrospective design based on data from a single center which may be subject of bias.
were recorded in low and high FF vitamin D3 groups. Wider implications of the findings: While the freeze-all strategy is a more
Main results and the role of chance: Age, number of eggs retrieved and practical way to schedule treatments, it has not shown better clinical results.
MII oocytes were comparable between Low FF Vitamin D3 Group (< 41 ng/ However, prospective randomized studies are needed to confirm the hypothesis
ml, n=152) and High FF Vitamin D3 groups (> 41 ng/ml, n= 148). % fertilization, and a cost-effectiveness analysis to determine its applicability.
% cleavage was significantly higher in low group than high vit D3 group ( 93.3 ± Trial registration number: NOT APPLICABLE
22.6 vs 86.1 ± 29.5% P: 0.0205). Overall Blastocyst formation remained similar
(50.3 ±37.9 vs. 54.1 ± 35.4% : P value 0.3740). However, top and good grade
blastocysts were significantly higher in low FF/vit. D3 group than in high FF/vit. P-256 The association of early-cleavage embryo morphokinetics
D3 group (Top: 34.4 ±0.4 vs 24.2 ±0.8%, P= 0.0183 and Good: 37.8 ± 0.69 vs and blastocyst formation rate does not vary by women age: a
21.4 ± 0.72, p= 0.0002). time-lapse monitoring study
Interestingly, poor quality blastocysts were significantly high in high FF/ vit. A. Gomes1, H. De Martin1,2, M. Fujji1, M. Conatti1, M. Nakano2,3,
D3 compared to FF/low vit. D3 group (45.0 ± 38.5 vs 26.3 ± 33.0 %, P value: T. Bonetti4,5, P. Monteleone2,6
< 0.0001). Thus, measurement of vit. D3 in follicular fluid has tremendous 1
Monteleone - Human reproduction center, Embryology, São Paulo, Brazil ;
potential to identify the embryonic development to blastocyst stage with top or 2
Universidade de São Paulo - USP, Gynecology Discipline - Obstetrics and
good quality so as to select the best embryo for transfer. It also helps enhance Gynecology Department, São Paulo, Brazil ;
the chances of getting a viable pregnancy resulting in live birth. 3
Monteleone - Human reproduction center, Clinical, São Paulo, Brazil ;
Limitations, reasons for caution: Since this study has very limited number 4
Monteleone - Human reproduction center, Scientific, São Paulo, Brazil ;
of cases, more multicentric studies should be carried out to endorse impact of 5
Universidade Federal de São Paulo, Gynecology Department, São Paulo, Brazil ;
vit. D3 on embryonic development. Hence, pre treatment of supplementation 6
Monteleone - Human reproduction center, Clinical director, São Paulo, Brazil
of Vit D3 also should be contemplated according to variability of sunlight all over
the world. Study question: Does the embryo morphokinetics, from early cleavage until
Wider implications of the findings: With changes in weather and carbon emis- blastocyst formation based on time-lapse monitoring (TLM), vary according to
sion with problems in ozone layers all over the world, it seems that natural synthesis women age?
of vit. D3 is disturbed. Hence, proper evaluation of this hormone should be judi- Summary answer: The time-range of embryos progressing from two to three-
ciously done to improve excellent embryo development and a viable pregnancy. cell (CC2) is associated to blastocyst formation and does not vary by women age
Trial registration number: not applicable What is known already: ART success depends on a number of factors and
one of the most significant is the women age. Reproductive capacity in women
P-255 A freeze-all strategy in a donor egg program declines with ageing, associated with a decrease in oocyte quantity and quality,
A.M. Quinteiro Retamar1, M. Miguens2, A. Coscia2, N. Allende2, which correlates with lower fertilization rates and poorer embryo development.
G. Vega Balbuena2, D. Acosta2, G. Fiszbajn1 Embryo selection is a crucial step in ART and the introduction of TLM provided
1
CEGYR, EGG DONATION, caba, Argentina ; additional noninvasive criteria for better embryo selection and embryonic kinet-
2
CEGYR, clinical, caba, Argentina ics markers starting from the first cleavage. Early cleavage has been described
as a key factor for embryo selection, however, there is studies evaluating the
Study question: To compare ongoing pregnancy and miscarriage rate after variation in these parameters according to maternal age.
blastocyst transfer in an oocyte donation program in patients with synchroniza- Study design, size, duration: This retrospective cohort study used prospec-
tion of donor-recipient cycle versus freeze-thawed strategy. tively collected data from 671 ICSI cycles from 486 patients between April/2018
Summary answer: A freeze-all strategy may be an advantage allowing to and September/2019 in a private reproductive medicine center. A total of 4724
schedule treatments without cancellation or delays, but the reproductive results mature (MII) oocytes were fertilized by ICSI and cultured in a time-lapse mon-
have not shown statistical difference. itoring system (Embryoscope®). The time-lapse monitoring parameters and

blastocyst formation rate (BlastR) were compared in groups according to women collaboration between our laboratory and the foreign banks we were able to
age (<38 years or ≥38 years). optimize the entire procedure.
Participants/materials, setting, methods: Early-cleavage parameters were Participants/materials, setting, methods: For each egg donation cycle we
analyzed and correlated to blastocyst formation in each group. The time between receive six oocytes. The kitazato protocol was utilized for oocytes warming.
two-cell and three-cell (cell cycle two – CC2), three-cell and five-cell (CC3), Banks can be asked to send 3 more oocytes if the survival rate was less than
five-cell and nine-cell (CC4) were calculated. The blastocysts were classified 50%. Rates of oocytes survival, fertilization, cleavage, clinical pregnancy and life
according to morphology by Gardner’s classification and those with trophoec- birth were reported, including CLBR. Embryo transfer and cryopreservation
toderm grade ≥3 and inner-cell mass A or B were considered top-quality. The were performed on day 3 or 5.
blastocyst rate (BlastR) was calculated by dividing the number of blastocysts Main results and the role of chance: From 2015 to 2019 a total of 900 egg
obtained by number of 2PN. donation cycles were performed. The oocyte survival rate is 81,0±19,3%. The
Main results and the role of chance: From the 4724 MII oocytes injected, fertilization rate is 70,0±25%. The cleavage rate is 91,0±14,2%. The percentage
3622 were normally fertilized (2PN). From those, 1635 came from women of pregnancy rate in the fresh transfer is 36% with a life birth rate of 26%.
<38 years (48.8%) and 1716 from women ≥38 years (51.2%). 271 2PN oocytes 30.5% of patients underwent ET also have cryopreserved embryos, thus they
were excluded from analysis due to missing data. The mean time of CC2 have the chance to have subsequent ETs.
(<38-years: 10.0±4.7 vs ≥38-years: 10.5±4.8; p=0.005), CC3 (<38-years: Considering the cumulative percentage, i.e. including ET with warmed
12.4±6.6 vs ≥38-years: 12.7±6.8; p=0.212) and CC4 (<38-years: 23.1±8.9 embryos, the pregnancy and the life birth rates increase significantly, up to 40%
vs ≥38-years: 23.4±9.0; p=0.300) did not clinically differed between age and 32% respectively (p <0.05 for both parameters).
groups. A total of 2159 embryos developed to blastocyst stage (64.5%) and These results indicate that cryopreserved embryo transfers contribute sub-
the BlastR did not clinically differed between <38-years group (66.4%) and stantially to the total success rate of an IVF cycle with egg donation.
>=38-years group (62.6%, p=0.023) as well. The BlastR varied according to Limitations, reasons for caution: Cumulative life birth rate has been cal-
CC2 and the higher BlastR occurred when embryos presented CC2 between culated on concluded IVF cycles, i.e. if life birth was achieved or when no more
8 and 13 hours for both groups (<38-years: 80.5% vs ≥38-years: 75.9%; vitrified embryos were available
p=0.007). For the top-quality blastocyst rate (TQ-BlastR) the same pattern Wider implications of the findings: Our findings confirmed the efficacy of
was observed and TQ-BlastR when CC2 was between 8 and 13 hours were the double vitrification-warming procedure (at oocytes and embryos stage) that
(<38-years: 59.9% vs ≥38-years: 49.5%; p<0.001). Despite of statistically sig- could be used for supernumerary embryos or to postpone embryo transfer
nificance observed in some parameters compared between groups <38-years when the endometrial pattern is suboptimal.
vs ≥38-years, those did not have clinical impact as the means are very close Trial registration number: not applicable
and the statistical differences occurred possibly by the high number of embryos
included in the analysis.
Limitations, reasons for caution: This retrospective study evaluated the P-258 Evaluation of the relation between the value of
association of CC2 and blastocyst formation according to women ages. The mitochondrial DNA (MitoScore) and euploid embryo quality
blastocyst choosing criteria was based on blastocyst morphology and did not M. Niag dos Santos Rocha1, D. Freitas2, R. Carvalho3, T. Oliveira3,
take CC2 into account. Then, we could not evaluate the impact of CC2 on C. Santos3, T. Abreu3, C. Teixeira2, A. Almeida2, L. Queiroz2,
pregnancy success. G. Coelho4
Wider implications of the findings: We can underline that time of CC2 1
Universidade Federal do Recôncavo da Bahia, Centro de Ciências da Saúde, Santo
predict blastocyst formation and it does not vary according to women age. Antônio de Jesus, Brazil ;
Although the pregnancy rate decreases in older women, our findings support 2
Insituto Valenciano de Infertilidade - IVI Salvador, IVF Lab, Salvador, Brazil ;
the hypothesis that the morphokinetics of embryos from the first cleavage to 3
Universidade Federal do Recôncavo da Bahia, Medical School, Santo Antônio de
the blastocyst formation does not vary with age. Jesus, Brazil ;
Insituto Valenciano de Infertilidade - IVI Salvador, Clinical Director, Salvador, Brazil
P-257 Two Labs-Two countries: five-years’ report with cumulative Study question: Can the copy number of Mitochondrial DNA (mDNA) –
live birth rate of the ovum donation program in an Italian MitoScore – be associated with quality of an euploid embryo?
University Hospital Summary answer: The evaluation of the number of copies of mitochondral
R. Picone1, F. Bertocci1, R. Fucci1, P. Falcone1, C. Giachini1, DNA (Mitoscore) does not correlate with the quality of the embryo.
E. Borrani1, M.E. Coccia1, G.M. Cito1, L. Badolato1, P. Evangelisti1, What is known already: Assisted reproduction has tried to identify and
F. Rizzello1, D. De Angelis1, C. Cozzi1 develop strategies that make it possible to increase the rates of embryonic
1
Careggi University Hospital, assisted reproduction technology, Florence, Italy implantation and, consequently, pregnancy. Implantation failure rates with mor-
phologically and chromosomally normal embryos show that there are other
Study question: Two labs-two countries’ for the egg donation program: how factors that affect embryonic quality and fertilization rate. Mitochondria and their
is it efficient in terms of cumulative life birth rate (CLBR)? DNA (mDNA) have been suggested as a potential marker of embryonic viability
Summary answer: Combining fresh/frozen embryo transfer in the same and chances of fertilization, since their copies number is related to the energy
couple, deriving from donor vitrified and travelling oocytes is an efficient way to supply of the embryo. It is known that a larger number of copies of mDNA in
improve significantly the CLBR. a blastocyst seem to be associated with a lower implantation rate.
What is known already: Vitrification of oocytes and embryos is the golden Study design, size, duration: This is a descriptive retrospective, observa-
standard used worldwide for the cryopreservation in IVF and fertility preserva- tional, cross-sectional study. A convenience sample was used, covering the anal-
tion programs. The IVF outcome, in terms of pregnancy and implantation rate, ysis of medical records of couples diagnosed with infertility, who underwent
are comparable with those obtained with fresh oocytes/embryos. Advances in treatment at an assisted reproduction clinic in Salvador, Bahia, Brazil, in total
reproductive techniques, i.e. the introduction of oocytes vitrification, has allowed convenience sample of 69 cycles, from a total of 69 patients. This work was
simplifying the donation process by the formation of egg banks. The extremely developed during the period from from August 2016 to December 2019.
low rate of egg donation by Italian women does not satisfy the current level of Participants/materials, setting, methods: This study evaluated 173
demand and over 90% of the IVF cycles with egg donation originated from foreign embryos which underwent preimplantation genetic testing for aneuploidy (PGT-
(mainly Spanish) gamete bank. A) and MitoScore testing (on euploid embryos) was performed by Igenomix.
Study design, size, duration: Retro-prospective study on our couples from Embryos were classified on the basis of embryo quality (EQ) utilizing ASEBIR
2015 to 2019. In our five-years’ experience we have been used the so-called criteria into three groups: Good (AA/AB/BA/BB), Fair (BC/CB), Poor (CC).
‘two country-two lab’ model meaning that the oocytes retrieval and the subse- Software Statistical analysis was used and the differences between three groups
quent vitrification is performed in foreign banks while their fertilization after was estimated by ANOVA test. In all cases, p < 0.05 was considered statistically
warming by ICSI and embryo transfer is made in our laboratory. Thanks to significant.

Main results and the role of chance: Data from a total of 69 cycles, of 69 Overall, 176 embryos were obtained and assessed for conventional morphology
women, With an average age of 36.31 (+ 5.29) years. Of the 173 embryos ana- showing a significantly higher morphological score on day 2 (7.4±2.3 vs 6.7±2.1;
lyzed, there were 115 (66.47%) embryos with Good EQ, 41 (23.70%) embryos p<0.01) and a higher percentage of top quality embryos (score≥8) (45.2% vs 26.1;
with Fair EQ and 17 (9.83%) embryos with Poor EQ. In embryo group with Good p<0.01) in MI group (n=84) compared to FA group (n=92). At variance, compa-
EQ, the average of Mitoscore value was 23,29 (+ 5.98); in group with Fair EQ, rable embryo morphokinetic parameters were observed in the study groups: time
the average of MitoScore value was 22,76 (+ 5.42); and in group with Poor EQ, of pronuclear appearance (tPNa), pronuclear fading (tPNf ), completion of cleav-
the average of MitoScore value was 24,12 (+ 6.07). There was no statistically age to two, three, four and eight cells (t2, t3, t4, t8), time between first and second
significant difference between the groups evaluated (p=0.7167). In addition, it was cytokinesis (P2) and between second and third cytokinesis (P3).
also observed that of the 173 embryos analyzed, 86 were transferred, in a total After ultrasound-guided ET, the implantation rate was 30% (10/30) in MI
of 63 transfer procedures (with one or two embryos). Of this total transfer, 29 group and 11% (3/28) in FA group, showing significant differences (p<0.05) and
procedures were implanted, which represents a 46.0% implantation rate. an ongoing pregnancy rate of 40% (8/20) in MI group and 5% (1/20) in FA group
Limitations, reasons for caution: The reduced number of embryos analyzed (p<0.01)Limitations, reasons for caution: The conclusions of this study
in this study does not allow establishing more precise correlations between may not be generalizable to other patient populations, time-lapse systems or
analyzed variables. This study will be complemented with new cycles to increase microscopy. The number of observations is limited and the conclusions should
the investigation about importance of MitoScore in prediction of embryo viability be verified in a prospective study on a larger number of embryos
and implantation rates. Wider implications of the findings: This is the first, preliminary evaluation
Wider implications of the findings: There is no association between of the influence of a multivitamin compound on oocyte morphological features
embryo quality (EQ) and embryo’s MitoScore in this study. This result is in and embryo morphokinetics. If confirmed on a larger sample size, our preliminary
agreement with evidences in literature. However, the knowledge of this value findings may help to a better oocyte and embryo selection in overweight patients
can be useful in the decision in order of embryonic transfer, when having Trial registration number: not applicable
embryos with similar EQ.
Trial registration number: Not applicable P-260 Post-warming culture duration and effect on live birth rates
in single, top-quality vitrified blastocyst transfers – A retrospective
analysis of 839 transfers.
P-259 Effect of Myo-Inositol and Alpha-Lipoic Acid on oocyte H. Premannandan1, S. Yasmin1, G. Sotirchou1, G. Srivastava1
morphology and embryo morphokinetics: a prospective 1
Homerton University Hospital, Women’s and Sexual Health, Homerton, United
preliminary analysis of 40 overweight patients undergoing ICSI
Kingdom
treatment
G. Gennarelli1, C. Paschero2, S. Canosa1, C. Benedetto1, A. Revelli1 Study question: Do live birth rates differ based on the post-warming culture
1
Turin University, Obstetrics & Gynaecology, Torino, Italy ; duration in top-quality, single vitrified blastocyst transfers in good prognosis women?
2
Turin University, Obstetrics & Gynecology, Torino, Italy Summary answer: Live birth rates were similar in top-quality, single vitrified
blastocyst transfers cultured ≤ 2 hours, between 2 to 3 hours and > 3 hours
Study question: May the administration of a multivitamin compound based post-warming.
on Myo-Inositol, Alpha-Lipoic Acid and Folic Acid before controlled ovarian What is known already: It is generally accepted that the post-warming culture
stimulation influence oocyte and embryo quality? duration should be a minimum of 2 hours before the transfer. However, the
Summary answer: The administration of Myo-Inositol, Alpha-Lipoic Acid and question of difference in live birth rates, when transferring a viable blastocyst
Folic Acid may affect both oocyte and embryo morphological features and repro- that was of top-quality at vitrification, before or after the recommended
ductive outcomes in overweight patients post-warming culture duration, has been not fully answered.
What is known already: Several clinical studies have demonstrated that both Study design, size, duration: Retrospective cohort study of 839 frozen
Myo-Inositol and α–Lipoic Acid are involved in mitochondrial respiratory chain blastocyst transfers in a single UK centre (March 2015 - November 2018). Post-
and energy metabolism having a positive effect oocyte quality and embryo devel- warming culture duration of ≤ 2 hours (Group A, n=208), between 2-3 hours
opment of women affected by PCOS. Therefore the administration in PCOS (Group B, n=396) and > 3 hours (Group C, n=235) were compared to live birth
patients may improve reproductive outcomes by both affecting metabolic path- rates. For power of 90% with 95% CI, demonstrating medium effect size, mini-
ways and reducing oxidative stress. Nevertheless, data are missing on the use mum 141 participants were needed in each group for statistical significance using
of Myo-Inositol in women defined poor responders undergoing ovarian stimu- Chi-Square (χ2) test of independence.
lation for ICSI cycle. Therefore, the aim of the present study was to provide Participants/materials, setting, methods: Non-smoker women with BMI
evidence on the effect of this compound in overweight patients less than 30, age ≤ 37 years at oocyte retrieval, having a single, top-quality
Study design, size, duration: This is a randomized prospective analysis of blastocyst at vitrification were included. All women had hormone replacement
40 patients aged 25-40 years, with 27≤BMI≤32, AMH≥0.1, FSH≤15 without therapy (HRT) for the preparation of the endometrium. Standard dose of
diagnosis of diabetes and autoimmune diseases undergoing ICSI treatment at micronized progesterone pessaries was used for luteal phase support. Outcome
our IVF Unit between 2018 and 2019. According to the randomization patients measure was live birth rates.
assumed either the multivitamin compound (MI group, n=20) or Folic Acid alone Main results and the role of chance: In total, 839 transfers were analysed.
(FA group, n=20) for three months before COS (i) The 3 groups of post-warming culture duration intervals yielded live birth
Participants/materials, setting, methods: Clinical characteristics and rates of 32.69%, 38.64% and 39.15% respectively. No statistical difference
reproductive outcomes (clinical pregnancy rate and miscarriage rate) of the between the groups was found [X2 (2, N = 839) = 2.534, p = 0.2817].
whole cohort of patients were recorded and analysed using t-test or chi-squared Post-warming culture duration does not impact the chance of live birth rates in
test as appropriate. Oocyte Zona Pellucida (ZP) and Meiotic Spindle (MS) mor- single, top-quality vitrified/warmed blastocyst transfers in good prognosis
phological features were examined using Polarized Light Microscopy whereas patients.Limitations, reasons for caution: Retrospective nature of the study.
embryo quality was assessed using the Integrated Morphology Cleavage Score The expansion of the blastocysts has not been considered in this study; however,
evaluated on day 2 (Holte 2007) integrated with morphokinetics annotation by only viable embryos were transferred.
Time-Lapse system. Statistical significance was set at p<0.05 Wider implications of the findings: In busy IVF units, top-quality vitrified
Main results and the role of chance: The clinical characteristics of the blastocysts can be transferred between 2 to 4 hours without any detrimental
patients and the outcome of their IVF cycles showed no statistically significant effect on the live birth rate.
differences. A total number of 324 oocytes were observed under PLM revealing Trial registration number: not applicable
a significantly higher zona retardance (2.1±0.6 vs 1.9±0.5; p<0.0001), area
(2895.3±574.4 vs 2574.8±491.1;p<0.00001) and thickness (5.3±1.4 vs 4.5±1.2; P-261 Post warming re-expansion of single top-quality blastocysts
p<0.05) and a significantly shorter meiotic spindle axis (11.8±1.8 vs 12.3±2.3) and the relationship with live birth rates (LBR) – A retrospective
in MI group (n=155) compared to FA group (n=169). analysis of 839 frozen embryo transfers

G. Srivastava1, S. .yasmin1, H. .premannandan1, G. Sotirchou1, R. effect on embryo metabolism, developmental competence and quality. Especially
Homburg1 the blastulation rate that’s much affected by high pH/osmolality.
1
Homerton University Hospital - E9 6SR, Fertility Department, London, United Study design, size, duration: A prospective randomized study included 1695
Kingdom MII sibling oocytes collected from 100 patients undergoing ICSI at private center
from August 2019 to January 2020. We used sibling oocytes splits between two
Study question: Do the live birth rates (LBR) differ based on the post-warming different dishes : GPS (life global, USA) and SPL (life science, Korea) with two
culture duration in top-quality, single blastocyst transfers in good progno- different oils : light mineral oil (LM)(Irvine, USA) and paraffin oil (PO)(Vitro life,
sis women? Sweden). All embryos cultured in dry incubators at the same conditions.
Summary answer: LBR differ significantly based on the re-expansion of vit- Participants/materials, setting, methods: Maternal age is ≤37 years old
rified top-quality blastocyst cultured post-warming for ≤ 2, 2 to 3 and ≥ 3 hours. with ≥ 8 MII oocytes. First, we split (211 MII) between GPS and SPL dishes
What is known already: Re-expansion of the vitrified blastocyst is accepted overlaid with LM, Other (435 MII) splits overlaid with PO, second: (176 MII)
to give an indication of its implantation potential and reproductive outcomes. siblings were cultured in GPS dishes and splits between LM and PO, (416 MII)
Failure to re-expand affects 25% of warmed blastocysts. Nonexpanded, viable, cultured in SPL dishes and splits between LM and PO. And, third: (457 MII) splits
blastocysts are deemed suitable for transfer within 4 hours of post-thaw culture were cultured in GPS overlaid with PO vs SPL overlaid with LM.
in many units across the world. Warming of another blastocyst is rarely justified Main results and the role of chance: Data collected and analyzed using
unless substantial damage to the trophectoderm is observed. statistical software, results considered significant if p value ≤ 0.05.There were
Study design, size, duration: Retrospective cohort study of 839 frozen no significant differences in fertilization, cleavage rates or high-quality day 3
embryo transfers conducted in single UK centre from March 2015 to November embryos in all groups. There was significant difference in blastulation rate
2018. The post-warming culture duration was divided into three groups: ≤ 2 (75%,60%) (p = 0.02) between GPS dishes vs SPL dishes overlaid with LM
hours (Group A, n1 = 61 n2 = 147), between 2-3 hours (Group B, n1 = 106, respectively. GPS dishes overlaid with PO showed significantly higher blastulation
n2 = 290) and ≥ 3 hours (Group C, n1 = 54, n2 = 181). LBR were compared rate (67% vs 47%) (p<0.001) and high-quality day 5 embryos (75%,48%)
between non-expanded (n1) and expanded (n2) group respectively. (p <0.001) than SPL overlaid with PO. On the other hand, there were no sig-
Participants/materials, setting, methods: Non-smoking women, with BMI nificance in SPL splits between LM vs PO overlay in any recorded embryological
l< than 30, age ≤ 37 years at oocyte retrieval, having a single, top-quality embryo parameters. In GPS splits between LM vs PO groups, the only significance was
at vitrification were included. All women had hormone replacement therapy in high quality day 5 embryos (51%,32%) (p = 0.01) respectively. the combined
(HRT) for the preparation of the endometrium. A standard dose of micronized system of GPS with PO vs SPL with LM showed significant increase in high quality
progesterone pessaries were used for luteal phase support. The outcome mea- day 5 embryos (75%,62%) (p = 0.003) respectively, and tend to be significant in
sure was the live birth rate. High quality day 3 embryos (75%,67%) (p = 0.06) respectively. So, media drop
Main results and the role of chance: In total, 839 transfers were analysed geometry of 3D dishes combined with paraffin oil lead to higher pre implantation
with non-expanded and expanded status with an enrollment ratio of approxi- embryonic development with higher morphological quality.
mately 1:2.5. The study was powered at 80% with a 95% confidence interval. Limitations, reasons for caution: Small sample size and the morphological
To demonstrate high effect size at least 143 total participants (n1+n2)were assessment would be better if combined with morphokinetics or PGT-A results,
needed for each of the time intervals respectively. especially aneuploidy, mosaicism and deletion vs addition in the chromosomes.
The proportion tests, two-sided Z-tests at 5% level of significance for each The study needs to be expanded to include ongoing pregnancy rate.
group confirmed that the differences in live birth rates between non-expanded Wider implications of the findings: If these findings backed with larger
and expanded status at different time intervals are statistically significant (Group prospective randomized trials to reach statistically powered results, the impact
A: non-expanded 21.31% vs expanded 37.41%, p = 0.024; Group B: non-ex- of dry conditions on embryonic culture media could be overcame by using 3D
panded 30.19% vs expanded 41.72%, p = 0.03; Group C: non-expanded 22.22% dishes overlaid with heavy oil.
vs expanded 44.20%, p = 0.003) Trial registration number: Not applicable
Limitations, reasons for caution: The study was retrospective in nature.
Wider implications of the findings: In busy units, top-quality, warmed, P-263 Validation of a deep convolutional neural network
viable, non-expanded blastocysts can be transferred between 2 to 4 hours with trained to assess whether an embryo meets criteria for biopsy/
acceptable LBR. However,re-expanded blastocysts increased LBR significantly. cryopreservation using a test set of PGT embryos
We recommend, if the blastocyst has not re-expanded within 4 hours, there I. Souter1, S. Vagios1, C. Sacha1, M.K. Kanakasabapathy-2,
can be a discussion with the patient to warm remaining blastocyst to improve LBR. P. Thirumalaraju2, C. Bormann1, H. Shafiee2
Massachusetts General Hospital-Harvard Medical School, Obstetrics/Gynecology-
Reproductive Endocrinology and Infertility, Boston-MA, U.S.A. ;
P-262 The effect of culture media microdroplet geometry and 2
Brigham & Women’s Hospital-Harvard Medical School, Medicine/Division of
different oil overlay on pre implantation embryonic development Engineering in Medicine, Boston, U.S.A.
in dry incubators: Prospective randomized study
M. Hozaien1, A. El shimy1, H. El khedr1, H. Zaki2 Study question: Can a deep convolutional neural network (CNN) decide
1
Ganin Fertility Center, IVF Laboratory, cairo, Egypt ; whether a Day-5 (D5) embryo, previously assisted hatched (AH) for preimplan-
2
Ganin fertility center, Clinic, Cairo, Egypt tation genetic testing (PGT), meets biopsy/cryopreservation criteria?
Summary answer: The CNN exhibited an outstanding performance in iden-
Study question: Do the culture media microdroplet geometry of different tifying, among AH embryos, D5 blastocysts that met developmental criteria for
culture dishes and oil overlay affect the pre implantation embryo development biopsy and cryopreservation.
in dry incubators? What is known already: Embryologists make procedural and disposition
Summary answer: Pre implantation embryonic development in dry incubators decisions based on embryo morphology. There is a high degree of variability in
could be affected significantly by the geometry of culture media microdroplet scoring embryos, likely due to the subjective nature of morphology grading. This
and the oil overlay used. leads to less precise decisions for identifying embryos that meet biopsy or freez-
What is known already: Commonly used embryo culture dishes are not ing criteria. This issue can be compounded when artificially hatching an embryo
developed especially for embryological purposes. There is a little work of phys- as this procedure allows blastocysts to escape from the zona at an earlier stage
ical embryonic requirements and culture platform role on human embryonic of development. A deep CNN has been developed using untested embryos to
development. Different drop geometries(2D/3D dishes)results in different sur- make biopsy and cryopreservation decisions1. Could this CNN be used to accu-
face areas, evaporation rates and osmolality of media microdroplets. Oil types rately assess embryos that have undergone AH for PGT?
with different viscosities, densities and water content used for media overlay Study design, size, duration: Retrospective cohort study of 1944 embryos
may result in changes of media evaporation rate, pH and osmolality especially derived from 224 PGT cycles that took place at an academic fertility center
in dry incubators. Minor changes in media’s pH or osmolality has a dramatic between 8/2014 and 12/2019.

Participants/materials, setting, methods: We evaluated the ability of a the Gardner’s classification system). Timing to morula (-2.4 hr), blastulation
CNN-based artificial intelligence platform, trained using untested embryos, to (-1.85hr) and blastocysts expansion (-1.94hr) were significantly (p< 0.001)
identify D5 embryos for biopsy and cryopreservation and compared it with shorter in collapsing embryos. Irrespective to blastocysts collapse dynamic and
highly-trained embryologists. All embryos were laser-AH on D3 to help facilitate frequency, embryos with TE graded A displayed an increased chance of a positive
trophectoderm biopsy at the blastocyst stage. The developmental criteria for KID score (p< 0.004, OR and 95% CI (A vs. B 0.1651 (0.0518, 0.5266), A vs
D5 biopsy and cryopreservation are the same in our center: ≥ grade 3 embryos C 0.2122 (0.0652, 0.6908), A vs D 0.0611 (0.0045, .0.8311)). We observed no
and grade C or better for inner cell mass and trophectoderm (Gardner grading significant effect from potential confounding factors such as patient age or stim-
system). ulation protocol
Main results and the role of chance: Deep CNN correctly identified which Limitations, reasons for caution: The retrospective design of the study
D5 embryos met criteria for biopsy and cryopreservation with sensitivity: 93.7% limits its value. Another limitation is presented by the fact that one embryologist
(95%CI: 91.6-95.3%), and specificity: 96.3% (95%CI: 95.1-97.3%), (n=1944). measured the blastocysts collapsing frequency and dynamics and therefore, no
The positive and negative predictive values of the CNN-based artificial intelli- inter-operator quality assurance.
gence platform were 93.4% (95%CI: 91.3-95.1%) and 96.5% (95%CI: 95.2- Wider implications of the findings: The study of blastocyst morphological
97.4%), respectively (n=1944). dynamics may help to improve the selection criteria of blastocyst with higher
Our system displayed an accuracy of 95.4% (95%CI: 94.3-96.5%) (n=1944). potential to implant especially for those patients with poor prognosis.
Limitations, reasons for caution: Images utilized were obtained using a Trial registration number: not applicable
single imaging platform (EmbryoscopeTM) at one timepoint and were annotated
using criteria from a single IVF program. Prospective trials should confirm our
findings prior to wider application of this system in clinical practice. P-265 For blastocyst frozen embryo transfer: prolonged post-thaw
Wider implications of the findings: Our results suggest that in embryology, culture may improve transfer outcomes
CNNs can be used to automate decision-making related to classification and S.K. Nguyen1, K.C. Do1, T.T.C. Bach1, A.T. Vu1, V.T. Phung1,
disposition of AH embryos with a high degree of accuracy. Future studies will T.L. Dinh1
utilize expanded datasets, additional time-points and clinically relevant patient/ 1
Buu Dien Hospital, ART center, Hanoi, Vietnam
cycle variables to enhance the power and accuracy of testing algorithms.
Trial registration number: NOT APPLICABLE Study question: In comparison with two hour post-thaw culture, are
the blastocyst transfer outcomes of prolonged culture (16 ± 1 hour) bet-
P-264 Using Time-lapse imaging (TLI) to assess blastocyst ter or not?
collapse dynamics and Trophectoderm (TE) quality as a marker of Summary answer: Prolonged post-thaw culture resulted in better blastocyst
implantation potential transfer outcomes: significantly higher implantation rate and significantly
R. Alhamdan1, L. Zujovic2, J. Hernandez Medrano3 increased clinical pregnancy rate.
1
School Of Human Development, Obs & Gyne, Nottingham, United Kingdom ; What is known already: Frozen blastocysts have been commonly transferred
2
Nurture Fertility, Director of embryology, Nottingham, United Kingdom ; two to four hours after thawing. However, another school of thought has advised
3
University of Nottingham, Child health and obstetric and gynaecology, that prolonged post-thaw culture of blastocysts for more than 20 hours could assist
Nottingham, United Kingdom in monitoring the post-thaw development of blastocysts, leading to better embryo
selection for transfer; thus, better outcomes ultimately. In addition, embryos that
Study question: Does the frequency and type of blastocyst collapsing impact underwent two freeze–thaw cycles may experience more stress than embryos that
implantation and wither blastocyst TE quality and embryo developmental kinetics underwent one cycle. Here, we examine the correlation between post-thaw culture
influence the type of blastocyst collapsing observed duration of blastocysts and the transfer outcomes. We also analyse attentively this
Summary answer: Frequency and type of blastocyst collapsing did not influ- correlation of embryos that have undergone two frozen-thaw cycles.
ence implantation potential. However, those blastocysts exhibit higher quality Study design, size, duration: A retrospective study: data transfer cycles
TE and a shorter time to form. from 01/2019-01/2020 were analyzed. Transfers of one or two blastocysts
What is known already: Few studies have examined the phenomenon of were included (good and fair grade only; poor quality blastocyst transfers were
blastocyst collapsing in mammals and their results appears to be contradictory. excluded). Patients were divided into two groups: group L (n=112) comprises
A study in mice reported that collapsing does not affect implantation when it patients who had their frozen blastocysts thawed in the previous afternoon then
is not extensive (≤ 20%). While another study in human, suggests that col- transferred the next morning (16 ± 1 hour culture). Group S (n=100) includes
lapsing embryos have lower implantation potential. This reduction in implan- cases of frozen blastocyst transfer two hour after thawing.
tation potential was present irrespective of whether the extent of collapsing Participants/materials, setting, methods: The majority of cases in each
pattern is week (≤50% separation of the TE) or strong (≥50%separation of group had their blastocysts vitrified on day 5 of development (one freeze-thaw
the TE). Therefore a more comprehensive understanding of the type and cycle). However, each group has a subgroup of embryos that had been vitrified
frequency of blastocyst collapsing and its connection to implantation potential on day 2, thawed, let grow until day 5, underwent one more freeze-thaw cycle
is required (two freeze-thaw cycles) (named L-twice: n=31 and S-twice: n=26) . No PGT
Study design, size, duration: A retrospective analysis including a total of involved; donations excluded. BetaHCG positive, implantation, clinical pregnancy
500 embryos after IVF/ICSI and fresh blastocyst transfer between 2017/2018, were registered. t-test and Chi-square test were applied.
in Nurture Fertility, The Fertility Partnership group, Nottingham, UK. Main results and the role of chance: The average maternal ages of group
Participants/materials, setting, methods: Blastocyst collapsing dynamics L and group S were comparable (29.32 ± 4.26 and 29.98 ± 4.16 years, respec-
were collected from the TLI (Embryoscope, Vitrolife). Exact traceability of trans- tively, p>0.05). The average numbers of transferred embryos per cycle of group
ferred embryos was included in the analysis. The association between blastocyst L and S were equal (1.32 ± 0.47 and 1.33 ±0.47, p>0.05). Group L (prolonged
collapsing dynamics and frequency (no collapse, ≥50% or <50% separation of culture) showed significantly better outcomes than group S (short culture): pos-
the surface of the TE from the ZP), zona pellucida thickness (µm), blastocyst itive BetaHCG rate was 73.21% vs 58.00% (p<0.05), CPR was 67.86% vs.
expansion (µm2), and KID score were assessed. Embryo developmental kinetics 53.00%, and IR was 60.81% vs. 46.62% (p<0.05), respectively. Multiple pregnancy
and TE quality were also compared. Potential confounding factors, e.g: maternal rate was comparable between two groups (13.92% for L group and 14.81% for
age and stimulation protocol were considered S group), possibly due to similar average numbers of transferred embryos
Main results and the role of chance: There was no significant difference between two groups. Notably, for blastocysts which had undergone two freeze-
in implantation potential based on the frequency or type of blastocyst collapse thaw cycles, the correlation remains the same: prolong culture had better out-
dynamics (no collapse, ≥50% or <50% separation of the surface of the TE from come. For L-twice, positive Beta-hCG rate, CPR and IR was 64.52%, 61.29%
the ZP). However, collapsing blastocysts significantly correlated with thinner ZP and 54.35%; while for S-twice, these rates were 57.69%, 57.69% and 52.63%,
(p =0.005). Blastocysts exhibiting ≥50% separation of the blastocoel cavity respectively. These data suggest that prolong culture may improve transfer out-
strongly correlated with the highest TE quality (graded as A in accordance with comes of frozen blastocyst transfer cycles.

Limitations, reasons for caution: This is a retrospective study. Further Trial registration number: not applicable
prospective, randomized clinical trial is required. The sizes of subgroup S-twice
and L-twice are small, extend analysis is required to collect larger sample size. P-267 Duration of exposure to EmbryoGlue (EG) may significantly
Wider implications of the findings: Previous reports on the impact of impact implantation and clinical pregnancy rates following
post-thaw culturing duration on transfer outcomes have been controversial. Our Embryo Transfer (ET).
findings suggest that longer culture may assist blastocysts to “recover” from R. Ebrahimi Ghaei1, C. Harrity2, C. Riordan2, C. Moran2, D. Keane2
freeze-thaw cycles. Further analysis of the impact of prolonged culture on the 1
University of Royal College of Surgeons in Ireland RCSI, Obstetrics & Gynaecology,
epigenetics of the embryos may be valuable.
Dublin, Ireland ;
ReproMed Ireland, Reproductive Medicine, Dublin, Ireland
P-266 Selecting spermatozoa with the highest chromatin integrity Study question: Does EG increase implantation and clinical pregnancy rates
in fresh transfer Assisted Reproductive Technology (ART) cycles and is the dura-
J. Stewart1, A. Parrella1, D. Tavares1, M.S. Wang1, M. Haddad1,
tion of exposure significant?
Z. Rosenwaks1, G.D. Palermo1
1
Summary answer: Embryos exposed to EG >30mins have significantly lower
Weill Cornell Medicine, The Ronald O. Perelman and Claudia Cohen Center for Implantation Rates (IR) and Clinical Pregnancy Rates (CPR) than those exposed
Reproductive Medicine, New York, U.S.A. <30 mins or not at all.
What is known already: Various modifications have been made to culture
Study question: What is the impact of selecting spermatozoa with the highest
media in the hope of increasing success rates, including the addition of Hyaluronan
chromatin integrity on ICSI outcomes?
(HA). EG (Vitrolife, Goteburg) is one such medium. The 2014 Cochrane review
Summary answer: We selected spermatozoa with the highest progressive
(Bontekoe et al) concluded moderate quality evidence of benefit when using
motility and chromatin integrity by microfluidic sperm selection (MFSS) and
HA enriched media prior to ET.
achieved superior implantation and delivery rates.
Vitrolife, recommends equilibration of embryos in EG between 10 mins – 4
What is known already: Sperm preparation methods aim at providing spec-
hrs prior to transfer. Studies including duration of exposure as a parameter have,
imens for insemination with the highest progressive motility independent of
to date, taken 10 mins as the cut off time. We could find no study which com-
phenotypic and genomic integrity. It has recently been recognized that a micro-
pared the effect of different durations of exposure to EG.
fluidics device yielded spermatozoa with the highest progressive motility as well
Study design, size, duration: A retrospective cohort study was designed to
as superior chromatin integrity. Here we compared two sperm selection meth-
determine if duration of exposure to EG influenced outcomes. Cycles from 01
ods: density gradient centrifugation (DGC) and MFSS.
January 2015 – 31 December 2019 were analysed.
Study design, size, duration: From October 2016 to January 2020, ejaculates
1859 cycles reached oocyte retrieval and were screened against the inclusion/
that were processed by DGC and MFSS for ICSI treatment from 8 consenting
exclusion criteria to identify relevant patients.
men were screening for DNA fragmentation by TUNEL. In addition, ejaculates
3 study groups were chosen
from 22 men were processed solely by MFSS for ICSI treatment. Semen param-
eters, chromatin integrity, embryo implantation, and pregnancy characteristics Control: Transfers without EG (138)
were compared. Group 1: Transfers with EG exposure <30mins prior to ET (170)
Participants/materials, setting, methods: Fresh ejaculated specimens from Group 2: Transfers with EG exposure for >30mins prior to ET (295)
consenting men were collected for standard semen analysis in accordance with
WHO 2010 criteria. DGC and MFSS were used to isolate motile spermatozoa Participants/materials, setting, methods: The study was performed in a
based on cell motility and fluid dynamics. Sperm chromatin fragmentation (SCF) single university-affiliated reproductive medicine centre. Inclusion criteria were
was assessed by terminal deoxynucleotidyl transferase dUTP nick end labeling IVF/ICSI cycles resulting in fresh blastocyst transfer.
(TUNEL) on at least 500 spermatozoa under a fluorescent microscope utilizing Cycles with elective freeze, cleavage transfer, failed fertilisation, or no oocytes/
a threshold of ≥15%. ICSI was performed in the standard fashion. embryos were excluded. From 1859 retrievals, 540 patients had 603 transfers
Main results and the role of chance: A total of 20 men (43±6 years) had of 809 blastocysts meeting the inclusion criteria during the study period.
the following average semen parameters: concentration of 18±15 x 106/mL, Primary outcome was IR, secondary outcome was Pregnancy Rate (PR).
30±18% motility, and 2.4±1% morphology. After DGC and MFSS, the sperm Statistical analysis was performed using Chi-square for proportions and t-test
concentration was 2.2±1 and 1.5±12.6 x106/mL, with 49±32% and 97.4±5% for means.
motility, respectively (P<0.0001). Main results and the role of chance: Control group, no EG (n=138):
The morphology of the raw sperm sample improved from 2.3±1% to 4.0±1%
after MFSS, while it remained at 2.6±1% after DGC. • Age: 37.0
In 8 men (43±6 years), the SCF in their raw sample was 23%, falling to 18% • Anti-Müllerian hormone (AMH): 15.0 pmol/l
after DCG selection and to 1.2% after MFSS (P < 0.0001). They underwent 20 • Number of embryos 160
ICSI cycles with their female partners, (37±3 years) with DGS sperm selection, • Mean number blastocyst per ET: 1.16
achieving a 58% fertilization rate (80/138). The implantation rate was 5% (2/41) • PR: 0.536
with an 11% clinical pregnancy rate (CPR) (2/18) and one pregnancy loss. • IR: 0.269
Subsequently, ICSI with MFSS achieved a fertilization rate of 65%, a 29% (5/17; • CPR: 0.312
P<0.01) implantation rate, and a 63% (5/8; P<0.001) CPR. An additional 22
men (43±7 years) underwent 28 ICSI cycles solely with MFSS due to poor
Group 1, EG exposure <30mins (n=170):
reproductive history. A 76% fertilization rate (203/266) and 54% (31/57) good
• Age: 37.3
quality embryos were achieved. The implantation rate was 26% (15/57) with a
• AMH: 13.8 pmol/l
50% CPR (13/26).
• Number of embryos 199
Limitations, reasons for caution: This is a preliminary study on a small
• Mean number blastocyst per ET: 1.16
number of subjects. Although we controlled for a concurrent female factor, this
• PR: 0.547
cannot be excluded with certainty. As this is a new method, the health of the
• IR: 0.367
resulting offspring would need to be evaluated to confirm the safety of the
• CPR: 0.429
technique.
Wider implications of the findings: According to our study, SCF appears
to be linked to the kinetic characteristics of the sperm cell. This novel microfluidic Group 2, EG exposure >30mins (n=295):
device may help to identify spermatozoa with the highest functional and genomic • Age: 37.4
integrity and ultimately be routinely used in ART treatments. • AMH: 14.8 pmol/l

• Number of embryos 450 maternal age and recurrent implantation failure but future RCT studies are nec-
• Mean number blastocyst per ET: 1.51 essary to evaluate impact of Embryo-glue on all patients.
• PR: 0.495 Trial registration number: Not applicable
• IR: 0.156
• CPR: 0.237
There was no difference between age (p=0.2) or AMH (p=0.79) between POSTER VIEWING SESSION
groups. Significantly more embryos were transferred in group 2 (p<0.0001).
ENDOMETRIOSIS, ENDOMETRIUM AND AND FALOPIAN
The difference in PR between all 3 groups was not significant (p=0.5), although
TUBE, AND BENIGN DISORDERS OF THE ENDOMETRIUM
this may be influenced by the number of embryos transferred. Both IR
AND FALLOPIAN TUBE
(p<0.0001) and CPR (p<0.0001) were significantly lower in the group with
extended exposure to EG.
Unfortunately, this was a retrospective cohort study rather than a prospective
randomised controlled trial. Therefore, the impact of bias cannot be excluded. P-269 Cumulative live birth rates after IVF-ICSI cycles in women
Limitations, reasons for caution: This is a retrospective study over 5 years, with endometriosis-associated infertility: prolonged GnRH-agonist
during which subtle changes may have occurred in laboratory practice and per- versus GnRH-antagonist protocols
sonnel. The potential impact of additional confounding factors cannot be
H. Wei1, Y. Chen1, X. Li1, Y. Lin1, R. Li1, C. Ma1
accounted for without randomisation.
1
Embryo quality was not included as a parameter, but exclusion of cleav- Peking University Third Hospital, Center for Human Reproductive Medicine,
age-stage transfers should limit this impact. Peking, China
Wider implications of the findings: Studies to date have shown either a
Study question: Are cumulative live birth rates similar in prolonged GnRH-
potential benefit or no benefit with the use of EG. Our study supports this
agonist and GnRH-antagonist protocols after IVF-ICSI cycles in women with
conclusion but also suggests that exposure to EG for >30mins may be
endometriosis-associated infertility including all subsequent frozen–thaw cycles
detrimental.
from the same oocyte retrieval?
Trial registration number: Not Applicable
Summary answer: No differences were found in cumulative live birth rates(-
CLBRs) after one complete IVF-ET cycles between prolonged GnRH-agonist
P-268 Embryo glue transfer medium for women >= 35 years of and GnRH-antagonist protocols in women with endometriosis-associated
age - outcome in fresh IVF/ICSI cycles infertility.
V. Tomic1, M. Sommergruber1 What is known already: Endometriosis is known to have an impact on fertility.
1
University hospital SALK Paracelsus Medical University, Gynaecology and Obstetrics, Women with endometriosis often require in vitro fertilization (IVF) to improve
Salzburg, Austria the chance of pregnancy. A reduced ovarian reserve was demonstrated in patients
with endometriosis. Multiple studies of the prolonged GnRH-agonist and GnRH-
Study question: Add-on embryo glue in single embryo transfer in the women antagonist protocols on pregnancy rate and live birth rate have yielded controver-
≥ 35 years of age, does it make a difference? sial findings. A 2006 systematic review of 27 RCTs showed that GnRH-antagonist
Summary answer: Results showed beneficial effect of embryo glue medium protocol has a significantly lower clinical pregnancy rate and live birth rates than
in women ≥ 35 years of age and those with recurrent implantation failure. those in GnRH-agonist long protocol. However, a 2016 systematic review of 73
What is known already: A Cochrane review of 6 RCTs by Bontekoe and al. RCTs concluded that these two protocols have equivalent live birth rates.
found higher live birth rate using embryo glue (OR 1.41) (Bontekoe et al., 2014). Study design, size, duration: A retrospective case-control study comprised
However, the incidence of multiple pregnancies was also increased significantly women with endometriosis who underwent IVF-ICSI cycles between January
(OR 1.86). 2015 to December 2016, at the Reproductive Medical Center of Peking
Afterwards, study by Signh et al.from 2017 and Fancovits et.al from 2015 did University Third Hospital. Propensity score matching (matching ratio 1:1, caliper
not found statistically significant difference in the clinical pregnancy rate with the width = 0.02) was used to create a comparable control group, including 303
use of embryo glue medium. However, the most of te studies are statistically women with prolonged GnRH-agonist protocol and 303 women with GnRH-
powerless because of a small sample size and/or different criteria, and studies antagonist protocol.
with a SET are currently lacking. Participants/materials, setting, methods: A retrospective case-control
Study design, size, duration: A retrospective cohort study on totally 680 study comprised women with endometriosis who underwent IVF-ICSI cycles
patients, divided into two groups undergoing IVF/ICSI, in the tertiary University between January 2015 to December 2016. Propensity score matching (matching
center between 2013-2019. ratio 1:1, caliper width = 0.02) was used to create a comparable control group,
Participants/materials, setting, methods: In total 295 patients in the study including 303 women with prolonged GnRH-agonist protocol and 303 women
group, embryos were transferred into 50μL of Embryo-glue for 15 minutes prior with GnRH-antagonist protocol.Cumulative live birth rates (CLBRs) upon the
to transfer and control group where embryo were transferred to conventional first complete IVF-ICSI cycles, including transfers of all resulting embryos was
blastocyste culture medium. Statistical analysis was performed by SPSS ver. 22 considered as a primary outcome measure.
for Windows. Main results and the role of chance: Baseline characteristics including
Main results and the role of chance: Clinical pregnancy rate in the study BMI, age, basal FSH, duration of infertility were comparable in the prolonged
group with the use of hyaluronan enriched medium (Embryo-glue) was sig- GnRH-agonist group and the GnRH-antagonist group. Patients in the pro-
nificantly higher than the control group (36.5% vs 28.3%, p<0.05). When we longed GnRH-agonist group had a significantly higher gonadotropin con-
analyzed subgroup of women with recurrent implantation failure, statistical sumption (3487.55 (1200-10425) vs. 2175.00 (600-5550)(IU), P<0.01) and
difference was even more powerful with p<0.01. However, our results did a longer time of stimulation (13.00(8.00-24.00) vs. 10.00(4.00-19.00)(days),
not found difference in the number of retrieved eggs, days of stimulation, total P<0.01) than that in the GnRH-antagonist group. There’s no significantly
dose of gonadotrophins, type of fertilization (IVF or ICSI), fertilization rate difference between two groups of the estradiol levels on hCG trigger day
or miscarriage rate. Multiple pregnancy rate was also similar between (7087.00(525.00-29976.00) vs. 5995.00(676.00-26075) (pmol/mL),
two groups. P=0.06). The number of oocytes retrieved per cycle and number of usable
Limitations, reasons for caution: Limitations of the current study include embryos achieved were comparable between two groups. The implantation
retrospective design and as such bias can not be excluded. Also our study rate in the GnRH-antagonist group after fresh ET was significantly lower
excluded women in favorable maternal age (younger then 35 years of age) so (28.5% vs. 34.9%, p =0.047) compared with prolonged GnRH-agonist group.
effect on embryo glue in this age group could not be commented. A GnRH-antagonist protocol appears to result in lower rates of clinical
Wider implications of the findings: Results of current study showed ben- pregnancy and live birth after fresh ET (40.8% vs. 46.5%, p = 0.22; 33.2%
eficial effect of Embryo-glue in selected group of patients, women in advanced vs. 39.1%, p = 0.19, respectively), but the difference of which did not reach

statistical significance.The cumulative clinical pregnancy rates and CLBRs Wider implications of the findings: These findings of different gene and
were comparable for the prolonged GnRH-agonist group and the GnRH- protein expression between different levels of EM may support the idea of a
antagonist group with 55.1% vs. 52.8%, (odds ratio(OR): 0.91; 95% CI: distinct pathophysiology between mild and severe EM and suggest the potential
0.66-1.25; P=0.57) and 45.5% vs. 43.6%, (OR: 0.92; 95% CI: 0.67-1.27; use of FN1 and GRN as new clinical biomarkers.
P=0.62), respectively. Trial registration number: not applicable
Limitations, reasons for caution: One limitation of this study is its ret-
rospective nature, however propensity score matching was selected to P-271 Effects of chronic endometritis on fertility and pregnancy
create control cohorts for a better matched analysis. The reproductive out- outcomes
comes in different pathological types of endometriosis were not analyzed, A. Morimune1, F. Kimura1, A. Nakamura1, J. Kitazawa1, T. Hanada1,
such as peritoneal endometriosis, deep infiltrating endometriosis or K. Hirata2, A. Takebayashi3, A. Takashima1, S. Kaku1, S. Moritani4,
endometrioma. R. Kushima4, T. Murakami1
Wider implications of the findings: A GnRH-antagonist protocol appears 1
Shiga University of Medical Science, Department of Obstetrics and Gynecology,
to result in lower rates of implantation, clinical pregnancy and live birth after
Shiga, Japan ;
fresh ET, suggesting that a GnRH- antagonist protocol might negatively impact 2
Goto Ladies Clinic, Division of Reproductive Medicine, Osaka, Japan ;
endometrial receptivity. However, we find similar CLBRs between the two 3
National Hospital Organization Higashi-ohmi General Medical Center,
groups in women with endometriosis-associated infertility.
Department of Obstetrics and Gynecology, Shiga, Japan ;
Shiga University of Medical Science, Department of Clinical Laboratory Medicine
and Division of Diagnostic Pathology, Shiga, Japan
P-270 Fibronectin (FN1) and Progranulin (GRN) expression in
eutopic endometrium – potential biomarkers among different Study question: Does chronic endometritis (CE) affect fertility and the course
stages of endometriosis (EM) of pregnancy in patients without implantation failure, habitual abortion, and
I. Holzer1, A. Machado Weber1, A. Marshall1, A. Freis1, diseases suspected to cause implantation failure?
T. Strowitzki1, A. Germeyer1 Summary answer: Patients with CE have lower pregnancy and live birth rates
1 and higher miscarriage and preterm birth rates.
University Hospital Heidelberg, Dept. of Gynaecological Endocrinology and Fertility
Disorders, Heidelberg, Germany What is known already: Several reports have demonstrated that CE is asso-
ciated with infertility and implantation failure. However, these clinical researches
have mainly involved patients with recurrent implantation failure (RIF). The
Study question: Does the local endometrial gene and protein expression of
subsequent pregnancy rate after the diagnosis of RIF is considered to be low
FN1 and GRN vary among different degrees of EM?
even without CE. Thus, RIF without CE cannot be used as the control group to
Summary answer: Significant differences in gene and protein expression were examine the effects of CE on fertility and pregnancy outcomes. In the present
identified not only between women with and without EM but also among dif- study, patients with RIF, recurrent pregnancy loss (RPL), and diseases suspected
ferent levels of the disease. to cause implantation failure were excluded, and the effects of CE on fertility
What is known already: Endometriosis is a common gynecological disease and pregnancy outcomes were determined.
affecting 10 – 15 % of women of reproductive age. However, molecular mech- Study design, size, duration: This retrospective study reviewed a registration
anisms and pathogenesis are still not completely understood. Due to the absence list of patients attending our hospital for infertility and histologically diagnosed
of a reliable clinical biomarker, the only viable method for the often delayed with the presence or absence of CE from September 2013 to December 2017
definitive diagnosis is laparoscopic surgery. FN1 is a glycoprotein, which is to identify study patients. Patients were then followed up based on their medical
involved in cell adhesion, migration and implantation. GRN, a protein with pro- charts for 1 year after the diagnosis of CE.
liferative and invasive properties, seems to be involved in embryogenesis. Our Participants/materials, setting, methods: Patients were treated with IVF for
objective was to investigate molecular mechanisms of these two genes and a year after the histological diagnosis of CE. Endometrium was collected 5–9 days
proteins among various stages of EM and analyze their potential as diagnostic after ovulation. The pregnancy and pregnancy outcome rates were analyzed. Those
markers. who received antibiotics for >7 days to treat CE, aged >40 years, or had RIF, RPL,
Study design, size, duration: This study analyzed eutopic endometrial tissue and a suspected cause of implantation failure were excluded from the study.
of women suffering from different stages of EM (n = 58). Our aim was to inves- Main results and the role of chance: A total of 40 non-CE and 44 CE
tigate changes in endometrium of women with EM compared to healthy endo- patients were finally analyzed.
metrium as well as among different levels of EM (minimal, mild, moderate) to Percentages of pregnancies and live births were as follows: 95% vs. 72.7%, P
detect potential new biomarkers for the disease. < 0.01; 85% and 45.4%, P < 0.0003 in the non-CE and CE groups, respectively.
Participants/materials, setting, methods: Endometrial biopsies were taken Percentages of miscarriages in pregnancy were 12.8% vs. 40.0%, P < 0.03,
from women undergoing a laparoscopic surgery for benign reason. The expres- whereas percentages of preterm births in ongoing pregnancy were 2.9% vs.
sion on mRNA level of FN1 and GRN was analyzed using real-time quantitative 23.8%, P < 0.03 in the non-CE and CE groups, respectively.
reverse transcription polymerase chain reaction (qRT-PCR). Protein expression In the logistic analysis, CE was a factor adversely affecting objective pregnancy
of the proteins was determined using Enzyme-linked immunosorbent assay variables associated with live births in all patients, miscarriage in pregnancy, and
(ELISA) as well as the Avidin-Biotin method to stain cryosections for preterm birth in ongoing pregnancy.
immunohistochemistry. Limitations, reasons for caution: This was a retrospective study; hence,
Main results and the role of chance: Significant differences in gene and inevitable limitations are observed. Patients who received antibiotic treatment
protein expression of both examined genes were identified not only in for CE were excluded. Patients who are seeking antibiotic treatment might have
eutopic endometrial tissue of women with EM compared to women without a long history of infertility treatment and/or might have more refractory infer-
EM but also among different levels of the disease (p < 0.05). The strongest tility. These patients were excluded only from CE.
mRNA expression of both FN1 and GRN was seen in women with minimal Wider implications of the findings: Pregnancy and live birth rates within a
EM. However, protein expression of FN1 and GRN was significantly reduced year after the diagnosis of CE were lower in patients undergoing IVF not diag-
in women with EM compared to women without EM. More precisely, the nosed with RIF, RPL, or a disease suspected to cause implantation failure. These
magnitude of the protein expression decreased with the severity of the findings suggest that patients undergoing IVF should be examined for CE.
disease (p < 0.05). While the protein expression of FN1 was mainly noticed Trial registration number: R2014-090
in stromal and endothelial cells, GRN was mostly present in stromal and
epithelial cells.
Limitations, reasons for caution: The main limitation of this study is its P-272 The pattern of vaginal/endometrial microbiome as a
small sample size. A larger study population would be needed to validate the predictor for outcome of in vitro fertilization (IVF) in patients
results. with or without repetitive implantation failure:a pilot study

A. Bernabeu1, B. Lledo2, M.C. Diaz3, F. Lozano2, A. Rodriguez- 1

Amsterdam UMC- Vrije Universiteit Amsterdam, Reproductive Medicine,
Arnedo3, R. Morales2, R. Bernabeu1 Amsterdam, The Netherlands ;
2
1
Instituto Bernabeu, Reproductive Medicine, Alicante, Spain ; Leiden University Medical Center, Biomedical Data Sciences, Leiden, The
2
IB Biotech, Molecular Laboratory, Alicante, Spain ; Netherlands ;
3
3
Instituto Bernabeu, IVF Lab, Alicante, Spain Monash University, Obstetrics and Gynaecology, Melbourne, Australia ;
4
Amsterdam UMC- University of Amsterdam, Reproductive Medicine, Amsterdam,
Study question: Is the pattern of certain vaginal/endometrial bacteria asso- The Netherlands
ciated with failure or success to achieve pregnancy after IVF in patients with or
without repetitive implantation failure? Study question: Does the fertility enhancing effect of tubal flushing during
Summary answer: The vaginal and endometrial microbiome pattern correlate hysterosalpingography (HSG) with oil-based contrast change over time?
with the pregnancy rate, being different between patients who suffer or not Summary answer: The hazard ratio for ongoing pregnancy of oil-versus
repetitive implantation failures. water-based contrast was 1.71 immediately after HSG, then decreased and
What is known already: Thanks to an improved technology in the last decade plateaued towards 1 after approximately two years.
as for instance, mass sequencing, a progress in the study of microbial commu- What is known already: HSG with oil-based contrast results in a 10% higher
nities was made. Successful embryo transfer depends on many factors, including absolute ongoing pregnancy rate after six months when compared to an HSG
the presence of microbial colonization of the upper genital tract. The vaginal with water-based contrast. In the long-term follow up study, this effect remained
microbiome plays an important role in the maintenance of the health of the present 3-5 years after the HSG, albeit that the difference was reduced to 5%
woman. Currently more and more patients are using assisted reproduction after additional IVF in both groups. The fertility enhancing mechanism of oil-based
techniques, which should be offered a multidisciplinary approach for diagnosis. contrast remains unclear. Both immunological effects on the endometrium and
The aim of this study is to investigate if the vaginal and endometrial microbiome peritoneum or a possible mechanical effect by dislodging debris or mucus plugs
in couples undergo ART could affect the pregnancy rate. from the fallopian tubes have been proposed, which suggests a temporary or a
Study design, size, duration: Prospective analysis including 48 patients (18- lasting effect, respectively.
50 years) recruited from May 2017-May 2019 (264 samples). Of these, 23 Study design, size, duration: We performed a secondary analysis of the
patients experienced unexplained repetitive implantation failure (RIF). All patients H2Oil (long term follow-up) study, a multicenter randomized controlled trial
performed PGT-A and single embryo cryotransfer. Vaginal samples were col- among 1,119 women evaluating the effectiveness of oil-based contrast versus
lected on the immediate pre-transfer cycle (artificial endometrial preparation) water-based contrast during HSG. Here, we investigate whether the fertility
(visit 1), on the day of the transfer (visit 2) and on the day of the pregnancy test enhancing effect of oil- versus water-based contrast on ongoing pregnancy
(visit 3). Endometrial samples were taken on the immediate pre-transfer cycle changed over time or remained stable.
(visit 1). Participants/materials, setting, methods: Women undergoing fertility
Participants/materials, setting, methods: DNA extraction was performed work-up were included in the H2Oil study. We used a Cox proportional hazards
using the PureLink Microbiome DNA Purification kit. Sequencing the rRNA16S model for time to pregnancy for a maximum of three years of follow-up, then
V3V4 region analysis, it was carried out according to Illumina Metagenomics evaluated whether the assumption of ‘no change of effect over time’ holds using
protocol using the NexteraXT library on the Miseq instrument. The bioinformatic plots and tests on the scaled Schoenfeld residuals and by assessing interactions
tools QIIME2, MicrobiomeAnalyst and Phyloseq have been used to determine between treatment and time. We repeated the analysis in a subgroup of women
the composition of the microbiome. who experienced pain during HSG (Visual-Analogue-Scale-score ≥6).
Main results and the role of chance: Regarding to the microbiome dynamics Main results and the role of chance: Data on 1107 couples were available.
during the cycle, we observed a decrease in α-diversity from the follicular to Of these, 550 couples had oil-based contrast and 557 had water-based contrast
luteal phase in the study group, in contrast to a stability of the pattern in the RIF at HSG. Ongoing pregnancy rates after 3 years were 77% and 71%, respectively.
group. As for endometrium and vagina microbiome, there were α-diversity Median follow up was 9-10 months (5th-95th percentile: <1-36). The hazard
differences (Shannon p=0.0139, Simpson p=0.046), being higher for endome- ratio for ongoing pregnancy of oil versus water over three years of follow up
trium; and also β-diversity was different (p=0.001). A greater relative abundance was 1.26 (95%CI: 1.10-1.44). On the scaled Schoenfeld residual plots, we found
was observed in vagina of Lactobacillus spp., (83.17% vs 84.82%,p<0.0001), a decrease of the hazard ratio that was linear with log-transformed time. After
Streptococcus spp. (1.59% vs 7.74%,p=0.014) and Ureaplasma spp. (0% vs. including an interaction with log-transformed time, we found that the hazard
0.89%,p=0.006). Besides, it was lower for Delftia spp. (0.95% vs 0%,p=0.0003), ratio immediately after HSG was 1.71 (95%CI: 1.27-2.31) and reduced to 1.06
Anaerobacillus spp. (1.59% vs 0%,p=0.0004), and Ralstonia spp. (3.17% vs (95%CI: 0.86-1.30) at approximately two years of follow up. There was no
0%,p=0.0006). Concerning RIF, we observed differences in α-diversity for the evidence for a change in hazard ratio over time in a subgroup of women who
study and RIF group (Shannon p =0.0206, Simpson p=0.0206). Moreover, we experienced pain during HSG.
were able to observe significant differences in β-diversity between both groups. Limitations, reasons for caution: This analysis was performed using data
Also, we found differences for Ralstonia spp., (0.09% RIF and 0.73% control, from women with unexplained or mild male infertility. The results should not be
p=0.0012). Finally, as for pregnancy rate, we found a difference between patients generalized to women with ovulatory dysfunction, women older than 39 years
that did not achieve[JCCF1] or achieved a pregnancy for Lactobacillus spp. (91% or women at high risk for tubal pathology based on their medical history.
vs 99%,p=0.0445 visit 1, 94.63% vs. 97.69%,p=0.0268 visit 2, 97.73% vs Wider implications of the findings: The effect of oil-versus water-based
99.74%,p=0.0492 visit 3) and L.reuteri (0.39% vs 0.17%,p=0.0397 visit 1, 0.15% contrast decreased in the first year after HSG to no effect after approximately
vs 0.30%, p=0.0491 visit 3). two years. In women who experienced pain during HSG, the effect might last
Limitations, reasons for caution: Future larger sample size studies are longer. Both results seem to favour the hypothesis that oil-based contrast may
warranted to corroborate our initial results. dislodge debris or mucus from fallopian tubes.
Wider implications of the findings: The lack of dynamism in the micro- Trial registration number: Netherlands Trial Register NTR 3270 (H2Oil
bioma pattern of RIF patients might reflect a impaired adaptation to endometrial study), NTR 6577 (H2Oil follow-up study), www.trialregister.nl
changes. A greater relative abundance of Lactobacillus spp. and L.reuteri is cor-
related with higher chances of pregnancy. P-274 Functional analysis of the histidine N-methyltransferase
Trial registration number: NCT03153787 SETD3 in endometriosis
M. Götte1, M. Poloczek1
1
Muenster University Hospital, Gynecology and Obstetrics, Münster/Westf., Germany
P-273 Is the fertility enhancing effect of hysterosalpingography
with oil-based contrast temporary or long-lasting? Study question: Does the novel actin methyltransferase SETD3 have an impact
N. Van Welie1, K. Rosielle1, K. Dreyer1, J. Van Rijswijk1, on invasive growth of endometriotic cells?
C.B. Lambalk1, N. Van Geloven2, V. Mijatovic1, B.W. Mol3, Summary answer: SETD3 affects invasive growth, cell motility and contrac-
R. Van Eekelen4 tility of endometriotic cells in vitro.

What is known already: Endometriosis is a common, benign and chronic Participants/materials, setting, methods: The QoL of study participants
disease in women of reproductive age that is characterized by invasive growth was assessed at the date before embryo transfer. Atosiban (a mixed oxytocin/
of endometrial tissue outside the uterus. The novel actin-specific histidine vasopressin V1A receptor antagonist) can be administered intravenously during
N-methyltransferaseprevents primary dystocia and modulates actin function as the embryo transfer. The GEE analysis was performed to assess the association
a prerequisite for cell motlilty, contractility and invasive growth. between QoL during IVF treatment and subsequent pregnancy outcomes, with
Study design, size, duration: This is an in vitro laboratory study on the adjustment for time-varying factors (e.g., age) across multiple embryo transfers
immortalized endometriotic cell line 12Z and primary endometriotic stroma for an individual person. The analysis was further stratified by the number of
cells. Cells were subjected to siRNA-mediated knockdown of SETD3 or a neg- embryo transfer cycle.
ative control siRNA, and the impact on cell motility, contractility, invasiveness, Main results and the role of chance: Compared to a control group of
cytoskeletal structure and gene expression were studied. non-endometriosis women, endometriosis patients had significantly lower QoL,
Participants/materials, setting, methods: Primary endometrial stroma as indicated by the mind/body and treatment environment domain scores, total
cells and 12Z human immortalized endometriotic cells were used for in vitro treatment scores, and total FertiQoL scores. FertiQoL scores were significantly
experiments. SETD3 was knocked down by siRNA transfection with two differ- associated with the success of pregnancy outcomes among non-endometriosis
ent SETD3 siRNAs. Gene expression analysis was performed by real-time qPCR. women; with a one unit increase in the emotional domain score, the probabilities
Changes at the protein level were analysed by Western blotting, ELISA and of ongoing pregnancy and live birth significantly increased by 2.4% and 2.6%,
immunofluorescence. Cell migration was investigated by scratch wound healing respectively (p < 0.05). In overall study subjects (without stratified by the number
assay. To assess cytoskeletal functions, a collagen contraction assay was per- of embryo transfer cycle), the use of atosiban was not significantly associated
formed. Invasive growth was assessed by Matrigel assay. with positive pregnancy outcomes, while among those requiring more than two
Main results and the role of chance: Knockdown of SETD3 was confirmed cycles, significantly increased pregnancy rates with atosiban use were found.
by qPCR and Western blotting. Only moderate changes in cytoskeletal element These results regarding the association between atosiban use and IVF pregnancy
gene expression were observed after SETD3 knockdown, whereas VEGF expres- outcomes are consistent in endometriosis and non-endometriosis groups.
sion was downregulated by >40% in both experimental systems (p<0.05, n>3). Limitations, reasons for caution: This is a study enrolled the patients from
At the functional level, SETD3 depletion resulted in a delay in cell motility in a single medical center. Only the QoL from the date before ET was measured,
scratch wound assays (p<0.05, n>5), in a reduction in invasive growth of 12Z while time-varying QoL data from different points of time during IVF treatment
cells by 50% (p<0.05, n>5) and in a reduced capability to contract collagen gels were not collected. The factors associated with the QoL of study patients were
(p<0.05, n>5). Immunofluorescence microscopy for actin revealed an altered not further explored.
cytoskeletal structure and cell morphology. Wider implications of the findings: FeriQoL is useful in clinical practice to
Limitations, reasons for caution: This is a transfection-based in vitro strudy, understand the QoL of women undergoing IVF treatment. Further developing
which needs to be confirmed in more complex experimental models. clinical strategies to improve these patients’ QoL would be of importance to
Wider implications of the findings: The presented data suggest that meth- enhance pregnancy outcomes. Oxytocin antagonists may enhance IVF pregnancy
ylation of beta-actin, as mediated by SETD3, contributes to the remodelling of rates, especially among the patients requiring more than two embryo trans-
the cytoskeleton that plays a key role in cell migration and invasion. A dysregu- fer cycles.
lation of SETD3 could play a role in the process of invasive growth in Trial registration number: nil
endometriosis.
Trial registration number: MedK P-276 Resveratrol supplementation enhances decidualization of
human endometrial stromal cells with concomitant decrease in
P-275 Generalized estimating equation analysis for assessing the cell proliferation
association between the quality of life and pregnancy outcomes of A.C. Mestre Citrinovitz1, T. Strowitzki1, A. Germeyer1
IVF treatment in patients with endometriosis-related infertility 1
Heidelberg University - Women’s Hospital, Department of Gynecologic Endocrinology
M.H. Wu1, H.T. Ou2, C.W. Lin1 and Fertility Disorders, Heidelberg, Germany
1
College of Medicine and Hospital- National Cheng Kung University, Obstetrics
and Gynecology, Tainan City, Taiwan R.O.C. ; Study question: Is resveratrol supplementation beneficial for human endo-
2
College of Medicine and Hospital- National Cheng Kung University, Pharmacy, metrial stromal cell in vitro decidualization?
Tainan City, Taiwan R.O.C. Summary answer: Resveratrol supplementation enhances, in a dose-depen-
dent manner, human endometrial stromal cell in vitro decidualization.
Study question: Are the quality of life (QoL) of endometriosis-related infer- What is known already: Decidualization is the differentiation of endometrial
tility women measured during IVF treatment related to the pregnancy outcomes stromal cells, a change needed to regulate trophoblast invasion and to support
following the treatment? pregnancy. Decidualization follows endometrial stromal cell proliferation and it
Summary answer: Improving the QoL during IVF treatment in women with has been described that cell cycle arrest contributes to a proper decidualization.
endometriosis-related infertility may increase subsequent pregnancy rates. Interestingly, resveratrol, a natural compound derived from grapes with antiox-
What is known already: Generalized estimating equation (GEE) analysis that idant properties, has been widely studied in relation to endometrial health.
accounts for the dependence of observations from same person can minimize However, little is known about the effect of resveratrol supplementation during
the potential biases commonly seen in longitudinal cohort studies, especially the decidualization.
studies of the IVF data that comprised a series of repeatedly measured pregnancy Study design, size, duration: This study was performed using primary human
outcomes and associated patient characteristics from multiple embryo transfer endometrial stromal cells (HESC). Endometrial biopsies from mid-late prolifer-
cycles in same study subject. From emotional aspect, psychological counseling ative-phase were obtained from healthy women (34.4 +/- 1.1 years old) after
for patients undergoing IVF treatment might improve their QoL and enhance written informed-consent that was obtained (approved protocol S-239/2005).
subsequent IVF pregnancy rates. In addition, oxytocin antagonist treatments HESC were decidualized in vitro with a decidualization cocktail containing
given before embryo transfer in the women with endometriosis may maximize medroxyprogesterone acetate, estradiol and 8-Bromo-cyclic adenosine mono-
the chance of success of pregnancy outcomes. phosphate. Pre-decidualized cells (three days) were further treated with the
Study design, size, duration: This longitudinal cohort study included 686 decidualization cocktail supplemented with different doses of resveratrol (0
women with 1,205 embryo transfers during 2012-2017. The QoL of study to 50 µM).
women during IVF treatment was measured by FertiQoL comprising a Core Participants/materials, setting, methods: Decidualization was evaluated
module (mind/body, emotional, relational, and social domains) and a Treatment by measuring prolactin (PRL) protein levels in cell culture supernatant and by
module (treatment environment and tolerability domains). The FertiQol scores measuring PRL and Insulin-like growth factor binding protein 1 (IGFBP1) mRNA
were computed and scored in the range of 0-100, with higher scores indicating levels by real time polymerase chain reaction (RT-PCR). Changes in the expres-
better QoL. The pregnancy outcomes of interest were chemical and ongoing sion of genes related to cell cycle progression (KI67, CCNA1, CCNB1, CCND1,
pregnancies, and live birth. CDC2, CDK2, CDK4, p53, p21) and cell proliferation were evaluated for the

25 µM dose of resveratrol by RT-PCR and by crystal violet proliferation assay by the antibiotic treatment, whereas the concentration of IL-6 was not signifi-
respectively. cantly altered. In the samples obtained after antibiotic treatment, secreted levels
Main results and the role of chance: Resveratrol supplementation of TNF-α (p=0.024), IL-1β (p=0.019), and IFN-γ (p=0.0071) were significantly
increased, in a dose-dependent manner, the expression levels of PRL and IGFBP1, lower in the patients in whom CE was cured than in the persistent CE patients.
indicating an enhanced in vitro decidualization (PRL protein levels, mU/l: 0µM Limitations, reasons for caution: The number of samples was small.
193±35, 6.25µM 226±38, 12.5µM 270±49**, 25µM 296±61***, 50µM However, there were still significant differences in TNF-α, IL-1β, and IFN-γ levels.
274±49***; PRL mRNA levels, fold change related to 0µM of resveratrol: 0µM We speculate that with an increased number of cases, a significant difference in
1.0±0.0, 6.25µM 1.4±0.2*, 12.5µM 1.9±0.2***, 25µM 1.8±0.2***, 50µM IL-6 may be observed.
1.9±0.2***; IGFBP1 mRNA levels, fold change related to 0µM of resveratrol: Wider implications of the findings: When CE was cured, secretion of
0µM 1.0±0.0, 6.25µM 1.4±0.2, 12.5µM 2.3±0.4, 25µM 2.4±0.6**, 50µM proinflammatory cytokines from endometrial stromal cells was normalized to
1.3±0.2**). Enhanced decidualization was accompanied by a decrease in cell the level of healthy patients. Thus, CE was related to the uterine microbiome
proliferation (25µM 4±1%* of reduced proliferation compared to 0µM) and by because antibiotic treatment was effective in suppressing proinflammatory cyto-
changes in the mRNA expression levels of key cell cycle regulators (mRNA kine secretion from endometrial stromal cells.
levels; fold change related to 0µM of resveratrol: CCNB1 0.65±0.07***, CCND1 Trial registration number: not applicable
0.44±0.09**, CDK2 0.72±0.07**, CDK4 0.71±0.03***, p53 0.53±0.06***, p21
0.61±0.07***). Resveratrol supplementation seems to enhance decidualization P-278 Vitamin D as an effective treatment to reduce cell
by reinforcing the effect of the decidualization cocktail, in particular by reinforcing proliferation and extracellular matrix formation in human uterine
the inhibition in cell proliferation. Statistical differences *p<0.05 **p<0.01; leiomyomas regardless MED12 mutation status
***p<0.001 compared to 0μM of resveratrol. A. Corachán1, M.G. Trejo1, M.C. Carbajo-García1,2, A. Faus1, J.
Limitations, reasons for caution: This study was performed in vitro using Escrig3, J. Monleón3, A. Pellicer1,4, I. Cervelló1, H. Ferrero1
primary HESC. Further studies are necessary to confirm the effect of resveratrol 1
IVI Foundation- IIS La Fe, Innovation, Valencia, Spain ;
during in vivo decidualization and to define the optimal dose and administration 2
Universidad de Valencia, Pediatrics- Obstetrics and Gynecology, Valencia, Spain ;
schedule for a beneficial outcome. Special caution needs to be taken in relation 3
Hospital Universitario y Politécnico La Fe, Obstetrics and Gynaecology Service,
to resveratrol action over cell proliferation
Valencia, Spain ;
Wider implications of the findings: Resveratrol seems to be an effective 4
IVIRMA Rome, Department of Reproductive Endocrinology and Infertility, Rome,
supplementation to reinforce hormone action during HESC decidualization. We
Italy
believe that further insights into resveratrol action and its interaction with estra-
diol and progesterone signaling pathways could facilitate the identification of
Study question: Could Vitamin D be an effective treatment to reduce the size
new therapeutic strategies for the improvement of women´s health.
of different molecular subtypes of human uterine leiomyoma?
Summary answer: Vitamin D inhibited cell proliferation, Wnt/β-catenin and
TGFβ pathways in human uterine leiomyoma primary cells from two molecular
P-277 The Effect of Antibiotic Treatment on the Secretion of subtypes: MED12-mutated and wild type.
Proinflammatory Cytokines in Endometrial Stromal Cells in What is known already: Uterine leiomyomas, the most common benign
Patients with Chronic Endometritis tumor of reproductive tract, are clinically and scientifically treated as a single
A. Nakamura1, F. Kimura1, J. Kitazawa1, A. Morimune1, entity despite the existence of four molecular subtypes, among which MED12
A. Takashima1, S. Kaku1, T. Murakami1 mutation is the most frequent genetic alteration. It has been suggested that
1
Gynecology, Obstetrics and Gynecology, Otsu, Japan Wnt/β-catenin and TGFβ pathways contribute to MED12-mutated leiomyoma
growth via cell proliferation and extracellular matrix (ECM) formation stimula-
Study question: Can administration of oral antibiotics in patients with chronic tion. In pursuit of new treatments, several studies propose Vitamin D (VitD) as
endometritis (CE) reverse the increased inflammation observed in the a safe therapeutic option to reduce leiomyomas. However, these studies do not
endometrium? examine the existing genetic heterogeneity, a crucial fact to consider in the
Summary answer: The secretion of proinflammatory cytokines was reduced development of an efficient treatment.
in the endometrial stromal cells of patients in whom CE was cured histologically Study design, size, duration: Prospective study in which human uterine
but not in non-cured patients. leiomyomas (UL) and matched myometrium (M) tissues (n=37) were analyzed
What is known already: CE is a mild, persistent inflammation of the endo- to identify MED12 mutations. The status of Wnt/β-catenin and TGFβ pathways
metrium, which is often asymptomatic but can cause infertility. CE is identified was determined in a paired design study in UL and M tissues depending on the
by the presence of plasma cells in the stromal layer of the endometrium. Plasma presence/absence of MED12 mutation. Similarly, the effect of VitD treatment
cell expression indicates the presence of a continuous immune response to any on proliferation and the above-mentioned pathways was evaluated in vitro in
constituent of the endometrium. An increased pregnancy rate in CE patients human uterine leiomyoma primary (HULP) cells depending on MED12 status
treated with antibiotics has been reported. We have previously reported that (n=3/group).
CE patients had higher levels of inflammatory cytokines in their endometrial Participants/materials, setting, methods: UL and M were collected from
stromal cells. However, the effects on endometrial characteristics of curing CE 37 patients (30-61 years) and MED12 mutations were identified by Sanger
with antibiotics are not yet known. sequencing in both tissues. The expression level of WNT4, β-Catenin (Wnt/
Study design, size, duration: Primary culture of endometrial stromal cells β-catenin pathway), TGFβ3, MMP9 (TGFβ pathway) in UL and M was determined
was performed. Thirty endometrial biopsied samples were obtained from 15 by quantitative PCR (qPCR). HULP cells were isolated and treated in vitro with/
women from August 2018 to March 2019. without 1000nm VitD (48h). To evaluate treatment effect, the expression of the
Participants/materials, setting, methods: The subjects were diagnosed above-mentioned genes along with PCNA (cell proliferation marker) was mea-
with CE during in vitro fertilization (IVF) treatment. From each patient, endo- sured by qPCR.
metrial biopsies were collected during the mid-secretory phase, both before and Main results and the role of chance: Sequencing data showed that 46% of
after antibiotic administration. Patients with low serum progesterone levels on UL presented MED12 mutations, while no mutations were detected in their
the day of examination were excluded. The endometrial stromal cells from these corresponding M. According to this, UL were divided into two groups depending
samples were used for primary cell culture. To evaluate the status of inflamma- on MED12 status: MED12-mutated or wild type (WT). The evaluation of Wnt/
tion, ELISA of the culture media was performed. β-catenin and TGFβ pathways in UL and M tissues showed that MED12-mutated
Main results and the role of chance: The numbers of patients with cured UL presented significant higher levels of WNT4 (fold regulation [fr]= 13.05,
CE and persistent CE were 8 and 7, respectively. p=0.02), β-Catenin (fr= 2.84, p=0.01), TGFβ3 (fr= 5.27, p=0.002) and MMP9
For ELISA, endometrial stromal cells were cultured for 14 days and the super- (fr=13.73,p=0.004) than their corresponding M, while no significant differences
natant was obtained. In the cured CE patients, secreted levels of TNF-α were found in WT UL. In vitro study demonstrated that VitD decreased the
(p=0.0094), IL-1β (p=0.031), and IFN-γ (p=0.0089) were significantly decreased expression of the cell proliferation marker PCNA in HULP cells from

MED12-mutated (fr= -1.7, p=0.05) and WT UL (fr= -2.8, p=0.01). Likewise, Wider implications of the findings: This study provides evidence that adeno-
the analysis of TGFβ pathway genes showed that VitD significantly inhibited myotic lesions could be a consequence of immune modifications and Notch1 acti-
TGFβ3 expression in HULP cells from MED12-mutated (fr= -3.1, p=0.02) vation. Hypothesis is that an increase in local inflammation lead to activation of
and WT UL Notch1pathway. NICD, induced in the nucleus, transcriptional activation of genes
(fr= -4, p=0.02). Accordingly, MMP9 expression decreased after VitD treatment related to EMT leading to migration of endometrial cells through myometrium.
in HULP cells from both MED12-mutated and WT UL (fr=-1.47 and -1.45, Trial registration number: NA
respectively). Regarding Wnt/β-catenin pathway, the expression of β-Catenin
was decreased in HULP cells from both MED12-mutated and WT UL (fr= -1.2 P-280 Relation between clinical profiles and adenomyosis
and -1.5, respectively) after VitD treatment. phenotypes assessed by magnetic resonance imaging
Limitations, reasons for caution: Considering the great variability that exists J. Oliveira1, M. Bourdon1, L. Marcellin1, C. Bordonne1, L. Maitrot
between patients, the limitation of this work includes the small sample size in Mantelet1, A.E. Millischer2, G. Plu Bureau1, P. Santulli1,
the in vitro study. Further studies are necessary to translate in vitro results into C. Chapron1
practical clinical applications. 1
University of Paris.- Faculty of Medicine-, Department of Obstetrics and
Wider implications of the findings: Notwithstanding the molecular differ- Gynecology II and Reproductive Medicine- Cochin University Hospital., Paris, France ;
ences found between MED12-mutated and WT UL, VitD could inhibit Wnt/β- 2
Centre de Radiologie Bachaumont, Centre de diagnostique médicale de Paris
catenin and TGFβ pathways in all HULP cells. These results suggest that VitD IMPC, Paris, France
would be an effective treatment to reduce tumor size via the reduction of cell
proliferation and ECM formation in different molecular subtypes of UL. Study question: Do adenomyosis phenotypes such as focal (in the outer
Trial registration number: not applicable myometrium) and diffuse adenomyosis, as diagnosed by magnetic resonance
imaging (MRI), have the same clinical characteristics?
Summary answer: Focal and diffuse adenomyosis phenotypes exhibit distinct
P-279 Aberrant expression of immune cells and Notch1 activation clinical profiles in terms of both women’s characteristics (age, gravity, previous
coincides with epithelial to mesenchymal transition in the surgery, endometriosis) and symptoms (menorrhagia, infertility).
development of adenomyosis in mice. What is known already: Uterine adenomyosis is characterized by the pres-
M. Bourdon1, P. Santulli1, L. Doridot2, M. Jeljeli3, C. Nicco2, ence of endometrial glands and stroma deep within the myometrium. Various
S. Chouzenoux2, C. Chène2, F. Batteux3, C. Chapron1 forms have been described, including focal adenomyosis(FOC-ADE), which
1
université Paris Centre, Gynecology and reproductive médecine, Paris, France ; corresponds to nodular lesions separated from the junctional zone in the outer
2
INSERM, Institut Cochin, Paris, France ; myometrium, and diffuse adenomyosis (DIF-ADE), which is characterized by
3
Université Paris Centre, Immunology, Paris, France endometrial implants scattered throughout the myometrium and enlargement
of the junctional zone. Although the pathogenesis of adenomyosis is not clearly
Study question: In a mouse model of adenomyosis, do innate and adaptive understood, several lines of evidence suggest that these two phenotypes could
immune cells and Notch1 pathway have any role in the development of have distinct origins. The clinical presentation of different forms of adenomyosis
adenomyosis? in patients warrants further investigation.
Summary answer: Notch1 activation and aberrant expression of innate and Study design, size, duration: This was an observational study. Non-pregnant
adaptive immune cells promote adenomyosis developpement through the epi- patients, aged between 18 and 42 years, who had undergone surgical exploration
thelial to mesenchymal transition (EMT). for benign gynecological conditions at our institution between May 2005 and
What is known already: EMT has been implicated in the development of May 2018 and with a preoperative uterine MRI were included in the study. Data
adenomyosis, along with a dysregulated innate and adaptive immune response. was prospectively collected by a standardized questionnaire regarding women’s
One hypothesis is that immune changes in uterus may promote endometrial histories and clinical symptoms, completed during a face-to-face interview con-
cells proliferation and migration to myometrium and EMT activation, leading to ducted by the surgeon in the month preceding the surgery.
adenomyosis lesions. Besides, inflammation is known to be a potential activator Participants/materials, setting, methods: 248 women had a radiological
of the Notch signaling pathways known to have capacity to favor the EMT diagnosis of adenomyosis assessed by MRI and signed informed consent. Based
process. on the MRI findings, the women were diagnosed as having FOC- and/or DIF-
Study design, size, duration: Adenomyosis was induced in 26 female CD1 adenomyosis. The women were allocated to two groups according to the ade-
mice by neonatal dosing of tamoxifen. Another 28 were neonatally dosed with- nomyosis phenotype (FOC-ADE versus DIF-ADE). Women exhibiting an
out tamoxifen (control group). These mice were sacrificed serially and their association of both the FOC- and the DIF-adenomyosis form were excluded.
tissue samples were subsequently evaluated. Main results and the role of chance: All up, following the MRI findings, 109
Participants/materials, setting, methods: Female CD-1mice with and with- women (44.0%) exhibited only FOC-ADE and 78 (31.5%) had only DIF-ADE.
out induced adenomyosis were sacrificed in batch at 30,60 and 90 days of age. The The women with FOC-ADE were significantly younger (mean of31.9 years of
depth of myometrial infiltration of endometrial tissues was assessed with histology. age ± 4.6 SD versus 33.8 ± 5.2 SD, respectively; p = 0.006), more often nulli-
Immune cells markers disturbance (CD45,Ly6C1,CD86,Arginine1,CD19,CD4, gravid ( 74.3%versus 33.3%; p ≤ 0.001), exhibited a higher rate of associated-
CD8) were analysed by RTqPCR. Notch1 and target genes (HEY1,HEY2,HES1,HES5) endometriosis (p= 0.001) especially deep infiltrating endometriosis (89.0%
were analysed by RTqPCR. The Notch intracellular domain (NICD) was measured versus 30.8%) and a higher rate of infertility (p=0.021) especially primary infer-
with western blot analysis. Analysis of EMT biomarkers was performed with tility (25.1% versus 9.0%) compared to the women in the DIF-ADE group.
RTqPCR (E-Cadherin,Vimentin,TGFb,SNAIL1,SLUG,Snail 3) and Western blot Moreover, the women exhibiting DIF-ADE had significantly more often a history
(E-Cadherin,Vimentin). of previous uterine surgery (21.8% versus 6.4%;p = 0.002) and menorrhagia
Main results and the role of chance: Aberrant expression of immune cells (79.5% versus 53.2%; p= 0.001) compared to the women with FOC-ADE. No
markers was observed in uteri from adenomyosis -mice over the development of differences in the pain scores (i.e., dysmenorrhea, non-cyclic pelvic pain, and
the disease. Expression of innate inflammatory cells markers, notably M1 macro- dyspareunia) were observed between the two groups.
phages and natural killer’s cells were increased from an early stage (Day30) com- Limitations, reasons for caution: The exclusive inclusion of surgical patients
pared to uteri from control mice, followed by an increase of CD4 T cells at day 60. as the limitation age could constitute possible selection bias. Moreover, the
Inversely, expression of CD19 B cells were significantly decreased during all studied women referred to our center may have suffered from particularly severe clinical
stage. Activated-Notch1 was also highly activated during adenomyosis development symptoms.
compared to control mice at day 30 and D60. Concomitantly, some markers for Wider implications of the findings: In clinical practice, these two forms of
EMT also was increased. This Notch activation could be related to the activation adenomyosis should be differentiated using an appropriate imaging work-up in
of EMT and migration of endometrial cells within the myometrium. order to search for associated endometriosis and to devise a tailored therapeutic
Limitations, reasons for caution: This study is limited by the use of an strategy. To better understand the pathogenesis, future mechanistic studies aimed
animal model and the lack of intervention. at better characterization of diffuse and focal adenomyosis are needed.

Trial registration number: not applicable Summary answer: RWC-TVS and SGV have similar accuracy in the diagnosis
of posterior DIE. However, RWC-TVS is more accurate in diagnosing presence
P-281 Regulatory effect of prohibitin on glucose metabolism of and characteristics of rectosigmoid endometriosis.
granulosa cell in endometriosis. What is known already: Surgical treatment of posterior DIE may be chal-
M. Jingqin1, C.Y. Zhong1, L. Gao1, J. Zhang1, J.Y. Zhang1, L. Dai1, lenging for surgeons and carry significant risks for patients. Preoperative assess-
Y.D. Mao1 ment by imaging of the location, characteristics and presence of posterior DIE
1 nodules is important in order to inform the patient on the possible treatments
First Affiliated Hospital of Nanjing Medical University, Clinical Center of Reproductive
and to provide informed consent to patients undergoing surgery. RWC-TVS and
Medicine, Nanjing, China
SGV consist of enhanced transvaginal ultrasound scans based on the introduction
Study question: What role does prohibitin play in glucose metabolism in of contrast into the rectum and vagina, respectively.
granulosa cells of patients with endometriosis? Study design, size, duration: This was a prospective comparative study,
Summary answer: Patients with endometriosis shows the high expression of enrolling 221 women with clinical suspicion of posterior DIE.
prohibitin to make up for glucose aerobic metabolism defects. Participants/materials, setting, methods: Patients with previous diagnosis
What is known already: The metabolic dysfunction in granulosa cells of of posterior DIE by magnetic resonance, computed tomography or laparoscopy
endometriosis patients has been examined by various groups. Prohibitin is ubiq- were excluded from the study. All patients underwent RWC-TVS and SVG,
uitously expressed, mainly presenting in mitochondria to maintain the membrane performed by two independent ultrasonographers, informed of the patients’
potential of mitochondrial, to facilitate in assembly of respiration chain. clinical history and symptoms but blinded to the other diagnostic exam. The
Meanwhile, prohibitin play a critical role in energy metabolism. presence of rectovaginal septum, rectosigmoid, uterosacral ligaments, and vagina
Study design, size, duration: In this study, glucose metabolic phenotype of endometriosis was investigated. Patients underwent laparoscopy within the fol-
granulosa cells were examined in vitro. The human granulosa cell line KGN were lowing 6 months. Imaging findings were compared with surgical and histological
transfected with lentivirus to knock down and over express the expression of results.
prohibitin. Afterwards, the glucose metabolism phenotype and cell vitality were Main results and the role of chance: In 89.6% (n=198/221) of the patients,
evaluated. posterior DIE was laparoscopically confirmed. The nodules were localized in
Participants/materials, setting, methods: The primary granulosa cells rectovaginal septum (36.4%), rectosigmoid (24.2%), uterosacral ligaments
were collected to test the glucose metabolism features as follows: lactate pro- (66.1%), and vagina (6.4%). There was no significant difference in the perfor-
duction (Lactate assay kit) glucose consumption (Glucose assay kit) and the mance of RWC-TVS and SVG in diagnosing posterior DIE (p=0.187). The accu-
expression of prohibitin and glucose metabolism related enzymes(q-PCR and racy of RWC-TVS in the detection of posterior DIE was 92.2%, sensitivity was
Western blotting). The KGN was transfected with lentivirus to knock down and 89.4%, specificity 95.0%, positive predictive value 94.6%, negative predictive
over express the expression of prohibitin. The mitochondrial function and cell value 90.1%, positive likelihood ratio 18.0 and negative likelihood ratio 0.1. For
proliferation were tested by ATP assay kit and CCK8 kit. SVG, the corresponding values were 88.8%, 82.8%, 94.9%, 94.3%, 84.4%, 16.3
Main results and the role of chance: In women with endometriosis, signifi- and 0.2, respectively. RWC-TVS had higher accuracy than SVG in diagnosing
cantly increased glucose consumption, lactate production, prohibitin expression presence of rectosigmoid endometriosis (sensitivity 91.2% vs 80.7%; specificity
and aberrant expression of glycolysis-related enzymes were found compared with 95.8% vs 89.0%; p<0.001), depth of penetration of endometriosis in the intes-
women who do not have endometriosis (P < 0.05). To determine whether pro- tinal muscularis propria (89.3% vs 79.5%; 90.2% vs 77.8%; p<0.001), and distance
hibitin regulated glucose metabolism by affecting the expression of glycolysis between the nodule and anal verge (91.3% vs 77.2%; 88.3% vs 74.2%; p<0.001).
related enzymes, the expression of enzymes were evaluated after lentivirus trans- There was no significant difference between the two techniques in diagnosing
fection in KGN. When prohibitin was down regulated in KGN, the mRNA expres- endometriotic nodules of utero-sacral ligaments (83.5% vs 82.1%, 82.4% vs
sion of HK1, ENO1α, PDHA, CS, SDHB and ATP5A were significantly decreased 79.1%; p=0.312) and rectovaginal septum (92.5% vs 88.2%, 87.8% vs 85.3%;
(P < 0.05). When prohibitin was over expressed, the the mRNA expression of p=0.431). RWC-TVS had lower sensitivity (75.0% vs 91.7%, p=0.031) but similar
Glut1, HK1, ENO1α, LDHA and PDHA were significantly increased (P < 0.05). specificity (98.1% vs 97.6%; p=0.453) than SVG in diagnosing vaginal
Meanwhile, the over-expression of prohibitin led to elevated glucose consumption endometriosis.
and lactate production (P < 0.05), whereas, the knock-down of prohibitin made Limitations, reasons for caution: A limitation of the study if that the diag-
no change. Additionally, cell viability and proliferation were significantly inhibited nostic performance of both techniques is depending on the experience of the
after the down regulation of prohibitin(P < 0.05). examiners. The surgeons were aware of the findings of these diagnostic exams.
Limitations, reasons for caution: More experiments are necessary to Both techniques cannot diagnose endometriotic nodules located above the
explore the specific molecule signal pathway involved with prohibitin to regulate rectosigmoid.
the glucose metabolism. Wider implications of the findings: This study showed that RWC-TVS and
Wider implications of the findings: Women with endometriosis exhibits SVG have similar performance in diagnosing deep posterior DIE. RWC-TVS is
aberrant glucose metabolism and high level expression of prohibitin in granulosa more accurate that SVG in assessing the presence and characteristics of recto-
cells. According the results, prohibitin may improve the glycolysis to make up sigmoid endometriosis.
the energy output deficiency resulting from mitochondrial dysfunction, to exert Trial registration number: Not applicable
the protective effect on gruanlosa cells in women with endometriosis.
Trial registration number: 81671438, 81730041 P-283 Three-dimensional rectal water contrast transvaginal
ultrasonography versus virtual colonoscopy for diagnosing
presence and characteristics of rectosigmoid endometriosis
P-282 Rectal water-contrast transvaginal ultrasonography versus F. Barra1, E. Biscaldi2, C. Scala3, V.G. Vellone4, C. Stabilini4,
sonovaginography for the diagnosis of posterior deep pelvic S. Ferrero1
endometriosis: a prospective comparative study 1
IRCCS Ospedale Policlinico San Martino - University of Genoa, Academic Unit of
F. Barra1, C. Scala2, V.G. Vellone3, C. Stabilini3, S. Ferrero1 Obstetrics and Gynecology, Genoa, Italy ;
1
IRCCS Ospedale Policlinico San Martino- University of Genoa, Academic Unit of 2
Galliera Hospital, Radiology, genova, Italy ;
Obstetrics and Gynecology, Genoa, Italy ; 3
Gaslini Institute, Unit of Obstetrics and Gynecology, Genoa, Italy ;
2
Gaslini Institute, Unit of Obstetrics and Gynecology, Genoa, Italy ; 4
IRCCS Ospedale Policlinico San Martino - University of Genoa, Department of
3
IRCCS Ospedale Policlinico San Martino- University of Genoa, Department of Surgical and Diagnostic Sciences DISC, Genoa, Italy
Surgical and Diagnostic Sciences DISC, Genoa, Italy
Study question: To compare the performance of three-dimensional rectal
Study question: To investigate the accuracy of rectal water-contrast trans- water contrast transvaginal ultrasonography (3D-RWC-TVS) and computed
vaginal ultrasonography (RWC-TVS), and sonovaginography (SVG) in patients tomographic colonography (CTC) in predicting the presence and characteristics
with clinical suspicion of posterior deep infiltrating endometriosis (DIE). of rectosigmoid endometriosis.

Summary answer: 3D-RWC-TVS and CTC have similar diagnostic accuracy mechanism involved in the disease-related infertility. Recently, the sequencing
for diagnosing rectosigmoid endometriosis. However, CTC is more accurate in of mRNAs (RNA-Seq) did not show differential expression in the transcripts of
ruling out the presence of sigmoid endometriotic nodules. infertile patients with endometriosis when compared to infertile and fertile con-
What is known already: A non-invasive diagnosis of bowel endometriosis trols during the implantation window. However, these findings do not exclude
is relevant to provide the patients information on the potential treatments (hor- the possibility of existing other molecular alterations with possible impact on
monal therapies or surgery) and to obtain adequate informed consent in case protein synthesis and, potentially, on the establishment of endometrial receptivity.
of surgery. Over the last 10 years, RWC-TVS and CTC based on the distention Study design, size, duration: In this descriptive study, data from RNA-Seq
of the rectosigmoid have been proposed with the aim to improve the diagnosis analysis of endometrial biopsies collected during the implantation window from
of deep infiltrating endometriosis. Both exams allow the evaluation of bowel 17 patients (3 infertile women with endometriosis I/II, 3 infertile women with
using a pseudoendoscopic view. endometriosis III/IV, 6 infertile controls and 5 fertile controls) were in silico
Study design, size, duration: This was a prospective comparative study analyzed by bioinformatics tools for variant discovery, functional mutations iden-
including patients who were referred to our institution for suspicion of recto- tification and selection of proteins possibly affected. A targeted study of the
sigmoid endometriosis between January 2018 to December 2019. alterations found was performed to understand the data in the disease’s context.
Participants/materials, setting, methods: Patients with symptoms sug- Participants/materials, setting, methods: Analyzes were performed in
gestive of rectosigmoid involvement by endometriosis were included in the study. the GATK (Genome Analysis Toolkit), following the best practices recommended
They underwent 3D-RWC-TVS performed by a sonographer skilled in the diag- for the discovery of variants in RNA-Seq data. Data were filtered according to
nosis of deep endometriosis. CTC was done within the following three months the coverage, gene region, variant function and description in NCBI databases.
by a radiologist expert in the diagnosis of deep endometriosis, blinded to the Subsequently, the variants were evaluated by Pathogenicity Predictors. The first
results of the previous ultrasonographic exam. Patients underwent surgery within predictor applied was the CADD, followed by the agreement of the predictors
the following six months. Imaging findings were compared with surgical and PROVEAN, SIFT and Polyphen 2 in classifying deleterious or possibly deleterious
pathologic results. variants.
Main results and the role of chance: Out of 68 women included, 37 (48.9; Main results and the role of chance: None of the variants found were
95% C.I. 38.2%-59.7%) had rectosigmoid nodules which required bowel surgery. common to other samples within the same group, as well, no mutation was
The main nodules were located on the sigmoid in 16 (43.2%) patients, on the repeated in patients with endometriosis, infertile controls and fertile controls.
rectosigmoid junction in 4 (10.8%) patients, on the upper rectum in 10 (27.0%) In the endometriosis group, 9 mutations predicted to be harmful were identified.
patients and on the lower rectum in 7 patients (18.9%). There was no significant Only one of the mutations (in the gene SLC2A9) had already been described as
difference in the accuracy of 3D-RWC-TVS and CTC in diagnosing the presence associated with clinical conditions (familial renal hypouricemia), with no evident
of rectosigmoid endometriotic nodules (p = 0.118). However, a subgroup anal- impact on the endometrium. However, when crossing the genes related to the
ysis demonstrated that CTC was more precise than 3D-RWC-TVS in diagnosing 9 mutations found with the keywords “endometriosis” and/or “endometrium”,
endometriosis located in sigmoid (p = 0.016). The presence of an endometrioma the gene CMKLR1 was associated either with the inflammatory response in
with diameter > 4 cm was positively statistically correlated to lack of identifica- endometriosis or with important endometrial processes for pregnancy
tion of sigmoid endometriotic nodules (phi coefficient 0.516; p=0.039) by per- establishment.
forming 3D-RWC-TVS. Both exams estimated similarly the largest diameter of Limitations, reasons for caution: The small sample size does not allow us
the main endometriotic nodule, independently of their location (p = 0.090). to state whether there are differential mutations among women with endome-
CTC was more accurate than 3D-RWC-TVS in estimating the distance between triosis compared to fertile and infertile controls. Moreover, the search for variants
the lower margin of the intestinal nodule and the anal verge (p = 0.030). There was performed on RNA-Seq data and only expressed transcripts were evaluated.
was no significant difference in the performance of 3D-RWC-TVS and CTC in Regulatory regions were not analyzed.
diagnosing multifocal disease (p = 1.000). Wider implications of the findings: Despite no pattern of mutation was
Limitations, reasons for caution: A limitation of both techniques is repre- found, we ponder the small sample size and the analysis on RNA-Seq data.
sented by the extensive experience of the radiologist and the gynecologist per- Considering the study’s purpose of screening and the CMKLR1 importance on
forming CTC and 3D-RWC-TVS, respectively, which may have influenced the endometrial modulation, it could be a candidate gene for further studies evalu-
performance of these techniques in ruling out the presence of rectosigmoid ating mutations in eutopic endometrium from endometriosis patients.
endometriosis. Trial registration number: Not applicable
Wider implications of the findings: 3D-RWC-TVS and CTC have similar
diagnostic performance for diagnosing rectosigmoid endometriosis, although P-285 Pretreatment with dienogest in women with endometriosis
3D-RWC-TVS is less accurate in detecting sigmoid nodules. CTC may be com- undergoing in vitro-fertilization after a previous failed cycle
bined with 3D-RWC-TVS because of the high diagnostic performance in detect- F. Barra1, C. Scala2, G. Evangelisti1, S. Ferrero1
ing rectosigmoid endometriosis and the ability to diagnose endometriotic nodules 1
IRCCS Ospedale Policlinico San Martino - University of Genoa, Academic Unit of
located above the rectosigmoid junction.
Obstetrics and Gynecology, Genoa, Italy ;
Gaslini Hospital, Unit of Obstetrics and Gynaecology, Genoa, Italy
P-284 In silico screening and variant analysis in the transcript Study question: To investigate the effect of dienogest (DNG) administered
profile of eutopic endometrium from infertile women with in women with endometriosis undergoing IVF after a previous failed cycle.
endometriosis and controls during the implantation window Summary answer: In women with endometriosis, particularly in those with
M. Da Broi1, J. Plaça2, R. Ferriani1, P. Navarro1 large endometriomas, implantation rate and clinical pregnancy rate can be
1
Ribeirão Preto School of Medicine- University of São Paulo, Department of improved by pretreatment with DNG.
Obstetrics and Gynecology, Ribeirão Preto, Brazil ; What is known already: It is generally assumed that diminished ovarian
2
Center for Medical Genomics- HCFMRP/USP, Department of Genetics, Ribeirão reserve, impaired endometrial receptivity and low quality of embryos may be
Preto, Brazil causes of the lower in vitro fertilization (IVF) outcomes of women with endo-
metriosis. The use of prolonged courses of hormone therapy may improve IVF
Study question: Is there a pattern of functional mutations in transcripts of outcome of patients with endometriosis.
the eutopic endometrium of infertile women with endometriosis during the Study design, size, duration: This was a retrospective analysis of a prospec-
implantation window that could be related to impaired endometrial receptivity? tive database collected between January 2016 to July 2019. Women who failed
Summary answer: No pattern of functional mutations was identified in the a previous IVF cycle and all subsequent embryo transfers, with concomitant
transcripts of the eutopic endometrium of infertile women with endometriosis diagnosis of endometriosis at transvaginal ultrasonography or magnetic reso-
during the implantation window. nance imaging were included in this study.
What is known already: The molecular and functional impairment of the Participants/materials, setting, methods: Inclusion criteria were age ≤40
eutopic endometrium in women with endometriosis is questioned as a possible years; basal FSH <14.0 IU/L and anti-Müllerian hormone (AMH) blood level

≥0.5 g/mL. Women who underwent previous surgical treatment of endome- irradiated mice (uterus lead shielded) with congenic bone marrow to allow
triosis were excluded. Following a failed IVF cycle, patients either directly under- tracking of donor cells in uterine tissue.
went IVF without receiving hormonal treatment or received a three-month DNG Main results and the role of chance: Mature macrophages were present
(2 mg daily) treatment before IVF. The choice of treatment was based on at all stages of the murine estrous cycle and expressed features indicating involve-
patients’ preference. ment in tissue remodelling and repair, including high levels of the IL-4 receptor,
Main results and the role of chance: 151 patients were included in the CSF-1 receptor, mannose receptor CD206 and protein Relmα (critical for
study, 88 (58.3%) underwent IVF without previous hormonal treatment and 63 remodelling and repair in other tissues). Accordingly, endometrial macrophages
(41.7%) received pretreatment with DNG. At baseline, there was no significant were more responsive to IL-4 than mucosal macrophages from other tissues
difference in age, average duration of infertility, antral follicle count, basal serum (p<0.001). Unlike other tissue macrophages, endometrial macrophages did not
FSH, AMH and presence of endometriomas between the two groups. The proliferate in situ but the substantial majority were differentiated from circulating
implantation rate and clinical pregnancy rate were significantly higher in the monocytes recruited into the endometrium in a manner dependent on the
DNG-treated group (39.7% and 33.3%) compared with the non-treated group chemokine receptor CCR2 (p<0.01). By 20 weeks post irradiation, turnover of
(23.8% and 18.2%; p=0.037 and p=0.033, respectively). The largest diameter macrophages in the uterus from blood monocytes was higher than in the intestine
and volume of endometriomas significantly decreased after pretreatment with (p<0.05), previously demonstrated to have one of the highest rates of replen-
DNG (-0.7 ± 0.8 cm and -3.1 ± 7.7 cm3, p<0.001 and p<0.001, respectively). ishment. Numbers of monocytes in the uterus were profoundly increased during
A subgroup analysis demonstrated that the use of DNG increased significantly estrus (receptivity period when estrogen is low; p<0.01), likely accounting for
the number of oocytes retrieved (6.1 ± 2.8 versus 5.3 ± 2.2, p=0.039), of 2 the high rate of turnover into macrophages. These recruited macrophages were
pronuclear (2PN) embryos (2.9 ± 1.6 versus 2.2 ± 1.8, p=0.015) and of blas- highly responsive to IL-4 and exhibited enhanced levels of CD206 and Relmα
tocysts (2.4 ± 1.6 versus 2.0 ± 1.4, p=0.049) obtained in patients with endo- compared to their tissue-resident counterparts (p<0.001). Preliminary data
metriomas with diameter ≥ 4 cm. suggest human endometrial macrophages are drastically expanded in Asherman’s
Limitations, reasons for caution: While baseline demographics and first syndrome and that these cells have immature properties suggesting dysregulated
IVF cycle stimulation parameters for the studied patients were comparable, differentiation.
the non-randomized allocation of subjects may mask a hidden bias in Limitations, reasons for caution: The remodelling events that occur
this study. throughout the murine estrous cycle differ from those in the human menstrual
Wider implications of the findings: IVF outcomes of women with endo- cycle in terms of endometrial shedding/menstruation which does not occur in
metriosis seem to be improved after pretreatment with DNG. In particular, this mice. Nonetheless, mouse models provide the opportunity to perform the in
progestin may give a benefit in patients with large endometriomas. vivo ontogeny and functional studies that are not possible in humans.
Trial registration number: Not applicable Wider implications of the findings: The estrous cycle is marked by dynamic
changes in populations of endometrial monocytes/macrophages that are likely
P-286 Dynamic properties of endometrial macrophages regulate to play a key role in tissue remodelling during the receptivity period in which
tissue homeostasis and disease in the uterus implantation can occur in homeostasis, but may contribute to pathology in
E. Mann1, N. Scott1, L. Lacerda Mariano2, P. Ruane3, M. Ingersoll2, fibrotic disease such as Asherman’s syndrome when dysregulated.
L. Mohiyiddeen4
1
University of Manchester, Lydia Becker Institute of Immunology and
P-287 Suppression of ovarian activity during co-administration
Inflammation, Manchester, United Kingdom ;
2 of the oral gonadotropin-releasing hormone receptor antagonist
Institut Pasteur, Immunology, Paris, France ;
3 relugolix, estradiol, and norethindrone acetate in healthy female
University of Manchester, Maternal and Fetal Health Research Centre,
volunteers
Manchester, United Kingdom ;
4
Manchester NHS Foundation Trust, Reproductive Medicine, Manchester, United I. Duijkers1, C. Klipping1, C. Draeger2, B. Schug3, R.S.
Kingdom Wedemeyer3, J. Li4, J.C. Arjona Ferreira4, E. Migoya4
1
dinox consultancy, Groningen, The Netherlands ;
Study question: Do macrophages contribute to tissue remodelling and repair 2
dinox GmbH, Berlin, Germany ;
in the uterus, and contribute to pathology in fibrosis/Asherman’s syndrome 3
SocraTec R&D GmbH, Oberursel, Germany ;
when their function is dysregulated? 4
Myovant Sciences Inc., Brisbane, U.S.A.
Summary answer: Monocytes recruited from the bloodstream generate
uterine macrophages with distinct roles in tissue repair and remodelling, and Study question: Does relugolix combination therapy (relugolix, oral gonado-
macrophages may be dysregulated in Asherman’s syndrome. tropin-releasing hormone (GnRH) receptor antagonist, 40mg, estradiol (E2) 1mg
What is known already: Dysregulated tissue degradation, repair and remod- and norethindrone acetate (NETA) 0.5mg) suppress ovarian activity and
elling and can lead to fibrosis and adhesions in the uterus (known as Asherman’s ovulation?
syndrome), causing reproductive dysfunction. In other tissues, macrophages Summary answer: Once daily oral administration of relugolix combination
(immune cells) are central to tissue remodelling and repair. However, the ontog- therapy for 84 days suppressed ovarian activity and inhibited ovulation in 100%
eny and functions of macrophages within the uterus are poorly defined, despite of subjects.
dysregulation of macrophage recruitment and function driving fibrosis at other What is known already: Relugolix competitively binds to the GnRH receptor,
mucosal sites. preventing the release of follicle-stimulating hormone (FSH) and luteinizing hor-
Study design, size, duration: Macrophages were characterized at different mone (LH), thereby decreasing the production of E2 and progesterone (P). By
stages of the estrous cycle in mice (proestrus, estrus, metestrus and diestrus), lowering systemic concentrations of E2 and P, oral administration of 40 mg
which reflects the hormone fluctuations that regulate tissue remodelling and relugolix monotherapy is effective in the treatment of symptoms associated with
receptivity to embryo implantation occurring in the human uterus (n=10-15 per uterine fibroids, however, is associated with hypoestrogenic symptoms and bone
estrous cycle stage). Differentiation of murine endometrial macrophages from mineral density loss. In current phase 3 trials in patients with uterine fibroids or
infiltrating blood monocytes in vivo was assessed using methods described below endometriosis, relugolix treatment is combined with 1 mg E2 and 0.5 mg NETA
(n=6-12 per time point). Macrophages were characterized from human endo- to mitigate hypoestrogenic side effects.
metrial biopsies, obtained following informed consent (n=5). Study design, size, duration: An open-label, single treatment-group clinical
Participants/materials, setting, methods: Macrophages were character- study in 70 women, 67 of which completed the study. The study included a
ised from uterine tissue (mouse and human) by multi-parameter flow cytometry pre-treatment cycle to assess ovulatory status, an 84-day treatment period to
following enzymatic digestion. Mice were staged using vaginal smear cytology to assess ovarian activity during combination treatment (co-administration of a
determine estrous cycle phases according to proportions of epithelial cells, relugolix 40mg tablet and an E2/NETA 1mg/0.5mg tablet (Activelle®)), and a
cornified cells and leucocytes. Ontogeny of murine endometrial macrophages post-treatment cycle to assess return of ovulation. Subjects visited the study
was defined at multiple time points 8 and 20 weeks after reconstitution of center every third (± 1) day during the entire study.

Participants/materials, setting, methods: The study, conducted in a clinical included endometrial thickness, implantation rate, clinical pregnancy rate, mis-
research center, included healthy premenopausal women 18-35 years of age. carriage rate, complications of treatment and obstetric/perinatal outcomes.
During the study visits ultrasound measurements of follicular growth and endo- Mantel-Haenszel odds ration with random effects was used as analysis method.
metrial thickness, and blood sampling for determination of serum E2, P, FSH and Main results and the role of chance: A total of 350 items were identified,
LH concentrations were performed. Ovarian activity was categorized using the after deduplication 266 titles were screened. 47 papers were included for detailed
Hoogland-Skouby score. Safety parameters and vaginal bleeding were also analysis with 31 studies included in the meta-analysis. Identified treatments
assessed. The primary endpoint was the proportion of subjects with Hoogland- included infusion of granulocyte colony stimulating factor (G-CSF); infusion of
Skouby scores <5 during the treatment period. platelet rich plasma (PRP); long term treatment with pentoxyfilline and tocoph-
Main results and the role of chance: Once-daily dosing with relugolix erol; pelvic flood bionic stimulation, growth hormone injections, high dose
combination therapy resulted in suppression of ovarian activity and inhibition of estrogen therapy, use of prostaglandins, addition of gonadotrophin releasing
ovulation in 100% of women as demonstrated by Hoogland-Skouby scores <5 hormone agonists (GNRH-a) as luteal phase support.
during the entire 84-day treatment period. In most subjects the largest follicular Live birth rate was reported by 6 studies with addition of GnRH-a to luteal phase
diameter remained below 10 mm throughout treatment, resulting in a Hoogland- showing an odds ratio (OR) of 8.80 [95% Confidence Intervals(CI) 2.44-31.73,
Skouby score of 1 (no ovarian activity). Mean and median E2 concentrations, P=0.0009, one study). Most significant increase in endometrial thickness was
reflecting both endogenous and exogenously-administered E2, were between noted with long treatment with pentoxifylline and tocopherol (mean difference
110 and 160 pmol/L (30 and 44 pg/mL). Serum P, FSH, and LH concentrations 2.40, 95% CI 2.15-2.65, P<0.00001, one study). Endometrial perfusion with PRP
were suppressed for the duration of treatment. Ovarian activity resumed after demonstrated highest chance of implantation (OR 4.92, 95% CI 1.78-13.58,
treatment discontinuation with ovulation occurring a mean of 23.5 days after P=0.002, two studies). Addition of GnRH-a to luteal support demonstrated
the last treatment day. Relugolix combination therapy was generally well toler- highest chance for clinical pregnancy (OR=3.76; 95% CI 1.51-9.36, P=0.004,
ated in premenopausal women over 84 days of treatment. one study). No complications or obstetric outcomes were reported.
Limitations, reasons for caution: Compliance regarding tablet intake could Based on the risk of bias, only one study rated as good with majority rated as
be lower in clinical practice than in the setting of a clinical trial, therefore these poor. Heterogeneity ranged from I2=0 (low) to >80% (high).
results may differ from those in the clinical setting. However, the robustness of Limitations, reasons for caution: Only 10 randomized controlled trials
the ovarian suppression limits the risk of escape ovulations due to missed med- were identified with the largest including only 135 patients. The quality of the
ication intake. studies was mainly poor to moderate. Studies reporting other therapies identified
Wider implications of the findings: Since relugolix combination therapy by the search did not have adequate data to carry out meta-analysis.
completely inhibited ovulation, the use of non-hormonal contraception during Wider implications of the findings: Our study highlights the lack of good
treatment may not be required for prevention of pregnancy. The rapid and quality evidence for different types of treatments for women with persistently
predictable return of ovulation after treatment discontinuation is advantageous thin endometrium. Addition of GnRH-a to luteal phase support shows some
for patients who wish to conceive after completion of treatment. promise however, larger studies are necessary to validate this finding. Other
Trial registration number: EudraCT 2018-004130-15 therapies require more well-designed RCTs to confirm their clinical benefit.
Trial registration number: PROSPERO registration number
P-288 Interventions for women with thin endometrium CRD42019154704.
undergoing assisted reproductive treatment (ART)- A systematic
review and meta-analysis. P-289 The effect of endometriosis in the chromosomal status
L. Polanski1, M. Baumgarten2, K. Jayaprakasan3 and developmental competence of cleavage-stage embryos.
1 Differences with embryos in the blastocyst stage.
Assisted Conception Unit, Guy’s and St Thomas’ NHS Foundation Trust, London,
United Kingdom ; P. Hernandez1, B. Gadea2, V. Lozoya3, V. Legidos4, M. Muñoz4
2 1
Addenbrooke’s Hospital, Obstetrics and Gynaecology, Cambridge, United Kingdom ; IVI Alicante, Local Research Department, Alicante, Spain ;
3 2
Royal Derby Hospital, Derby Fertility Unit, Derby, United Kingdom IVI Alicante, IVF Laboratory, Alicante, Spain ;
3
IVI Foundation, Research Support and Management UAGI, Alicante, Spain ;
Study question: What interventions, if any, have been shown to improve ART 4
IVI Alicante, Gynecologist, Alicante, Spain
outcomes in women with thin endometrium defined as <7mm despite routine
treatments? Study question: Is there any difference in the PGT-A results between tro-
Summary answer: A number of interventions have been identified. Study phectoderm and blastomere biopsies from patients with endometriosis com-
quality was rated as poor and live birth rates were reported by only 6 studies. pared to control patients?
What is known already: Normal endometrial receptivity is essential for Summary answer: Unlike PGT-A results in blastocyst, the number or euploid
pregnancy establishment and normal development. Endometrial thickness <7 cleavage-stage embryos is lower in endometriosis patients but it does not affect
mm is associated with a reduced chance of pregnancy in ART, an increased risk embryo developmental arrest.
of miscarriage and adverse obstetric outcomes. The aetiology of thin endome- What is known already: Endometriosis affects around 30-50% of infertile
trium remains uncertain, however damage of the endometrial basal layer is the women undergoing fertility treatment and negatively impacts on their chances of
likely underlying cause. Prevalence of thin endometrium is reported in 2.4%- 8.5% successful IVF outcomes. Among the underlying mechanism responsible for the
of women undergoing ART, fresh or frozen cycles. Many interventions have been poor outcomes, oocyte meiotic spindle alterations and chromosomal instabilities,
proposed to improve endometrial thickness and reproductive outcomes in subsequently resulting in embryo aneuploidies, have been suggested. The only data
these women. available for human embryo chromosomal analysis suggested that endometriosis
Study design, size, duration: Design- systematic review and Meta-analysis does not impact on aneuploidy rates but only embryos reaching the blastocyst
We identified 31 studies, comprising 2421 women, that compared treatments stage were included in the analysis. Therefore, aneuploidies leading to embryonic
for thin endometrium in fresh or frozen embryo transfer cycles. Randomized developmental arrest before the blastocyst stage may not have been detected.
controlled trials (RCTs), case controlled studies and cohort studies with a control Study design, size, duration: Multicenter retrospective cohort study to
population were included. Medline, Embase and CENTRAL databases have been evaluate the chromosomal status of both cleavage-stage and blastocyst embryos
systematically searched until 08.11.2019 using terms (thin OR atrophic AND from patients with endometriosis (n=507) compared with disease-free control
endometri*) AND (assisted conception OR reproduction OR in vitro fertili*(IVF) women (n= 7969). Women aged 18-42 years old undergoing IVF with preim-
OR intracytoplasmic sperm injection(ICSI) OR frozen embryo). plantation genetic testing for aneuploidies (PGT-A) in IVIRMA Clinics between
Participants/materials, setting, methods: Infertile women undergoing 2012 and 2019 were included. Both, developmental competence and PGT-A
fresh or frozen autologous embryo transfer where endometrial thickness as results were also examined in the cleavage-stage biopsied embryos to ascertain
defined by the authors was thin (<8mm) were included in the analysis. Control the effect of endometriosis on aneuploidy-related embryonic arrest.
populations included women with no intervention or previous cycle data (for Participants/materials, setting, methods: Presence of endometriosis was
endometrial thickness). The main outcome was live birth, secondary outcomes evidenced at the time of abdominal surgery or after pelvic ultrasound or NMR.

Severe male factors and patients with chromosomopathy in previous embryos samples were histologically assessed by an expert pathologist blinded to the
or pregnancies were excluded. Both D3 and D5-7 biopsies were analyzed using treatment.
comparative genomic hybridization (CGH) arrays or next generation sequencing Participants/materials, setting, methods: The study was conducted in a
(NGS). For the statistical analysis, chi-square test for categorical variables or single center in Sweden. Eligible were healthy women opting for a LNG-IUS,
Student’s t-test for quantitative data were applied to compare baseline charac- aged 18-43, with normal menstrual cycles. The paired endometrial samples were
teristics between groups. stained with hematoxylin and eosin and assessment involved classifying biopsies
Main results and the role of chance: Two thousand seventy-two embryos according to (1) primary diagnosis (benign, hyperplastic or malignant), (2)
from patients with endometriosis were analyzed and 32008 were included in the whether appearances represented a physiological state or not and if physiolog-
control group. Among them, 9049 were cleavage-stage embryos (943 and 8106 ical, record the stage of the menstrual cycle and (3) the presence or absence
embryos in endometriosis and control group, respectively) and 25031 were of PAEC.
blastocyst (1129 and 23902 blastocysts from patients with endometriosis and Main results and the role of chance: This is the first study assessing endo-
control women). The mean number of euploid embryos from patients with metrial safety and presence of PAEC after long-term treatment with a PRM with
endometriosis (1.04; CI: 0.93-1.15) tends to be lower than in the control group subsequent placement of an LNG-IUS with no prior endometrial shedding. There
(1.17; CI: 1.14-1.2). We only found significant differences in the number of were no differences in baseline characteristics between the groups. Nine paired
euploid embryos when results from D3-biopsies were considered (0.94 vs. 1.18; biopsies in the mifepristone group and eight paired biopsies in the comparator
p=0.0084 in endometriosis vs. control group). Although no significant differences group were included in the histological analysis after study drop-outs, loss of
were shown, a similar trend was observed when ploidy rates from D3 embryos samples and failure in endometrial retrieval. All endometrial biopsies at baseline
were analyzed (euploid rates: 0.18 vs. 0.21 in endometriosis vs. control group; revealed a normal histological appearance. Endometrial biopsies after three
p= 0.08). In D3-biopsied embryos, we also analyzed the ability of blastocyst months with the LNG-IUS in situ showed no PAEC in either group and as
formation and its relationship with their ploidy status. Similar frequencies of expected, they were consistent with progestin effects using the LNG-IUS. While
embryo arrest before reaching the blastocyst stage were found in both groups PAEC are now well described, considered benign, and seem reversible after
(20.13 % vs. 22.66 % in endometriosis and control group; p=0.09). Focusing on cessation of treatment, the mechanism of their development and their long-term
the PGT-A results, data showed no differences in the ploidy status when arrested significance remain unknown. This constitutes the reason for the clinical recom-
embryos from both patients’ groups were compared (OR: 1.12; (0.51-2.02)). mendation of treatment cessation after three months of continuous PRM use
Limitations, reasons for caution: Although the study groups were as homo- for medical treatment of uterine fibroids and endometrial shedding after PRM
geneous as possible, results should be taken into caution due to the retrospective treatment to ensure that PAEC disappears before commencing any long-acting
nature of the study. Multivariate regression models to avoid confounding vari- contraceptive method. This could though constitute a draw-back in women with
ables are still pending. Differences in ploidies according to the severity of the uterine fibroids or endometriosis. Additionally, treatment gaps in fertile women
disease have not been evaluated. can result in unplanned pregnancies.
Wider implications of the findings: Further research is needed to firmly Limitations, reasons for caution: Since this is a pilot study and no prior
convey no association between endometriosis and embryo aneuploidy rates. similar studies have been conducted, no specific power calculation was per-
Prospective and well-designed studies stratifying PGT-A of women with different formed for the secondary outcome reported. Even though the sample size was
stages of endometriosis are required. Moreover, association between aneuploidy small, these data still provide us with unique findings concerning the safety of
and embryo developmental arrest should be considered in the analysis. endometrial morphology following this treatment regimen.
Trial registration number: Not Applicable Wider implications of the findings: Our results suggest that it is safe to
proceed with a long-acting progestin such as the LNG-IUS immediately after
P-290 Effects of the levonorgestrel intrauterine system on long-term PRM treatment for uterine fibroids or endometriosis. However, the
the endometrium after long-term exposure to mifepristone: endometrial safety with this or similar treatment regimens needs to be further
secondary outcomes of a randomized controlled trial explored in larger data settings.
Trial registration number: ClinicalTrials.gov NCT01931657
K. Papaikonomou1, F. Caroline1, A.R. Williams2, K. Gemzell
Danielsson1
1 P-291 The impact of chronic endometritis on endometrial
Karolinska Institutet and Karolinska University Hospital, Department of Women’s
receptivity array and pregnancy rates
and Children’s Health- Division of Obstetrics and Gynecology, Stockholm, Sweden ;
2
The Royal Infirmary of Edinburgh- The University of Edinburgh, Division of K. Kuroda1, T. Horikawa1, A. Moriyama1, K. Nakao1, H. Juen1,
Pathology, Edinburgh, United Kingdom S. Takamizawa1, Y. Ojiro1, K. Nakagawa1, R. Sugiyama1
1
Sugiyama Clinic Shinjuku, Division of Reproductive Medicine and Implantation
Study question: Is direct placement of a levonorgestrel intrauterine system Research, Tokyo, Japan
(LNG-IUS), after long-term treatment with the progesterone receptor modu-
lator (PRM) mifepristone, a safe alternative regarding the endometrium? Study question: To identify the effects of chronic endometritis (CE) on endo-
Summary answer: LNG-IUS placed immediately following long-term treat- metrial receptivity array (ERA) outcomes.
ment with a PRM without any prior endometrial shedding may represent a safe Summary answer: Most patients with CE have ‘non-receptive’ endometrium
treatment regimen for the endometrium. in the ERA test.
What is known already: Clinical trials on long-term administration of mife- What is known already: ERA is a novel tool for the objective identification
pristone have demonstrated its potential use for contraception, medical treat- of the personal window of implantation (WOI). CE is a chronic localized inflam-
ment of uterine fibroids, endometriosis, improvement of bleeding patterns in matory condition of the endometrium with the CD138-positive plasma cells,
women using progesterone only contraceptives and optimization of in vitro yet essentially asymptomatic and undetectable by common infertility testing.
fertilization treatment. Ulipristal acetate, another PRM, is licensed for treatment Recovery from CE can lead to the improvement of pregnancy outcomes in IVF
of uterine fibroids and more PRMs are under development. However, continuous treatment. CE is associated with decreased expression of decidual markers.
administration of PRMs is associated with a risk of developing a characteristic Therefore, CE may have adverse effects on endometrium decidualization and
endometrial morphology described as progesterone receptor modulator asso- formation of an optimal WOI; however, the clinical impact of CE on the ERA
ciated endometrial changes (PAEC). Although accumulating evidence implies test has not been established.
that this histological entity is benign, its long-term safety profile remains uncertain. Study design, size, duration: A retrospective cross-sectional study was
Study design, size, duration: We report the secondary outcome from a performed between August 2018 and October 2019 on 66 infertile women who
double-blinded randomized controlled trial conducted from November 2009 underwent histological examinations and endometrial sampling for ERA testing.
to January 2015. Eligible women were randomized to 50mg mifepristone (n=29) The study protocol was approved by the Ethics Committee of Sugiyama Clinic.
or the comparator (vitamin B) (n=29) for two months, followed by insertion Participants/materials, setting, methods: Of 66 women, we recruited 56
of the LNG-IUS 52mg. Endometrial biopsies were obtained at baseline and who underwent ERA testing and immunohistochemistry (IHC) of CD138 within
three months after placement of the LNG-IUS, with the device in situ. Paired 3 months. We divided the subjects into 3 groups as follows: women without CE

(non-CE group, n=26), women with untreated CE at the time of ERA (CE group, Study design, size, duration: This study was a basic genomic analysis of
n=12), and women successfully treated for CE before ERA (cured-CE group, human endometrial biopsies taken from eight healthy fertile women in the secre-
n=18). CE diagnosis was defined as ≥5 CD138-positive cells per 10 nonover- tory phase.
lapping random stromal areas visualized at 400 ×magnification. Participants/materials, setting, methods: PBF containing a mixture of
Main results and the role of chance: In the non-CE, CE, and cured-CE phosal 50PG, glycerol and deionized water was prepared using a probe sonicator.
groups, the number of CD138-positive plasma cells per 10 random stromal In vitro model, human endometrial cell were into four groups. The control group
areas within 3 months before or after ERA testing were 0.5 ± 0.9, 29.9 ± 31.3, was incubated only with DMEM medium containing antibiotics and 10% FBS and
and 0.9 ± 1.0, respectively. The difference in number of CD 138-positive cells the experimental groups co-cultured with sperm, co-cultured with sperm and
between each group was found to be statistically significant (p < 0.001). clomiphene citrate, co-cultured with sperm and nano-clomiphene citrate were
Regarding ERA results, the rate of ‘receptive’ endometrium in the non-CE and incubated for 24 h and the next day, cells were harvested for q-PCR.
cured-CE groups were 57.7% (15/26) and 55.6% (10/18), respectively; however, Main results and the role of chance: The optimized PBF contained Phosal
in the CE group, the ‘receptive’ endometrium rate was 8.3% (1/12), which was 50PG/glycerol in a 2:8 ratios (w/w) and mean particle size of Nano drug used
significantly lower than the other 2 groups (p < 0.001). In the CE group, ‘non-re- were 67±9.0 nm, the release of CC from Nano emulsion was slightly faster in
ceptive’ endometrium was observed in most patients, including ‘late or post-re- the first 24 hours; during this period, 29% of CC was released. After 120 hours
ceptive’ in 58.4% of patients, on 5 days after the initiation of progesterone 76% of CC was released. The mean mRNA level of MUC1 gene was significantly
administration. When CE was recognized, it was treated with antibiotic therapy increased in CC and CC / PBF group compared to control (P <0.001). The
prior to ET in all patients. The clinical pregnancy rates in the first ET at the time mean mRNA level of PGR gene was significantly increased in the sperm-treated
designated by ERA results in non-CE, CE, and cured-CE groups were 66.7% group (P <0.05) and in the sperm and PBF group compared to CC (P <0.001).
(14/21), 16.7% (2/12), and 50.0% (8/16), respectively (p = 0.022). Despite The mean mRNA level of VEGF gene in sperm and PBF group was significantly
completion of CE treatment, poor pregnancy rates were observed in the increased (P <0.05) compared to control and CC / PBF was significantly
CE group. increased (P <0.001) compared to CC and control.
Limitations, reasons for caution: In limitations, a retrospective study and Limitations, reasons for caution: The main limitation of this study is a low
the small sample size are the main sources of bias. number of human endometrial samples
Wider implications of the findings: WOI may be negatively affected by Wider implications of the findings: Formulation sustained release of clo-
CE. And the patients may achieve optimal WOI after proper CE treatment. miphene citrate increased its targeting efficiency and improved the impact of
Therefore, screening and appropriate treatment for CE should be considered the CC on the serum levels of estradiol and expression of genes involve implan-
prior to ERA testing. Moreover, ERA outcomes in patients with CE may be tation. A new Phosal-Based Formulation (PBF) was introduced to decrease side
unreliable. effect of clomiphene citrate on endometrium.
Trial registration number: not applicable Trial registration number: 6736
P-292 Design and evaluation of a novel nanodrug delivery P-293 Use of a Vaginal Probiotic Suppository and Antibiotics in
system for reducing the side effects of clomiphene citrate on the the Treatment of endometrial microbiota.
endometrium D. Kadogami1, Y. Nakaoka1, Y. Morimoto2
M. Ajdary1, R. Aflatoonian2, F. Keyhanfar3, F. Amjadi4, Z. Zandieh , 1
IVF Namba Clinic, medical office, Osaka, Japan ;
zandie.dvm@gmail.com4, M. Ashrafi , ashrafi.m@iums.ac.ir5, M. 2
HORAC Grandfront Osaka Clinic, medical office, Osaka, Japan
Mehdizadeh , mehdizadeh.m@iums.ac.ir6
1
Cellular and Molecular Research Center- Iran University of Medical Sciences- Study question: What is an effective treatment for endometrial microbiota?
Tehran- Iran, Cellular and Molecular Research Center- Iran University of Medical Summary answer: The combination use of vaginal probiotic suppository and
Sciences, Tehran, Iran ; antibiotics was effective for the treatment of endometrial microbiota.
2
2.Department of Endocrinology and Female Infertility- Reproductive Biomedicine What is known already: The development of next-generation sequencing (NGS)
Research Center- Royan Institute for Reproductive Biomedicine- Tehran- Iran, 2. technologies has made it possible to quantify subdivided bacteria using the variable
Department of Endocrinology and Female Infertility- Reproductive Biomedicine region of the 16S rRNA gene, enabling a more comprehensive assessment of the
Research Center-, ; microbiota. Through this innovation, various studies have focused on the relation-
3
Pharmacology Department- Tehran University of Medical Sciences- Hemmat ships between the female genital microbiota and pregnancy. Some authors evaluated
Pardis- Tehran- Iran, Pharmacology Department- Tehran University of Medical the endometrial microbiota of IVF patients using NGS and showed that the group
Sciences- Hemmat Pardis- Tehran- Iran, tehran, Iran ; with 90% or higher Lactobacillus(LD) had significantly higher implantation rates and
4
Department of Anatomical Science- Iran University of Medical Sciences- Tehran- pregnancy rates than those with less than 90% Lactobacillus(NLD) . Therefore, new
Iran, Department of Anatomical Science- Iran University of Medical Sciences- strategies are needed to improve pregnancy rates in patients with NLD.
Tehran- Iran, tehran, Iran ; Study design, size, duration: Randomized controlled trial in our clinic,
5
Obstetrics and Gynecology Department- School of Medicine- Iran University of between December 2018 to October 2019.
Medical Science- Tehran- Iran, Obstetrics and Gynecology Department- School of Evaluation of the endometrial microbiota was performed in 224 patients with
Medicine- Iran University of Medical Science- Tehran- Iran, tehran, Iran ; RIF. NLD cases were randomly assigned to treatment groups that combine
6
Cellular and Molecular Research Center- Faculty of Advanced Technologies in Lactoferrin(oral prebiotic), Lebenin(oral probiotic), inVag(vaginal probiotic sup-
Medicine- Department of Anatomy- Iran University of Medical Sciences- Tehran- pository) and Metronidazole in various ways. Re-analysis was performed after
Iran, Cellular and Molecular Research Center- Faculty of Advanced Technologies in treatments and results were presented as healing rates. The healing rate was
Medicine- Depart, defined as the rate of cured cases among treated cases.
Participants/materials, setting, methods: The treatment protocol was
Study question: Is there a change in the expression of genes involved in determined as follows. Protocol A was Lactoferrin(300mg/day for 30days) and
implantation in cells treated with clomiphene citrate compared to clomiphene Lebenin(3g/day for 30days), B was Metronidazole(vaginal suppository 250mg,
citrate loaded in PBF? only once and oral 750mg/day for 7days), C combined A and B, D was
Summary answer: A significant difference was found between genes involved Lactoferrin(600mg/day for 30days) and Lebenin(3g/day for 30days) and
in the implantation of cells treated with clomiphene citrate compared to clomi- Metronidazole (vaginal suppository 250mg and oral 750mg/day for 7days),
phene citrate loaded in PBF. E was inVag(1capsule/day for 7days), F was inVag (1capsule/day for 7days) and
What is known already: Growing evidence suggests that a disorder of endo- Metronidazole(vaginal suppository 250mg and oral 750mg/day for 7days).
metrial receptivity may contribute to the adverse reproductive outcomes in the Main results and the role of chance: Evaluation of the endometrial microbiota
stimulation of ovulation by clomiphene citrate (PCOs). And by developing a was performed in 224 patients with RIF. LD was found in 114 patients (50.9%),
slow-release property in lipophilic drugs, Phospholipids can prevent the damage whereas NLD was found in 110 patients (49.1%). There were no significant dif-
of lipophilic drugs to non-target tissues. ferences in patient backgrounds between groups. In NLD cases, Gardnerella was

the most abundant bacteria, and Atopobium, Streptococcus, and Prevotella were P-295 Sonographic Quantification of Myometrial Conditions in
detected abundantly, after excluding Lactobacillus and Bifidobacterium. Adenomyosis Patients with Computer Assisted Image Analysis-A
About protocol A, B and C, the healing rate was only 30%, and there were no Pilot Study
significant differences in healing rate between treatment groups (P = 0.91 and S.Y. Chou1, L. Buo Jia1, C. Yi-En1, C. Cindy1, C. Ching Hui1,
P = 0.60, respectively). Especially in A, there were no cured cases among patients C. Chi-Huang1
those Lactobacillus proportion was less than 70% before treatment. These results 1
Taipei Medical University Hospital, Reproductive medicine and science, Taipei,
suggested that oral prebiotics and probiotics alone were not sufficient to improve Taiwan R.O.C.
the endometrial microbiota and that antibiotics were necessary. Between C and
D, there were no significant difference in healing rate (33.3% versus 32.5%, Study question: We examined the diagnostic feasibility of using artificial neural
respectively, P = 1.00). network (ANN)-analyzed sonographic gray-scale histograms to assess myome-
However, higher healing rate was reported in E compared with D(32.5% trial conditions based on serum cancer antigen 125 (Ca-125) levels in patients
versus 43.6%, respectively, P = 0.39). Further, F had a significantly higher healing with adenomyosis.
rate compared with D(32.5% versus 72.7%, respectively, P = 0.02). Summary answer: By analyzing the gray scale histograms of the myometrium
Throughout this study, no side effects that were likely related to the medication on ultrasonographic images, this quantitative approach may help in the initial
were observed. diagnosis of adenomyosis.
Limitations, reasons for caution: Assessment of the endometrial microbiota What is known already: The only definitive method for the diagnosis
requires transvaginal collection of endometrial fluid using a catheter tube, which of adenomyosis is to perform a hysterectomy combined with histological
may cause contamination. Moreover, although NGS is useful for quantifying examination. However, there is no universal agreement on the exact histo-
bacteria, it only measures the bacterial 16S rRNA gene and does not guarantee logical criteria, and the diagnostic methodcannot be made prior to invasive
bacterial viability. surgery.
Wider implications of the findings: The endometrial microbiota was eval- The accuracy of using only a serum Ca-125 assay to diagnose adenomyosis
uated in patients with RIF, and 49.1% of cases had less than 90% Lactobacillus . remains limited, Ca-125 can help in the initial screenings of women with possible
In such cases, probiotics combined with vaginal probiotic suppository and anti- adenomyosis
biotics proved to be highly effective. In our future studies, we will examine Ultrasonography technology is a quick, safe, easy to use, and inexpensive modal-
pregnancy rates after treatment of the endometrial microbiota. ity that makes it possible to recognize adenomyosis without surgery. However,
Trial registration number: UMIN-CTR　000038582 there are no standard diagnostic imaging criteria for adenomyosis.
Study design, size, duration: This was a retrospective observational study
P-294 Surgical management of deep endometriosis and in vitro of 26 female patients attending Taipei Medical University Hospital, Taiwan for
fertilization outcomes: A systematic review and meta-analysis infertility between September 1, 2018 and March 31, 2019.The research protocol
B. Woolnough1, C. Qianqian Wu1, T. Vause1, H. Shenassa1 of this study was approved by the Institutional Review Board of Taipei Medical
1
University of Ottawa, Obstetrics and Gynecology, Ottawa, Canada University Hospital (IRB number: N201711084).
Participants/materials, setting, methods: A total 54 ultrasonographic
Study question: In women with deep endometriosis (DE), does surgery fol- images of the myometrium were divided into three groups based on serum
lowed by in vitro fertilization (IVF) improve fertility outcomes compared to Ca-125 levels:15 images from patients with Ca-125 values <35 U/ml, 19 images
first-line IVF? from patients with Ca-125 values between 35 and 100 U/ml, and 20 images
Summary answer: Considering limited evidence, surgical management of DE from patients with Ca-125 values >100 U/ml. Each patient with the same
before IVF is associated with improved pregnancy rates but similar live birth machine and settings, and sonographic gray-scale histograms of the endome-
rates when compared with first-line IVF. trium were analyzed with ImageJ image processing software using the
What is known already: Surgical management of DE in infertile women prior ANN model.
to in vitro fertilization (IVF) is controversial. Surgeries are often invasive and may Main results and the role of chance: ANN classification was used to char-
cause significant complications. Recent evidence suggests that surgery may acterize the sonographic data. Although the median signal intensity score of the
improve IVF outcomes, however studies have been small and underpowered. Ca-125>100 group (2333811) was higher than that of the Ca-125<35 group
Study design, size, duration: We conducted a systematic review and (1652460) and the 35≤Ca-125≤100 group (1915953), Kruska-Wallis statistical
meta-analysis, which identified studies including a total of 538 women. We analyses indicated no significant difference among groups (p=0.55).
searched MEDLINE, EMBASE and the Cochrane Library (inception to October The Youden index is a measure for evaluating the effectiveness of a biomarker.
2019) for studies comparing fertility outcomes in women with DE with and The cut-points of signal intensity scores among the three groups according to
without surgical treatment prior to IVF. the Youden index and their sensitivity and specificity in predicting adenomyosis
Participants/materials, setting, methods: The study population included probability are shown. For the comparison between Ca-125<35 and 35≤Ca-
all identified women with deep endometriosis who received treatment of either 125≤100, the sensitivity and specificity were 36.8% and 86.7%, respectively; for
surgery followed by IVF or IVF alone. Study selection, data extraction and quality the comparison between Ca-125<35 and Ca-125>100, the sensitivity and spec-
assessment were conducted independently by 2 reviewers. Meta-analysis was ificity were 40% and 93.3%, respectively; for the comparison between 35≤Ca-
conducted using a random effects model. 125≤100 and Ca-125>100, the sensitivity and specificity were 35% and 84.2%,
Main results and the role of chance: Four studies meeting the inclusion respectively.
criteria were identified, one prospective and three retrospective cohort studies. This quantitative approach had a high specificity, which might be useful in the
Three of these were able to be included in the quantitative meta-analysis. Clinical early diagnosis of adenomyosis.
pregnancy rate was significantly higher in the surgery group (risk ratio [RR] = Limitations, reasons for caution: This study was highly operator-dependent
1.41; 95% CI = 1.03 to 1.95; P = 0.03; I2 = 25%). Live birth rates were compa- and did not detract from the observational experience required in ultrasono-
rable between the groups (RR = 1.60; 95% CI = 0.97 to 2.62; P = 0.06; I2 = graphic examinations.
48%). The total number of oocytes retrieved, gonadotropin dose used, and The diagnosis was made by ultrasonographic examination, with no histopatho-
miscarriage rates were also similar between the two groups. logical confirmation, which may limit the accuracy of our findings.
Limitations, reasons for caution: The retrospective nature of the included The small sample size and irregular distribution of baseline characteristics may
studies permits introduction of bias, limiting the conclusions that can be made. have influenced the results.
Wider implications of the findings: The potential benefit of surgery must Wider implications of the findings: In our study, not only the size and
be weighed against risk, including potential reduction in ovarian reserve. While volume of the uterus but also its contents and textures were accessed quanti-
this study suggests similar outcomes, well-designed and appropriately-powered tatively by analyzing the gray-scale histogram of the myometrium. Our approach
randomized controlled trials are needed to determine if there is any clinical is of great assistance in the initial screening of women with suspected
advantage to surgical management of DE prior to IVF. adenomyosis.
Trial registration number: not applicable Trial registration number: N201711084

P-296 3D-Saline Contrast Sonohysterography in detecting uterine (N=40) or recurrent miscarriage (RM) (N=40) originating from natural concep-
anomalies and pathologies among sub-fertile women and women tion, without any other infertility aetiology. Following diagnosis of CE, patients
with AUB were randomized into two groups, one receiving oral treatment and the other
D. Al-Jaroudi1, F. Khan1, M. Al-Tannir2, I. Al Fayyad2 receiving a combination of oral antibiotics and intrauterine infusion, stratified by
1 the referral diagnosis. Enrolment of patients was performed from March 2017
King Fahad Medical City, Women’s Specialized Hospital, Riyadh, Saudi Arabia ;
2 till February 2019.
King Fahad Medical City, Research center, Riyadh, Saudi Arabia
Participants/materials, setting, methods: Diagnosis of CE and treatment
Study question: Does 3D-SCSH detect both congenital uterine anomalies evaluation were performed by employing hysteroscopic investigation, endome-
and uterine pathologies in sub-fertile and AUB patients? trial biopsy, along with histological analysis and microbiological culture. Patients
Summary answer: 3D-SCSH detects both congenital uterine anomalies and following successful treatment proceeded with pursuing a pregnancy via natural
uterine pathologies in sub-fertile and AUB patients and is a tool to be used when conception or IVF, according to the original referral diagnosis. Patients presenting
suspecting uterine pathology. with RIF proceeded with a single IVF cycle. Patients presenting with RM were
What is known already: Uterine cavity anomalies could be a contributing invited to conceive naturally over the course of six months. The study was
cause of subfertility in women, and abnormal uterine findings are reported in as conducted in a single IVF center.
many as 50% of women with recurrent implantation failure. Main results and the role of chance: The combined oral and intrauterine
Study design, size, duration: A prospective cohort study was conducted administration of antibiotics provided statistically significant improved results
over three years. Total number of 3D-TVS performed was 2,222. 1477 women regarding efficiency of treatment when compared to oral antibiotic administration
were from the sub-fertile group and 745 patients from AUB group. only (34/40vs25/40; RR:1.36; 95%CI:1.04-1.79; p=0.04). It should be men-
Participants/materials, setting, methods: A prospective cohort study was tioned that the number of treatment days was also statistically significantly
conducted over three years. The women were recruited from the outpatient increased (29.93vs15.00; p<0.001) in the combination group. Regarding the IVF
department and Reproductive Endocrinology and Infertility Medicine arm no statistically significant difference was indicated between the combination
Department. All women of reproductive age group, either with abnormal uterine group (13/18) and the oral administration (9/13) regarding neither clinical preg-
bleeding (AUB) or sub-fertile underwent 3D-TVS. If initial 3D-TVS detected any nancy (CP) (13/18vs9/13; RR:1.04; 95%CI:0.66-1.66; p=0.85) nor live-birth
uterine abnormality, then the patient was booked for 3D-SCSH. Uterine anom- (LB) (13/18vs8/13; RR:1.17; 95%CI:0.70-1.96; p=0.70). Intention-to-treat (ITT)
alies were recorded according to the new ESHRE/ESGE classification of uterine analysis did not alter the results regarding neither CP (RR:1.18; 95%CI:0.71-1.97;
anomalies and compared in the two groups. p=0.75) nor LB (RR:1.63; 95%CI:0.87-3.04; p=0.20). Regarding the natural con-
Main results and the role of chance: Results: Total number of 3D-TVS ception arm no statistically significant difference was presented between the
performed was 2,222. 1477 women were from the sub-fertile group and 745 combination group (11/17) and the oral administration group (7/12) regarding
patients from AUB group. Total of 330 women required 3D-SCSH, of whom neither CP (11/17vs7/12; RR:1.12; 95%CI:0.66-1.66; p=0.85) nor LB
215 (65%) were sub-fertile women, and 115 (35%) were AUB. Uterine anomalies (10/17vs7/12; RR:1.00; 95%CI:0.54-1.87; p=0.98). ITT did not alter the results
were found in 15 (7%) in the sub-fertile group compared to 2 (1.7%) In the AUB regarding neither CP (RR:1.49; 95%CI: 0.80-2.78; p=0.34) nor LB (RR:1.35;
group. The partial septate uterus was the most common finding among the 95%CI 0.73-2.50; p=0.52). Pooling of data presented a marginally statistically
sub-fertile group 7 (3.3%) versus 2 (1.7%) with dysmorphic uterus in the AUB significant improvement in the combination group compared to oral administra-
group. Uterine polyps were the commonest uterine pathology in both groups. tion regarding both CP (25/40vs16/40; RR:1.56; 95%CI:1.00-2.45; p=0.05),
Conclusion: 3D-SCSH may detect congenital uterine anomalies in sub-fertile and and LB (24/40vs15/40; RR:1.58; 95%CI:1.00-2.05;p=0.05).
AUB patients. This diagnostic accuracy may support the adoption of 3D-SCSH as Limitations, reasons for caution: The small sample size along with the fact
a routine investigation in the evaluation of sub-fertile women prior to IVF. that the study was carried out in a single center are limitations for the study.
Limitations, reasons for caution: NA Moreover, since CE is regarded as a Sexually Transmitted Disease the lack of
Wider implications of the findings: The prevalence of uterine anomalies follow-up regarding the male partner stands as a reason for caution.
among sub-fertile women was found to be higher in comparison to women with Wider implications of the findings: Results indicate that intrauterine anti-
AUB. 3D-TVS and 3D-SCSH are accurate, and safe in determining the prevalence biotic infusion may hold the key to a long-term effective treatment, extending
of congenital uterine anomalies in sub-fertile women. to establishing a favorable basis for achieving pregnancy for patients diagnosed
Trial registration number: NA with CE and subsequent infertility. This study establishes preliminary grounds
for conducting research focused on a pharmaceutically developed drug appro-
priate for endometrial administration.
P-297 Introducing intrauterine antibiotic infusion as a means Trial registration number: Not applicable
towards eradicating chronic endometritis and restoring
reproductive dynamics: A randomized pilot study P-298 Expression of claudin 3 and caludin 4 in deep endometriosis
K. Sfakianoudis1, M. Simopoulou2, E. Maziotis2, A. Rapani2, M. Mendonça Carneiro1,2,3, P. Vidigal4, L.P. Costa3, C. Moreira1,
P. Tzonis1, N. Nitsos1, P. Giannelou2, S. Grigoriadis2, P. Tsioulou2, T. Moreira1, V. Campos1, P. Gouvea3, T. Pardini1, A. Silva Filho1,
M. Koutsilieris2, K. Pantos1 I. Ávila3
1
1
Genesis, Assisted Conception Unit, Chalandri- Athens, Greece ; Universidade Federal de Minas Gerais, Obstetrics and Gynecology, Belo Horizonte,
2
National and Kapodistrian University of Athens, Physiology, Athens, Greece Brazil ;
2
Rede Mater Dei de Saúde, Center for Human Reproduction, Belo Horizonte,
Study question: Does intrauterine antibiotic infusion in combination with per Brazil ;
os administration in patients with Chronic Endometritis (CE) enhance treatment 3
Biocor Instituto, Endometriosis Multidisciplinary team, Belo Horizonte, Brazil ;
and fertility status? 4
Universidade Federal de Minas Gerais, Pathology, Belo Horizonte, Brazil
Summary answer: The combination of oral administration and intrauterine
infusion of antibiotics provided enhanced treatment results when compared to Study question: To evaluate the association between immunoexpression of
oral antibiotic administration for CE patients. claudin 3(CLND-3) and 4 (CLDN-4) with clinical, surgical and biochemical data
What is known already: An infectious endometrial environment caused by in women with deep endometriosis (DE).
certain pathologies could compromise pregnancy rates for patients pursuing Summary answer: DE expresses both CLDN-3 and CLDN-4. Expression
fertility treatment. In the case of persistent endometrial inflammation, a diagnosis was weak and not influenced by clinical parameters. CLDN-3 expression was
of CE may be established. Oral antibiotic administration constitutes the gold significantly higher and related to disease extension.
standard approach for treating CE cases and has been confirmed to contribute What is known already: DE is characterized by lesions which extend more
to a clearance rate, albeit without adequate treatment rate. than 5 mm underneath the peritoneum and is responsible for painful symptoms.
Study design, size, duration: Women included in the present study were Numerous molecular abnormalities have been related to DE pathogenesis includ-
referred for CE investigation following either repeated implantation failure (RIF) ing immunological, inflammatory, genetic and hormonal alterations. Claudins are

the main junctional proteins between epithelial cells and may be may play a role Participants/materials, setting, methods: Endometrial and blood profiling
in tissue remodeling and invasion. CLND-3 and CLDN-4 have been studied in was carried out in all participants in a hormone substituted cycle using estradiol
endometrial hyperplasia and cancer with growing evidence showing a possible and progesterone and sampled after five days of progesterone treatment.
contribution in their pathogenesis and thus prompted studies to assess their Endometrial biopsies were analysed by histology, immune cell profiling (CD56,
possible role in endometriosis. CLDN3 and 4 have not been studied in DE so far. CD16, CD163, CD138, regulatory T cells), and gene expression (ERA® test,
Study design, size, duration: women undergoing surgical treatment of DE Igenomix, Spain). The vaginal microbiome was analysed using a NGS technology
at a tertiary center (Biocor) with confirmed histological diagnosis from (ArtPRED®, The Netherlands). Blood tests included estradiol, progesterone,
August/2011 to Decemer/2016 comprised the study group whereas the control prolactin, TSH, anti-phospholipid screening, and vitamin D.
group included women undergoing surgery for other benign gynecological con- Main results and the role of chance: Patients with RIF demonstrated dif-
ditions (pelvic organ prolapse or uterine fibroids) who gave their consent to ferent individual endometrial profiles, indicating that a ‘one treatment fits all’
participate. The study protocol was approved by the local ethics committee. approach is inappropriate. Compared with controls, women with RIF demon-
Participants/materials, setting, methods: Samples were otained during strated a higher prevalence of chronic endometritis as defined by cells staining
surgery in both groups and prepared for IHC using antibodies for both CLDN-3 positive for CD138 by immunohistochemistry (25% vs. 3% in the control cohort,
and CLDN-4. IHC expression was considered present (+) or absent (-) and p = 0.004), a lower mean level of vitamin D (43 nmol/L vs. 51 nmol/L;
intensity classified into three categories: weak (+), moderate (++) and strong P = 0.04), and a borderline lower mean mid-luteal phase progesterone (39
(+++ and ++++). IHC was performed with the Streptavidin-Biotin protocol nmol/L vs. 44 nmol/L; P = 0.08). Compared with controls, women with RIF
using antibodies against CLDN3 and CLDN4. The data was analysed using the appeared to have a higher prevalence of a vaginal microbiome reported to be
Z test for proportions, Fisher’s exact chi-square test and an ordered logistic predictive of implantation (P = 0.04). While the RIF cohort was slightly older
regression. than the controls (mean age 33.8 years vs. 30.2 years P = 0.0001) no differences
Main results and the role of chance: This case-control study involved between the two groups were observed with regard to AMH, BMI, anti-phos-
immunohistochemical evaluation (IHC) of 48 samples from 36 patients under- pholipid antibody prevalence, mean mid-luteal estradiol, prolactin, TSH levels,
going surgical endometriosis treatment at a tertiary center by a multidiscplinary number of CD56 cells, CD16 cells, regulatory T cells or macrophages present
endometrisis team.Thirty-five (73%) subjects had DE diagnosis. CLDN-3 expres- in the mid-luteal endometrial biopsy.
sion was positive in 60% of DE and 80% of CLDN-4 samples. Expression was Limitations, reasons for caution: The tests performed in the study were
higher in the intestines (CLDN-3: p 0.016; CLDN-4: p 0.000). In other sites a taken in a standardised hormone substituted cycle and results should be inter-
weak intensity expression predominated, both from CLDN-3 (75.4%) and preted in that clinical context. The control cohort consisted of women referred
CLDN-4 (73.8%). Expressions of CLDN-3 and CLDN-4 were not associated for fertility treatment with male factor infertility or tubal factor, hence not rep-
with preoperative hormone use, body mass index (BMI), vitamin D, and serum resenting a proven fertile population.
125 (CA125) antigen. CLDN-3 showed more positivity in the more advanced Wider implications of the findings: Diagnosing the endometrium in women
stages of endometriosis (moderate and severe). CLDN-3 expression was sig- with RIF permits targeted rather than blind empirical interventions. Relative
nificantly higher than in controls (p < 0.001) Vitamin D deficiency, lower mid-luteal progesterone, and chronic endometritis
Limitations, reasons for caution: Although DE lesions expressed both are ready targets for treatment. Understanding the role and treatment of an
CLDN 3 and 4, the sample is small and comes form a single center. Claudin 3 unfavourable vaginal microbiome needs further investigation.
and 4 expression was studied only by IHC. Trial registration number: REG-117-2017
Wider implications of the findings: CLDN-3 expression was significantly
higher than in controls and showed more positivity in the more advanced stages
P-300 Differences in the effects of metformin treatment on
of endometriosis (moderate and severe). The role of CLDN3 in DE remains to
viability and migration of endometrial cancer cells in a normal or
be established.
high glucose environment
Trial registration number: NOT APPLICABLE
C. Lange1, A. Machado Weber1, T. Strowitzki1, A. Germeyer1
1
University Women’s Hospital Heidelberg, Department of Gynecological Endocrinology
P-299 Assessing the endometrium in recurrent implantation and Fertility Disorders, Heidelberg, Germany
failure (RIF) – a prospective controlled cohort study
M. Hviid1, T. Hallager2, G. Persson3, L. Grupe2, K. Birch Petersen4, Study question: Does metformin treatment have effects on cell viability and
T.V. Hviid3, N.S. Macklon4 migration of endometrial cancer cells and do these effects vary in a normal or
1
Zealand University Hospital, Department of Gynaecology- Obstetrics- and high glucose environment?
Fertility, Roskilde, Denmark ; Summary answer: Metformin decreased cellular viability and migration in
2
Zealand University Hospital, Pathology Department, Roskilde, Denmark ; endometrial cancer cell lines. However, variation of glucose levels had no effect
3
Zealand University Hospital, Department of Clinical Biochemistry, Roskilde, on cellular viability and migration.
Denmark ; What is known already: Endometrial cancer (EC) is one of the most common
4
Zealand University Hospital, Department of Gynaecology- Obstetrics and Fertility, gynecological malignancies and can occur in an estrogen-dependent (type I,
Roskilde, Denmark 70 – 80 %) or a more aggressive estrogen-independent form (type II). High
estrogen levels, e.g. due to metabolic syndrome and associated polycystic ovary
Study question: What markers of endometrial dysfunction can be identified syndrome (PCOS), or hyperglycemia, e.g. in due to diabetes, increase the risk
among women experiencing RIF and do they differ compared to control patients? of developing EC and lead to higher mortality, especially in obese women.
Summary answer: Women with RIF reveal a range of endometrial dysfunc- Metformin, an anti-hyperglycemic and insulin sensitizing drug, is used in the
tions different in prevalence from control patients. treatment of type II diabetes mellitus. However, anti-cancer effects have been
What is known already: The endometrial factor is recognized as a cause of observed in several studies, but detailed information is lacking for EC.
RIF. However, management remains largely empirical and of questionable effi- Study design, size, duration: In the present in vitro study, EC cells were
cacy. There is a need to guide therapy by making an endometrial diagnosis where cultured in normal (NG; 5.5 mmol/L, equivalent to 100 mg/dL) or high (HG;
possible. 17.0 mmol/L, equivalent to 306 mg/dL) glucose media (supplemented with 10
Study design, size, duration: Prospective controlled cohort study. Between nmol/L ß-estradiol) and treated with metformin (0.1–20.0 mmol/L) over a
November 2017 – September 2019, 86 women with a history of RIF (defined period of 7 d. Cells were subsequently analyzed for changes in cell viability and
as failure to conceive after at least three transfers of high quality embryos), migration rates. Untreated cells served as controls. Data was obtained from at
referred to a University Hospital-based ‘Implantation Clinic’, and 37 women least three independent experiments.
with no history of RIF underwent timed endometrial diagnostic profiling in a Participants/materials, setting, methods: The study was carried out with
hormone substituted cycle. Samples were collected throughout the span the two different EC cell lines HEC-1A (type II) and Ishikawa (type I). The analysis
of a year. of cellular viability was based on mitochondrial dehydrogenase activity in the

MTT assay. The migration of the cells was analyzed by changes in scratch area additioned with 18 mg/ml of nicotine) were all able to down-regulate 1.8-2
in the wound healing assay. folds the HOXa10, mucin-1 and β3 integrin expression when compared to
Main results and the role of chance: A dose-dependent decrease of cellular benzo-a-pyrene, cotinine and nicotine ditartate metabolites. E-cigarette liquids
viability was observed for HEC-1A and Ishikawa cells after metformin treatment, increased the relative expression of 2.7 fold of the pro-apoptotic gene FAS
irrespective of glucose levels. At a concentration ≤0.1 mmol/L, viability remained if compared to the induction generated by nicotine metabolites. E-cig liquids
≥80% compared to untreated controls, both in a normal and high glucose envi- treatments resulted in higher percentage of cell death after 1 hour incubation
ronment. Viability dropped to ≤10% at concentrations >5.0 mmol/L (HEC-1A) (54.4% Vs 31.3%) if compared to nicotine metabolites. After 3 hours of
and 10.0 mmol/L (Ishikawa) under any tested condition. IC50 values for Ishikawa treatments the rate o fcell death was 93.8% for e-cig liquids vs 64.8% for
cells were 5.10 mmol/L (NG; IC90: 44.4 mmol/L) and 3.90 mmol/L (HG; IC90: nicotine metabolites.
33.7 mmol/L). For HEC-1A cells, an IC50 value of 0.75 mmol/L (NG; IC90: 6.83 Limitations, reasons for caution: This study was conducted in vitro. Results
mmol/L) was established. The wound healing assay revealed that the ability to obtained should be possibly confirmed in vivo in the context of the whole human
migrate into the scratch area decreased with increasing metformin concentra- endometrial tissue. Longer incubation to test long-term effects were not possible
tions, but independent of glucose levels. For HEC-1A control cells, 70% (NG) due to high cell death observed after 3 hours exposure.
and 56% (HG) of the scratch area was covered with cells 24 h after wounding Wider implications of the findings: This study demonstrates that e-ciga-
of the cell monolayer. After treatment with 0.5 mmol/L metformin, the wounded rettes liquids have the same or even higher detrimental effect on the receptivity
area was invaded by 53.4% (NG) and 45.4% (HG), respectively, and only by of endometrium than nicotine metabolites.
17.7% (NG) and 20.8% (HG) at 5.0 mmol/L metformin. The same effect was Trial registration number: na
observed with Ishikawa cells, although migration was generally lower. Metformin
at 5.0 mmol/L even led to a small enlargement of the wound area by 2.6% (HG). P-302 Combined approach to treating endometriosis-associated
Limitations, reasons for caution: The results are obtained in an in vitro study infertility
with human cancer cell lines, and thus cannot be easily extrapolated to patients. N. Artymuk1, L. Danilova1, O. Zotova2
Wider implications of the findings: Results suggest that metformin treat- 1
Kemerovo State Medical University, Obstetrics and Gynecology Department,
ment can lower the migration ability of EC cells, especially of the more aggressive Kemerovo, Russia C.I.S. ;
type II. Therefore, metformin may also be able to prevent EC development in 2
Kemerovo Regional Clinical Hospital, Gynecological Department, Kemerovo,
patients at high risk, e.g. with high estrogen levels (metabolic syndrome and Russia C.I.S.
PCOS), hyperglycemia (type II diabetes) or obesity.
Trial registration number: not applicable Study question: To comparatively evaluate the effectiveness and compliance
of a combined approach to treatment of patients with endometriosis-associated
P-301 E-cigarettes smoke impairs endometrial receptivity infertility using gestagen and a gonadotropin-releasing hormone agonist.
A. La Marca1, S. Sacchi1 Summary answer: The use of gestagen and GnRH agonist in the combined
1
University of Modena and Reggio Emilia- and Clinica Eugin, Mother-Infant therapy of patients with endometriosis-associated infertility and pelvic pain has
Department, Modena, Italy similar efficacy, but different compliance.
What is known already: Endometriosis affects approximately 10% of
Study question: Are e-cigarette (e-cig) liquids and nicotine metabolites able reproductive-age women and at least one third of women with infertility.
to alter the receptivity of stromal endometrial cells (ESC)? Prevalence of endometriosis has increased up to 50 % in women with infer-
Summary answer: Different e-cigarette liquids and nicotine metabolites tility. Many studies have now shown that medical treatment for endometriosis
affect ESC receptivity and viability with different efficiency by down- does not improve fertility. Hormonal treatment of endometriosis cannot
regulating implantation marker genes expression and promoting apoptosis improve fertility and, therefore, their use only leads to delayed pregnancy.
of ESC In accordance with the recommendations of the CNGOF-HAS, NICE, ESHRE
What is known already: E-cigarettes are instruments that transform a solution guidelines, suppression of ovarian function should not be prescribed to
of propylene glycol and nicotine in a nicotine-containing aerosol by heating. improve fertility. Nevertheless, a combined approach in the treatment of
Studies show that in e-cigarette not-combustion products a number of potentially endometriosis-associated infertility may have advantages in patients with
toxic substances are present. While diseases in heavy cigarette smokers are well severe pelvic pain.
documented, the effect of e-cigarettes has been less studied. It has been shown Study design, size, duration: This was a randomized, open-label, mono-
that e-cigaretees may induce acute endothelial dysfunction and promote vascular centric trial. The study included 150 patients with histologically confirmed
oxidative stress. Many women are nowadays preferring this way of somoking. endometriosis-associated infertility, by excluding other causes of infertility.
However only few works have analyzed effects of e-cigarette on fertility with Patients from Group 1 (n=75) after surgical treatment (ablation and/or exci-
results that are indicating a possible detrimental effect of these substances on sion) has been administered the GnRH agonist (buserelin or diphereline) at a
fertility dose of 3.75 mg once every 28 days for 3 months. Patients from Group 2
Study design, size, duration: Endometrium sample biopsies were col- (n=75) prescribed gestagen (dienogest) at a dose of 2 mg daily for 6 months
lected during the proliferative phase of the menstrual cycle from patients continuously.
undergoing diagnostic hysteroscopy, dissected in smaller pieces then Participants/materials, setting, methods: All patients (n=150) underwent
digested with collagenase. Primary cultured ESC were cultured until con- laparoscopy and surgical treatment of endometriosis (ablation and / or excision).
fluence. ESC were then treated for 1 and 3 hours with different doses of Almost half of patients with endometriosis-associated infertility from groups I
e-cigs liquids or nicotine metabolites (benzo-a-pyrene, cotinine and nicotine and II (45.3% and 45.3%, respectively) had stages III and IY of the disease. The
ditartrate) predominant localization of endometriosis in infertile patients of both groups
Participants/materials, setting, methods: Primary cultures of endometrial was the pelvic peritoneum (60%), the sacro-uterine ligaments (88%) and the
cells were adopted as experimental model (n=7). Treated ESC were processed ovary (44%). After medication therapy, primary and secondary outcomes were
for total RNA extraction. Gene expression of HOXa10, mucin-1, β3 integrin assessed.
(implantation marker genes) and FAS (pro-apoptotic gene) normalized by β-actin Main results and the role of chance: The combined treatment of endo-
reference gene, was evaluated by RT-qPCR. Reactions were repeated in triplicate. metriosis-associated infertility via surgery, followed by the administration of both
Cells viability was evaluated by Trypan blue exclusion assay. Statistical analysis the GnRH agonist and dienogest, was highly similar effective in reducing the
was performed with Kruskal-Wallis test followed by the Dunn-Bonferroni’s test symptoms of endometriosis, such as pelvic pain - OR 1,95 [1,01- 3,77] (p=0,051),
(P < 0.05). dysmenorrhea - OR 1,14 [0,56-2,29] (p=0,720), dyspareunia - OR 1,55 [0,77-
Main results and the role of chance: All the treatment tested showed 3,10] (p=0,218), and bleeding - OR 1,44 [0,54- 3,81]. The spontaneous preg-
that e-cigarette liquids and nicotine metabolites are able to significantly nancy rate in both groups was similar, respectively, OR - 0.948 [0.49-1.80]
down-regulate the expression of implantation marker genes although with (p = 0.870). The fetal heartbeat registration rate at 12 weeks and live births did
different efficiency. E-cig liquids tested (propylene glycol and propylene glycol not differ statistically significantly between the Groups, however, patients who

were administered aGnRH had a tendency to increase the frequency of non-de- Y. Dogra1, N. Singh1
veloping pregnancy - in 17.3% and 9.3% - OR 2.04 [0, 76-5.43], and a lower 1
All India Institute of Medical Sciences, Division of Reproductive Medicine-
frequency of live births (p = 0.489). Clinically significant side effects were Department of Obs and Gynaecology, New Delhi, India
observed in 92% (69) of patients who were prescribed aGnRH and 16% (12)
of women who were administered dienogest - OR 60.375 [21.387-170.441] (p Study question: Does platelet-rich plasma (PRP) alters the endometrial milieu
<0.001). The most significant side effects in Group I were: hot flashes (82.7% making it receptive to an embryo in women with persistent thin endometrium
(62)), decreased libido (74.7% (56)) and sweating (60.0% (45)), in Group II - due to varied aetiology?
decreased libido (16 % (12)). Summary answer: PRP enhances the endometrial thickness (EMT) significantly
Limitations, reasons for caution: The trial is limited in sample size, in both fresh and FET cycles in thin endometrium associated with tuberculo-
single-center, no registration number. Further extensive randomized multicenter sis(TB), PCOS and Diminished Ovarian Reserve (DOR).
studies are required to confirm the benefits of administering gestagens in What is known already: Autologous PRP increases endometrial thickness
combination therapy for patients with endometriosis-associated infertility and during Frozen thawed Embryo transfer (FET) in women with previous cancelled
pelvic pain. cycles due to refractory thin endometrium despite hormone replacement ther-
Wider implications of the findings: Both approaches to the management apy (HRT). Limited literature is available regarding the effect of PRP in women
of patients with endometriosis-associated infertility and pain can be successfully with persistent thin endometrium planned for Fresh IVF-ET cycles as well as in
used, however, the administration of gestagen has advantages due to the lower thin endometrium due to different etiological factors. The role of PRP specific
incidence of side effects. to underlying etiology of thin endometrium has not been studied earlier.
Trial registration number: No Study design, size, duration: A prospective interventional cohort study was
conducted at Reproductive Medicine Centre at a tertiary care institute. Women with
P-303 Green tea extract administration reduces MDA level, TNFa persistent thin endometrium despite HRT were enrolled in the study during a period
expression and endometriotic implants area in mice from December 2018 to December 2019. Eighteen women underwent 21 cycles of
Y. Trisetiyono1, S.T. Hidayat1, N. Pramono1, W. Widjiati2 intrauterine PRP administration during fresh and frozen embryo transfer cycles.
1
Autologous venous blood was used to prepare PRP using double centrifuge technique .
Diponegoro University, Department of Obstetrics and Gynecology, Semarang,
Participants/materials, setting, methods: Women who were declined
Indonesia ;
2
fresh IVF-ET cycles because of persistent thin endometrium were selected. PRP
Airlangga University, 2Department of Embryology, Surabaya, Indonesia
was administered during unstimulated cycle on day 8/9 and repeated after 48
hours until ET reached 7 mm. Women who achieved adequate ET were only
Study question: Can the green tea extract (GTE) decrease oxidative stress,
enrolled. During fresh IVF, if endometrium was inadequate on day 8, PRP was
inflammation, angiogenesis, and reduce the extent of endometriosis implants
infused and repeated after 48 hours if required. Women with history of prior
in mice?
cancelled cycles were also enrolled for PRP during FET.
Summary answer: GTE administration resulted in lower malondialdehyde
Main results and the role of chance: Out of 18 patients, 11 had tubal factor
(MDA), tumor necrosing factor-alpha (TNF-a), and smaller endometriotic
with either hysteroscopic or laparoscopic evidence of TB, 4 patients had DOR,
implant, but no effect on vascular endothelial growth factor (VEGF).
and 3 had PCOS as cause of infertility. Out of 21 PRP cycles in 18 patients, 12
What is known already: Several studies showed GTE contains catechins,
had fresh IVF and 9 had FET. Data obtained was analysed using STATA (version
which is a powerful antioxidant, may decrease MDA titer. It also had anti-inflam-
12.0). Mean age was 31.81±3.5 years. Mean baseline ET was 4.76±0.72 mm.
matory action by regulating the expression of inflammatory cytokines and
Mean ET after HRT was 5.45±0.53 mm. During Fresh IVF, mean ET before PRP
enzymes. Other studies in mice also found that GTE inhibited microvessel for-
was 5.87 ± 0.86 mm and after PRP was 7.2 ± 0.5mm (t -7.737, df 11, p-value
mation in endometriotic implants by selectively suppressing VEGF. Some previous
0.000) with average increase of 1mm. During FET cycles, mean ET before PRP
studies also revealed the GTE administration on endometriosis mice induced
was 5.5±1.02 mm and after PRP was 7.2 ± 0.75mm (t -4.703, df 8, p-value
regression of this disease.
0.002) with average increase of 0.9mm. There was no significant difference
Study design, size, duration: This is an experimental study in mice. The size
between clinical pregnancy rate (CPR) and implantation rate (IR) in fresh vs FET
of the sample was obtained by using the equation of resources formula (21
cycles(25% and 13.7% vs 11.1% and 4.5%, p=0.6). The average increase in ET
mice). This research had been done in 3 months (October-December 2017).
after first PRP was significant in both TB and DOR (1mm and 1mm, p < 0.05)
Participants/materials, setting, methods: Twenty-one Balb/c female mice
as compared to PCOS (0.7mm, p= 0.07). There was no significant difference in
were divided into the control (C) group consisted of 7 untreated endometriosis
CPR among three etiological factors (23% vs 20% vs 0%, p=0.99).
mice model, the second group injected with 1 mg/kg BW leuprolide acetate
Limitations, reasons for caution: The limitation of present study is its small
(LA), and the third group given with 200 mg/kg BW/day GTE. MDA level was
sample size. However, the study is currently being endeavoured, and these are
measured by spectrophotometry, TNF-a, and VEGF expression by IHC, whereas
the interim results of the same. Further, more extensive studies in broader
the area of the endometriotic implants was measured using computerize tracing.
horizons are required to assess the efficacy of PRP on thin endometrium of
Main results and the role of chance: Compared to MDA serum in the C
various pathophysiologies.
group, both GTE and LA administration to endometriosis mice resulted in a
Wider implications of the findings: The present study favours the role of
significantly lower level (p=0.000), but they did not affect to VEGF expression
PRP in women who were deferred to undergo IVF or to opt for surrogacy
(p=0.123). Only GTE has a significant effect of lowering TNF-a expression com-
because of their refractory thin endometrium. PRP should be administered to
pared to the C group (p=0.04). Finally, the area of the endometriotic implants
improve the reproductive outcome only in the selected group of population
in the GTE and LA groups was smaller than the C group (p=0.008).
with a persistent thin endometrium.
Limitations, reasons for caution: The treatment given to each group in dif-
Trial registration number: CTRI/2018/12/016799
ferent ways that may affect the pharmacological treatment and the body’s response
of the mice. The study also did not combine GTE and LA, so it did not answer P-305 Microbial roadmap along the menstrual cycle: a new
whether GTE can strengthen the therapeutic effect of LA on endometriosis in mice. approach to identify active microorganism in the uterus
Wider implications of the findings: The overall results of this study show
A. Sola-Leyva1,2, N.M. Molina1,2, E. Andrés-León3, J. Plaza-Díaz4,
that GTE has a good effect on endometriosis disease. Thus, further investigation
M.J. Sáez-Lara1, J. Fontes-Jiménez2,5, J. Mozas-Moreno2,5,
is needed to be conducted to consider green tea as herbal medicine that can be
B. Romero-Guadix2,5, S. Ruiz-Durán2,5, L. Martínez-Navarro2,5,
used as an alternative therapy for endometriosis.
S. Altmäe1,2,6
University of Granada- Faculty of Sciences, Biochemistry and Molecular Biology I,
Granada, Spain ;
P-304 Autologous platelet-rich plasma optimizes endometrial 2
Instituto de Investigación Biosanitaria de Granada, ibs.GRANADA, Granada, Spain ;
thickness in women with refractory thin endometrium of varied 3
Instituto de Parasitología y Biomedicina “López-Neyra” CSIC, Unidad de
aetiology during fresh and frozen-thawed embryo transfer cycles Bioinformática, Granada, Spain ;

4
University of Granada- Faculty of Pharmacy, Biochemistry and Molecular Biology medical treatments such as GnRH agonists and oral contraceptives, are often
II, Granada, Spain ; associated with high recurrence rates and side effects such as hot flushes, epi-
5
HU Virgen de las Nieves, U. Reproducción- UGC Laboratorio clínico y UGC sodes of depression and iatrogenically induced osteoporosis. The identification
Obstetricia y Ginecología, Granada, Spain ; of new compound candidates for endometriosis treatment is needed.
6
Competence Centre on Health Technologies, Biotechnology Company, Tartu, Estonia Endometriosis is an estrogen-dependent disease. ER is a nuclear receptor that
controls the expression of estrogen responsive genes containing estrogen-re-
Study question: Does endometrium harbour functionally active microorgan- sponsive elements (EREs). Hsp90 is essential for ER binding to the EREs.
isms and whether these microbes differ between proliferative and mid- Study design, size, duration: Patient recruitment was carried out at the Sixth
secretory phases? Hospital of Sun Yat-sen University. Ectopic endometrium (endometriotic tissue,
Summary answer: Endometrium possesses active microorganisms, and the n=12) was collected from patients with ovarian endometriotic cysts undergoing
composition and metabolic function of these microorganisms change along the laparoscopy from January 2018 to January 2019.
menstrual cycle. Participants/materials, setting, methods: Primary cultured human endo-
What is known already: Endometrium harbours its unique microbial com- metriotic stromal cells were seeded and incubated with 10nM, 100nM, 1μM and
position, however, as all the microbiome studies are based on microbial DNA 10μM 17-AAG for 24 h. Cell viability was tested by MTT assay. Cell proliferation
sequence analysis, there is no knowledge whether these detected bacterial was detected by a colorimetric BrdU incorporation assay. Caspase-3 activity was
sequences reflect functionally active/alive bacteria within the uterus. Additionally, assessed by Caspase-Glo luminescent-based assays. Transcriptional activity of
it is not clear whether there are microbial changes along the menstrual cycle, as ER was measured by transient transfection of an ERE-tk-Luc reporter construct
a number of studies have detected no microbiome differences between different that contained the consensus ERE sequences. Luciferase activity was performed
menstrual cycle phases, while other studies have noticed differences. Thus, to evaluate the activity of ER.
whether endometrium harbours alive microbes and whether there are changes Main results and the role of chance: 17-AAG inhibited the viability of
along the menstrual phase remains an open issue. human primary endometriotic stromal cells in a dose-dependent manner from
Study design, size, duration: RNA-seq data from 14 endometrial paired the concentration of 10nM. The growth inhibition by 17-AAG started at 100nM
samples from healthy women, 7 samples from the mid-secretory phase and 7 and increased up to 10 μM. The caspase-3 activities were enhanced 2-, 8-, and
samples from the consecutive proliferative phase were analysed for the microbial 11-folds in endometriotic stromal cells after 100nM, 1μM and 10μM 17-AAG
RNA sequences. treatment for 24h. Similarly, transcriptional activity of ER were inhibited signifi-
Participants/materials, setting, methods: The raw RNA sequencing data cantly by 17-AAG in a dose-dependent manner.
was converted into FASTQ format using SRA Toolkit. In order to eliminate rRNA Limitations, reasons for caution: An experimental mouse model should
reads SortMeRNA software was used. R package MetagenomeSeq was used for be established to evaluate the therapeutic effect of 17-AAG. Further molecular
metataxonomic microorganismal analyses and HUMAnN2 algorithm was used mechanism of 17-AAG on ER transcriptional function also need to be explored.
for functional metagenome. The microbial maps were created using Kraken Wider implications of the findings: HSP90 inhibitor controls the develop-
metagenomic sequence classification. With this approach we were able to iden- ment of endometriosis by repressing transcriptional activity of ER. HSP90 inhib-
tify and map the ‘active’ microorganisms within the samples. itor could provide a valuable tool for better understanding of estrogen signaling,
Main results and the role of chance: With our study approach we were as well as for potential clinical use against estrogen-related diseases of women,
able to map the existing functionally active microorganisms in the endometrium including endometriosis.
along the menstrual cycle. Microbes such as bacteria, fungi, viruses and archaea Trial registration number: not applicable
were identified. At the taxonomical level, after stringent multiple correction,
significant differences in the microorganismal composition between proliferative
and mid-secretory phases were detected. Further, these microbes were involved P-307 Pharmaceutical targets of endometriosis for medical
in different metabolic functions, where amino acid and carbohydrate metabolisms treatment: a systematic review
were more active in the proliferative phase while lipid and energy metabolisms S.W. Hung1, R.Z. Zhang1, C.C. Wang1
were more active in the secretory phase. 1
The Chinese University of Hong Kong, Department of Obstetrics & Gynaecology,
Limitations, reasons for caution: These first pilot study results should be Hong Kong, Hong Kong
confirmed in a bigger sample size.
Wider implications of the findings: Our study confirms the presence of Study question: Is there any targeted therapy available for treatment of
‘active’ bacteria, fungi, viruses and archaea in the human uterus. We detected endometriosis?
significant differences in the microbial composition and the involved metabolic Summary answer: Approaches through specific-target, multi-targets, or a
functions between the proliferative and mid-secretory phases. Our study findings combination hold their promises to treat endometriosis and manage the com-
support the hypothesis that endometrial microbiome plays role in the endome- plexity of pathophysiology.
trial functions. What is known already: Growth and development of the endometriotic
Trial registration number: not applicable cells in ectopic sites can be promoted by multiple pathways, including prolifera-
tion, anti-apoptosis, migration, invasion, immune escape, inflammation, oxidative
P-306 Heat Shock Protein 90 Inhibitor 17-AAG Inhibits stress, angiogenesis, and disrupted autophagy. Endometriosis causes a negative
Proliferation of Human Endometriotic Stromal Cells by impact on life quality, productivity and infertility. Current medical treatments for
Repressing Estrogen Receptor Transcriptional Activity. endometriosis are rather empirical and incurable. Endometriosis surgery is often
J. Li1, P. Chen2, X. Liang1 performed to improve infertility. Effective medication for specific molecular and
1
the Sixth Affiliated Hospital- Sun Yat-sen University, Reproductive Medicine signalling pathways is limited.
Research Centre, Guangzhou- Guangdong, China ; Study design, size, duration: We systematically reviewed literatures that
2
the First Affiliated Hospital- Sun Yat-sen University, Department of Pharmacy, focused on pharmaceuticals targeting specific molecular and signalling pathways
Guangzhou, China in endometriosis. Selected publications included both animal and human studies;
in-vivo or in-vitro. We gathered high quality research studies of 150 publications
Study question: HSP90 is known to be associated with estrogen receptor to summarise the results.
(ER) and regulates ER mediated cell proliferation. Whether 17-AAG, a HSP90 Participants/materials, setting, methods: Literature search was per-
inhibitor, suppresses the growth of endometriotic cells is still unknown. formed using PubMed, Medline and Google Scholar for original articles that were
Summary answer: 17-AAG could control the progress of endometriotic written in English and Chinese. PRISM flowchart and Endnote were used for
stromal cells in vitro via inhibiting transcriptional function of the estrogen better management of the searches. We assessed the study quality by critically
receptor. apprising their appropriateness to the research objective and outcome repro-
What is known already: The prevalence of endometriosis is between 2% ducibility. We presented the data narratively to update the research progress
and 10% in the general population and 50% in the infertile population. Current on the medical treatment in endometriosis.

Main results and the role of chance: Akt, ERK, MAPK, MTor, NF-κB, P13K, compared to the non-PGT group: 32% (95%CI: 23%-40%) versus 11% (95%CI:
ROCK, ROS, TNF, Wnt, VEGF, estrogen, cytokines and their upstream and 7%-15%). As expected, more patients in the PGT-M group ceased their hormonal
downstream molecules are the key aberrant signalling and regulatory pathways contraception late: 64% (95%CI: 55%-73%) versus 2% (95%CI: 0%-4%) in the
promoting growth and development of endometriotic cells and tissues. non-PGT group. Average duration of hormonal contraceptive use was longer
Pharmaceuticals targeting these molecules were identified, amongst which in the PGT-M group than in the non-PGT group (10,6 years SD 4,3 versus 9,3
Western medicines focus on pain relief and anti-proliferation, such as hormonal years SD 4,5, p=0,02). To determine which factors were significant independent
(e.g. Dienogest, Aromatase inhibitors) targets on ER stress, estrogen receptors, predictors of thin endometrium, a multivariate logistic regression analysis was
or TNFα to inhibit NFκB; as well as non-hormonal (e.g. Pentoxifylline, Sunitinib) performed including the following variables: PGT-M yes/no, duration of hor-
targets on VEGF and VEGFR for anti-angiogenesis. Natural products and Chinese monal contraceptive use (years), late cessation yes/no, age, BMI, parity, previous
Medicines aim to reduce chronic pain, resolve infertility and improve quality of curettage and number of dominant follicles. Only late cessation (OR: 5,1,
life with few side-effects. They mainly regulate blood flow through targeting on 95%CI:1,5 – 16,7) and duration of hormonal contraceptive use (OR 1,1, 95%CI:
superoxide anion and catalase as radical scavengers for anti-oxidation (e.g. 1,0-1,3) remained significant independent predictors. The odds ratio for PGT-M
EGCG, Glycyrrhizae Radix, Ligusticum Rhizoma) and cytokines and TNFα for to predict a thin endometrium changed from 3.8 (95%CI:2,2-6,7) in an univariable
anti-inflammation (e.g. Genistein, Angelicae Sinensis Radix, Bupleuri Radix). analysis to 1,1 (95%CI: 0,3-3,3) in the multivariable analysis, suggesting that the
Chinese Medicine decoctions often contain many individual herbs, so multiple difference in prevalence of thin endometrium between PGT-M and non-PGT
targeting on several pathways with synergistic efficacy and better outcomes. patients could be attributed mainly to differences in prior contraceptive use.
Limitations, reasons for caution: Non-hormonal treatments are mostly Limitations, reasons for caution: The study is retrospective, using self-re-
under clinical trials or only studied in in-vitro or animal model. There are lack of ported data. Different types of hormonal contraceptives were reported (e.g. levo-
experimental and clinical evidences to support its effectiveness and safety of norgestrel IUD, oral combined contraceptives) which possibly exert different effects
TCM and natural products in alleviating the pathogenesis of endometriosis, when on the endometrium, however, study size was too small to make separate analyses.
it is compared to western medicine. Wider implications of the findings: This study provides further insight to
Wider implications of the findings: New drugs to treat endometriosis and a possible contributor to the occurrence of thin endometrium: prior hormonal
disease related symptoms and complications, with high efficacy, reduced off contraceptive use. Future studies should provide more information on its clinical
targets and other side effects, can be further developed based on knowledge of relevance, to determine whether PGT-M patients can be reassured or should
molecular targets. A detailed protein-protein network helps with target decon- be advised to stop hormonal contraceptives in advance of their treatment.
volution for therapeutically known compounds. Trial registration number: not applicable
P-309 High pregnancy rate with customized embryo transfer
P-308 Do PGT-M patients have a higher prevalence of thin after endometrial receptivity assessment using a transcriptomic
endometrial lining and is this explained by prior prolonged approach in patients candidate for eggs donation program
hormonal contraceptive use? D. Haouzi1, F. Entezami2, A. Ferriere-Hoa3, C. Mauries3, A. Gala3,
I. Homminga1, E. Ter Meer1, H. Groen1, A. Cantineau1, A. Hoek1 E. Vintejoux3, C. Brunet3, C. Vincens3, S. Bringer-Deutsch3,
1
University of Groningen-University Medical Center Groningen, Obstetrics and S. Brouillet3, S. Hamamah3
Gynecology, Groningen, The Netherlands 1
INSERM U1203, IRMB- Hôpital St-Eloi- CHRU Montpellier, MONTPELLIER,
France ;
Study question: Is thin endometrial lining on the day of hCG triggering more 2
American Hospital of Paris, IVF department, Neuilly-Sur-Seine, France ;
prevalent in PGT-M patients than in non-PGT patients, and related to differences 3
Arnaud de Villeneuve hospital- CHU Montpellier, ART/PGD department,
in previous hormonal contraceptive use? Montpellier, France
Summary answer: Endometrial lining <8mm is more prevalent in PGT-M
patients compared to non-PGT patients, and this is related to prolonged hor- Study question: The aim of this study was to optimize pregnancy outcome
monal contraceptive use up until IVF-treatment. using customized embryo transfer according to the evaluation of endometrial
What is known already: Thin endometrial lining has been associated with receptivity of patients included in eggs donation program.
lower pregnancy rates in multiple studies. We seemed to notice an increased Summary answer: Individual evaluation of endometrial receptivity allows a
prevalence of thin endometrial lining in patients attending our clinic for pre-im- customized embryo transfer (cET) according the endometrium receptivity day
plantation genetic testing for monogenetic disease (PGT-M). We hypothesized and improves pregnancy outcome in oocyte donation program.
this could be due to a prolonged use of hormonal contraceptives up until the What is known already: Many approaches for human endometrial receptivity
start of the PGT-IVF/ICSI treatment, which is typical for this patient group, as including microarray has been previously reported. However, efficiency of the
they usually want avoid spontaneous pregnancies. Though it is well known that tests according to the clinical results is still debated. The aim of this study is to
endometrial thinning occurs during the use of e.g. oral contraceptives, little is evaluate the endometrial receptivity using a transcriptomic approach in eggs
known about a possible lingering of this effect after cessation. donation program. This approach consists for screening 11 specific genes of the
Study design, size, duration: A retrospective cohort study was performed, receptivity window.
including all PGT-M patients attending the University Medical Centre Groningen, Study design, size, duration: Endometrial biopsies are performed during
between 2009 and 2018. For each PGT-M patient, two non-PGT IVF/ICSI the implantation windows 5-9 days after progesterone administration under
patients (indication male factor, tubal factor or unexplained subfertility) were hormone replacement therapy. Then, the transfer strategy consists in performing
included, matched for age and treatment period. Prevalence of thin endometrium cET of blastocysts according the endometrium receptivity day identified using
(<8mm) on the day of hCG triggering was compared between PGT-M and the Win-Test. Therefore, frozen day 2/3 embryos were transferred 72/48 hours
non-PGT patients. Hormonal contraceptive use (both duration and cessation before this specific receptivity day, respectively. When the endometrial sample
period) was compared between both groups. was defined as non-receptive, a second evaluation was performed later, accord-
Participants/materials, setting, methods: 122 PGT-M patients and 240 ing to transcriptomic result.
non-PGT patients were included. Only first ovarian hyperstimulation IVF/ICSI Participants/materials, setting, methods: 45 patients in eggs-donation
cycles were included. Patients with endometriosis, PCOS or uterine abnormal- program due to an advanced age (n=36) or premature ovarian failure (n=9)
ities were excluded. Data on history of hormonal contraceptive use prior to the were included. RNAs from biopsies were extracted and the Win-Test gene
treatment was obtained from patient questionnaires, both duration (years) and expression was assessed by qRT-PCR. Positive pregnancy test was defined as at
cessation period were included. Cessation categories were <1 year (late cessa- least two consecutive positive b-hCG serum concentration. Clinical pregnancy,
tion) or >1 year (early cessation) before treatment. Endometrial thickness was implantation rate and live birth rate were recorded after cET according to the
measured on day of HCG triggering. transcriptomic results.
Main results and the role of chance: Thin endometrium (< 8 mm) on the Main results and the role of chance: Analyses of endometrial receptivity
day of hCG triggering was found significantly more often in the PGT-M group, status (n=108 biopsies) in patients in oocyte donation program (age mean

± SD: 41.2 ± 3.8 years) revealed a strong inter-patient variability of the moment P-311 Expression of non-classical Human Leukocyte Antigen (HLA)-G
of the opening of the receptivity window within the implantation window. and -F in the endometrium in relation to immune cell infiltration in
Majority of patients (84%) present a delay of their receptivity window compared women experiencing recurrent implantation failure (RIF)
to the classical timing for blastocyst transfer (16% at Pg+5/Pg+6). This delay H. Papuchova1,2,3, M. Hviid Saxtorph2,4, T. Hallager2,5, I. Engberg
was mainly between 1 (40%) to 2 days (33%), or more (11%). Then, a cET can Jepsen2,4, G. Persson1,2, J.O. Eriksen2,5, L. Larsen Grupe2,5,
to be perform during a subsequent cycle according the endometrium receptivity N. Macklon2,6, T.V. Hviid1,2,3
day and in the respect of the synchronization of the fœto-maternal dialogue. 1
Zealand University Hospital, Department of Clinical Biochemistry- Centre for
Using this strategy, the positive b-hCG and clinical pregnancy rate per patient Immune Regulation and Reproductive Immunology CIRRI, Roskilde, Denmark ;
were 73.3% and 60%, respectively. The implantation and live birth rates after 2
Zealand University Hospital, The ReproHealth Research Consortium, Roskilde,
cET were 50% and 48.9%, respectively. Denmark ;
Limitations, reasons for caution: The benefits of this strategy will be ana- 3
University of Copenhagen, Department of Clinical Medicine, Copenhagen,
lyzed in a large cohort of patients in eggs/embryos donation program Denmark ;
Wider implications of the findings: This finding demonstrated that cus- 4
Zealand University Hospital, Department of Gynaecology- Obstetrics and Fertility,
tomized embryo transfer according to the specific cycle day where endometrium Roskilde/Køge, Denmark ;
is receptive improves both implantation rate and live birth rate in patients in 5
Zealand University Hospital, Department of Pathology, Roskilde, Denmark ;
oocyte donation program. 6
London Women’s Clinic, London Women’s Clinic, London, United Kingdom
Trial registration number: ID: NCT04192396
Study question: Does HLA-F and HLA-G expression correlate with the infil-
P-310 Increased expression of fucosyltransferase 4 (FUT4) mRNA tration of CD56+, CD16+, FOXP3+ or CD163+ immune cells in endometrial
in eutopic endometrium is a specific and sensitive marker of biopsies from women with RIF?
endometriosis Summary answer: Endometrial HLA-G expression of women experiencing RIF
J. Malejczyk1, M. Żeberkiewicz1, A. Hyc1, A. Iwan1, is correlated with immune cell infiltration (CD16+ and CD56+). HLA-F expression
A. Zwierzchowska2, E. Barcz2, A. Ścieżyńska1 is correlated with anti-inflammatory macrophage infiltration (CD163+).
1
Medical University of Warsaw, Department of Histology and Embryology, What is known already: HLA class Ib have a prominent immunomodulatory
Warsaw, Poland ; role at the fetal-maternal interface, promoting immune tolerance of fetal anti-
2
Medical University of Warsaw, 1st Department of Obstetrics and Gynaecology, gens. HLA-G-positive trophoblasts are in a close contact with decidualized
Warsaw, Poland endometrium during pregnancy, and HLA-G expression in the endometrium has
been identified as prognostic for achieving pregnancy. Evidence of HLA-F expres-
Study question: Are the levels of fucosyltransferase 4 (FUT4) mRNA in the sion in the endometrium and its interactions with immunoglobulin-like transcript
eutopic endometrium associated with endometriosis? (ILT) receptors expressed on Natural Killer (NK) cells, T cells and macrophages
Summary answer: FUT4 mRNA expression is significantly increased in the have been elucidated. These interactions between endometrial HLA class Ib and
eutopic endometrium from women with endometriosis and may serve as a immune cells could have a determinant immunosuppressive function required
diagnostic marker of endometriosis. for successful embryo implantation.
What is known already: Retrograde flow of shed endometrial cell may play Study design, size, duration: Endometrial biopsies were obtained from all
a role in development of endometriosis. It is postulated that survival of endo- participants in an estrogen/progesterone substituted cycle. Samples were collected
metriotic cells in the peritoneum is partially related to their decreased apoptosis after five days of progesterone treatment. In total, 86 women with RIF and 37
and increased invasiveness. In particular, development of endometriotic lesions controls attending a fertility clinic were recruited between November 2017 and
may be related to epithelial precursor/stem cells characterized by expression September 2019. The patients were divided into four groups based on the infertility
of stage specific embryonic antigen 1 (SSEA-1/CD15). Expression of SSEA-1 is cause: unknown, female factor, male factor and both male and female factor.
related to activity of FUT4. Preliminary data are presented including 51 women experiencing RIF.
Study design, size, duration: This control-case study involved endometrial Participants/materials, setting, methods: Immunohistochemical proce-
and endometrioid cyst samples collected by aspiration biopsy or laparoscopic dures were automatically performed on an Autostainer on paraffin-embedded
surgery from 86 women with and without endometriosis. sections using anti-human antibodies for HLA-F, HLA-G, CD56, CD16, CD163
Participants/materials, setting, methods: The study was conducted at a and FOXP3 with corresponding technical staining controls. Randomized and
university research institution. FUT4 mRNA were examined in eutopic endome- blinded analysis were performed using Tissue IA image analysis software (Digital
trium tissue samples from 58 women with endometriosis (mean age 32.4 years) Image Hub). Algorithms were developed for the above-mentioned antibodies.
and eutopic endometrium from 28 control women (mean age 36 years) by means HLA-F and HLA-G expression were studied by flow cytometry on fresh endo-
of quantitative reverse-transcription polymerase chain reaction (qRT-PCR). metrial samples. Statistical analysis based on Spearman correlation was per-
Main results and the role of chance: Expression of FUT4 mRNA in eutopic formed using SPSS version 25.
endometrium from women with endometriosis and controls did not differ in Main results and the role of chance: HLA-F and HLA-G membrane expres-
relation to phase of the menstrual cycle. The relative levels of FUT4 mRNA were sion on endometrial cells were confirmed by flow cytometry analysis on freshly
significantly increased in eutopic endometrium from endometriosis patients as isolated endometrial stromal cells from endometrial biopsies. Using immunohis-
compared to control women (P<0.0001). ROC analysis of FUT4 mRNA expres- tochemistry, the preliminary results show that the percentage of HLA-G-positive
sion in eutopic endometrium showed high statistical significance (AUC = 0.90, cells in the endometrial glands was positively correlated with the percentage of
95%CI = 0.81-0.99, P<0.0001) thus indicating endometrial FUT4 mRNA to be CD16-positive cells per analyzed area (p<0.05). As well as the semi-quantitative
a specific marker of endometriosis. Expression of FUT4 mRNA in eutopic endo- HLA-G H-score (staining intensity score) was positively correlated with per-
metrium was correlated with endometriosis severity (rs=0.5579, P<0.0001). centage of CD16-positive cells within the analyzed area (p<0.05). Moreover, in
There were no differences in endometrial FUT4 mRNA expression regarding a patient subgroup with no female factor identified, both percentage of HLA-
endometriotic lesion location and patients’ infertility status. G-positive cells and HLA-G H-score were positively correlated with percentage
Limitations, reasons for caution: The study was performed on a limited of CD16-positive, CD56-positive and CD163-positive cells per analyzed area
group of patients and controls. The results should be confirmed by a replicate (all p<0.05). Interestingly, all three HLA-F variables (number of positive cells
study on different patients’ cohorts. per mm2, percentage of positive cells per analyzed area and H-score) in both
Wider implications of the findings: The present results strongly suggest endometrial glands and stroma were positively correlated with anti-inflammatory
that expression of FUT4 mRNA in eutopic endometrium is a specific marker macrophages per mm2 and their percentage per analyzed area (identified by
suitable for the low-invasive diagnosis of endometriosis. It is possible that FUT4 CD163, p<0.01 and p<0.005 for glands and stroma respectively). The number
activity plays a role in development and persistence of endometriotic lesions of HLA-F-positive cells per mm2 and HLA-F score were also negatively cor-
and may potentially be a therapeutic target for endometriosis. related with the number of regulatory T cells per mm2 (identified by FOXP3,
Trial registration number: not applicable all p<0.05).

Limitations, reasons for caution: The data presented are preliminary data Wider implications of the findings: Vaginal microbiota seems to be asso-
based on 51 women out of 123 included in the study. Correlations indicate the ciated with the outcome of a frozen embryo transfer cycle. This association
relationship between two variables; however, influences of other variables should might be independent from the phase of the menstrual cycle. Asymptomatic
be taken into consideration. No corrections for multiple comparisons were chlamydial infections detected by 16s sequencing in patients undergoing IVF
performed because the character of the study is primarily exploratory. needs further investigation.
Wider implications of the findings: In the current preliminary analysis, we Trial registration number: clinicaltrials.gov ID: NCT03507673).
show interesting trends indicating links between NK cell, regulatory T cell and
macrophage markers with HLA class Ib molecules of potential importance for P-313 Prevalence of T-shape uterus among porous women based
establishing fetal-maternal tolerance at implantation and in early pregnancy. on Congenital Uterine Malformations by Experts (CUME) criteria
Trial registration number: not applicable A. Seyhan1, S. Ertas1, B. Urman1
1
American Hospital, Reproductive Endocrinology and Infertility, Istanbul, Turkey
P-312 16s sequencing of vaginal microbiota in patients undergoing
frozen embryo transfer following IVF Study question: What is the prevalence of T-shape uterus among parous
K. Neumann1, A. Masuch2, S. Graspeuntner3, L. Kirchhoff3, women based on CUME criteria?
A. Schultze-Mosgau2, M. Depenbusch2, J. Rupp3, G. Griesinger2 Summary answer: Parous women who conceived spontaneously have a low
1
prevalance of T-shape uterus.
University Hospital of Schleswig-Holstein- Luebeck- Schleswig-Holstein- Germany,
What is known already: Both prevalence of T shape uterus in parous women
Department of Gynecological Endocrinology and Reproductive Medicine, Lübeck,
and its effect on reproductive/obstetric outcome is unknown.
Germany ;
2
Study design, size, duration: Prospective cohort study including 238 repro-
ductive age group of women in who conceived spontaneously and had at least
Department of Reproductive Medicine and Gynecological Endocrinology, Luebeck,
one live birth. Participants were recruited from a contraception clinic between
Germany ;
3
January 2017 and December 2019.
Participants/materials, setting, methods: Participants underwent 3D
Department of Infectious Diseases and Microbiology, Luebeck, Germany
transvaginal ultrasound in the late luteal phase of menstrual cycle. The ESHRE/
ESGE classification of congenital anomalies of the female genital tract was used
Study question: Does 16s sequencing of vaginal microbiota predict the out- for the description of abnormal findings. The diagnosis of T-shape uterus was
come of a frozen embryo transfer cycle? Does the phase of the menstrual cycle based on the definition ofCUME criteria.The following measurements were taken
impact these microbiota? In the coronal view; 1)lateral indentation depth, 2) lateral indentation angle, 3)
Summary answer: Vaginal microbiota can contribute to pregnancy prediction T-angle. Uterine cavity volume was measured by Virtual Organ Computer-aided
which seems to be independent from the phase of the menstrual cycle during AnaLysis (VOCAL™) program.
which sampling was done. Main results and the role of chance: The mean (± standard deviation) age
What is known already: Recent studies have shown conflicting data on an was 35.7±5.9 years. Mean parity was 1.88 (min 1 max 7) and time to pregnancy
association of certain vaginal microbiota with the outcome of a fresh cycle of in (TTP) was between 1- 12 months. Mean antral follicle count was 12.2±9.5 and
vitro fertilization (IVF) (Koedooder et al. 2019, Haahr et al. 2019). These studies, 10.5 % had at least one ultrasound feature of adenomyosis. Nine patients (3%)
however, analyzed fresh embryo transfer cycles only and did not account for were diagnosed with congenital uterine abnormality of which 5 (2.1%) had partial
intra-individual variation, e.g. the impact of the sampling time-point within the septate uterus, 2 (0.8%) had a hemiuterus and 1 (0.4%) had a Tshape uterus and
menstrual cycle. 1 had borderline T-shape uterus.
Study design, size, duration: Prospective, clinical cohort study of patients Overall mean uterine volume, lateral indentation depth, lateral indentation
undergoing a frozen embryo transfer cycle (spontaneous or programmed) fol- angle and T-angle was 4 ±2.3 ml, 2.95±,1.16mm, 154.7 ±9.9°and
lowing IVF. Sample collection has started in 5/2018 and is still ongoing. The study 73.5±9.9°respectively. Six women fulfilled one criterion and 1 patient met 2
was prospectively registered (clinicaltrials.gov ID: NCT03507673). criteria with lateral indentation depth of 8.4mm and lateral indentation angle of
Participants/materials, setting, methods: Vaginal swabs were collected 128°. The women with T shaped uterus had uterine volume, lateral indentation
on day of planning of embryo transfer (in the late follicular phase day 13 – 15 depth, lateral indentation angle and T-angle of 2.62 ml, 7mm, 124°and 38°respec-
of the menstrual cycle), on day of embryo transfer (i.e. after 2 to 6 days of tively. Her TTP was 7 months and she had given birth to a 3450 gr child at 39.
progesterone administration) and on day of pregnancy test (10 – 14 days after Limitations, reasons for caution: The prevalence of T-shape uterus in a
embryo transfer). 16s ribosomal RNA sequencing was used for longitudinal population of parous women who conceived spontaneously is very low. The
investigation of vaginal microbiota which were classified according to previous management of this condition can be determined following comparative studies
reported outcome criteria. showing the prevalance among parous, recurrent miscarriage and infertilite
Main results and the role of chance: 16s sequencing of the first collected patients.
sample from the end of the follicular phase (n=56) of the frozen embryo transfer Wider implications of the findings: If women with infertility and recurrent
cycle revealed a statistically significant reduction in species richness (p=0.011) misccariage are shown to have higher prevalance of T shape uterus, possible
and in the chao 1 index (p=0.0045) for patients achieving a positive pregnancy benefit of surgical management may be claimed.
test (n=22) versus patients having a negative test (n=34). In patients having a Trial registration number: 2018.052.IRB1.008
sample available for analysis from all three time-points (n=13), a tendency
towards an increased relative read count for Lactobacillus crispatus for patients
P-314 Is endometrial gene expression profile affected in women
achieving a positive pregnancy test vs. patients having a negative test was
with endometriosis? a meta-analysis
observed. Additionally, a decreased alpha-diversity was observed for patients
with a positive pregnancy test. These differences were observed independent E. Vargas1, E. García-Moreno2, A. Salumets3, F.J. Esteban1,
from the phase of the menstrual cycle during which sampling was performed; S. Altmäe4
1
however, statistical significance was not reached yet for this subgroup. Surprisingly, University of Jaen, Department of Experimental Biology, Jaen, Spain ;
2
n=3 asymptomatic patients undergoing infertility treatment were diagnosed with Puerta del Mar University Hospital, Immunology Unit, Cadiz, Spain ;
3
a relative chlamydial abundance. None of these patients having a positive relative Competence Centre on Health Technologies, Competence Centre on Health
chlamydia read could achieve a pregnancy. Technologies, Tartu, Estonia ;
4
Limitations, reasons for caution: This study is ongoing, a higher number University of Granada, Department of Biochemistry and Molecular Biology,
of patients are needed for robust conclusions. Additionally, no stratification of Granada, Spain
subgroups (e.g. age, protocol of frozen embryo transfer cycle, etc.) or differen-
tiation of previously reported community state types of bacteria have been Study question: Do women with endometriosis have a different endometrial
conducted so far. gene expression profile at the time of embryo implantation than healthy women?

Summary answer: This meta-analysis identified no single genes to be differ- Study design, size, duration: This was a prospective observational cross-sectional
entially regulated in endometriosis, while apoptosis, immune response and study conducted over a period of 12 months. We included 50 consecutive women
wound healing pathways seem to be dysregulated. who were all examined by two operators (A and B) during their clinical attendance.
What is known already: ~50% of women with endometriosis suffer from Participants/materials, setting, methods: The study was carried out in a
infertility. Some studies suggest impaired endometrial receptivity in these wom- specialist endometriosis centre. We included all consecutive women who had
en,while other studies claim no impact of endometriosis in endometrial recep- ultrasound scans performed independently by two experienced operators during
tivity. The studies analysing endometrial gene expression profiles in endometriosis the same visit to the clinic. The outcomes of interest were the inter- and intraob-
have been performed in small sample size with low statistical power. We set out server reproducibility for the detection of endometriotic lesions. We also
to systematically gather published studies comparing endometrial gene expres- assessed repeatability of the measurements of lesion size.
sion signatures in women with endometriosis vs. healthy women. Based on the Main results and the role of chance: There was a good level of agreement
obtained data we conducted meta-analysis of differentially expressed genes in between operator A and operator B in detecting the presence of pelvic endo-
order to raise the analysis power and thus clarify whether endometriosis have metriotic lesions (k = 0.72). There was a very good level of agreement between
an impact on endometrial receptivity at molecular level. operators in identifying endometriotic cysts (k = 0.88) and a good level of
Study design, size, duration: A systematic literature search was performed agreement in identifying endometriotic nodules (k = 0.61). The inter- and
until 16th of January 2020 aiming to identify studies assessing the mid-secretory intraobserver repeatability of measuring endometriotic cysts was excellent (intra-
endometrial gene expression profile, i.e. transcriptome of women with endome- class correlation (ICC) ≥ 0.98). There was good interobserver measurement
triosis vs. healthy controls during the window of implantation. The eligible studies repeatability for bowel nodules (ICC 0.88), but the results for nodules in the
were further explored to extract the differentially expressed gene lists and the posterior compartment were poor (ICC 0.41). The intraobserver repeatability
associated fold changes and we then performed a meta-analysis using the robust for nodule size measurements was good for both operators (ICC ≥ 0.86).
rank aggregation (RRA) method to screen for highly representative genes. Limitations, reasons for caution: There was insufficient data to perform a
Participants/materials, setting, methods: The systematic literature search separate analysis for nodule size in the anterior compartment. All examinations
was conducted following the PRISMA criteria and included Pubmed, Cochrane, were performed within a specialised unit with a high prevalence of deep endo-
Google Scholar and Web of Science. A meta-analysis was performed on the metriosis. Our findings may not apply to operators without intensive ultrasound
selected studies to extract the differentially expressed genes described during training in the diagnosis of pelvic endometriosis.
the mid-secretory phase in women with endometriosis vs. healthy women using Wider implications of the findings: These findings are important because
the RRA method. In total endometrial transcriptome data of 128 women (84 ultrasound has been widely accepted as the first line investigation for the diag-
patients and 44 controls) were analysed in our meta-analysis. nosis of pelvic endometriosis, which often determines the need for future inves-
Main results and the role of chance: Within the systematic literature search, tigations and treatment. The detection and measurement of bowel nodules is
7 studies were eligible for the quantitative meta-analysis gathering transcriptome essential for anticipation of surgical risk and planning surgical excision.
data in total from 128 women of the mid-secretory phase of the menstrual cycle. Trial registration number: Not applicable
It is the first time when such a number of women have been analysed in order
to assess the impact of endometriosis on endometrial transcriptome profiles. P-316 Incidence of the Low Anterior Syndorme (LARS) after
The transcriptome data from 128 women covered a total number of 1100 dif- Colorectal Surgery for Deep Endometriosis: an International
ferentially expressed transcripts (644 up-regulated and 456 down-regulated), Multicentric Study
whose relevance was interrogated using the RRA method. After stringent mul- A. Bokor1, G. Hudelist2, B. Dauser3, R. Brubel1, J.J. Tuech5,
tiple correction, our meta-analysis did not detect any significantly differentially H. Roman4
expressed genes in the mid-secretory endometrium of patients vs. controls. 1
Semmelweis University, Dept.OB/GYN, Budapest, Hungary ;
However, when analysing the molecular pathways involved in the identified gene 2
Hospital St. John of God Vienna- Austria, Gynecology, Vienna, Austria ;
lists, we found that apoptosis, immune response and wound healing pathways 3
Hospital St. John of God- Vienna- Austria, Department of General Surgery,
were dysregulated among women with endometriosis.
Vienna, Austria ;
Limitations, reasons for caution: Most of the studies included into the 4
Centre d’endométriose Clinique Tivoli-Ducos Bordeaux- France, Endometriosis
meta-analysis are relatively small, using different platforms for measuring gene
Centre, Bordeaux, France ;
expression together with the low number of samples which might contribute 5
Department of surgery, Rouen University Hospital F-76000, France
to a bias.
Wider implications of the findings: The current meta-analysis supports
Study question: Is there a difference between the incidence of the Low
the hypothesis that a single gene is not dysregulated in endometriosis, rather is
Anterior Syndrome (LARS) after segmental resection or transanal disc excision
a cohort of genes that lead to dysregulation of specific molecular pathways.
performed for the treatment of colorectal deep endometriosis (DE).
Summary answer: LARS is not more frequent after nerve and vessel spearing
segmental resection (NVSSR) when compared to a more conservative approach,
P-315 The intra- and interobserver reproducibility of pelvic the Rouen-technique disc excision.
ultrasound for the detection and measurement of endometriotic What is known already: Several studies show a significant drop in pain scores
lesions and improved fertility outcomes in women following surgical resection of colorectal
E. Bean1, P. Chaggar1, N. Thanatsis1, W. Dooley1, C. Bottomley1, endometriosis. However, there is inceasing evidence that intermediate and long-
D. Jurkovic1 term bowel dysfunction may occur as a consequence of radical surgery for rectal
1
University College London Hospital, Women’s Health, London, United Kingdom DE. Typical symptoms include constipation, feeling of incomplete evacuation,
clustering of stools and urgency. This is described in the colorectal surgical literature
Study question: What is the interobserver and intraobserver reproducibility as LARS. Previous studies demonstrated statistically significant differences in func-
of pelvic ultrasound for the detection of endometriotic lesions? tional outcomes in favour of the conservative surgical approach, i.e. resection of
Summary answer: Pelvic ultrasound is highly reproducible for the detection endometriotic tissue with preservation of the luminal structure of the rectal wall.
of pelvic endometriotic lesions Study design, size, duration: A total of 205 patients, operated before the
What is known already: Transvaginal ultrasound has been widely adopted 1st of January 2018, were enrolled in this study. One hundred and thirty-nine
as the first-line assessment for the diagnosis and assessment of pelvic endome- patients with nerve and vessel sparing segmental resection (NVSSR) from the
triosis. Severity of endometriosis as assessed by ultrasound has been shown to University Hospital Rouen, France (n=52); Semmelweis University Budapest,
have good concordance with laparoscopy (kappa 0.79). The reproducibility of Hungary (n=50) and Hospital of St.John of God Vienna, Austria (n=37) as well
transvaginal ultrasound for assessment of ovarian mobility and pouch of Douglas as 66 patients operated at the University Hospital Rouen, France using the
obliteration using the ‘sliding sign’ has already been described in the literature. Rouen-technique disc excision (RTDE) were included.
However, there is no available data in the literature to demonstrate the intraob- Participants/materials, setting, methods: All patients underwent low
server repeatability of measurements for endometriotic cysts and nodules. rectal resection with a resection line lower than 7cm from anal verge.

Gastrointestinal functional outcomes of the two procedures were compared logistic regression model, adjusted on adenomyosis and endometriosis pheno-
using the validated LARS questionnaire. types, the association with endometriosis in women affected with adenomyosis
The median follow-up time was 46±11 months. As a secondary outcome, the significantly increase the risk of miscarriage (OR=3,198, 95% CI [1,1-9,65]). The
surgical sequelae and fertility results were examined. Statistical analysis was done risk was significantly higher in case of deep infiltrated endometriosis (OR 4,37,
using Fisher’s Exact and Pearson Chi Square test. 95% CI [1,32-14,53]).
Main results and the role of chance: We found no statistically significant Limitations, reasons for caution: The exclusive inclusion of patients from
difference between the incidence of the LARS (31.7% and 37.9% respectively) our referral center could constitute a possible selection bias, as those women
among patients operated using RTDE when compared to women treated by may have suffered from particularly severe forms of adenomyosis and/or
NVSSR (p=0.4). The occurence of LARS was positively associated with the use endometriosis.
of protective ileo-or colostomy (p=0.02). A higher rate of severe complications Wider implications of the findings: We reported in this study that endo-
was observed in RTDE (19.7%) compared to patients who underwent NVSSR metriosis associated with adenomyosis is associated with higher previous spon-
(9.0%, p=0.029). taneous miscarriage rate as compared to women with isolated adenomyosis.
Limitations, reasons for caution: Since digestive complaints may also be This study opens the doors to future, more mechanistic studies to establish the
present presurgically, evaluation of digestive complaints should have been exact link between endometriosis/adenomyosis and spontaneous miscar-
recorded pre- and post surgery as well. riage rates.
Although this study has important limitations due to its retrospective design, Trial registration number: not applicable
it highlights important factors that affects outcomes in patients undergoing col-
orectal resection for the treatment of DE. P-318 Alteration in gene expression of H19 lncRNA and IGF2
Wider implications of the findings: LARS is not more frequent after NVSSR in endometrial tissues of women with endometriosis
when compared to a more conservative approach RTDE. S. Kamrani1,2, E. Amirchaghmaghi3,4, K. Ghaedi1,5,
The findings of this study may help guide treatment strategies aimed at pro- M. Shahhoseini2,6,7
viding patients with colorectal DE an optimal surgical therapy. To confirm our 1
Department of Cell and Molecular Biology and Microbiology- Faculty of Biological
findings prospective studies are required.
Science and Technology- University of Isfahan- Isfahan- Iran., Department of Cell
and Molecular Biology and Microbiology, Isfahan, Iran ;
2
Department of Genetics- Reproductive Biomedicine Research Center- Royan
P-317 Increased rate of spontaneous miscarriages in women
Institute for Reproductive Biomedicine- ACECR- Tehran- Iran., Department of
affected with adenomyosis associated with endometriosis
Genetics, Tehran, Iran ;
B. Pham1, M. Bourdon1, L. Marcellin1, C. Maignien1, C. Chapron1, 3
Department of Regenerative Biomedicine- Cell Science Research Center- Royan
P. Santulli1 Institute for Stem Cell Biology and Technology- ACECR- Tehran- Iran., Department
1
Hopital Cochin Port-Royal, Gynecology and reproductive medecine, Paris, France of Regenerative Biomedicine- Cell Science Research Center-, Tehran, Iran ;
4
Department of Endocrinology and Female Infertility- Reproductive Biomedicine
Study question: Among women with a radiological diagnosis of adenomyosis, Research Center- Royan Institute for Reproductive Biomedicine- ACECR- Tehran-
are spontaneous miscarriages higher in women with endometriosis when com- Iran., Department of Endocrinology and Female Infertility- Reproductive
pared with endometriosis-free controls? Biomedicine Research Cent, ;
Summary answer: Endometriosis-affected women display a significantly higher 5
Department of Cellular Biotechnology- Cell Science Research Center- Royan
rate of previous spontaneous miscarriages than endometriosis-free controls in Institute for Biotechnology- Isfahan- Iran., Department of Cellular Biotechnology-
a population of adenomyosis affected women. Cell Science Research Center, Isfahan, Iran ;
What is known already: Endometriosis and adenomyosis are common, estro- 6
Department of Cell and Molecular Biology- School of Biology- College of
gen-dependent, inflammatory disorders in women of reproductive age. Those Science- University of Tehran- Iran., Department of Cell and Molecular Biology-
diseases could be responsible for pain and fertility alteration. Endometriosis and School of Biology, Tehran, Iran ;
adenomyosis are associated in a large part of women. In endometriosis, an 7
Department of Reproductive Epidemiology Research Center- Royan Institute for
association with an increased incidence of miscarriage was demonstrated com- Reproductive Biomedicine- ACECR- Tehran- Iran, Department of Reproductive
pared to endometriosis-free women. The association between adenomyosis and Epidemiology Research Center, Tehran, Iran
miscarriages has also been shown. When endometriosis and adenomyosis are
associated, we do not know what is their share of responsibility in the occurrence Study question: Is gene expression profile of H19 lncRNA and IGF2 different
of miscarriages. in endometrial tissues of women with endometriosis in compare to normal
Study design, size, duration: An observational study nested in a cohort of endometrium?
women aged between 18-42 years who had undergone surgical exploration for Summary answer: Decreased H19 and IGF2 expression was showed in
benign gynecological conditions or who underwent assisted-reproductive-tech- eutopic and ectopic endometriual tissues of women with endometriosis.
nology at our institution between May-2005 and May-2018. Only women who What is known already: Endometriosis is one of the most common diseases
presented adenomyosis lesions visualized at uterine-MRI were retained for this in female reproductive system. Several factors, including genetic and epigenetic
study. Women who had never been pregnant before were excluded. The con- factors are suggested to be involved in its pathogenesis. Adhesion and proliferation
trol-group included patients presenting adenomyosis without lesions of endo- of endometrial tissue, are essential elements in the pathogenesis of endometriosis.
metriosis and the study-group patients with adenomyosis associated with Thus growth factors such as insulin-like growth factor-2 (IGF2) may be involved
endometriosis. as inducers of cellular proliferation. IGF2 gene is among the most regulated genes
Participants/materials, setting, methods: Data were collected preop- in endometriosis that play important roles in regulating growth and differentiation
eratively using a structured questionnaire. Among women who previously of endometrial cells. Another factor is imprinted lncRNA H19. It is known to be
conceived, the type and number of the different previous first trimester preg- involved in the regulation of cell proliferation and differentiation.
nancies outcomes were studied. Previous history of miscarriage was studied Study design, size, duration: In this case-control study, 5 endometriotic
according to the phenotype of adenomyosis (focal and/or diffuse lesions) and lesions (ectopic), 5 endometrial samples (eutopic) of women with endometriosis
the phenotype of endometriosis (superficial, ovarian and/or deep infiltrating and 10 endometrial control samples were analysed. Control samples were
lesions). obtained from women who had no evidence of endometriosis during laparos-
Main results and the role of chance: 214 pregnancies in ‘Adenomyosis copy in Royan Institute. Ectopic samples were obtained during laparoscopy
associated with endometriosis’ group and 53 pregnancies in ‘Adenomyosis- surgery. Control and eutopic endometrial samples were obtained by pipelle. All
affected women without endometriosis’ group were analysed. Previous miscar- women signed the informed consent form and did not receive any hormonal
riage rate was significantly higher in adenomyosis affected women with treatments during the last three months.
endometriosis compared to the controls (61/214 [28.5%] vs. 6/53 [11.3%], Participants/materials, setting, methods: After endometrial tissues col-
respectively; p = 0.009). After multivariable generalized estimating equation lection, RNA extraction and cDNA synthesis were done. Real-time PCR

technique was used for quantitative gene expression of H19 lncRNA and IGF2. detected in eutopic endometrial samples compared to controls, while this dif-
Gene expression data were analyzed based on 2 –ΔΔCT to estimate the relative ference was not statistically significant (P>0.05). In addition, the binding of ICER
fold change value. One-Way ANOVA was used for data analysis. P value less to CYP19 promoter II was statistically decreased in eutopic samples compared
than 0.05 was considered statistically significant. to the control endometrial group (P=0.005).
Main results and the role of chance: Gene expression profile of H19 and IGF2 Limitations, reasons for caution: Larger sample size for the confirmation
was decreased in eutopic and ectopic endometrial lesions compared with control of these data is needed. Epigenetic evaluation of ectopic endometrial tissues
samples. These decreases were not statistically significant (p>0.05). In addition, gene especially in CYP19 promoter II is recommended.
expression levels of these genes were lower in ectopic lesions in compare to eutopic Wider implications of the findings: The overexpression of CREM in the endo-
samples in endometriosis women. These differences were not significant. metriosis tissue samples and decreased binding of the inhibitory isoform of CREM
Limitations, reasons for caution: The main limitations of this study is the (ICER) to the CYP19 prompter II may contribute to the pathogenesis of endome-
small sample size. For getting more information, we are going to study these triosis via its regulatory role in the expression of estrogen biosynthesis enzymes.
genes in a large number of women with and without endometriosis. Trial registration number: not applicable
Wider implications of the findings: It seems altered gene expression of
H19 and IGF2 could play role in the development of endometriosis, because of P-320 Oral dydrogesterone compared to intravaginal micronized
their roles in regulatory pathways of cellular proliferation. After increasing sample progesterone for endometrial preparation in frozen-thawed
size, epigenetic analysis will be performed on the samples. embryo transfer cycles: preliminary results of a randomized
Trial registration number: - controlled trial
M. Cavagna1, L.C. Mattos de Macedo1, S. Rolim2, A. Dzik1, N.
P-319 Increased expression of CREM is associated with decreased Donadio1, H.S. Marçon1, M.D. Vasconcelos1, B. Merquires de
binding of its inhibitor (ICER) to the promoter region of CYP19 Araujo1, S.M. Franceschini Cinquetti1, A. Mantovani Fonseca1,
aromatase gene in endometriosis A.R. Rocha1
S. Kalantari1,2, A. Saadat Varnosfaderani1,2, F. Ramezanali3, E. 1
CRSM Hospital Perola Byington, Human Reproduction, Sao Paulo, Brazil ;
Amirchaghmaghi3,4, M. Shahhoseini5,6,7 2
Faculdade de Ciências Médicas da Santa Casa de Sao Paulo, Obstetrics and
1
Reproductive Biomedicine Research Center- Royan Institute for Reproductive Gynecology, Sao Paulo, Brazil
Biomedicine- ACECR., Department of Genetics, Tehran, Iran ;
2
Faculty of Basic Sciences and Advanced Technologies in Biology- University of Study question: Can we achieve similar reproductive outcomes assessed by ultra-
Science and Culture, Department of Cell and Molecular Biology Science, Tehran, sound scan at 12 weeks of gestation using oral dydrogesterone or vaginal micronized
Iran ; progesterone for endometrial preparation in frozen-thawed embryo transfer (FET)?
3
Reproductive Biomedicine Research Center- Royan Institute for Reproductive Summary answer: At present, our preliminary results are suggesting that
Biomedicine, Department of Endocrinology and Female Infertility, Tehran, Iran ; reproductive outcomes of FET are similar with both methods of progesterone
4
Cell Science Research Center- Royan Institute for Stem Cell Biology and supplementation (oral dydrogesterone versus vaginal progesterone).
Technology- ACECR, Department of Regenerative Biomedicine, Tehran, Iran ; What is known already: In recent investigations, it has been demonstrated
5
Reproductive Biomedicine Research Center- Royan Institute for Reproductive that oral dydrogesterone for luteal phase supplementation in fresh embryo trans-
Biomedicine- ACECR, Department of Genetics, Tehran, Iran ; fer is as effective as micronized progesterone regarding reproductive outcomes.
6
School of Biology- College of Science- University of Tehran, Department of Cell Nevertheless, nothing is known about FET cycles.
and Molecular Biology, Tehran, Iran ; Study design, size, duration: We are performing a prospective randomized
7
Royan Institute for Reproductive Biomedicine- ACECR, Reproductive Epidemiology study comparing the use of oral dydrogesterone and intravaginal micronized
Research Center, Tehran, Iran progesterone for endometrial preparation in FET cycles.
Participants/materials, setting, methods: So far, 44 patients were ran-
Study question: Is there any association between gene expression of CREM domly selected to receive either oral dydrogesterone, 40 mg/daily (DDG group,
and binding of its inhibitory isoform ICER to CYP19 promoter II (PII) in women n=22) or vaginal micronized progesterone, 800 mg/daily (VMP group, n=22),
with endometriosis? following endometrial preparation with transdermal estradiol. Patients who did
Summary answer: Overexpression of CREM was detected in the ectopic not reach an endometrial thickness ≥ 7 mm or who developed a dominant follicle
endometrium and the binding of ICER to CYP19 promoter II in eutopic tissues were excluded of the study. The main outcome was a viable ongoing pregnancy
was decreased. at 12 weeks of gestation assessed by ultrasound scans.
What is known already: Endometriosis is an estrogen-dependent disease. Main results and the role of chance: The median age of the patients was
The synthesis of estrogen depends on the expression of the aromatase enzyme 34 years (DDG group) and 32 years (VMP group). The median endometrial
(CYP19 gene). cAMP response element modulator (CREM) is a member of the thickness on the day of progesterone administration was 8.7 mm in DDG group
bZIP family of transcription factors that has an important role in the regulation and 9.2 mm in VMP group. In DDG group, one embryo transfer was performed
of aromatase gene expression. Inducible cAMP early repressor (ICER) is one of in 50% (n=11) of patients, two embryos were transferred in 40.9% (n=7) of
CREM inhibitory isoforms that represses cAMP-induced transcription. patients and three embryos were transferred in 9.1% (n=2) of patients. In VPM
Study design, size, duration: Endometriotic biopsies (ectopic), endometrial group, one embryo transfer was carried out in 36.4% (n=8) of patients and two
samples (eutopic) of women with endometriosis and endometrial control sam- embryos were transferred in 63.6% (n=14) of patients. Blastocyst stage transfer
ples were collected. Control samples were obtained from women who had no was performed in 81.8% (n=18) in DDG group and in 54.5% (n=12) in VMP
evidence of endometriosis during laparoscopy. Ectopic biopsies from endome- group. A positive pregnancy test performed 14 days after FET was achieved in
triosis women were obtained through laparoscopy while eutopic samples and 45.5% (n=10) in both groups. Ongoing pregnancy at 12 weeks of gestation was
control endometrial samples were collected by pipelle. Twelve endometrial observed in 36.4% (n=8) in DDG group and in 40.9% (n=9) in VMP group.
samples from each group were used for gene expression analysis of CREM, while Limitations, reasons for caution: The investigation is ongoing and these
6 control and 5 eutopic samples used for ChIP assay. are preliminary data. The number of patients is still small and the data must be
Participants/materials, setting, methods: Total RNA extraction and cDNA confirmed with larger number of patients and analysis of the reproductive out-
synthesis were done. Real-time PCR technique was used for quantitative gene expres- comes. At the present moment, no statistical analysis was performed and live-
sion of CREM. One-way ANOVA was used for data analysis. For protein-DNA birth rate was not still assessed.
interaction analysis, soluble chromatin was extracted and chromatin immunoprecip- Wider implications of the findings: The use of oral dydrogesterone may
itation (ChIP) coupled with Real-time PCR was performed to quantify the binding be a more patient-friendly approach for endometrial preparation in FET, avoiding
of ICER to CYP19 promoter II in control and eutopic endometrial samples. undesirable side effects and discomfort resulting from vaginal administration and
Main results and the role of chance: Gene expression level of CREM was providing similar reproductive outcomes.
statistically increased in ectopic lesions compared with control (P=0.001) and Trial registration number: UTN:U1111-1247-1845 – Registro Brasileiro de
eutopic (P=0.007) samples. Increased expression of the CREM gene was also Ensaios Clínicos

P-321 Intrauterine Granulocyte-Colony Stimulating Factor (IU Study question: Does PRP improve outcome of assisted conception in women
G-CSF) to improve the suboptimal endometrium: Is it worth a with unexplained recurrent implantation failure (RIF) or thin endometrium?
try? Summary answer: The use of PRP in women with unexplained RIF and thin
S. Ahuja1, Z. El Mahdi1, V. Shaikly1, M. Ranieri1, M. Taranissi1 endometrium has shown significant increase in chemical and clinical pregnancy
1 rates through assisted conception.
Assisted Reproduction and Gynaecology Centre, Assisted Conception, London, United
What is known already: PRP is the concentrate obtained by centrifugation
Kingdom
of the patient’s own whole blood. Platelets have granules containing numerous
Study question: Does intrauterine instillation of G-CSF improve the endo- proteins, several growth factors, and cytokines. In ex-vivo studies, activated PRP
metrial thickness in women with a sub-optimal endometrium (< 7 mm) going has been shown to promote the migration of endometrial stromal fibroblasts,
through an embryo transfer cycle? endometrial mesenchymal stem cells and bone marrow-derived mesenchymal
Summary answer: In this retrospective study of 32 women, the endometrium stem cells. This has formed the basis for in vivo use of autologous PRP to pro-
improved to over 7 mm in 71.8 % of cases following IU G-CSF. mote endometrial regeneration.It is also theorized that another mechanism of
What is known already: The thin endometrium is a challenge to treat in assisted PRP action is via anti-inflammatory processes involved in receptivity of the endo-
conception. Interventions to improve the endometrium include hysteroscopy to metrium and trophoblasts placentation. This would suggest that PRP could
diagnose and treat intrauterine pathology, estrogen tablets/patches, sildenafil improve implantation and clinical outcomes.
citrate orally/vaginally, vitamin E and l-arginine. Surrogacy is a last resort in women Study design, size, duration: This is a systematic review of available literature
where the endometrial factor shows no improvement, using the above strategies. including case series, cohort studies and randomised controlled trials (RCTs) on
G-CSF is a cytokine reported to improve the thin endometrium and implantation. the role of PRP in improving endometrial receptivity in assisted conception. The
There are few published studies that have reported a positive impact of IU databases were searched from inception to 2019. We then performed a
G-CSF on thin endometrium. This study adds to the existing literature. meta-analysis, which comprised of four prospective RCTs comparing use of PRP
Study design, size, duration: We report a retrospective study of 32 women intrauterine infusion or no treatment in women with unexplained RIF or thin
who were identified to have a sub-optimal endometrium of < 7 mm during the endometrium. There are 330 patients included in the meta-analysis.
treatment cycle over a period of 18 months. All the women were additionally Participants/materials, setting, methods: This is a systematic review of
on sildenafil citrate, aspirin and clexane to improve the endometrium. The available literature including case series, cohort studies and randomised controlled
women were counselled regarding the options of trying IU-GCSF versus cycle trials (RCTs) on the role of PRP in improving endometrial receptivity in assisted
cancellation versus proceeding with the transfer if the endometrium was con- conception. The databases were searched from inception to 2019. We then per-
sidered sub-optimal but suitable. formed a meta-analysis, which comprised of four prospective RCTs comparing
Participants/materials, setting, methods: Patients were selected for the use of PRP intrauterine infusion or no treatment in women with unexplained RIF
intervention if their endometrium was below 7 mm in late stimulation (1-2 days or thin endometrium. There are 330 patients included in the meta-analysis.
before HCG trigger) or in the late estrogenic phase (day 10-12) in a medicated Main results and the role of chance: The systematic review of current
frozen embryo replacement cycle with sufficient serum estrogen levels. Following literature suggests a benefit of PRP intrauterine infusion in improving assisted
informed consent, 30MU in 1 ml G-CSF was instilled into the uterus. The endo- conception outcomes. We performed a meta-analysis of available RCTs. Our
metrium was rechecked after 48 hours. A second dose was repeated if the inclusion criteria for the meta-analysis were: i) a prospective randomized con-
endometrium was still sub-optimal. trolled trial comparing PRP intrauterine infusion to controls in assisted concep-
Main results and the role of chance: Of the 32 patients, age 30-45 years, tion, ii) participants were characterized as having thin endometrium or RIF, and
who received IU G-CSF, 20 women were having fresh and 12 women were iii) all patients had a negative hysteroscopy to rule out uterine factors.
having frozen embryo transfer cycles. 7 patients had a history of uterine pathol- 134 relevant studies were identified. 92 were excluded as they did not meet the
ogy. The mean increase in endometrium was 1.23 mm with a range of 0 -3.7 inclusion criteria. 30 studies were reviewed for full text and 4 were included in
mm. The endometrium increased to > 7 mm in 71.8 % of cases. The transfer the meta-analysis.
was cancelled in 2 cases due to no change in the endometrium (6.6%). There The chemical and clinical pregnancy rate was reported in all 4 trials. The
were 8 pregnancies-3 have had livebirths, one of which was a set of twins, and chemical pregnancy rate from pooled data showed a significant difference in
one of them is about 30 weeks pregnant. 4 women had 1st trimester miscarriages. favour of PRP treatment with odds ratio of 2.14 (OR: 2.14, 95% CI: 1.49 – 3.07;
Only 1 woman complained of bone pain for about 1 week post IU G-CSF and P<0.0001). Similarly, the clinical pregnancy rate from pooled data showed that
her pregnancy sadly ended in a miscarriage. there was a significant difference between the two groups (OR: 2.58, 95% CI:
The women in this study comprised of patients with complex fertility problems 1.75 - 3.78; P <0.0001).
and previous treatment failures. Therefore, the positive and negative outcomes Two trials reported on change to endometrial thickness and miscarriage rate.
could have been due to other confounding actors such as embryo quality in the No significant difference was found for change of endometrial thickness or mis-
cycle rather than just the impact of the G-CSF on implantation. carriage rate, between the two groups.
Limitations, reasons for caution: The findings of this small retrospective Limitations, reasons for caution: The sample size in each RCT was relatively
study with no matched control, limit the conclusions that can be drawn. small and there was a lack of standardization in the PRP preparation. No trial
Furthermore, the women included in the study had complex fertility problems reported measures to avoid performance or detection bias. These factors will
that could have contributed to the success/failure of the cycle independent of impact the strength of evidence available.
endometrium thickness. Wider implications of the findings: These results support the evolving role
Wider implications of the findings: Treating the sub-optimal endometrium for PRP in improving endometrial receptivity in clinical practice. Nevertheless,
is one of the most challenging situations in assisted conception. This study sup- multi-centre RCTs with larger sample sizes are still needed to further evaluate
ports the evidence that IU G-CSF may have a positive impact. However, further the effectiveness of this intervention.
research is required to establish its benefits and safety, as an option for patients Trial registration number: Not Applicable
before they consider alternative options like surrogacy.
Trial registration number: Not Applicable P-323 Prevalence of chronic endometritis varies more by
diagnostic method than by reproductive history
M. Scarrone1, S. Bordoli1, V. Vanni2, L. Quaranta3, F. Pasi4,
P-322 Role of platelet rich plasma (PRP) in improving endometrial C. Patruno2, G. Campo2, V. Canti5, A. Quartucci2, M. Castiglioni4,
receptivity: A systematic review and meta-analysis. M. Candiani4, E. Papaleo3, P. Rovere Querini1, P. Giardina3
H.S. Naseem1, M. Lokman1, F. Soxibova2, L. Mohiyiddeen1, 1
Vita-Salute San Raffaele University, Internal and specialistic medicine, Milan, Italy ;
Y. Abdallah1 2
Vita-Salute San Raffaele University, IVF Unit, Milano, Italy ;
1
Manchester University NHS foundation Trust, Reproductive Medicine, 3
IRCCS San Raffaele Hospital, IVF Unit, Milan, Italy ;
Manchester, United Kingdom ; 4
IRCCS San Raffaele Hospital, Obstetrics and Gynecology, Milan, Italy ;
2
Medical University Sofia, Medical school, Bulgaria, Bulgaria 5
IRCCS San Raffaele Hospital, Internal and specialistic medicine, Milan, Italy

Study question: Does prevalence of chronic endometritis (CE) vary by repro- What is known already: Asymptomatic chronic endometritis (CE), diagnosed
ductive history or by diagnostic method used? by the presence of CD138-positive stromal cells on immunohistochemistry
Summary answer: CE is equally represented in patients affected by different (IHC), has been associated with recurrent implantation failure (RIF) and recurrent
types of reproductive failure. The different methods used for the diagnosis pregnancy loss (RPL). CE can be treated with antibiotics and has been linked
strongly influence its prevalence. with dysbiosis of the vaginal and endometrial microbiome. CD138 (syndecan 1)
What is known already: CE is an asymptomatic inflammation of the endo- is a selectively expressed heparan sulphate proteoglycan and putative cell surface
metrium mainly caused by bacteria. CE diagnosis is non-standardized and can marker of plasma cells. However, CD138 expression occurs in non-immune
be based on immunohistochemistry (IHC) for stromal endometrial CD138+ cells and has been associated with proliferation, apoptosis and angiogenesis. The
plasma cells or on endometrial cultures/molecular diagnosis for non-cultivable cycle-dependency and cellular distribution of CD138 expression in the endo-
microorganisms. CE might be associated with impaired receptivity and repro- metrium has not yet been analysed comprehensively.
ductive failure, thus emerging as a potential factor in the pathogenesis of unex- Study design, size, duration: Observational cohort study of (i) 27 prolifer-
plained infertility, repeated implantation failure (RIF) and recurrent pregnancy ative and 29 luteal phase endometrial samples and 3 paired proliferative/luteal
loss (RPL). CE prevalence is inconsistent among studies, possibly due to different phase endometrial samples; (ii) interrogation of published single-cell RNA-seq
diagnostic methods and vaginal contamination of cultures. data derived from 6 midluteal endometrial biopsies (GSE127918).
Study design, size, duration: We performed a prospective study of patients Participants/materials, setting, methods: Timed endometrial biopsies
referred to the hysteroscopic service of the San Raffaele Hospital (Milan, Italy) were obtained from local biobanks from control and recurrent reproductive
between May 2018 and December 2019 for diagnostic hysteroscopy and CE failure patients. Endometrial SDC1 (CD138) transcript levels were quantified by
diagnosis as a second-level investigation for unexplained infertility (n=15), RIF RT-qPCR and cellular distribution examined by single-molecule in-situ hybrid-
(n=28) or RPL (n=44). We stratified CE prevalence by diagnostic method in ization (smISH). Temporo-spatial expression of CD138 at protein level was
addition to reproductive history. examined by IHC followed by digital image analysis and deconvolution for quan-
Participants/materials, setting, methods: Patients underwent endometrial tification. Dual staining with Ki67 was performed to delineate CD138-positive
culture and concomitant vaginal and cervical cultures to account for possible proliferating cells. Single-cell transcriptomics data were interrogated to examine
contamination during sampling. A subgroup of n=43 samples were also subjected cell type-specific SDC1 expression.
to immunohistochemistry for CD138+ plasmacytes. Diagnosis of CE was done Main results and the role of chance: Whole tissue RT-qPCR demonstrated
in the presence of either a) positive endometrial culture/molecular bacterial non-immune SDC1 expression across the menstrual cycle with no significant
DNA amplification or b) >=5 CD138+ plasmacytes in 20 high-power fields. difference in transcript levels across cycle phases. Analysis of midluteal scRNA-
Affected patients were treated with targeted antibiotic therapy and re-tested seq data showed enriched SDC1 mRNA expression in endometrial epithelial
with endometrial culture to assess CE regression. cells, although transcripts were also detectable in stromal, immune and endo-
Main results and the role of chance: N=50 out of 87 cultures resulted thelial cell populations. The cellular distribution of SDC1 transcripts was con-
positive (57.5%). In only 2 out of 50 cases, the same pathogen was isolated in firmed by smISH. IHC of proliferative endometrium demonstrated very high
both endometrial and vaginal cultures - with a concordance rate of 4% - sug- stromal CD138 expression (>200 CD138-positive stromal cells/10mm2) in
gesting that vaginal contamination is infrequent. IHC was positive in 12 out of 26/27 samples. Expression was significantly reduced in luteal samples (Mann
43 cases (27.9%). Concordant results between culture and IHC were observed Whitney U-Test; P<0.005). This transition was also demonstrated across the
in 25 cases, with a matching accuracy of 58,1%. Cases with contradictory results paired proliferative/luteal samples. Dual IHC demonstrated a mean co-locali-
mostly showed positive bacterial cultures but negative immunohistochemical sation of 30% of CD138-positive stromal cells with the proliferation marker
analysis (n=16 out of 18, 88.9%) while only n=2 out of 18 (11.1%) resulted Ki67 across the menstrual cycle, suggesting that CD138 is not merely marking
positive at IHC but negative at culture. Of the n=22 affected patients who have proliferating cells. H-scoring of the glandular endometrium demonstrated a sim-
completed treatment and control cultures at the time of data analysis, n=9 ilar drop in expression from the proliferative to luteal phase (Mean H-Score
resulted negative (40.9%), n=6 (27.3%) tested positive for the same pathogen 197.51 vs. 104.56; Student T-test; P<0.05).
whereas n=7 (31.8%) tested positive for a different pathogen. We observed Limitations, reasons for caution: The marked reduction in CD138 immu-
similar prevalence of CE among groups of patients stratified by reproductive noreactivity upon transition from the proliferative to luteal phase was not
history as defined by RIF (n=17/28, 60.7%), RPL (n=23/45, 52.1%) or unex- reflected by a corresponding decrease in SDC1 mRNA levels in whole biopsies.
plained infertility (n=10/15, 66.7%). This discrepancy is likely due to post-transcriptional regulation of CD138 how-
Limitations, reasons for caution: The currently small sample size is the ever, further work is required to elucidate this.
main limitation of our study. Further prospective data collection and inclusion Wider implications of the findings: Detection of CD138-positive stromal
of pregnancy outcomes will be of interest. cells is used increasingly for the diagnosis of CE. It is important that CD138 is
Wider implications of the findings: Endometrial cultures are frequently assessed in the luteal phase to prevent over-diagnosis, that non-immune cell
positive in patients with reproductive failure. Our study shows that vaginal/ expression is acknowledged, and its clinical importance evaluated.
cervical contamination has marginal role in this finding. However, only 58.1% of Trial registration number: Not applicable
culture-positive patients show the presence of plasmacytes. Whether bacteria
mediate endometrial dysfunction in the absence of plasmacytes-mediated inflam- P-325 Presence of focal adenomyosis lesion based on MRI is
mation needs further investigation. associated with infertility issues
Trial registration number: not applicable L. Melka1, M. Bourdon1, J. Oliveira1, J. Cormier1, L. Marcellin1,
C. Maignien1, C. Bordonne2, A.E. Millischer3, G. Plu-Bureau1,
P-324 The importance of timing in detection of asymptomatic P. Santulli1, C. Chapron1
CD138 diagnosed chronic endometritis 1
Université de Paris- Faculté de Médecine- Paris- France, clinician, Paris, France ;
J. Odendaal1,2, K. Fishwick1, Q. Hussain1, J. Thornton1, K. Larsen1, 2
Université de Paris- Faculté de Médecine- Paris- France, Radiology, Paris, France ;
P. Vrljicak1, E. Lucas1, J.J. Brosens1,2, S. Quenby1,2 3
Centre de Radiologie Bachaumont- IMPC- Paris- France, radiology, paris, France
1
University of Warwick, Division of Biomedical Sciences, Coventry, United
Kingdom ; Study question: Is there an association between adenomyosis and infertility,
2
University Hospitals Coventry and Warwickshire, Biomedical Research Unit, according to the adenomyosis phenotype as diagnosed by magnetic resonance
Coventry, United Kingdom imaging (MRI)?
Summary answer: In the study population, the presence of focal adenomyosis
Study question: How does the expression of CD138 alter across the men- was associated with primary infertility.
strual cycle? What is known already: Adenomyosis is characterized by the presence of
Summary answer: CD138 is normally expressed, at mRNA and protein level, endometrial glands and stroma deep within the myometrium. This disease exhib-
in glands and stroma during the proliferative phase but expression decreases its various clinical presentations notably infertility, which prevalence and causes
markedly during the luteal phase. are still unknown. In addition, various forms have been described, including focal

(Foc-ADE) and diffuse adenomyosis (Dif-ADE), and several evidences suggest parts:one was fixed in 10% formaldehyde for histopathological analysis, the other
that those two forms could be considered as distinct entities, which could make piece was fixed in 2.5% glutaraldehyde for scanning electron microscopy (SEM) and
the analysis of adenomyosis-related infertility even more difficult. As a matter the third part in PBS was taken for gene expression detection using qPCR analysis.
of fact, the association between the distinct adenomyosis phenotypes (focal Main results and the role of chance: Of 42 samples 33 were obtained from
and/or diffuse forms) and infertility has not yet been investigated. women in the natural menstrual cycle and 9 samples were taken after oocyte
Study design, size, duration: This was a cross-sectional study using data retrieval, moreover 9 women also had been sampled in the natural cycle prior
prospectively collected in all non-pregnant patients aged between 18 and 42 to ovarian stimulation start. The expression of URG in edometrium cells was
years, surgically explored for benign gynecological conditions at our institution higher in stimulated cycles (n=7), but statistically it wasn’t significantly different
between May 2005 and May 2018. For each patient, a standardized questionnaire (SSD) (p>0,05), the expression pattern of the DRG was the same in natural and
was completed during a face-to-face interview conducted by the surgeon during stimulated cycles. Histopathological analysis of each sample also was similar and
the month preceding surgery. Only women with uterine MRIs performed by a showed type of tissue structure appropriate early secretion.
senior radiologist during the preoperative work-up were retained for this study. According to the results of SEM, all samples were divided into 3 groups: (A)
Participants/materials, setting, methods: A uterine-MRI was performed no signs of pinopodes formation (n=31), (B) increased number of ciliary cells
in 496 women operated for a benign gynecological disease. Among them, 248 without pinopodes formation (n=9) and (C) the presence of pinopodes in the
women did not present adenomyosis lesions and 248 women had a radiological samples (n=2). Group A had pattern with very low expression of URG and
diagnosis of adenomyosis. According to MRIfindings, women were diagnosed higher DRG. Group B samples demonstrated higher expression of URG and
with Foc-ADE and/or Dif-ADE forms. Three groups were compared: the non-in- high expression of some DRG. And group C samples showed high level expres-
fertile women group (n=361), the ‘primary infertility’ group (n=84) and the sion of URG and low expression of DRG similar to expression profile during
‘secondary infertility’ group (n=51). Univariate and multiple analysis were per- receptive phase; that showed a SSD calculated by Chi-square on levels of gene
formed to determine primary and secondary infertility-associated factors. expression relative to groups A and B (p<0,001).
Main results and the role of chance: The presence of focal adenomyosis was Limitations, reasons for caution: Tracking changes in endometrial mor-
significantly associated with primary infertility. Diffuse adenomyosis was not found phology using SEM and specific genes expression is a useful tool in predicting of
to be associated with infertility, whether at the primary or at the secondary stage. embryo implantation. But these indicators are very variable depending on the
The distribution of other benign gynecological diseases, notably endometriosis or individual characteristics of the patient as well as the selected drugs and dosages
leiomyomas, was not significantly different between the groups. After a multinomial correctness in the cycles of ovarian stimulation.
regression model including notably the women’s age and the presence of endo- Wider implications of the findings: A study of the morphology and gene
metriosis or leiomyoma, the presence of focal adenomyosis was identified as an expression of endometrial samples obtained from donor is the basis for assessing
independent associated factor of primary infertility (1.9; 95% CI: 1.1 - 3.3). the status of the endometrium in patients with infertility, which allows us to more
Limitations, reasons for caution: Inclusion of only surgical patients may con- accurately understand the problem of implantation failure and adjust treatment
stitute a possible selection bias. In addition, it cannot be excluded that other non– in the right direction
explored causes of infertility were present among the infertile women-group. Trial registration number: Not applicable
Wider implications of the findings: This study opens the door to future
clinical and basic studies aiming at better characterizing focal adenomyosis and P-327 Bowel endometriosis-related infertility: assisted
its infertility-related physiopathology reproductive technology (ART) outcomes are favorable in women
Trial registration number: NA with no prior history of surgery for endometriosis
C. Maignien1, P. Santulli1, M. Bourdon1, L. Marcellin1, K. Pocate-
P-326 The correlation between gene expression in endometrial Cheriet2, J. Gonnot1, V. Blanchet1, C. Chapron1
cells and the formation of pinopodes 1
Assistance Publique-Hôpitaux de Paris AP-HP- Centre Hospitalier Universitaire
I. Ilyin1, O. Parnitskaya2, N. Kazachkova3, J. Gontar3, Y. Gerevich1, CHU Cochin, Department of Gynaecology Obstetrics II and Reproductive
E. Kapustin1, N. Buderatskaya2, S. Lavrynenko2, Y. Lakhno3 Medicine, Paris, France ;
1 2
Medical Centre IGR, ART department, Kyiv, Ukraine ; Assistance Publique-Hôpitaux de Paris AP-HP- Centre Hospitalier Universitaire
2
Medical Centre IGR, Embryology, Kyiv, Ukraine ; CHU Cochin, Department of Reproductive Biology, Paris, France
3
Medical Centre IGR, Diagnostic Laboratory, Kyiv, Ukraine
Study question: To evaluate the ART cumulative live birth rates (CLBR) in a cohort
Study question: Do different patterns of gene expression affect morphology of bowel endometriosis (OSIS) patients with no prior history of surgery for OSIS
of the endometrial tissue in natural or stimulated cycles? Summary answer: ART CLBR are excellent (67.3%) in bowel OSIS patients
Summary answer: A specific pattern of gene expression contributes to the for- with no prior history of surgery. No bowel complications occurred during ovarian
mation of a certain endometrial morphology, such as the appearance of pinopodes. stimulation or pregnancy
What is known already: Well known that recurrent implantation failure is What is known already: Bowel OSIS-related infertility is a therapeutic chal-
tightly related with endometrial cells function. Endometrial structure and recep- lenge, with 2 main treatment modalities: surgery and ART, each with different
tivity are result of gene expression in certain period of menstrual cycle. Many risks of complications. Numerous reports have been published about ART preg-
investigations indicated main genes of endometrial receptivity regulation in sam- nancy rates after bowel surgery, but data are currently lacking on ART outcomes
ples of infertile women. But it is equally important to understand how genes are in bowel OSIS affected-women who reverted to ART first. Moreover, the deter-
expressed in the known morphology of endometrial tissue in fertile young minant factors of pregnancy chances in this specific population remain unclear
women, such as oocyte donors in the natural menstrual cycle and during ovarian Study design, size, duration: Observational cohort study on 101 consecutive
stimulation. Since there is an opinion that hormonal stimulation is ahead of the bowel OSIS patients without a prior history of surgery for OSIS, who underwent
endometrium development in comparison with the natural cycle. IVF-ICSI treatment at Cochin-Port-Royal Hospital between January 2013 and
Study design, size, duration: The study was performed in the Medical Centre December 2018. Diagnosis of OSIS was based on published imaging criteria
IGR from March 2018 to December 2019. It involved 42 endometrial samples using transvaginal sonography and magnetic resonance imaging. Patients with a
obtained from 32 oocyte donors whose mean age was 27.9±2.8 (range 22-32 prior history of surgery for OSIS were excluded
years). The parts of tissue sample were examined for detection of pinopodes, Participants/materials, setting, methods: The main outcome measure
tissue structure and expression of following genes generally upregulated during was CLBR. Miscarriage rates, perinatal outcomes and bowel complications were
receptive phase (URG) PAEP,GPX3, TAGLN,EDNRB, CLU, HABP2,LMOD1, IMPA2, also studied. We compared the characteristics of women who became pregnant
GAS, FXYD2 and such downregulated genes (DRG) as HLA-DOB, CTNNA2, and those who did not, using univariable and multivariable analysis, to identify
CAPN6, MAP2K6, NDRG2,SORD. determinant factors of fertility outcome
Participants/materials, setting, methods: The whole endometrial tissue sample Main results and the role of chance: One hundred and one bowel OSIS
was received by pipelle biopsy from 32 women on the mid-luteal phase of the natural patients underwent 176 ART cycles. The mean number of deep infiltrating endo-
menstrual cycle or on the day of oocyte pick up. Each biopsy was separated into three metriosis lesions per patient was 3 ± 0.9, with a mean number of bowel lesions

of 1.3 ± 0.6. Seventy-three percent of the patients had associated endometri- Limitations, reasons for caution: retrospective study, selection bias
omas, and 88.1% had associated adenomyosis. Overall, the cumulative live birth (Two referral centers of endometriosis surgery), difference between the
rate was 67.3%. No bowel complication occurred during ART cycles or preg- 2 groups ( 3 for 1 pairing after PSM)
nancies. Regarding perinatal outcomes, cesarean sections were performed in Wider implications of the findings: Pregnancy and live birth rates were
38.2% of the pregnancies; there were 7.4% preterm births (< 37 WG) and 10.3% significantly higher in the surgery first group compared to the IVF first group,
low birthweight (< 2500 g). Using multivariate analysis, AMH levels <2 ng/mL especially for the subgroup of patients with good prognostic factors (AMH>
(p < 0.001) and antral follicle count < 10 (p = 0.006) were the only factors 2 ng / mL, age <35 years and no adenomyosis).
associated with negative ART outcomes This study highlights a good live birth rate after surgery for the spontaneous
Limitations, reasons for caution: The diagnosis of OSIS was based on pregnancy, with a median time of 6 months.
imaging rather than surgery. This limitation is however inherent to the design of Trial registration number: not applicable
the studies on OSIS patients reverting to ART first
Wider implications of the findings: CLBR are excellent in bowel endome-
triosis patients undergoing ART first, with low risks of complications. Low ovarian
POSTER VIEWING SESSION
reserve seems to be the main prognostic factor of ART CLBR. These data urge
the clinician to carefully weight the pros and cons before systematically referring ETHICS AND LAW
infertile bowel OSIS patients to radical surgery
P-328 Deep endometriosis with digestive involvement :

Comparison of the surgical strategy and ATR P-329 Analysis of the outcomes chosen by european patients for
their surplus embryos, taking into account all possible options
R. Goldstein1, S. Bendifallah2, S. Touleimat3, H. Roman3, F. Krief4,
available in Spain
M. Ballester5, E. Darai1,2, 2. Bendifallah.S(1
1 M. Lopez-Teijon1, V. Walker1, C. Castello1, B. Marques1, S. Novo1,
Department of Gynaecology and Obstetrics, Tenon University Hospital, Assistance
F. Garcia1, A. Garcia-Faura1
Publique des Hôpitaux de Paris ;
1
2
INSERM UMRS 707, Epidemiology, Information Systems ; Institut Marques, Reproductive Medicine Service, Barcelona, Spain
3
Expert Center in the Diagnosis and Multidisciplinary Management of
Study question: What outcome do patients choose for their surplus frozen
Endometriosis, Department of Gynecology and Obstetrics, Rouen University
embryos?
Hospital, Rouen ;
4 Summary answer: Only 13,3% of patients who have surplus embryos make
Hôpital Jean verdier, Department of gynecology-obstetricics, Bondy ;
5 an active choice about their final outcome. 86,7% don’t respond, or keep the
Hôpital Diaconesse, Department of gynecology-obstetricics, Paris ;
6 embryos for themselves.
Hôpitaux de Saint Maurice, Department of gynecology-obstetricics, Saint Maurice
What is known already: Fertility clinics continue to accumulate frozen
Study question: To compare the fertility results of two therapeutic strategies embryos. Deciding what to do with surplus embryos can be difficult for patients
for colorectal endometriosis associated-infertility :surgery followed by sponta- and can create significant emotional conflict.
neous conception+ / - IVF (In vitro fertilization) versus IVF in first intention. It’s not that patients have an irresponsible attitude towards their embryos; rather
Summary answer: For women with colorectal endometriosis associated-in- it appears to be the long-reaching implications that their decision might have.
fertility, first-line surgery seems to be a good option, especially for patients with Patients don’t want to give the embryos to other women for fear their children
good prognostic factors. might one day meet their siblings; they don’t want to destroy them because it
What is known already: surgery for deep endometriosis improved fertility. makes them feel bad, and they don’t want to donate them to research because
Regarding endometriosis with digestive involvement, data in the literature are they are frightened of what might be done to them.
scarce. Their is no consensus for the management of those patients. . Regarding Study design, size, duration: In accordance with Spanish law, patients sign
endometriosis with digestive involvement, data in the literature are scarce. Their a consent form confirming their desired outcome for their surplus embryos. All
is no consensus for the management of those patients. options are open to them, and every 2 years after freezing the patients receive
Study design, size, duration: Retrospective cohort study conducted in hos- a letter asking them to reconfirm or alter their previous decision.
pital Tenon in Paris and in the University hospital of Rouen, two referral centers We assessed the informed consents from 7607 patients doing IVF (own eggs
in endometriosis. Patients: 159 patients were inclued between January 2010 and and donor eggs) between 2012 and 2016 with surplus frozen embryos. Patient
December 2016 follow-on replies up to 2018 were analysed.
Participants/materials, setting, methods: The patients were matched after Participants/materials, setting, methods: Patient consent forms and
using a propensity score (PSM). The propensity score was developed using a replies were analysed and the chosen outcome for the embryos noted. We
logistic regression model, with the following covariates : age, anti-mullerian hor- compared the replies given by patients from different european countries and
mone (AMH) and the presence of adenomyosis.113 patients were included in also the decisions made by patients regarding embryos made from their own
the surgery first group and 46 patients in the IVF first group. Patients follow-up gametes or from donor gametes.
was done for a period of at least 24 months after surgery or the first IVF cycle. Main results and the role of chance:
Main results and the role of chance: After PMS, The pregnancy rate was 1. B. 41% wished to maintain the embryos for their own use. 90% of these
76% (86/113) in the primary surgery group (including 33.7% (29/86) after spon- patients had completed their family, they didn’t want any more chil-
taneous conception and 66.3% (57/86) after IVF), versus 30.4% (14/46) in the dren but they still preferred to keep their surplus embryos.
IVF first group. The live birth rate was 50.4% (57/113) in the primary surgery C. 2% destroyed the embryos
group (including 45.6% (26/57) after spontaneous conception and 54.4% D. 3% gave them away to others
(31/57) after IVF), versus 23.9% (11/46) in the IVF first group. Overall the 182 E. 3% donated them to research
cycles performed, the cumulative rates of live birth after the first IVF cycle in
the surgery first group compared to the IVF first group were 29.6% versus 15.2%, There were no significant differences in the results according to the biological
for the second cycle 44.2% versus 28.6% and for the third cycle 69.6% versus origin of the embryos. There were though notable differences between the
40.5%.For patients oriented towards spontaneous conception attempt after options chosen by patients from countries in close geographical proximity, which
surgery, the pregnancy rate was 69% (29/42) and the live birth rate 61.9% highlights the idiosyncrasies of each culture.
(26/42), with a median time of 6 months [IQR: 5-11 months] for pregnancy.
Pregnancy and live birth rates were significantly higher in the surgery first group Ireland: A 45%, B 45%, C 1,5%, D 7%, E 1,5%
compared to the IVF first group, especially for the subgroup of patients with good UK: A 63%, B 29%, C 3%, D 3%, E 2%.
prognostic factors (AMH> 2 ng / mL, age <35 years and no adenomyosis). Germany: A 36%, B 51%, C 6,5%, D 6%, E 0,5%.

France: A 61%, B 25%, C 7%, D 4%, E 3%. collection, transport and taking responsibility of the biological material, acknowl-
Spain: A 56%, B 33%, C 2%, D 3%, E 6%. edgment of samples arrival, acknowledgement of any adverse event occurred.
Limitations, reasons for caution: SIERR will evaluate the validity of these
Limitations, reasons for caution: The study did not differentiate between modules through several pilot SIERR affiliated ART centres in order to evaluate
replies from couples or single patients. Therefore in the majority of cases it is a the concrete application and usefulness of the forms.
joint decision made by the couple, and maybe if just one partner was asked to Wider implications of the findings: Transfer of cryopreserved samples
decide the results would have been different. between centers implies effective and appropriate on time communication in order
Wider implications of the findings: When couples finish their reproduc- to establish the approaches of transport itself, in terms of ethical, legal and practical
tive project their surplus embryos create a problem they do not know how management. It is increasingly essential to share a homogeneous procedure that
to resolve. guarantees the management of cryopreserved gametes and embryos.
If they don’t answer the letters sent to them, embryos are considered aban- Trial registration number: not applicable
doned. Spanish law allows fertility clinics to choose the outcome for these
embryos, that can include entering the embryo-adoption program. P-331 PGT-A in any Egg Donation
Trial registration number: not applicable A. Anastasi1, F. Capodanno1, S. Rossi1, A. Gallinelli1
1
AUSL di Ferrara - Ospedale del Delta di Lagosanto, Centro di Fisiopatologia della
P-330 Import/export of cryopreserved gametes and embryos Riproduzione Umana, Ferrara, Italy
between European centers: what should be done, which forms,
which protocols, which responsibilities: an Italian experience Study question: Why Pre implantation Genetic Test for Aneuploid diseases
C. Garello1,15, B. Dal Canto2,15, E. Baldi3,15, M. Cervi4,15, (PGT-A) should be routinely performed in Egg Donation (ED)?
S. Dabizzi5,15, D. Farace6,15, D. Giacchetta7,15, M. Menegazzo8,15, Summary answer: Only euploid embryos should be routinely transferred
M.G. Minasi9,15, M. Oneta10,15, L. Rienzi11,15, G. Taliani12,15, after ED. Following a stressful and expensive ED, embryo transfer with
D. Paoli13,15, L. De Santis14,15, On behalf of SIERR15 euploid-blastocyst should be strongly recommended.
1
LIVET s.r.l., IVF Unit, Torino, Italy ; What is known already: Data from EIM for ESHRE, suggest that 39 Countries
2
Zucchi Clinical Institutes, Biogenesi Reproductive Medicine Center, Monza, Italy ; and 1.279 Institutions perform ART, with a total of 56.516 ED. No informations
3
University of Florence, Experimental and Clinical Medicine, Florence, Italy ; about ED following embryo-transfer with euploid-embryos are available. Even if
4
Healthcare “Friuli Occidentale”, IVF Unit, Sacile PN, Italy ; egg-donors are young, aneuploidy increases in women older than 26. A slightly
5
Careggi Hospital, Unit of Andrology- Female Endocrinology and Gender increased prevalence of embryo aneuploidy is reported in young patients,
Incongruence, Florence, Italy ; with >40% aneuploidy in women 23 years and less. Aneuploidy can not be excluded
6
University of Rome Tor Vergata, Department of System Medicine, Rome, Italy ; in young egg donors.
7
San Paolo Hospital, Laboratory of Seminology and male gametes cryopreservation, IVF Centers discourage PGT-A following ED, without any scientifically reason.
Milan, Italy ; We speculate that PGT-A in ED is not perfomed with the questionable aim to
8
University Hospital, Andrology and Reproductive Medicine, Padova, Italy ; guarantee fresh and following frozen-embryo-transfer.
9
European Hospital, Center for Reproductive Medicine, Rome, Italy ; Study design, size, duration: To perform PGT in all ED, offering to the patients
10
Sacco Clinical Sciences Institute, Reproductive Unit Department of Obstetric and the higher chances to transfer an euploid embryo. It would be advisable to set a
Gynaecology, Milan, Italy ; duration of 2 years in order to evaluate the cost-effective of this approach.
11
Clinica Valle Giulia, Genera Centres for Reproductive Medicine, Roma, Italy ; Participants/materials, setting, methods: The are 39 Countries and 1.279
12
Sapienza of Rome University, Umberto I Hospital, Rome, Italy ; Institutions offering ART services, 56.516 treatment cycles with egg donation
13
Sapienza University of Rome, Laboratory of Seminology-Sperm Bank “Loredana (ED) where the study could be conducted.
Gandini”, Rome, Italy ; Main results and the role of chance: The main goal is to establish a clinical
14
San Raffaele Scientific Institute, Ob/Gyn IVF UNIT, Milan, Italy ; setting that it offer to the couples submitted to ED the highest chances to transfer
15
SIERR, Italian Society of Embryology- Reproduction and Research, Italy, Italy one euploid embryo, optimizing the costs of the procedure. The second objec-
tive is the cost-effective evaluation of the policy we suggest: cost-effectiveness
Study question: is standardization of National and International procedures analysis is an hot topic in the debate about clinical application of PGT-A, but we
and forms for cryopreserved samples transfer sufficient to meet quality, trace- assume that the high cost of new techniques for the genetic screening such as
ability and safety requirements? the Next Generation Sequencing (NGS), could be surely covered by high number
Summary answer: Indications on shipping between tissue institutions and of couples seeking PGT after ED. Recent studies suggest a positive cost-effective
legal responsibilities are important points and a working protocol with shared ratio only in women older than 35 years old. However, no data are available
transport forms has been defined. regarding cost-effectiveness of PGT-A in younger patients or in those submitted
What is known already: In Italy, the transport of cryopreserved biological to ED. In our view, PGT-A could allow to optimize the cost of ED, allowing to
material is dealt exhaustively by several Legislative Decree (No. 191/2007;16/2010 the couples to pay for euploid embryos, and not for the aneuploid ones. The
and D Lgs 10 October 2012) which transpose European Directives. Given the question we should make us in term of costs is the following: how much any
nature of their application, the transport of reproductive cells has peculiar quality aneuploid embryo cost respect to any euploid one?
and safety requirements that must be applied univocally, minimizing the chance of The unique way to support couples hopeful having euploid gametes is PGT-A,
error. In order to standardize the transborder shipping procedure to meet the especially in ED where financial and psychological investment of infertile women
quality, traceability and safety criteria for cells and tissues, it is appropriate to estab- are consistent.
lish a unified process using the same tools, forms and communication channels. Limitations, reasons for caution: PGT-A following ED could flow in the indis-
Study design, size, duration: The Italian Society of Embryology, Reproduction criminate employment of this technique, with a consequent uncontrolled business.
and Research (SIERR) has defined a working protocol with shared forms in order to PGT-A following ED is difficult to adopt because of financial, social and orga-
distribute them nationally and internationally. This is necessary in light of the increas- nizational reasons.
ingly widespread movement of biological samples between the various countries. Wider implications of the findings: Optimization of financial and psycho-
Participants/materials, setting, methods: A working group has been created logical investment of couples submitted to ED.
by SIERR. This “FOCUS Group” was constituted by representatives from Italian To get more data about aneuplody frequency in egg donors.
ART centres and Sperm Banks who worked together to define joint procedural Trial registration number: not applicable
steps and create specific forms to support movement of cryopreserved samples.
Main results and the role of chance: The FOCUS Group identified the critical P-332 Ethical concern regarding the assisted reproduction
steps in the communication procedures between centres and created the related techniques: a qualitative interview study with professionals,
forms: patient authorisation, request from the recipient centre, critical checks patients, donors and general population
carried out by both sending and recipient centres, start of samples transfer, J. Guijarro1, R. Nuñez-Calonge2

1
Hospital Virgen de la Luz- Cuenca, Obstetricia y Ginecología, Cuenca, Spain ; and how it could remedy gender inequity and enhance reproductive autonomy.
2
Duncan, Ethics and reproduction, Madrid, Spain So far, the morally relevant connection between ectogenesis and the fetus as a
beneficiary of treatment is largely overlooked. Ectogenesis could provide easier
Study question: Is there a different ethical consideration regarding the assisted access to the fetus and thus contribute to the allocation of a patient status to
reproduction techniques of professionals compared to patients, donors and the fetus.
general population? Study design, size, duration: A literature study of the scientific literature
Summary answer: There is a different moral consideration towards the prob- was performed to inventory the state-of-the-art of artificial womb technology
lems derived from assisted reproduction techniques, being higher in physicians and a literature study of the respective bioethical literature was performed to
and laboratory personnel and lower in donors. inventory the main ethical arguments. Next, ethical problems in prenatal med-
What is known already: Assisted reproduction techniques (ART) have icine were conjoined with normative argumentation to arrive at substantiated
opened numerous ethical questions different from religious or legal approaches, ethical judgements with regards to the ethics of ectogenesis and decision making
involving not only the professionals who perform them but also patients and in prenatal medicine.
donors. Additionally, many ethical challenges generated by new scientific devel- Participants/materials, setting, methods: Literature study, conceptual
opments in this field generate widely differing opinions. analysis, normative analysis.
To date, there are no studies comparing the concerns of the different moral Main results and the role of chance: Suggestions have been made that
agents involved in these processes and whether the ethical issues raised are the ectogenesis could make it easier for clinicians to fix fetal defects by allowing to
same in all these groups. treat the fetus outside the woman’s body, inside the artificial womb. Combined
Study design, size, duration: The plan was set-up as an electronic qualitative with social expectations about maternal responsibilities, this may put pressure
survey study and took place between September and December 2018. on pregnant women to undergo prenatal interventions for the benefit of the
Participants/materials, setting, methods: Data were collected from 123 future child. However, despite an expected better fetal accessibility, concerns
voluntarily participating assisted reproductive centres (including clinicians, embry- for the pregnant woman’s wellbeing still remain in case of surgical removal and
ologists, nurses and administrative staff ) and 110 non-professionals (oocyte and physical translocation of the fetus to the artificial uterus. While a pregnant
sperm donors, patients and general population). Participants filled out an anon- woman may have obligations of beneficence towards the fetus, she may nev-
ymous questionnaire asking for their socio-demographic characteristics and 100 ertheless refuse interventions, not only because of the value of autonomy, but
questions based on the main ethical problems. After a depuration process by also that of bodily integrity. An argument can be made that a focus on fetal
means of Factorial Analysis a definitive questionnaire with 41 items was obtained. beneficence should not disregard potential cases of autonomy violation as
Main results and the role of chance: Based on the interviews, we articulated connected with a person’s life goals and personal interests (like the woman’s
four main topics describing participants views regarding the ethical aspects of wish to carry the fetus in the womb to birth). As such, ectogenesis aided fetal
assisted reproduction: solidarity, morality, religiosity and confidence. Comparing treatment could induce higher tensions between a clinician’s obligations towards
the average of scores in the four issues according to the professional relationship the pregnant woman and the future child. It is defended here that ectogenesis
with assisted reproduction it was concluded that the professional / non-profes- would not circumvent the key ethical concerns that come with in utero fetal
sional dichotomy is the one that occupies more extreme values: the professionals treatment.
display a greater preoccupation by the morality, a greater confidence and a more Limitations, reasons for caution: Human ectogenesis is still hypothetical,
restrictive attitude, without influence by religiosity. Conversely, non-professionals especially the variant where the offspring’s development happens from concep-
present a more solidary and mistrustful profile, with greater disregard for moral tion to birth completely outside a woman’s uterus. This paper focuses on partial
issues. Within the group of professionals, doctors have a greater moral concern ectogenesis, meaning that ‘ectogenesis’ will generally refer to the transfer of a
than the rest and a less supportive attitude. partially developed fetus to an artificial womb for further development.
People without a direct relationship with reproduction have a more supportive Wider implications of the findings: The availability of ectogenesis aided
and distrustful profile, similar to the patients, but clearly present a greater indif- fetal treatment could open the door for routine directive counselling for fetal
ference for moral problems. Donors are the group that presents less concern benefit. With the advent of artificial womb technology, it is timely to inquire
for ethical or moral aspects and greater confidence in medical decisions. whether actual clinical codes of practice are sufficiently fine-grained to provide
Limitations, reasons for caution: The non-existence of religious differences ethical guidance to the future practice of prenatal medicine.
in the surveyed population may have led to a bias in some answers. Trial registration number: BOF19/24J/066
A possible effect of social desirability in the answers by the patients should
be considered.
Wider implications of the findings: The differences in systems of values P-334 The ethical implications of the framing of egg donation on
demonstrate the need to have a thorough discussion on the specific meaning of Belgian fertility clinic websites
the ethics terms. L. Jacxsens1, C. Coveny2, L. Culley3, T. Goethals1, C. Herbrand4,
The personal profile can be useful in clinical practice when dealing with the S. Lafuente5, V. Pavone6, G. Pennings7, V. Provoost7, C. Weis3,
patient in an individualized manner. N. Hudson3
Ghent University, Bioethics Institute Ghent, Ghent, Belgium ;
2
Loughborough University, Department of Social Sciences, Loughborough, United
P-333 Artificial womb technology and the ethics of prenatal Kingdom ;
medicine 3
De Montfort University, Centre for Reproduction Research, Leichester, United
S. Segers1 Kingdom ;
4
1
Bioethics Institute Ghent - Ghent University, Department of Philosophy and Moral De Montfort University, Centre for Reproduction Research, Leicester, United
Sciences, Gent, Belgium Kingdom ;
5
Goethe-Universität Frankfurt am Main, Faculty of Social Sciences, Frankfurt am
Study question: How would artificial womb technology affect how obligations Main, Germany ;
6
towards the future child and obligations towards the expecting mother are bal- Consejo Superior de Investigaciones Cientificas, Institute of Public Goods and
anced against each other? Policies, Madrid, Spain ;
7
Summary answer: Artificial womb technology could facilitate prenatal inter- Ghent University, Bio-ethics, Ghent, Belgium
ventions and may exacerbate potential conflicts between directive treatment rec-
ommendations and the pregnant woman’s autonomous decision to the contrary. Study question: How is egg donation framed on the websites of fertility clinics
What is known already: Current developments in artificial womb technology in Belgium and what are the ethical implications of this framing?
have rekindled the expectation that human ectogenesis may come within reach. Summary answer: Donors were presented as mothers under 37; recipients
As yet, the ethics debate has largely focused on the link with abortion, how this as heterosexual couples with a medical need. Overall, egg donation was framed
technology may provide an alternative for surrogacy or uterus transplantation as a medical act.

What is known already: How information is presented or framed can signifi- Participants/materials, setting, methods: Literature research; normative
cantly influence the interpretation and decisions of the reader. Therefore, the analysis
framing of egg donation in information for prospective donors is ethically import- The method that is used to bring empirical data (as found in literature research)
ant. In the literature, the most important ethical debates concerning egg donation and normative ethics together is the ‘Wide Reflective Equilibrium’, the most
deal with the topics of commercial egg donation, postmenopausal pregnancies commonly used method in bioethics.
and anonymity. Studies (mostly conducted in the United States) have shown that Main results and the role of chance: Personal autonomy is a very highly
the framing of gamete donation is gendered and that altruism seems to play an valued principle within biomedical ethics, yet it has been criticized for ignoring
important role. This is the first frame analysis of the Belgian context. the fact that people are essentially relational, interconnected beings, that people’s
Study design, size, duration: This study is part of the ESRC-funded ‘EDNA’ identities are shaped by those with whom they interact and that one person’s
project that aims to explore the social, political, economic and moral configura- decisions have implications for others. Feminist scholars have therefore intro-
tion of egg donation in the United Kingdom, Belgium and Spain. In this study we duced the concept of relational autonomy, which emphasizes the importance
only focused on Belgium. of input from the web of people a patient is connected with in medical decision
Participants/materials, setting, methods: We analysed the websites of making. While one should be very wary of this concept being abused to sideline
all 18 Belgian fertility clinics that perform egg donation. Frame analysis and con- autonomous choice, posthumous sperm donation may be one case in which a
tent analysis were used. The websites of clinics in Flanders and Wallonia were relational approach can be well defended. In first instance, this is because genetic
analysed separately. material is concerned, over which people can be said to have shared ‘ownership’,
Main results and the role of chance: On Belgian clinic websites egg rather than individual ownership (and thus decisional authority). Secondly, the
donation is presented as a service that suffers from scarcity in donors. Images benefits for the donor are dubious, whereas the donor’s action can cause sig-
found on clinic websites were mainly in the style of commercial advertisement nificant distress and grievance in his family. Also, the benefits for the recipients
images, with the exception of a small number of websites where ‘scientific’ are of a different order than in the case of organ donation. Sharing the decisional
images were used. Overall, the donor was presented as someone who is authority between family members therefore seems appropriate.
(preferably) a mother, younger than 37 years, and highly motivated, typically Limitations, reasons for caution: What does not follow from this reasoning
because she donates to or for (in the case of cross donation) family or friends. is that family members would have decisional authority over posthumous sperm
The recipients were framed as heterosexual couples who are in need of eggs retrieval and use in the absence of advance directives by the deceased. In this
due to medical reasons. In general, egg donation was framed as a medical act. case the family would seek a benefit at the expense of the deceased’s autonomy,
Information relating to risks mainly dealt with physical aspects whilst psycho- instead of avoiding harm.
logical risks were less frequently mentioned. Minor risks (such as temporary Wider implications of the findings: Although personal autonomy is and should
physical discomfort) and more serious risks (e.g. ovarian hyperstimulation be a central concept in reproductive ethics, incorporating relational autonomy can
syndrome) received equal coverage. All websites where compensation was lead to new perspectives on how to deal with reproductive decision making.
mentioned made it clear that this is not a payment and only one website Trial registration number: N/A
mentioned the amount of compensation. On the Flemish websites expressions
of clinic preference for anonymous donation were present whereas this was
not found on the Walloon websites.
Limitations, reasons for caution: Clinic websites are the first step in a POSTER VIEWING SESSION
process of information provision that continues in the clinic. The current analysis IMPLANTATION AND EARLY PREGNANCY
is limited to one country. In a further phase, these data will be combined with
data from the two other countries (UK and Spain) involved in the EDNA-project
Wider implications of the findings: The presentation of donor or recipient
profiles is ethically important since people who do not relate to these profiles
might feel excluded, thus dismissing egg donation as an appropriate practice. P-336 Anterior gradient protein 3 and S100P levels in the
The framing of risks has ethical implications for the donors’ informed consent. reproductive tract mucosa may play an important role in human
Trial registration number: ES/N010604/1 fertility
N. Tempest1,2, E. Batchelor2, J. Drury2, D. Hapangama1,2
P-335 Relational autonomy in postmortem sperm retrieval and 1
Liverpool Women’s Hospital, Obstetrics and Gynaecology, Liverpool, United Kingdom ;
donation 2
University of Liverpool, Department of Women`s and Children`s Health- Institute
H. Mertes1 of Translational MedicineLiverpool, Liverpool, United Kingdom
1
Bioethics Institute Ghent BIG Ghent University, Department of Philosphy and Moral
Sciences, Gent, Belgium Study question: Do women with implantation problems have altered levels
of two endometrial calcium binding proteins (Anterior grade protein 3 (AGR3)
Study question: Should relational autonomy be the standard instead of per- and S100P) when compared with women of proven fertility?
sonal autonomy in the case of postmortem sperm retrieval and donation? Summary answer: Significantly higher AGR3 and S100P immunostaining was
Summary answer: As genetic material ‘belongs’ to several family members observed in cilia of luminal epithelium (LE) of women with implantation problems
and the disadvantages of the procedure fall disproportionately on family mem- compared with women of proven fertility.
bers, a relational approach should be pursued. What is known already: Calcium signalling pathways are of vital importance
What is known already: Recently, a study was published pleading to include the to embryo implantation. Calcium-binding proteins such as S100P, Anterior gra-
option of postmortem sperm donation in organ donor programmes in order to dient protein 2 (AGR2) and Anterior gradient protein 3 (AGR3) participate in
increase the availability of donor sperm. This is technically feasible and if one relies these pathways. AGR2 and S100P have previously been shown to be involved
on the analogy with organ donation, one could argue that it is the deceased’s right to in endometriosis and endometrial cancer and S100P demonstrates highest endo-
decide how his tissues are used after passing away. At the same time, several legal metrial levels during the window of implantation (WOI). Endometrial expression
cases have been brought forward in which family members request control over and the functional relevance of AGR3 is unknown, but it may play an important
reproductive material of a deceased person, some of which were successful. role in the regulation of ciliary beat frequency, thus in the LE of the endometrium
Study design, size, duration: This is a normative analysis in which (a) different may play an important functional role in embryo transfer and fertility.
arguments from the literature and from legal cases are inventoried, compared Study design, size, duration: This observational study examined luteinising
and scrutinized; (b) the analogy with posthumous organ donation is dismantled, hormone timed human endometrial samples taken in the WOI from 53 women
looking for semantic and structural (in)consistencies and (c) an exploration is (10 recurrent miscarriage (RM), 13 recurrent implantation failure (RIF), 30 con-
made of how the concept of relational autonomy might be operationalized in trol). The women underwent gynaecological procedures at a UK tertiary referral
this case and whether this would lead to better or worse outcomes for the centre. The women did not have any known endometrial pathology and had not
different stakeholders. been on hormone treatment for 3 months prior to the procedure.

Participants/materials, setting, methods: Immunohistochemistry was delivered at a significantly earlier gestational age, 36.6 ± 3.2 vs. 37.3 ± 2.9 weeks,
used to confirm the presence of and immuno-staining patterns of AGR3 and p=0.02. A trend for a higher rate of cesarean delivery was noted in the IVF
S100P in human endometrium during the WOI. Immuno-reactivity was analysed group, 69.5% vs. 60.7%, p=0.07, while birth weight was non-significant between
with a quick score that quantified immunostaining in the different endometrial the groups. A significantly higher rate of neonates in the control group experi-
areas at the cellular level. enced an adverse neonatal outcome. No difference in placental mal-perfusion
Main results and the role of chance: In total, 53 patients (10 RM, 13 RIF, lesions was demonstrated between the groups. However, a significantly higher
30 control) with an average age of 39 years were included in the study. The rate of villitis of unknown etiology (VUE) was demonstrated in the IVF group,
women in the RM group had an average of 4 previous miscarriages, while women 16.1% vs. 8.3%, p=0.007. A sub-analysis of stimulated cycles, frozen embryo
in the RIF group had an average of 8 previous failed embryo transfers and the transfer cycles and controls, demonstrated a significantly higher rate of VUE
control group on average had 2 previous children. among stimulated cycles as compared to controls (21.0% vs. 8.3%, p=0.006),
The epithelial cell sub-region and subcellular location of S100P immunoreac- with no significance between stimulated and frozen cycles, or frozen cycles and
tivity was different in the healthy fertile controls when compared with the RM controls.
and RIF samples. Nuclear S100P was seen in control samples whereas RM/RIF Limitations, reasons for caution: Despite a departmental protocol neces-
samples showed no evidence of nuclear S100P immunoreactivity in the WOI. sitating placental examination in complicated pregnancies, the study cohort
Cilia of the LE of the RM/RIF samples demonstrated significantly higher (p<0.01) probably represents more severe cases of FGR and PE, for which placental
S100P immuno staining when compared with the control samples. examination was performed. In addition, certain data was unavailable and
Similar differences were observed in AGR3 immunoreactivity of the ciliated LE underpowered for sub-analysis, such as IVF cycle characteristics (stimulated/
cells, where significantly higher immune-scores were demonstrated by RM/RIF frozen).
samples (p<0.01). Wider implications of the findings: PE and FGR are common among IVF
This observed higher immunoreactivity for 2 calcium binding proteins in the pregnancies, although neonatal outcome is more favorable in this group. Placental
LE cilia of women with a history of reproductive failure suggests their inaccurate examination demonstrated a higher rate of VUE in IVF pregnancies, pointing to
subcellular location associated pathophysiology for these conditions. The nuclear an additional immunological pathogenesis, thus contributing to our knowledge
localisation of S100P may allow transcriptional regulatory function that is nec- regarding the excess risk for placental disorders in IVF pregnancies.
essary for implantation of a viable pregnancy. Trial registration number: N/A
Limitations, reasons for caution: This study employs only immunohisto-
chemistry and although we have confirmed gene expression with qPCR no P-338 Proteomic characterization of endometrium in IVF-ET
functional studies have been conducted to confirm a role of the 2 calcium binding patients with repeated implantation failure
proteins in the implantation process. C. Wang1, H.Y. Zhang1, Z.J. Cheng1, Y. Feng1, W.J. Zhou1,
Wider implications of the findings: We describe the presence of epithelial M.Y. Jiang1, Y. Zhou1, X.Y. Fei1
AGR3 in the human endometrium and demonstrate differential AGR3 and S100P 1
Hangzhou Women’s Hospital Hangzhou Maternity and Child Health Care Hospital,
in the WOI in women with a history of reproductive failure. These proteins may
Reproductive Medicine Center, Hangzhou, China
have a functional role in fertility. Further work is thus warranted to assess their
utility as a diagnostic/therapeutic targets.
Study question: Repeated implantation failure (RIF) seriously affects the clinical
pregnancy rate of in vitro fertilization-embryo transfer (IVF-ET) treatment, while
the pathogenesis of RIF was still unknown.
P-337 Placental disorders of pregnancy and In-Vitro Fertilization – Summary answer: We highlighted antithrombin-III and corticosteroid-binding
does placental histological examination explain the excess risk? globulin as RIF-related proteins by screening the RIF patients’ endometrium proteins.
H. Ganer Herman1, L. Tamayev2, O. Feldstein2, M. Bustan2, What is known already: Assisted reproductive technology has developed
Z. Rachmiel2, L. Schreiber3, A. Raziel1, B. Jacob2, M. Kovo2 for more than 40 years, but there is no significant breakthrough in pregnancy
1
Edith Wolfson Medical Center, In Vitro Fertilization, Holon, Israel ; rate. RIF is a difficult problem in IVF-ET treatment. The uterine environment,
2
Edith Wolfson Medical Center, Obstetrics and Gynecology, Holon, Israel ; especially the endometrial receptivity during the implantation window, has a
3
Edith Wolfson Medical Center, Pathology, Holon, Israel great influence on the pregnancy of such patients.
Study design, size, duration: In this study, we collected the endometrium of
Study question: Assessment of clinical characteristics of in vitro fertilization IVF-ET patients in our hospital during 2016 to 2019. According to the pregnancy
(IVF) pregnancies complicated by fetal growth retardation (FGR) and preeclamp- outcomes, we finally gained 40 RIF cases. According to the patients’ age, infertile
sia (PE), and the correlation to histopathological placental findings. factors, and visiting time, we matched our RIF cases with 40 pregnant cases.
Summary answer: Neonatal outcome is relatively favorable in placental dis- Participants/materials, setting, methods: All the female patients were
eases in IVF pregnancies. Placental villitis is more common in IVF pregnancies, aged under 40. We performed iTRAQ-2D LC-MS/MS between RIF and pregnant
pointing to an additive immunological etiology. group. Clinical data were analyzed using paired t test.
What is known already: Assisted reproduction is correlated with an increased Main results and the role of chance: Fifty-five higher expressed proteins
risk for FGR and PE, independent of the higher incidence of risk factors among (> 1.50 times, P < 0.05) and 27 lower expressed proteins (< 0.67 times,
IVF patients. FGR and PE, termed “the placental diseases of pregnancy”, share P < 0.05) were found significant changed by iTRAQ-2D LC-MS/MS in RIF
a common pathogenesis of ischemia, with a higher rate of vascular mal-perfusion patients. Furthermore, the most different categories: negative regulation of
placental lesions. Villitis of unknown etiology is an additional placental finding in hydrolase activity, blood particles, and enzyme inhibitor activity were found in
FGR, assumed to reflect an immunological pathogenesis. the biological process, cell composition and molecular function through GO
Study design, size, duration: This was a retrospective cohort of 1114 deliv- analysis with hypergeometric test. We also found that the differential proteins
eries during an 11-year period. were mostly in the immune system, and mRNA translation through KEGG anal-
Participants/materials, setting, methods: The study was performed in a ysis, while most of them were enriched in the primary immunodeficiency. At
tertiary hospital. The cohort included deliveries with a diagnosis of FGR and/ last, we discovered that these RIF-related proteins were clustered mostly in the
or PE, with a singleton live birth at 22 weeks of gestation or more, whose pla- post-translational modification, protein turnover, chaperones; transduction,
centas were sent to histological examination. We compared obstetric, neonatal ribosome structure and biogenesis; and function unknown using the EggNOG
and placental characteristics between IVF conceived pregnancies and controls. database. The most repeated proteins were antithrombin-III and corticoste-
Main results and the role of chance: A total of 1114 deliveries with a roid-binding globulin, in the different categories we found by GO analysis, which
diagnosis of placental related pregnancy complications were included - 105 con- related to hypercoagulable state and lower progesterone level, respectively.
ceived with IVF (IVF group) and 1009 non-IVF conceived (control group). Patients However, we found higher level of D-Dimer and lower level of progesterone
in the IVF group were older, and of lesser parity and gravidity. The rate of dia- in RIF patients with no significant differences (P > 0.05).
betes mellitus and chronic hypertension was significantly higher among IVF Limitations, reasons for caution: Most of the proteins that screened by
patients, and the rate of smokers significantly lower. Patients in the IVF group iTRAQ-2D LC-MS/MS were lack of commercialized ELISA Kit or antibody, and

we could only compare paired RIF and pregnant cases’ clinical data to reveal the P-340 Can AUTOLOGOUS PLATELET RICH PLASMA treatment
hypercoagulable state and lower progesterone level of RIP patients. More cases improve pregnancy outcome in ivf/et cases : a case control study
of clinical data were needed. done at Teerthanker Mahaveer Medical College & RC
Wider implications of the findings: Our results might give a new sight into P. Singh1
the implantation-related molecular, moreover, also provides a new experimental 1
TMMC&RC, OBS & GYNAE / IVF, MORADABAD, India
basis to understand the pathogenesis of RIF.
Trial registration number: Project supported by the Medicine and Health Study question: Can AUTOLOGOUS PLATELET RICH PLASMA treatment
Science Technology project of Zhejiang province,China(2017KY550), the Natural improve pregnancy outcome in ivf/et cases ?
Science Foundation of Zhejiang province,China (LQ18H040009). Summary answer: the use of autologous PRP holds promise in the treatment
of women with suboptimal ET and vascularity for embryo transfer.
What is known already: Poor endometrium is a cause of treatment faliure
P-339 Circ-CCNB1 inhibits trophoblast cells proliferation and in majority of cases of assisted reproduction .refractory endometrium is seen
inasion by sponging miR-223 to regulate SIAH1 during embryo in many cases which does not respond to various treatments e.g. vaginal silnafil
implantation , esrtogen,androgens,low dose aspirin etc.to overcome this refractory endome-
D. Wang1, L. Xie1, M. Jin1, Y. Zhou1, A. Liu1 trium PRP treatment was found effective .
1
School of Medicine- Zhejiang University, Reproductive endocrinology, Hangzhou, China Study design, size, duration: CASE CONTROL STUDY
18 MONTHS
Study question: Whether circRNAs regulate trophoblasts function and the 168 PATIENTS WERE INCLUDED IN STUDY.
tentative molecular mechanisms in aberrant implantation environment remains Participants/materials, setting, methods: Intrauterine instillation of autol-
unknown. ogous PRP was done in 168 women,101 did not received aprp treatment due
Summary answer: circ-CCNB1 regulates trophoblast cell proliferation and to various reasons,taken as controls between 22 and 45 years, over 18 months,
invasion through sponging miR-223 to increase SIAH1 expression and inhibit with suboptimal endometrial growth, and patients with repeated cycle cancel-
CCNB1 nuclear translocation. lations, in addition to vaginal sildenafil ,Estradiol valerate etc . Embryo transfer
What is known already: Ectopic pregnancy (EP) has been recognized as a model was performed when the endometrium reached an optimal pattern in terms of
to investigate the signaling and pathways regulated by trophoblast cells during thickness, appearance, and vascularity.
embryo implantation. Circular RNAs (circRNAs) have been reported to exert Main results and the role of chance: The mean pre-PRP endometrial thick-
important regulatory effects on trophoblasts function and embryo development. ness (ET) was 3.5 mm which significantly increased to 7.18 mm post-PRP. There
Study design, size, duration: To study the tentative molecular mechanisms was a significant increase in vascularity, seen by the number of vascular signals
in the occurrence of EP, five trophoblast cells samples from EP and five tropho- seen on Power Doppler, reaching the zones 3 of the endometrium. The positive
blast cells samples from normal pregenancy were used to analyze the differential beta Human Chorionic Gonadotropin (hCG) rate was 70.93% and the clinical
expression of circRNAs by high-throughput sequencing. To determine the effect pregnancy rate was 42.31%. A total of 47 delivered healthy babies ,11 women
of circ-CCNB1 on trophoblast cells function, JEG-3 and HTR-8 cells were trans- are in the second trimester, 12 are in the first trimester with a healthy intrauterine
fected with overexpression of circ-CCNB1 vectors or empty vectors for 24 h pregnancy, one patient had an ectopic gestation, 22 had missed abortions.
or 48 h in vitro. Limitations, reasons for caution: Small sample size
Participants/materials, setting, methods: Through high-throughput Wider implications of the findings: Patients with refractory endometrium
sequencing, we found the decreased expression of (circRNA CyclinB1) circ- ,especially in younger age groups seems to be benifitted by this technique.
CCNB1 in the patients with EP. Subsequently, overexpression of circ-CCNB1 Trial registration number: NA
vectors, CCK-8, transwell, immunofluorescence, quantitative PCR, western blot,
RNA pull-down and dual-luciferase reporter assays were used to investigate the
P-341 Impact of uterine contractility on pregnancy success rate
effect of circ-CCNB1 on the function of trophoblast cells.
during in-vitro fertilisation: assessment by quantitative ultrasound
Main results and the role of chance: In vitro study demonstrated that
imaging
overexpression of circ-Ccnb1 slightly inhibited JEG-3 cell proliferation and sig-
nificantly inhibited HTR-8 cell proliferation after 24h and 48h treatment. C. Rees1, C. Blank1, N. Kuijsters2, Y. Huang2, F. Sammali2, H. Van
Moreover, transfection of circ-Ccnb1 overexpression vectors lead to a significant Vliet1, P. De Sutter3, M. Mischi2, B.C. Schoot1
1
decrease of cell invasion in JEG-3 and HTR-8 cells. Furthermore, miR-233 was Catharina Hospital Eindhoven, Obstetrics and Gynaecology, Eindhoven, The
predicted and demonstrated to interact with circ-Ccnb1 in trophoblast cells. Netherlands ;
2
We found miR-233 mimics treatment decreased the expression levels of circ- Eindhoven University of Technology, Signal Processing Systems, Eindhoven, The
Ccnb1, whereas miR-233 inhibitor significantly enhanced the expression levels Netherlands ;
3
of circ-Ccnb1 in trophoblast cells. Meanwhile, miR-233 inhibitor treatment sup- Ghent University Hospital, Reproductive Medicine, Ghent, Belgium
pressed cell proliferation and invasion in HTR-8 cells. Interestingly, we found
miR-233 dramatically increased cyclin related genes CCNB1, Thymidine kinase Study question: Can assessment of uterine activity in patients with successful
(TK1) Krüppel like factor-4 (KLF4) and nuclear factor kappa B (NF-κB) expres- in-vitro fertilisation (IVF) versus patients with unsuccessful IVF treatment be used
sion levels, while significantly decreased the expression levels of E3 ubiquitin to predict success of embryo implantation?
ligase, seven in absentia homolog-1 (SIAH1) and F-box and WD-40 domain Summary answer: Measuring uterine contractility prior to embryo implan-
protein 7 (FBXW7), and apoptotic factor caspase-3. SIAH1 was testified to be tation in IVF patients is able to predict chance of pregnancy with 93.8% accuracy.
directly targeted by miR-233, which could be activated by circ-Ccnb1 treatment What is known already: During the menstrual cycle, the natural contraction
in trophoblast cells. In addition, we also found circ-Ccnb1 increased CCNB1 pattern of the uterus is affected by changes in hormones. Abnormal uterine
expression levels and suppressed CCNB1 nuclear translocation in HTR-8 cells. contraction activity is known to influence the outcome of fertility treatment.
Limitations, reasons for caution: The investigation of the molecular mech- Recently, an automated measurement tool to characterise these movements
anism of circ-CCNB1 regulating trophoblast cells function was limited in tro- has been developed, but has not yet been widely applied to patients receiving
phoblast cells in vitro. It’s necessary to testify this mechanism in the EP animal fertility treatment.
models in future. Study design, size, duration: A single-centre prospective study included 16
Wider implications of the findings: These results demonstrated an import- IVF/ICSI patients between January 2017 and July 2018. Participants underwent
ant molecular mechanism of circ-CCNB1 regulating trophoblast cell proliferation three quantitative 2D-transvaginal ultrasound measurements prior during three
and invasion through sponging miR-223 to increase SIAH1 and inhibit CCNB1 phases of IVF treatment: follicle stimulation (FS), one hour before embryo trans-
nuclear translocation, which might provide a novel pathophysiological charac- fer (ET1), and five to seven days after ET (ET5-7).
teristic during embryo implantation. Participants/materials, setting, methods: A dedicated speckle tracking
Trial registration number: 20180192 algorithm was implemented to extract frequency and amplitude-related features

of uterine motion in relation to ongoing implantation at 6 weeks up to 11 weeks endometrial thickness on the day of recombinant-hCG between two groups
pregnancy. Machine learning and random forest models were used to create a (p>0.05).
prediction model based on uterine motion characteristics prior to ET in order When we compared the results of mNC-FET cycles, OPR was 68.7% in MVP
to suggest a subsequent chance of pregnancy. gel and 71.6% in dydrogesterone group, respectively (p=0.706). Biochemical -
Main results and the role of chance: Overall, frequency and amplitude of clinical pregnancy rates and biochemical - clinical miscarriage rates were also com-
uterine significantly decreased during the phases of the IVF/ICSI treatment parable between two groups (p=0.382, p=0.547, p=1.0, p=1.0 respectively).
showing FS as the most active phase followed by ET1 and ET5-7. Women with A significantly higher patient tolerability score was present in the dydroges-
successful implantation/pregnancy showed significantly higher uterine contrac- terone arm (4,13±0,97 vs 3,43±1,17, p=0.001).
tion frequency compared to those without in all phases. Conversely, uterine Vaginal discharge (76,9% vs. 12,3%, p=0.001), vaginal irritation (44,6% vs.
contraction amplitude was significantly lower (p = 0.037) in women at ET5-7 in 1,6%, p=0.001) and interference with coitus (50% vs. 1,6%, p=0.001) were
case of with ongoing implantation/pregnancy. Analysing uterine contraction significantly higher in MVP gel compared to dydrogesterone group. On the other
frequency and coordination prior to ET was able to predict chance of pregnancy hand, mastalgia (36,9 % vs. 4,6%, p= 0.001), headache (13,8% vs. 0%, p=0.003),
after ET with a sensitivity of 85.7%, specificity of 100% and an accuracy of 93.8%. dizziness (9,4% vs. 0%, p=0.013), flatulence (32,8% vs. 12,3%, p=0.005) and
Limitations, reasons for caution: The small sample size limits statistical somnolence (38,5% vs. 1,5%, p=0.001) were also significantly higher in the
power of this study. Research should be continued with larger patient cohorts dydrogesterone group compared to MVP gel.
to confirm the results. Additionally, not all uterine motion characteristics were Limitations, reasons for caution: In order to avoid any possible selection
taken into account, in future research the full characterisation of the uterine bias, which can result from compromised clinical features, only good prognosis
activity during IVF treatment should be completed. patients with top and good blastocysts were included in the study.
Wider implications of the findings: This study is a first step to objectively Wider implications of the findings: When choosing the form of proges-
and non-invasively assess uterine contraction characteristics during IVF cycles, terone supplementation, effectiveness of treatment, as well as side effects,
and introduces a new method to predict chance of IVF success. In future, it may patients’ preference and convenience should be considered. Dydrogesterone,
boost IVF success rates by for example introducing specific uterine motion mod- as an alternative oral preparation for LPS, constitutes an effective option, espe-
ulators prior to ET. cially for patients suffering from side effects of vaginal progesterone.
Trial registration number: NL5035 Trial registration number: NCT04124913
P-342 Oral Dydrogesterone vs. Micronized Vaginal P-343 Embryos versus endometrium: who is to blame for the
Progesterone(MVP) Gel for Luteal Phase Support (LPS) in Frozen- impaired implantation rate in morbidly obese women?
Thawed Single Blastocyst Transfer in Good Prognosis Patients I. Antonova1, T. Milachich1, I. Bochev1, M. Yunakova2,
G. Özer1, K.B. Yuksel1, O.S. Yucel Cicek2, S. Kahraman1 M. Magunska2, A. Shterev2
1
Istanbul Şişli Memorial Hospital, Memorial Hospital- ART and Reproductive 1
SAGBAL “DR. SHTEREV” EOOD, Embryology, Sofia, Bulgaria ;
2
Genetics Center- Istanbul., Istanbul, Turkey ; SAGBAL “DR. Shterev” EOOD, Ivf, Sofia, Bulgaria
2
Kocaeli Universıty School of Medicine, Department of Gynecology and Obstetrics,
Kocaeli, Turkey Study question: What is the major reason for the reduced implantation and
clinical pregnancy rates in cases with obese women who have undergone ART
Study question: Is oral dydrogesterone an effective option for luteal phase treatment?
support in modified natural cycle frozen -thawed embryo transfers (mNC-FET) Summary answer: Our results suggest that the endometrial receptiveness is
when compared to MVP gel? adversely affected by higher proportions of the body fat and leads to difficulties
Summary answer: In mNC-FET cycles, oral dydrogesterone provides an in pregnancy achievement.
effective option compared to MVP gel with similar ongoing pregnancy rates and What is known already: Obesity has become one of the greatest public health
with fewer intolerable side effects. challenges of the last few decades. Its prevalence has tripled in many countries of
What is known already: Administration of progesterone for LPS could be the WHO European Region since the 1980s, and the numbers of those affected
oral, intramuscular, vaginal and subcutaneous with each route having various continue to rise at an alarming rate. Overweight female patients undergoing fertility
bioavailability, side effects and tolerability profiles. The optimal form of proges- treatment face many complications. Obesity drastically increases gonadotrophin
terone supplementation has not been determined from available data. Since consumption and is linked with lower embryo implantation and higher miscarriage
bioavailability of oral micronized progesterone is <10 % and is related to drows- rate as well. Elevated body mass index (BMI) is associated with complicated obstet-
iness, flushing, and nausea, it is not commonly used for LPS. However, dydro- ric anamnesis such as risk of pre-eclampsia and gestational diabetes.
gesterone, which is an optical isomer of progesterone, has better bioavailability Study design, size, duration: This is retrospective analysis from hospital data-
and progestogenic activity and causes endometrial transformation at a dose of base for 5-year period. This study includes 3599 fresh cycles using own gametes
10–20 fold lower than that of oral micronized progesterone. and additional group of 343 patients with donor oocytes as a control group. For
Study design, size, duration: This was a randomized, single center, parallel the purpose of the investigation, all patients were divided into 4 groups according
controlled trial conducted at Istanbul Memorial Hospital IVF and Reproductive the international BMI WHO classification: underweight (BMI<18.49) group A
Genetics Centre, Istanbul, Turkey between January and August 2019. A total of (n=336); normal range (BMI 18.5-24.9), group B (n=2291); overweight (BMI
134 women, aged below 38, were assigned randomly in a ratio of 1:1, based on 25-29.9), group C (n=639) overweight and D obese (BMI>30) (n=333).
a computer-generated list to receive oral dydrogesterone (n=67) or MVP (n=67) Participants/materials, setting, methods: In order to establish possible impact
for LPS in mNC-FET cycle. of excess weight on oocyte and embryo quality, we compared average number of
Participants/materials, setting, methods: The exclusion criteria were a his- the retrieved oocytes, fertilization rate, embryo quality at day 3 and blastocyst rate
tory of ≥2 unsuccessful cycles and early pregnancy losses, uterine malformation, between the groups. For the examination of the linkage between obesity and endo-
severe endometriosis, azoospermia and women with endocrine or metabolic metrial receptivity, we measured main outcomes such as pregnancy and implantation
disorders rates and compared the results with a control donor-oocyte recipient group.
The primary outcome was ongoing pregnancy rate (OPR). Secondary out- Chi-squared analysis was used to compare results.
comes were clinical pregnancy and miscarriage rates, patients’ satisfaction and Main results and the role of chance: All four groups were homogeneous
tolerability of oral and vaginal progesterone. A questionnaire was developed to according the average number of the retrieved oocytes (5.97 vs 6.11 vs 5.99 vs
compare side effects and satisfaction profiles. Data was collected through tele- 6.25; p: NS) and regarding the fertilization rate: 73.5% for group A, 74.4% Group
phone interviews on the day of pregnancy test. B, 73.6% group C and 73.3% for group D respectively.
Main results and the role of chance: There was no significant difference At cleavage stage we measured the embryo quality in groups and they showed
in demographic features such as female age, body mass index, AMH levels and similar proportions of good, fair and bad embryos. Day 5 blastocyst rate was
fresh cycle characteristics including the number of retrieved oocytes, mature slightly higher in normal BMI group B 45%), but not significant referred to other
oocytes, ICSI-fertilized oocytes, the number of vitrified blastocysts and groups (A - 42% vs C - 42% vs D - 40%).

After clinical pregnancy rates (CPR) measurement groups A and B appeared and 6.06pmol/mL in serum, whereas the median testosterone level was
comparable results (34.6% vs 35.7%). In overweight group (C) CPR was lower, 0.34pmol/g and 0.86pmol/mL in tissue and serum, respectively.
but not significantly (31.5%); (P = 0.0895). Statistical difference was found in Among corticosteroids, median 11-deoxycortisol level was 0.32 pmol/g in
obese group D where CPR dropped to 28.9% (P = 0.0392). In the control group tissue and 0.64 pmol/mL in serum. Androsterone in tissue was below quantifi-
of 343 patients using ovum donation the results were even more demonstrative cation limit (<3.26pmol/mL) and median serum level was 0.60 pmol/mL.
where CPR was 56% for underweight women (n=33); 52.8% for normal range No statistically significant differences were found in steroid tissue concen-
(n=208); 50% in overweight (n=70) and only 17.4% in the obese group (n=32) . trations between cases and controls. Mean serum 11-deoxycortisol levels were
Limitations, reasons for caution: This is observational retrospective study higher in women who conceived compared with those who did not (1.38±1.24
and it is limited by the subgroups size. More detailed studies are required to vs. 0.45±0.23 pmol/mL, p=0.04). The opposite was seen for androsterone,
achieve a deeper understanding and to shed light into the finer aspects of this whose mean serum levels were lower in women who conceived compared with
complicated implantation process. women who did not (0.47±0.23 vs 0.72±0.10 pmol/mL, p=0.02).
Wider implications of the findings: Our survey shows that the endome- Tissue and blood concentrations were not correlated for estrogens, indicative
trium receptiveness is the one that is negatively influenced by excess fat cells. of local steroid metabolism of these steroids, but they were significantly cor-
This could be a good predictive model for implantation capability as there is a related for most progestogens, androgens and corticosteroids.
clear cut-off range in BMI values. Weight-loss concealing before fertility treatment Limitations, reasons for caution: Even though cases and controls were
could be beneficial for the successful outcome. matched for several important variables, it cannot be excluded that other mater-
Trial registration number: not applicable nal factors or embryo aneuploidy could have led to implantation failure, as there
is heterogeneity in IVF/ICSI treatment indication in the (small) group of not-preg-
P-344 Endometrial steroid metabolism (intracrinology) in relation nant women.
to pregnancy outcome in women undergoing IVF/ICSI Wider implications of the findings: Further clarification of the complex
interplay between endometrial steroidogenesis, the tissue availability of ste-
L. Brentjens1, J. Den Hartog1, B. Delvoux1, M. Häkkinen2, J.
roidogenic enzymes, and that of the steroid hormone receptors in women with
Koskivuori2, P. Koskimies2, S. Auriola2, N. Van Hoogenhuijze3, H.
multiple failed IVF/ICSI cycles is necessary to unravel a possible role in the
Torrance3, F. Broekmans3, SCRaTCH-2 study group(4), R. Van
implantation process.
Golde1, A. Romano1
1
Trial registration number: NTR5342
Maastricht University Medical Centre - GROW Maastricht University, Obstetrics &
Gynaecology, Maastricht, The Netherlands ;
2 P-345 Impaired myeloid-derived suppressor cells are associated
University of Eastern Finland, School of Pharmacy, Kuopio, Finland ;
3 with recurrent implantation failure
Universitair Medisch Centrum Utrecht, Divisie Vrouw en Baby - Gynaecologie &
Voortplantingsgeneeskunde, Utrecht, The Netherlands ; H. Jiang1, Y. Cao1, K. Wang1, K. Bi1, Z. Lu1, P. Guo1, C. Li1
4 1
Amsterdam Medisch Centrum, F. Mol, Jeroen Bosch Ziekenhuis the First Affiliated Hospital of Anhui Medical University, Obstetrics and Gynecology,
Hefei, China
Study question: Does the endometrial steroid profile (estrogens, progesto-
gens, androgens and corticosteroids) differ between receptive and non-receptive Study question: Studies have reported that myeloid-derived suppressor cells
endometrium? (MDSCs) contribute to maintain pregnancy. The aim of this case–control study
Summary answer: The levels of steroids were similar in the endometrium was to test whether there is a dysregulation of peripheral MDSCs in recurrent
of women who did and did not achieve a clinical pregnancy through IVF/ICSI. implantation failure (RIF).
What is known already: Implantation failure of high-quality embryos is a Summary answer: This study indicated that the dysregulation of MDSCs
main concern in IVF/ICSI treatment. We previously reported that in IVF/ICSI might impaired maternal-fetal immune balance thus result in RIF.
patients, blood sex-steroid concentrations do not reflect their corresponding What is known already: Recurrent implantation failure (RIF) is diagnosed
concentrations in endometrial tissue. This is in line with the concept that blood when women experienced 3 or more frozen or fresh cycles with being trans-
steroids (and precursors) are locally converted to bioactive metabolites and vice ferred high-quality embryos and failed to obtain a clinical pregnancy. Myeloid-
versa, leading to a specific intra-tissue steroid milieu. The endometrium regulates derived suppressor cells (MDSCs) represent a heterogeneous cell group with
local steroid concentrations by expressing steroid-metabolising enzymes, or attributes in myeloid origin, immature state, and immunosuppressive function.
‘intracrinology’. Studies indicate that alterations in intracrinology might modulate Study design, size, duration: In our study, We collected 26 RIF cases and
endometrial receptivity. We hypothesize that the endometrial steroid profile 30 controls 2019. Then we completed the experiments in two months.
during the window of implantation differs between pregnant and non-pregnant Participants/materials, setting, methods: 26 RIF patients and 30 controls
IVF/ICSI patients. were recruited. Flow cytometry was applied to characterize polymorphonuclear
Study design, size, duration: Case-control study of 40 patients that were (PMN)-MDSCs, monocytic-MDSCs (M-MDSCs), effector T cells (Teffs) and
recruited in the SCRaTCH study (NTR 5342), a randomised trial exploring regulatory T cells (Tregs) in blood. ELISA was used to define MDSCs correlative
whether ‘endometrial scratching’ in patients with a previous IVF/ICSI cycle failure cytokines and chemokines in serum from all patients.
affects pregnancy outcome in a subsequent IVF/ICSI cycle. For the present Main results and the role of chance: Compared with controls, RIF patients
investigation, 20 endometrial biopsies from women who became clinically preg- showed significant reductions of blood PMN-MDSCs, M-MDSCs, Tregs and NO
nant after fresh embryo transfer were compared with 20 endometrial biopsies production by PMN-MDSCs, whereas the expression of ζ chain on CD4+T cell
of women that did not conceive after fresh embryo transfer. receptor (TCR) and CD8+TCR displayed a remarkable upregulation in RIF patients.
Participants/materials, setting, methods: Endometrial biopsies and serum Moreover, RIF patients presented a lower concentrations of serum chemokine
were obtained at LH+ 5-8 days (urinary test) in a natural cycle. In their next (C-C motif ) ligand (CCL) 5 and transforming growth factor (TGF)-β than those
cycle, subjects underwent IVF/ICSI with fresh embryo transfer. Cases (negative from controls. Furthermore, the level of TCR ζ chain on CD4+ and CD8+Teffs
pregnancy test, n=20) and controls (clinically pregnant, n=20) were selected was negatively correlated not only with the percentage of PMN-MDSCs, but also
and matched for primary vs. secondary infertility, embryo quality and age. Steroid with the amount of NO produced by PMN-MDSCs. The frequency of PMN-
concentration was determined in endometrial ‘scratched’ tissue homogenates MDSCs had positive correlations with the concentration of CCL5 and TGF-β.
and serum (liquid chromatography mass spectrometry). Statistics was computed Limitations, reasons for caution: Consistent with previous observations,
with Pearson Correlation and unpaired Student’s t-test. results showed Treg failure in patients with RIF. However, the proportion of
Main results and the role of chance: Steroids were measured in 40 endo- Treg in this study is independent of the proportion of MDSC. This may be
metrial biopsies and 16 serum samples. Median estrone level was 0.23 pmol/g because hormones can also induce Tregs.
in tissue and 0.29 pmol/mL in serum. Median estradiol level was 0.37 pmol/g Wider implications of the findings: our study discovers the role of MDSCs
in tissue and 0.45 pmol/mL in serum. Tissue progesterone (P4) median was and their related mediators in RIF. Therefore, targeting these cells could provide
51.32pmol/g; P4 in serum was above quantification limit (33.3pmol/mL). With new treatment methods in the future.
regard to androgens, median androstenedione level was 2.02pmol/g in tissue Trial registration number: 1808085QH273

P-346 Validation of an in vitro model to study the interactions implantation, and potentially lead to novel markers or predictors of implantation
between endometrial cells, uterine natural killer cells and failure.
embryonic signals during human implantation. Trial registration number: NA
A. Almansa1, C.S. King2, P. Brighton2, G. Tiscornia1,
G. Hartshorne2, J. Muter2, M. Barragan1, J. Brosens2, R. Vassena1 P-347 Patients with repeated implantation failure and Natural
1
Clinica Eugin, Eugin, Barcelona, Spain ; Killer cells immaturity: the efficiency of the endometrial
2
Warwick Medical School, University of Warwick University Hospital Coventry and scratching strategy
Warwickshire, Covnetry, United Kingdom F. Lamazou1, P. Oger1, P. Meicler1, G. Kerbrat1, G. Soudre1,
E. Genauzeau2, V. Maget1, X. Guettier1, L. Delaroche2, N. Ledee3
Study question: What is the effect of human embryo conditioned media 1
RAMSAY SANTE Hôpital Privé de Parly 2, Reproductive Medecine, Le Chesnay,
obtained from blastocysts of different quality on uterine natural killer (uNK) cell France ;
activity during implantation? 2
Centre de Biologie Medicale Hôpital Prive de Parly 2, ART laboratory, Le Chesnay,
Summary answer: Conditioned medium from blastocysts that failed to give France ;
rise to biochemical pregnancy diminished uterine NK mediated clearance of 3
MatriceLab innove pépinière paris santé cochin hôpital cochin paris, ART Center,
senescent endometrial cells in vitro. Paris, France
What is known already: Embryo implantation is a regulated process involving
synchronous communication between the embryo, endometrium and immune Study question: Is the scratching an efficient treatment for patients with history
system. Recent studies demonstrate the emergence of acute cellular senescence of repeated implantation failure (RIF) associated with a uterine Natural Killer
during decidualization. Senescent cells irreversibly exit the cell cycle and remain (uNK) cells immaturity?
biologically active, but may be detrimental to endometrial function if left Summary answer: The endometrial scratching performed in the mid luteal
unchecked. uNKs infiltrate the endometrium and are present in high numbers phase may increase live birth rates in patients with RIF and uNK cells immaturity
during implantation where they neutralize senescence by selectively targeting What is known already: Twenty to 25% of RIF patients have been described
and eliminate the senescent decidual cell population. Here we validate a model to have uNK cells immaturity. When performed in the mid-luteal phase, the
for the uNK clearance of senescence, and apply it to investigate interactions endometrial scratching stimulates the local expression of IL-15 in the mid-luteal
between different endometrial cell types involved in this process. phase of the following cycle. The IL-15 secretion allows the uNK cells maturity
Study design, size, duration: Basic research study. We co-cultured decid- which is essential for an effective implantation since they regulate local angio-
ualized human endometrial stromal cells (EnSCs) from 5 women with human genesis and invasion of spiral arteries.
uNK cells in the presence embryo-conditioned media (ECM) obtained from Study design, size, duration: This is a single-center observational study
cultured human embryos that were transferred as part of IVF treatment. performed from January 2016 to March 2019. The institutional review board
Transferred embryos were grouped depending on whether they resulted in a approved this study and the patients’consent to the use of their data was prior
positive biochemical pregnancy or not. Non-conditioned culture media was obtained.
used as control. The effect of conditioned media on uNK cell clearance was Participants/materials, setting, methods: Patients with RIF history (>4
measured. top blastocysts) and normal explorations were proposed an endometrial biopsy
Participants/materials, setting, methods: EnSCs were cultured from 5 to analyse their uterine immune profile. The endometrial immune profile doc-
different endometrial biopsies. uNK were isolated by MACS targeting CD56, umented the ratio of IL-15/Fn-14 mRNA and the IL-18/TWEAK mRNA ratio.
and purity and viability assessed by flow cytometry. Stromal cells were decidu- Patients with an uNK cells immaturity were included. A scratching in the mid-lu-
alised for 6 days with 8-bromo-cAMP and medroxyprogesterone acetate, before teal phase was performed before a new fresh or a frozen-thawed blastocyst
uNKs were added. After 2 days of co-culture, levels of senescence associat- transfer in the following cycle. The main endpoints were the implantation and
ed-β-Galactosidase (SA-β-Gal) and senescent-specific genes DIO2 and CLU were live birth rates.
used as direct indicators of uNK clearance. Effects of ECM were assessed by Main results and the role of chance: Sixty-nine RIF patients with an uNK
inclusion at day 6. cells immaturity on their uterine immune profile were included in this study. They
Main results and the role of chance: uNK cells isolated from endometrial were 35.1 ± 1.2 [LD1] years old and had a median AMH level of 2.6 ng/ml
biopsies were round, granulated and stained positively for CD56. Characterization [1,2-4.2]. Forty-four patients (64%) received an endometrial scratching in the
by flow cytometry revealed high purity (83.5%) and viability (87.4%). As mid-luteal phase followed by an embryo transfer. The mean number of blasto-
expected, we observed the emergence of decidual senescence in EnSCs by cysts transferred was 2.2 [1-2]. The implantation rate was 40% and 54.4% of
increased SA-β-Gal, and expression of DIO2 and CLU. The addition of uNK cells these couples achieved a live birth. The rate of spontaneous abortion
in co-cultures effectively eliminated cellular senescence by significantly reducing reached 11.2%.
SA-β-Gal (p=0.038), DIO2 (p=0.046) and CLU (p=0.021). Markers for non-se- Limitations, reasons for caution: Despite the bias inherent to a retrospec-
nescent, mature decidual cells (SCARA5 and CXCL14) were unaffected. This tive study and the small number of patients included, these preliminary encour-
demonstrates the specific uNK-mediated targeting and elimination of senescent aging results call a prospective randomized controlled trial with a higher number
decidual cells. The effects of embryo-derived signals on uNK function were of patients.
assessed by inclusion of ECM at day 6. Exposure to non-conditioned media, or Wider implications of the findings: This study suggests that the uterine
media from embryos that resulted in positive biochemical pregnancies had no immune profile can select a population for who the endometrial scratching can
effect on uNK cell mediated senescence clearance. However, on evaluation of be an efficient strategy in RIF situation with an uNK cells immaturity to improve
SA-β-Gal we detected a significant loss of clearance activity when the system the implantation and live birth rates.
was cultured with ECM from embryos that failed to establish pregnancies Trial registration number: non applicable
(p=0.015). We propose that embryonic signals from poor quality embryos inhibit
uNK function. This demonstrates our model can detect uNK cell biological
P-348 The remaining cell-free DNA from noninvasive
activity in response to embryo-derived stimuli and could be useful as an improved
preimplantation genetic test (NIPGT-A) could be used to
model to help understand human implantation
identify single nucleotide polymorphisms (SNPs) related to the
Limitations, reasons for caution: This study was conducted with biopsies
implantation process.
from a limited number of patients; higher numbers of samples will need to be
characterized to confirm our findings. Furthermore, our model lacks the endo- J.G. Franco Jr., M.D.1,2, L.D. Vagnini2, C.G. Petersen1,2, A. Renzi2, M.
metrial epithelial component involved in apposition of the embryo to the Canas2, B. Petersen1,2, A.L. Mauri1,2, M. Mattila1, J. Ricci1, A.
endometrium. Nicoletti1, C. Zamara1, A.H. Oliani3, F. Dieamant1,2, R. Baruffi1,2,
Wider implications of the findings: This newly developed model will allow J.B.A. Oliveira1,2
1
us to better characterize the interactions between immune cells, endometrium Centre for Human Reproduction Prof. Franco Jr, Research, Ribeirao Preto, Brazil ;
2
and the implanting embryos. This will provide further insights into human embryo Paulista Center for Diagnosis Research and Training, Research, Ribeirao Preto, Brazil ;

3
Sao Jose do Rio Preto School of Medicine FAMERP, Research, Sao Jose do Rio
Preto, Brazil P-348 Table 1. Main results of SNPs genotyping
Study question: Could the remaining cell-free DNA from the same sample SAMPLE DNA LIF VEGF TP53 MMP9
used for NIPGT-A diagnosis also identify SNPs related to the embryonic implan- ID [ng/uL] (G>T) (C>T) (C>G) (A>G)
tation process?
Summary answer: Regardless of the cell-free DNA concentration after the 1 10.4 * CC CC GG
amplification process used by NIPGT-A technology, the success rate of geno-
typing was 75%.
2 13.4 * TT GG *
What is known already: NIPGT-A is a new technology that uses cell-free 3 13.3 GG CC * *
DNA present in the spent culture medium of human blastocyst, which reflects 4 18.2 TT * * AA
its ploidy status. This DNA is an important source of information for the embryo.
Since there is a gene pattern signature of endometrial receptivity, several SNPs 5 10.2 * CC GG GG
such as those found in the LIF, VEGF, TP53 and MMP9 genes have been used as 6 9.7 TT * GG *
markers of the implantation process. However, there are no reports on the
possibility of identifying SNPs in embryos using remaining cell-free DNA in cul- 7 47.0 TT TT GG AA
ture medium. 8 16.8 GG CT GG GG
Study design, size, duration: This prospective cohort study included 23
9 11.7 GG TT * *
samples of remaining cell-free DNA obtained in culture medium on day 5 after
embryo culture during NIPGT-A technique (Yikon Genomics). A total of 13 10 37.0 TT TT CC AG
patients participated in this study after the couple’s informed consent. 11 24.0 TT TT CC AA
Participants/materials, setting, methods: Cell-free DNA evaluation used
the amplified DNA obtained after NIPGT-A technique (Yikon Genomics) and 12 27.0 TT TT GG GG
quantified by Qubit fluorometer (Thermo Fisher Scientific). SNPs were evaluated 13 31.0 TT TT GG GG
by real-time polymerase chain reaction (PCR) using individual TaqMan® SNP
genotyping assays (Thermo Fisher Scientific) for each SNP (LIF rs929271, TP53
14 25.0 TT TT * AA
rs1042522, VEGF rs3025010, MMP9 rs17576) and TaqPath™ ProAmp™ Master 15 92.0 TT CT CC AA
Mix (Thermo Fisher Scientific), following the manufacturer’s instructions, on a 16 32.0 TT TT CC *
StepOnePlus™ Realtime PCR System (Thermo Fisher Scientific).
Main results and the role of chance: All cell-free DNA samples in the 17 24.0 TT TT * AA
culture medium had at least one SNP identified regardless of their quantification 18 32.0 TT CT CC AA
(Table 1). It was observed that of all 92 genotyping reactions performed, 69
were properly amplified, leading to an accuracy of 75%.
19 15.8 * CT * *
Limitations, reasons for caution: Clinical validation studies are underway 20 9.7 * * CC *
to determine the predictive value of this methodology. 21 24.6 TT TT GG GG
Wider implications of the findings: This study showed a novel system
(NIPGT- A/SNPs) for embryo tracking. The studied SNPs involved in the implan- 22 28.6 GG CC GG AA
tation process were successfully amplified and genotyped with the remaining 23 9.8 GG TT * *
cell-free DNA of culture medium after NIPGT-A. In the future, NIPGT-A/SNPs
dual evaluation could be an additional tool for embryo selection *genotyping failure
P-349 Serum beta human chorionic gonadotropin (β-hCG) levels The study explores the correlation between positive early serum β-hCG
13 to 14 days after embryo transfer (ET) and the predictability of (mUI/ml) and pregnancy outcome in terms of: biochemical pregnancy (BP), early
pregnancy outcome in IVF cycles pregnancy loss (EPL), ongoing pregnancy (OP) and live birth (LB) rate.
L. Luque1, N. Ruiz2, Á. Linares3, J. Bartolomé3, J.A. Ortíz3, Participants/materials, setting, methods: Women who carried out β-hCG
R. Bernabéu4 determination in blood 13/14 days post egg retrieval in fresh and frozen transfer,
1
Instituto Bernabeu Albacete, Reproductive Medicine, ALBACETE, Spain ; respectively. Blood levels were measured using enzyme-linked fluorescent immu-
2
noassay (ELFA), with Minividas from Biomérieux analyzer. Own and donor egg
Instituto Bernabeu Albacete, Embriologist, Albacete, Spain ;
cycles were included. The link between variables of IVF-cycle and serum β-hCG
3
Instituto Bernabeu Albacete, Embriologist, Albacete, Spain ; was evaluated using U-Mann-Whitney or Kruskal-Wallis. The β-hCG value as a
4
Instituto Bernabeu, Reporoductive Medicine, Alicante, Spain predictor of the clinical results of IVF was evaluated by a binary logistic regression
(SPSSv20.0).
Study question: Is the first β-hCG value predictive of pregnancy outcome in Main results and the role of chance: The mean patient age was 39.1 years.
IVF cycles? Average blood levels of β-hCG were 201.74 ± 339.26 mIU/mL. The total clinical
Summary answer: Pregnancy viability can be predicted by the levels of early rates were: 28.1% BP, 33.7% EPL, 71.9 OP y 42.9 % LR. Statistically significant
β-hCG in IVF cycles. differences were found between β-hCG levels in fresh embryo cycles (170.6)
What is known already: There is an international agreement to consider> and frozen (228.7) (p=0.006). No statistically significant differences were seen
5 mUI / ml of β-hCG in blood as the positive predictive value after 10-15 days between β-hCG values in own egg cycles or donor (p=0.954) nor with natural
post-transfer, this being the standardized reference level for a pregnancy diag- endometrial preparation or with transdermal estrogen or oral and vaginal pro-
nosis. The subsequent evolution is very variable according to the initial value. gesterone in frozen transfers (p=0.099). No link was found between β-hCG
Levels past 50 mIU / ml are considered to have a good prognosis. Low levels levels and the day of embryo development. With regards to clinical prognosis,
are related, in most cases, with non-evolutionary pregnancies. There is a growing a direct relation was found between β-hCG levels and rates of BP(OR=0.981,
interest in establishing ranges that can be related to the subsequent evolution IC 95% [0.974-0.988], p<0.001), OP (OR=1.02, IC 95% [1.012-1.027], p<0.001)
of pregnancy. y LB ido vivo (OR=1.003, IC 95% [1.001-1.005], p<0.001). Confounding vari-
Study design, size, duration: Retrospective evaluation of a cohort of women ables were introduced to avoid bias: maternal age, cycle of own or donor egg,
with a positive result of serum β-hCG in IVF-ET cycles in a private fertility center. endometrial thickness, natural or HRT cycle, number of embryos transferred
The sample includes 196 cycles performed between July 2017 and January 2020. and fresh or frozen embryos. When analyzed in quartile the value of β-hCG,

1
the rate of LB was: first quartile (hCG38.5) 6.5%, second (hCG: 35.5-120-8) Reproduction Clinic Tokyo, Department of reproductive medicine, Tokyo, Japan ;
2
47.4%, third (120.8-214.5) 54.1%, fourth (>214.5) of74.3% (p<0.001). Reproduction Clinic Osaka, Department of reproductive medicine, Osaka, Japan
Limitations, reasons for caution: Larger prospective studies including
homogeneous cohorts are needed in order to corroborate our initial results. Study question: Is the new ERPeakSM endometrial receptivity test useful for
Wider implications of the findings: Early serum β-hCG values constitute patients with recurrent implantation failure (RIF)?
a potential biomarker for pregnancy outcome in assisted reproduction IVF cycles. Summary answer: A displaced window of implantation detected by ERPeakSM
Trial registration number: Not applicable test was frequently observed in RIF patients. Personalized embryo transfer after
ERPeakSM testing may improve pregnancy outcomes.
P-350 Endometrial assessment and personalised plan in the What is known already: Lack of synchronization between an embryo and
treatment of patients with unexplained repeated embryo the timing of endometrial receptivity is thought to be a cause of RIF. Therefore,
implantation failure: initial outcomes from a dedicated correctly identifying the window of implantation (WOI) is essential for maxi-
Implantation Clinic mizing the effectiveness of assisted reproduction treatments. While the wide-
spread endometrial receptivity assay (ERA) uses microarray analysis to
M. Rahmati1, A. Ripanelli1, M. Ollila1, S. Nair1, A. Gill1, R. Balet1,
determine the transcriptomic profile of 238 genes, the ERPeakSM test analyses
E. Kuan1, K. Ahuja1, N. Macklon1
48 genes by RT-qPCR, a methodology that has been demonstrated to have
1
London Women’s Clinic, Reproductive Medicine, London, United Kingdom the highest sensitivity, widest dynamic range and least bias for gene expression
analysis. This is the first report of clinical outcomes using ERPeakSM testing for
Study question: Can targeted treatment based on endometrial assessment RIF patients.
produce satisfactory outcomes in women who have suffered unexplained Study design, size, duration: A retrospective review was performed for
repeated embryo implantation failure? 137 patients who underwent ERPeakSM testing in our clinic between April and
Summary answer: A personalised treatment plan based on endometrial diag- October 2019. A total of 119 patients under 45 years old, who had 2 or more
nostics leads to good clinical outcomes and may represent an improvement on failed embryo transfers and underwent personalized embryo transfer (pET) after
current largely empirical management approaches. ERPeakSM testing, participated in this study. A hormone replacement cycle had
What is known already: Unexplained recurrent embryo implantation failure been performed for all patients. The first day of progesterone administration
(RIF) represents a major challenge in reproductive medicine. The current man- was defined as P+0.
agement of RIF is largely based on empirical treatments aim at improving endo- Participants/materials, setting, methods: An endometrial biopsy was
metrial receptivity. However, the emergence of tests of different aspects of
performed on day P+5 in an HRT cycle. After the ERPeakSM test result was given
endometrial function provides the opportunity to direct therapies to address
as receptive, pre-receptive or post-receptive, pET was performed in a subse-
underlying pathologies.
quent cycle on the day where the ERPeakSM test indicated optimal receptivity.
Study design, size, duration: A prospective cohort follow up study analysed In receptive cases, we also considered embryonic developmental speed to set
the endometrial testing results and clinical outcomes achieved from a referral the day of transfer: blastocyst grade 3,4,5 and 6 were transferred on day P+5,
clinic set up in June 2018, dedicated to the investigation and treatment of patients P+5.5, P+6.0 and P+6.5, respectively.
with unexplained repeated embryo implantation failure, who had at least three Main results and the role of chance: Of 119 RIF patients (average age,
high quality blastocysts transferred without success. 38.8 years), ERPeakSM testing showed a shifted WOI result in 50 patients (42.0%)
Participants/materials, setting, methods: Timed endometrial biopsy was and a receptive (R) result in 69 patients (58.0%). In the shifted WOI group, 66.0%
performed after 5 days of luteal support in a hormone substituted cycle. Biopsies (33/50) indicated a pre-receptive state and 34.0% (17/50) resulted in post-re-
were subject to endometrial dating by gene expression (ERA test, iGenomix, ceptive state. After pET for shifted WOI patients, we found that the pregnancy
Valencia) and to endometrial immune profiling including the recruitment and rate and implantation rate were similar between shifted WOI and R patients
activation of the uterine Natural Killer cells (MLI test, Matrice Lab Innove, Paris). (46.0% vs. 41.5% and 23.8% vs. 19.3%, respectively), which is consistent with
Based on the results, a management plan focusing on optimizing endometrial previous studies of pET based on the ERA test. pET for shifted WOI patients
preparation and embryo transfer was proposed to the referring clinician. showed similar pregnancy rate (42.9%, 47.4% and 47.1%) and implantation rate
Main results and the role of chance: 170 patients were referred to our (20.0%, 28.6% and 21.9%) stratified by patients’ age (<38, 39-41, 42-45 years
unit. 112 patients with an average of 4.3 blastocysts previously transferred old), respectively. Among R patients, 16 patients received pET in consideration
underwent investigation. 91% of the patients tested revealed at least one with embryonic developmental speed and 53 patients without such consider-
abnormal test outcome. For 70% of the patients, the ERA test showed a ation. the pregnancy rate and implantation rate of the former group were higher
non-receptive endometrium. For 75% of the patients, the MLI test found an (62.5% vs. 41.5% and 34.5% vs. 19.3%) compared to the latter, although differ-
endometrial immune dysregulation. No correlation was observed between ences were not statistically significant.
the results of the two test profiles. The outcome of the first attempt imple- Limitations, reasons for caution: There are several limitations, including
menting a personalised plan showed a pregnancy rate of 52% and an ongoing small sample size and lack of control group to compare clinical outcomes among
pregnancy rate of 37% after the first trimester, per transferred blastocyst. RIF patients who are not treated by pET with the ERPeakSM test. Embryos were
Those who did not achieve a subsequent implantation showed a higher prev- selected for transfer by morphology alone, rather than chromosomal screening,
alence of normal endometrial profiling compared to those who conceived on which may have affected the clinical outcome.
the implemented treatment plan. Wider implications of the findings: A shifted WOI detected by ERPeakSM
Limitations, reasons for caution: In order to assess the added value of was frequently observed in RIF patients. pET for shifted WOI patients after
personalised a treatment plan, outcomes need to be compared prospectively ERPeakSM testing was consistent with ERA for pregnancy outcomes regardless
with a matched control group who are managed without implementation of a of patients’ age. pET for R patients in consideration with embryonic develop-
personalised plan. mental speed may also improve pregnancy outcomes. Further studies are
Wider implications of the findings: Our study suggests that treatment required.
based on endometrial diagnostics might improve outcomes compared with Trial registration number: not applicable
empirical management. This approach provides the opportunity to carry out
randomized controlled trials of targeted rather than blind interventions. P-352 Retrospective evaluation of diagnostic usefulness of
Trial registration number: Not Applicable endometrial lymphocyte subpopulations quantification: influence
in endometrial receptivity and in pregnancy outcome in the
P-351 First clinical outcomes after personalized embryo setting of assisted reproduction techniques
transfer using the new endometrial receptivity test in recurrent D. Valero Hervás1, J. Sarasa2, M. Enciso2, L. Agúndez1, J. Aizpurua3
implantation failure patients 1
iGLS, Immunology, Alicante, Spain ;
Y. Ohara1, H. Matsubayashi1,2, K. Yamaguchi1,2, K. Kitaya2, 2
iGLS, Research and Development, Alicante, Spain ;
M. Doshida1, T. Takeuchi1, T. Ishikawa1,2 3
IVF Spain, Gynaecology, Alicante, Spain

Study question: Is the distribution of lymphocyte populations in the endo- Summary answer: A DNA extraction protocol designed for low-biomass
metrium associated with endometrial receptivity and the appearance of early samples allows to increase intra-uterine microbiota profiles quality.
complications during pregnancy? What is known already: In recent years, the view of a sterile uterus is being
Summary answer: Endometrial lymphocyte populations are associated with challenged, there is a growing interest in understanding the potential role of
endometrial receptivity status and can be a useful indicator for the early diagnosis microbiota of the female upper reproductive tract in processes leading to a
of adverse reproductive outcomes. healthy pregnancy. Indeed, several publications have shown preliminary associ-
What is known already: During pregnancy, a series of crucial adaptations ations between the intra-uterine microbiota and the outcome of pregnancy.
occur in the maternal immune system, aimed at ensuring correct embryo implan- However, samples containing low microbial biomass, such as intra-uterine sam-
tation and development, while avoiding recognition of the embryo as foreign ples, are especially susceptible to contamination by DNA and/or cross-contam-
tissue that could lead to its rejection. Recent studies have shown that distur- ination between samples, which can easily dominate the obtained microbiota
bances in this balance may lead to pregnancy complications, so the achievement profiles and lead to erroneous interpretation of results.
of a successful pregnancy requires maternal and fetal cells coexistence in a tightly Study design, size, duration: Bacterial DNA from intra-uterine biological
regulated balance. In this study, the different immune populations’ profiles pres- replicate samples (n=38x2) was extracted using two methods: automated
ent in the endometrium and their association with different infertility conditions extraction using a standardized kit and manual extraction in a room specially
have been evaluated. designed to handle low-biomass samples. Technical and procedural negative
Study design, size, duration: This is a retrospective study analysing immune controls were taken along the process (n=33).
populations distribution in endometrial tissue and assisted reproduction tech- Participants/materials, setting, methods: Intra-uterine samples were
niques (ART) outcomes of patients undergoing immune status assessment under retrieved using embryo transfer catheters with a sterile bridging tube, flash-frozen
different clinical suspicions (recurrent implantation failure or/and miscarriage in liquid nitrogen and stored at -80C. DNA was isolated using a Tecan Nucleic
and primary infertility). Immune profiles were analysed in 239 patients between Acid Extraction-Platform (MagAttract PowerMicrobiome DNA/RNA kit
October 2018 and December 2019. These profiles were compared according [Qiagen]) or using a low-biomass-optimized manual extraction (Allprep DNA/
to the day of the endometrial cycle in which the sample was obtained and the RNA kit [Qiagen]). Microbiota profiles were obtained by 16S rRNA amplicon
receptivity status measured by ER Map®. sequencing (V4) using the gold-standard DADA2 pipeline, and analysed using
Participants/materials, setting, methods: Patients referred for endome- constrained principal coordinates analysis at genus level (cPCoA).
trial function evaluation were included in the study. Biopsy samples were obtained Main results and the role of chance: Biological replicate samples (N=38x2)
in hormone replacement therapy (HRT). ImMap® test consisting on the evalu- of the intra-uterine microbiota were subjected to two extraction protocols to
ation of the percentage of different Natural Killer (NK) cell phenotypes, Th1, evaluate the importance of protocol optimization for these low-biomass samples.
Th2, Th17 and regulatory T lymphocytes, and B1a lymphocytes by means of The extraction protocol explained 27% of the variation in intra-uterine micro-
flow cytometry was performed in all cases. The profiles identified were evaluated biota profiles (n=76, db-RDA, R2=0.27, adjP=0.0002). Microbial diversity was
according to the available values of normality stated in the scientific literature. higher with the DNA extraction protocol optimized for low-biomass samples
Main results and the role of chance: Our study showed differences in as compared to the standard protocol (n=76, Simpson diversity index, Wilcoxon
population dynamics depending on the day of the cycle in which the biopsy was rank-sum, r=0.0028, P=3.61E-09). We tracked, in the negative controls of each
performed. The samples obtained between days P4+5 and P4+6 showed sig- protocol, a total of 119 genera as contaminants, 13 of which were common to
nificant enrichment in NK cells compared to those taken further away from the both protocols. We thus identified 90 genera of trustworthy biological sampling
theoretical implantation window (WOI) (T-test: 35.67% vs 26.37%; p=0.016). origin (non-contaminants) in the standard protocol, compared to 136 in the
These differences were confirmed analysing exclusively those endometria clas- optimized protocol. These genera were used to calculate the amount of sequenc-
sified as receptive using ER Map® technology (T-test: 36.02% vs 25.59%; ing data generated per biological sample that is of trustworthy origin. With the
p=0.039), and a decrease in the Tc cells was also identified (T-test: 54.97% vs standard protocol, only 21% of the intra-uterine samples achieved the minimum
48.82%; p=0.042). Furthermore, the receptivity stratification also showed that threshold of 1000 trustworthy sequencing reads, while 79% of the biological
the lack of this predominance of the NK cells is associated with a post-receptive samples passed that threshold with the optimized protocol.
endometrial state (T-test: 36.39% vs 11.47%; p=0.028). Similarly, a receptive Limitations, reasons for caution: The negative controls used in this analysis
endometrium would be characterized by a lower proportion of the lymphoid focused on the evaluation of contaminants during DNA extraction and sequenc-
group characterized by CD45 (Man-Whitney test: p=0.0001), compared with ing. The addition of negative controls for the sampling procedure (empty sam-
those non-receptive. In the context of the clinical outcomes, the results show pling devices) would give additional information on potential DNA contaminants
that in women with a WOI between P4+5 and P4+6 who suffered from early in devices used for sampling.
abortions, had a lower proportion of activated NK cells when compared to Wider implications of the findings: Applying 16S rRNA amplicon
those women who did not develop any adverse effects (p=0.034). sequencing on low-biomass intra-uterine samples is a two-edged sword since
Limitations, reasons for caution: Our study reveals differences in immune its efficiency for detecting microbial DNA also makes it highly sensitive to
populations dynamics with potential usefulness in predicting pregnancy evo- contaminant DNA and to cross-contamination. Ignoring this drawback can
lution. However, the context and type of sample evaluated hinders the estab- lead to erroneous interpretation of microbiota profiles and their role in human
lishment of normality ranges, so to understand the extent of its usefulness it reproductive health.
would be necessary to carry out clinical trials including healthy con- Trial registration number: NCT03105453
trolled women.
Wider implications of the findings: The application of ImMap® assessment P-354 Placental morphology remains unaffected by addition of
as a routine tool for identifying abnormal immune profiles would lead to the GM-CSF during pre-implantation development
optimal and appropriate medical intervention, in order to prevent or minimize V. Nordhoff1, L. Ogoniak1, T. Pock1, K. Schulte2, S. Schlatt1,
the appearance of adverse outcomes and improve effectively the ART outcomes, M. Boiani3
and the life-quality of couples with a reproductive desire. 1
University Hospital of Muenster, Centre of Reproductive Medicine and Andrology,
Trial registration number: Not applicable Münster, Germany ;
2
University of Muenster, Central Animal Facility of the Faculty of Medicine,
P-353 DNA extraction protocol optimized for low-biomass Münster, Germany ;
samples are essential for accurate profiling of the intra-uterine 3
University of Münster, Max Planck Institute for Molecular Biomedicine, Münster,
microbiota. Germany
J. Centelles Lodeiro1
1
KU Leuven, VIB-KU Leuven Center for Microbiology, Leuven, Belgium Study question: Does GM-CSF supplemented during pre-implantation devel-
opment have an influence on fetal growth or placenta development?
Study question: Does the DNA extraction protocol affect the accuracy of Summary answer: GM-CSF had an effect on fetal weight but placenta mor-
low biomass intra-uterine microbiota profiling? phology was not affected by different concentrations of GM-CSF.

What is known already: The growth factor granulocyte macrophage colo- aim to identify WOI transcriptomic signatures with clinical significance associated
ny-stimulating factor (GM-CSF) is used in ART for preimplantation embryo to different infertility conditions and ART outcomes.
cultivation to increase survival of embryos up to week 12, especially in women Study design, size, duration: This is a retrospective study analysing tran-
with previous miscarriages. Naturally, GM-CSF is produced by the endometrium scriptomic signatures, infertility conditions and ART outcomes of 1095 patients
during early gestation. The corresponding receptors can be found in the tropho- undergoing endometrial receptivity assessment by ER Map® between March
blast cells of the early embryo. After implantation trophoblast cells establish the 2016 and January 2020. Results obtained when embryo transfers were scheduled
connection of the embryo with the maternal tissue and the placenta forms. The on the moment of endometrial receptivity (WOI timeframe) were analysed.
mature placenta consists of the fetal layers labyrinth and spongiotrophoblast and Prediction models of gene expression profiles of patients with different infertility
the maternal decidua. conditions and ART outcomes were evaluated.
Study design, size, duration: Female mice (B6C3F1, 5-10 weeks old) were Participants/materials, setting, methods: A control group of healthy
superovulated by injection of 5 IU PMSG and 10 IU hCG and mated to males egg-donors and patients referred for ER Map® analysis for a variety of reasons
(C57BL/6J). Zygotes were isolated on day 0.5 post coitum, pooled and randomly including implantation failure (RIF) and recurrent miscarriage (RM) were included
allocated to KSOM(aa) with 0, 2 and 10 ng/mL mGM-CSF. Medium refreshment in the study. ER Map® gene expression analysis of endometrial biopsies obtained
was applied on day 2.5. Embryos were transferred to foster mothers and the in HRT cycles at P4+5.5 was performed by qRT-PCR (Biomark-HD, Fluidigm).
developing fetuses and placentas were dissected at day 13.5 of gestation. Transcriptomic profiles from donors, different infertility conditions (RIF, RM)
Participants/materials, setting, methods: In total 48 placentas and fetuses and ART outcomes (successful implantation, biochemical pregnancy and clinical
could be collected of which 41 placentas were appropriate for analysis: 13 pla- pregnancy) were evaluated by discriminant analysis.
centas in the control group (KSOM(aa)), 15 in the group with 2 ng/mL GM-CSF Main results and the role of chance: We have identified WOI transcrip-
and 13 in the group with 10 ng/mL GM-CSF. Tissues were collected to measure tomic signatures predictive of recurrent miscarriage (RM) and implantation failure
fetus weight and length and placenta weight. Placentas were embedded, cut and (RIF). In the RM group the overall accuracy prediction in the training set is 100%,
stained by PAS-reaction. Area and diameter of the single placenta layers were all 37 RM patients and 9 controls were classified in their respective groups.
measured from 6 sections each. Leave-one-out cross-validation presented a positive predictive value (PPV) of
Main results and the role of chance: Fetal weight was significantly higher after 86.67%. In the RIF group the overall accuracy prediction in the training set is
culture in medium containing 2 ng/mL GM-CSF compared to the control medium 99.2%, only 2 out of 235 RIF patients were misclassified as controls. Leave-one-
(mean values: 0ng/mL=0.17g; 2ng/mL=0.21g; 10ng/mL=0.18g; P-value <0.05). out cross-validation presented a PPV of 99.57%.
No difference was found between placenta weight in different concentrations of Analysis of ART outcomes in our cohort showed that 81.64% (894/1095) of
GM-CSF (mean values: 0 ng/mL=0.115 g; 2ng/mL=0.122 g; 10ng/mL=0.122 g). patients achieved implantation, of which 9.96% (89/894) had a biochemical
Values for placenta layer areas and diameters were similar in each group, as well as pregnancy and 73.52% (805/1095) continued to have clinical pregnancies.
the whole placental area (mean values: 0 ng/mL=10.01mm2 g; 2ng/mL=10.49mm2 Analysis of the gene expression profiles allowed the identification of a WOI
g; 10ng/mL=10.65mm2 g) and diameter (mean values: 0 ng/mL=2.05mm2; 2ng/ signature predictive of successful outcome (clinical pregnancy) and two WOI
mL=2.19mm2 g; 10ng/mL=2.29mm2). A significant correlation was found between signatures predictive of negative reproductive outcomes (implantation failure
body length and weight of the fetuses (R2 = 0.3869). Additionally, fetus length and and biochemical pregnancy). The group of samples identified to present a pos-
placenta weight were associated as was placenta area and placenta weight. Whole itive outcome signature had an implantation rate of 91.95%, a biochemical preg-
placenta area and also the single layers (labyrinth, spongiotrophoblast and decidua) nancy rate of 3.43% and a clinical pregnancy rate of 88.52%. These rates are
and their proportions were similar among all groups. significantly different (p<0.001) than the ones obtained in the group of samples
Limitations, reasons for caution: A limitation of this study is the use of an identified to present negative outcomes signatures.
animal model and the number of examined placentas. Data cannot be transferred Limitations, reasons for caution: ER Map® test can serve as a valuable tool
1:1 to the human situation. Further studies should investigate the effect of to improve the diagnosis of endometrial function and anticipate ART outcomes
GM-CSF during several developmental steps in the development of either the in sub-fertile patients, however, other types of investigations aimed to determine
embryo or the fetus. the therapeutic options for patients at risk of negative reproductive outcomes
Wider implications of the findings: GM-CSF had an influence on fetus will also be necessary.
weight but not on placenta weight or morphology. Although it seems that this Wider implications of the findings: The ability of ER Map® for the pre-
growth factor has no major influence during culture, still our data create aware- diction of cases at risk of negative reproductive outcomes offers a comprehen-
ness for the general use of growth factors in ART culture media. sive method for the diagnosis of endometrial function and infertility. This tool
Trial registration number: not applicable may open new research lines to understand endometrial pathology and develop
new treatment strategies.
P-355 Evaluation of ER Map® test to predict endometrial
transcriptomic signatures associated to different infertility P-356 Whole-genome sequencing of embryos from pregnancy loss
conditions and ART outcomes. identifies putatively detrimental genomic variants
J. Sarasa Marcuello1, J. Aizpurua2, B. Rodriguez-Estrada1, I. S. Bonaiuto1, I. Di Biase2, V. Aleotti3, A. Ravaei3, G. Damaggio1,
Jurado1, M. Ferrández1, A.B. Climent3, M. Enciso1 M. Chierici4, P. Madhuri5, A. Qasim5, C. Furlanello4, E. Garrison6,
1
iGLS, Research and Devolopment, Alicante, Spain ; N. Soranzo7, A. Capalbo8, R. Michele3, S. Di Biase2, V. Colonna1
2
IVF Spain, Reproductive Medicine, Alicante, Spain ; 1
Consiglio Nazionale delle Ricerche, Istituto di Genetica e Biofisica, Napoli, Italy ;
3
IVF Spain, Statistics, Alicante, Spain 2
Merigen Research s.r.l., Research & Development, Naples, Italy ;
3
University of Ferrara, Department of Environmental- Biological and
Study question: Is it possible to identify endometrial transcriptomic signatures Pharmaceutical Sciences and Technologies, Ferrara, Italy ;
with clinical significance using the molecular tool ER Map®? 4
Fondazione Bruno Kessler, Predictive Models for Biomedicine & Environment,
Summary answer: ER Map® gene expression analysis is able to predict infer- Trento, Italy ;
tility conditions and ART outcomes with clinical significance. 5
Monash University Malaysia, School of Science, Jalan Lagoon Selatan, Malaysia ;
What is known already: Transcriptomic analyses have been proposed in 6
University of California Santa Cruz, UC Santa Cruz Genomics Institute, Santa
many fields of medicine as powerful tools to identify pathological conditions. In Cruz, U.S.A. ;
reproductive medicine, gene expression analysis of endometrial tissue has been 7
Wellcome Sanger Institute, Human Genetics, Hinxton, United Kingdom ;
recognized as an effective approach for endometrial receptivity evaluation. ER 8
Igenomix, Igenomix, Marostica, Italy
Map® is a molecular diagnostic tool able to accurately predict the window of
implantation (WOI) by analysing the expression profile of 40 genes by Study question: Do whole-genome sequences of embryos from recurrent
high-throughput RT-qPCR. Other conditions of altered endometrial function pregnancy loss clarify the impact of genetic variants of small-size and/or located
beyond receptivity could be responsible of failed ART results. In this study we in non-coding regulatory regions?

Summary answer: Whole-genome analysis of embryonic DNA from preg- Study question: Can assessment of pH at the surface of the endometrium of
nancy loss successfully identifies putatively detrimental genomic variants the uterine fundus distinguish the condition of chronic endometritis(CE) from
What is known already: Pregnancy Loss (PL), the spontaneous demise the normal condition?
of pregnancy before 24 weeks of gestation, occurs in 10-15% of pregnancies. Summary answer: Assessment of the pH at the surface of the endometrium
PL is often the result of chromosomal aneuploidies of the gametes but it can of the uterine fundus is thought to be an excellent novel marker of chronic
also have non-random genetic causes like small mutations (SNPs and indels), endometritis.
both de-novo or inherited from parents. Comparative genomic hybridization What is known already: CE is a state of persistent inflamation in the endo-
(CGH) detects variants of several thousand base pairs while targeted resequenc- metrium caused by common bacterial pathogens. It is reported to be associated
ing resolves point mutations. Both are currently the most accurate methods for with reproductive failure. Endometrial plasma cells (CD138 immunostainined
the genetic analysis of PL, but are not sensitive to small variants (CGH), or do cells) are accepted as the standard criterion to establish a diagnosis of CE.
not target those located in non-coding regulatory regions (targeted Recently, sequencing of the 16S rebosomal RNA revealed that lactobacillus
resequencing). species are abundant in the uterine cavity in women without CE, and much less
Study design, size, duration: Our aim is to identify small-size genetic variants abundant in CE patients. However, the effects of lactobacillus in the endome-
likely to cause PL using a predictive model integrating whole-genome sequence trium are unknown. Uterine fluid pH in both CE and non-CE is reported to be
data with functional annotations and gene networks relevant to embryonic devel- around 7.2 (Moreno et al.,2016).
opment. Seventy women, mostly European (82.7%) diagnosed with first (n=39, Study design, size, duration: We tried to determine whether assessment
av.age 28.9 ) or recurrent (n=31, av.age 39.0) miscarriage were recruited by the of the pH of the endometrium could be a new marker of chronic endotmetritis.
University of Ferrara from 2017 to 2019. Ethical committee approval is from Asymptomatic women (N=56) with recurrent implantation failure or recurrent
Emilia-Romagna CE/FE (#170475). miscarriage were recruited. Chronic endometritis was diagnosed using CD138
Participants/materials, setting, methods: Fetal DNA extracted immunostaining, and the uterine microbioma by using sequensing of the 16S
from chorionic villi was used to exclude samples with aneuploidies using ribosomal RNA gene. We checked uterine pH in both CE and non-CE. This
both CGH and shallow sequencing of random genomic regions. Euploid sam- study was performed from June 1 until December 31,2019.
ples were whole-genome sequenced at high-coverage. Variants were called Participants/materials, setting, methods: Participants were recruited from
against the reference genome GRCh38 with Freebayes. Variants were anno- the infertility clinic of Tokeidai Memorial Hospital. Plasma cells in the stroma of
tated with a custom script that integrates information from Ensembl99 with endometrium were indentified by means of immunohistochemistry staining.
publicly available manually curated lists of genes associated with embryonic Sequencing of 16S rebosomal RNA gene in the endometrial fluid was done for
development, miscarriages, lethality, cell cycle. The code is available on gitHub detecting bacteria. Endometrial surface pH was examined directly at the endo-
(ezcn/grep) metrial surface using a Portable pH Meter (Japan Asch Inc., Japan) that has been
Main results and the role of chance: We understood the requirements to utilized for the esophageal mucosa of esophagitis ( Journal of Gastroenterology,
scale-up the project and obtained initial results from the analysis of these genomic Japan, 1990).
sequences. We determined that shallow sequencing of random genomic regions Main results and the role of chance: We tried to examine some markers
is more efficient compared to CGH in detecting aneuploidies in samples with of CE in a total of 56 patients with reproductive failure who gave informed
poor quality DNA. We estimated that 20% of collected samples are suitable for consent. We first screened for CE using our criteria following 2 criteria reported
sequencing, the rest presenting aneuploidies or quality issues and maternal con- by Liu et al.(Fertil Steril, 2019), >5.15 CD138 stained cell/10mm2 or <85%
tamination. Sequenced samples have on average 4M high-quality variable sites lactobacillus in the Endometrial Microbioma Test (Varinos Inc., Japan). The endo-
that were annotated with information on gene content and functional conse- metrial pHs of the follicular phase and luteal phase in non-CE were
quences. In the autosomes 4.3k variants are ranked as having a high deleterious 5.34±1.00(mean±SD)(n=8) and 5.43±0.97(n=13), respectively. This difference
impact according to Ensembl, 69.1k moderate, and 111k low. After filtering was not significant. The endometrial pH of CE, 6.23±0.65(n=35), was signifi-
based on the combination of several criteria, such as allele frequency in the cantly different from that of non-CE, 5.61±0.74 (n=21)(P<0.01). The endome-
general population, homozygosity, impact of the allele with consequences we trial pH of CE (CD138 cell number>5.15/10 mm2) was 6.39±0.58(n=18),
identify a number of putatively detrimental mutations. Three samples carry two which was also significantly different from that of non-CE(CD138 cell number
homozygous missense mutations in the same exon of the AHNAK2 cancer-re- <5.15/10mm2), which was 5.41±0.79 (n=20)(p<0.01). The endometrial pH
lated gene, which codes for a cytoplasmic nucleoprotein. Five samples share the of CE(lactobacillus<85%),was 6.67±0.39(n=17), whcih was significantly different
same stop gain mutation in the RPTOR gene, a component of a signaling pathway from that of non-CE(lactobacillus >85%), which was 5.58±0.59 (n=15)(p<0.01).
that regulates cell growth and survival, and autophagy in response to nutrient Limitations, reasons for caution: As this study was performed in only 56
and hormonal signals. female patients with infertility or recurrent miscarriage, a larger number of
Limitations, reasons for caution: While encouraging, these first results patients need to be studied for precise evaluation of the usefulluness of the
needs to be corroborated with a more comprehensive analysis that fully imple- dendometrial pH.
ment a predictive model. Results will be validated in a biobank of embryonic Wider implications of the findings: The endometrial pH was usually acidic,
sequence data from PL. unlike in from Mereno et al.’s report. The endometrial pH of CE was significantly
Wider implications of the findings: We demonstrated that whole-genome higher than that of non-CE. The endometrial lactobacillus is associated with the
sequencing can help to clarify the causes of RPL. The pilot study described here endometrial surface pH. Therefore, endometrial pH is thought to be an excellent
has discovered plausible candidate RPL-associated variants and provides essential novel CE maker.
indications for the realization of a larger study. Trial registration number: R1919
P-357 Assessment of pH at the surface of the endometrium of the P-358 Modelling embryo implantation in vitro using a three-
uterine fundus is a novel marker of chronic endometritis dimensional co-culture system
T. Endo1, T. Baba2, E. Itabashi3, H. Honnma4, H. Henmi5, U. Ikeda5, C. Hill1, D. Hapangama1, M. Phelan2
T. Kiya6, T. Saito7, A. Azumaguchi8, M. Fujii3 1
University of Liverpool, Women’s and Children’s Health, Liverpool, United
1
Sapporo Medical University, Obs & Gynae Dept., Sapporo- Hokkaido, Japan ; Kingdom ;
2
2
Sapporo Medical University, Obstetrics and Gynecology, Sapporo, Japan ; University of Liverpool, Institute of Integrative Biology, Liverpool, United Kingdom
3
Tokeidai Memorial Hospital, Obstetrics and Gynecology, Sapporo, Japan ;
4
Sapporo ART Clinic, Obstetrics and Gynecology, Sapporo, Japan ; Study question: Can we construct a 3D in vitro model of endometrial embryo
5
Tonan Hospital, Obstetrics and Gynecology, Sapporo, Japan ; implantation to study early human pregnancy establishment?
6
ENA Aazabu ART Clinic, Obstetics and Gynecology, Sapporo, Japan ; Summary answer: We were able to develop a 3D human endometrial co-cul-
7
Sapporo Medical University, Obstetriics and Gynecology, Sapporo, Japan ; ture model that accepted surrogate embryo implantation to simulate early human
8
Sapporo Endometrium Research, Obstetrics and Gynecology, Sapporo, Japan pregnancy.

What is known already: The human and non-human primate endometrium Study design, size, duration: This was a prospective study in a single repro-
is unique amongst mammals in its ability to decidualise, shed and regenerate. ductive centre. The infertile patients included in this study underwent IVF treat-
Furthermore, the cellular architecture of human endometrium is found only in ment between June 2016 and December 2017.
higher primates. These fundamental differences make it difficult to model early Participants/materials, setting, methods: A cohort of 13,977 consecutive
pregnancy establishment in routinely studied laboratory animals. Therefore, women undergoing transvaginal ultrasound scan on Day 27–29 after ET were
novel models of human embryo-endometrium interactions are needed to fully included. Ultrasound measurements and clinical characteristics were recorded.
define the factors involved in successful implantation. The first trimester pregnancy outcome of these women was noted at 12 weeks
Study design, size, duration: A 3D in vitro model of the endometrium was of gestation. The cases of first year were used to generate a training sample
established using endometrial epithelial cell line Ishikawa and a telomerase (7,261) and a simple scoring system was derived from this. The cases of second
immortalised human endometrial stromal cell line embedded in a collagen hydro- year were used as a verification sample (6,716).
gel. The model was expanded to include primary human endometrial epithelial Main results and the role of chance: There were 12,051 cases with an
and stromal cells isolated from freshly collected endometrial biopsies from ongoing pregnancy and 1,926 cases had EPL. Maternal age (MA), gestational sac
healthy women not on hormonal medication. Embryo implantation was simu- diameter (GSD), crown-rump length (CRL), embryonic heart rate (HR), yolk
lated by attachement of Jeg3 trophoblast spheroids over a ten day period. sac diameter (YSD) and endometrium thickness (EM) on transfer day finally
Participants/materials, setting, methods: Liverpool Women’s Hospital entered the scoring system after stepwise screening. Points associated with each
is a tertiary referral centre affiliated to the University Hospital Research Centre. category of each risk factor were computed and the risks associated with point
Samples of endometrium was harvested from women undergoing benign gynae- totals were determined according to the “The Framingham Study risk score
cological procedures after obtaining informed consent. Surrogate embryo system”.
implantation in 3D culture models was assessed by immunohistochemistry. The The scoring system gave an area under the curve (AUC) of 0.884 (95% confi-
metabolomic signature of early pregnancy was assessed by NMR analysis of dence interval 0.870-0.899) in the training set. The score totals range from point
conditioned medium from endometrium-embryo co-cultures. -8 to 14. When point 5 was chosen as the cutoff value, the predicting risk of
Main results and the role of chance: We initially employed human cell lines EPL was 30.03%, with a sensitivity of 66.87%, specificity of 98.11%, positive
to develop a 3D construct containing epithelial and stromal cells, and a variety predictive value (PPV) of 84.44% and negative predictive value (NPV) of 95.07%
of protocol optimisations were necessary to replicate the different cycle phases in the test set, which performed with a sensitivity of 64.69%, specificity of
and decidualisation process of the endometrium. The optimised conditions for 98.78%, PPV of 89.87% and NPV of 93.62% in the test set. 94.01% cases of the
cell lines were further adjusted for primary cells, which demonstrated an training sample could be correctly predicted as EPL and in the verification sam-
enhanced capacity for self-organisation and remodelling of the collagen matrix. ples, 93.91% of the cases were correctly predicted.
Successful decidualisation of the 3D constructs in the presence of oestrogen Limitations, reasons for caution: This scoring system was used for easy
and progesterone was confirmed by ELISA and immunohistochemistry. Jeg3 and quick prediction of EPL with false positive and false negative results and the
spheroids were show to attach to the 3D constructs and interact with the epi- miscarriage outcome cannot be changed by this system. Smoking variable was
thelial and stromal elements. Preliminary analysis of the metabolome defined by a potential omission in the data collection and might further improve the pre-
NMR analysis of the co-culture conditioned medium identified metabolites dictive performance if included.
unique to embryo implantation. Wider implications of the findings: This simple scoring system incorporates
Limitations, reasons for caution: Only the Jeg3 choriocarcinoma cell line readily available data that are routinely collected in clinical practice and does not
was used as a surrogate embryo model, which does not fully capture all stages rely on complex data entry. The findings could be easily incorporated into early
of blastocyst implantation and invasion (e.g. syncytialisation). Future work should pregnancy care and effectively guide subsequent medical plans.
incorporate other cell lines and primary cells to model all elements of trophoblast Trial registration number: not applicable
adhesion, invasion and migration.
Wider implications of the findings: Modelling human embryo-endometrial
P-360 The incidence of monozygonic twins in assisted
interactions in vitro helps to widen our understanding of implantation. Such
reproduction technology -a retrospective study
information will prove invaluable in many areas of reproductive medicine, such
as increasing the success of IVF treatment and defining the causes of recurrent X. Feng1, P. Liu1
1
miscarriage. Center for Reproductive Medicine, Department of Obstetrics and Gynecology-
Trial registration number: not applicable Peking University Third Hospital, Beijing, China
P-359 A simple scoring system for prediction of early pregnancy Study question: does the incidence of MZT (monozygonic twin) in assisted
loss developed by following 13,977 infertile patients after in vitro reproduction techonology higher and what are the risk factors?
fertilization Summary answer: this retrospective research shows that blastocyst culture
Y. Ouyang1, P. Cai2, X. Li1 significantly increased the risk of monozygonic twins. However, the age, ICSI or
1
Reproductive and genetic hospital of Citic-XIangya, Imaging Department, assisted hatching have no signifcant impact.
Changsha, China ; What is known already: It is known that the incidence of monozygonic twin
2
Central South University, Institute of Reproductive and Stem cell engineering, (MZT) in natural conception is about 0.4%. Previous report has shown that the
Changsha, China risk of MZT has doubled after assisted reproductive technology. MZT can lead
to miscarrige, increase the incidence of preterm birth and other adverse neonatal
Study question: What is the performance of a simple scoring system to predict outcomes. Therefore, researchers have been take much attention to the inci-
early pregnancy loss (EPL) after in vitro fertilization-embryo transfer (IVF-ET)? dence and reasons of MZT in assisted reproduction and found that maternal
Summary answer: A simple scoring system using prospectively collected age, ICSI and blastocyst tranplantion may be associated with the happening
routine data could be easily used in early pregnancy care and effectively guide of MZT.
subsequent medical plans. Study design, size, duration: we conducted a retrospective study in our
What is known already: The first routine ultrasound scan is commonly center based on data from 2014 to 2018. In all, 41538 patients that got pregnancy
arranged on day 28 after ET in most reproductive centers to determine the after IVF were include in this study.
location and viability of the embryo. The mental pressure associated with a Participants/materials, setting, methods: there were 20048 fresh cyclyes
pregnancy of uncertain viability are significant in patients undergoing IVF-ET, and 11885 frozen cycles included. We analyzed different clinical data (maternal
thus, a rapid and accurate method to predict the possibility of early miscarriage age and stimulation strategy) and IVF strategy such as type of embryos, insem-
would be helpful for determining subsequent treatment. Previous mathematical ination method, embryo stage at time of ET. All pregnance were confirmed by
models have combined individual risk factors with reasonable performance. detection of fetal heart activity . The identification of MZT is identified when
However, a simple scoring system that can be easily implemented in clinical more than one fetal poles was visualized in one gestational sac via trans-vaginal
practice has not been described for the prediction of EPL after IVF-ET. ultrasound.For the statistical analysis, SPSS software was used.

Main results and the role of chance: In our clinic center, there were 59 Limitations, reasons for caution: The embryo transfer was done in case
patients who were MZT from 12060 patient who transferred D3 fresh of the emdometrium thickness not less then 6 mm.
embryos(0.49%). In addition, 9 patients were MZT from 590 patients who trans- Wider implications of the findings: It is not clear how the intrauterine
ferred fresh blastocyst(1.52%). These two groups have significant difference. In administration of PRP acts to affect endometrial thickness. Results from studies
frozen cycles, there were 32 patients who got MZT in 5042 patients who received on the role of endometrial thickness on implantation and live births are contra-
D3 embryo transfer(0.63%)and 145 patients who got MZT in 7085 ones who dictory. There is the urge for well-designed randomized studies to improve our
received blastocyst embryo transfer(1.96%).The incidence of MZT was signifi- knowledge on PRP in basic science.
cantly lower in D3 ET cycles than that in blastocyst ET cycles. In addition, the Trial registration number: No number
incidence of MZT in patients who were more than 35 years old have no signif-
icant difference with those who wre no more than 35 years old (fresh cycles: P-362 New predictors of early impaired placentation preceding
0.66% vs 0.47; frozen cyclyes:1.17% vs 0.86%). Moreover,we analyzed the cor- miscarriage before 10 weeks of gestation in IVF pregnancies: a
relation of insemination with MZT and found that there were no difference no prospective study
matter whether ICSI were used or not(0.66% vs 0.57% in ICSI). At last, the
A. Ben-Haroush1, H. Hamutal2, A. Wertheimer1, K. Tenenbaum-
manupulation of assisted hybradation have no impact on MZT incidence( 1.68%
Gavish3, Y. Shufaro1, A. Idelson3
vs 1.23% for AH ).
1
Limitations, reasons for caution: . It should be noticed that more data from Beilinson, IVF and Infertility Unit, Petach - Tikva, Israel ;
2
randomized trial were required. TeleMarpe Ltd, Tel Aviv, Tel Aviv, Israel ;
3
Wider implications of the findings: From the data we can found that Rabin Medical Center, Obstetrics and Gynecology Ultrasound Unit, Petach Tikva,
blasotcyst culture and transfer have been identifed to significantly influence the Israel
incidence of MZT. Therefore patients should be noticed of the risk of blastocyst
cutlure and tranfer. Study question: We prospectively evaluated early first-trimester gestational
Trial registration number: no sac and fetal biometry with maternal blood PP13, to predict impaired placenta-
tion prior to miscarriage before GA 10 weeks in IVF pregnancies.
Summary answer: In IVF pregnancies with live embryo at first ultrasound
P-361 Clinical applications of platelet-rich plasma in poor scan, high UtA-PI/CRL and PP13/CRL ratios may indicate impaired placentation
endometrium patients in IVF practice preceded early pregnancy loss.
O. Feskov1, Y. Zhylkova2, A. Feskova3, O. Blazhko4, O. Yegunkova5 What is known already: Fetal loss during the first trimester occurs in 10-20
1
Centre of Human Reproduction Sana-Med, IVF, Kharkiv, Ukraine ; percent of pregnancies. In a previous study we showed that small CRL in the
2
Center of Reproductive Medicine “Clinic of professor Feskov”, IVF Department, early first trimester may predict early fetal loss in IVF pregnancies; fetal losse
Kharkiv, Ukraine ; rate was 17.2% in pregnancies with CRL ≤10 percentile compared to 6.6% in
3
Kharkiv National Medical University, Department of Obstetrics and Gynaecology, the appropriate to GA CRL group (P=0.005). Placental protein 13 (PP13), is
Kharkiv, Ukraine ; exclusively expressed in placenta from a very early stage of pregnancy and can
4
Feskov Human Reproduction Group, Surrogacy Department, Kharkiv, Ukraine ; be detected in the maternal blood from the fifth week of gestation. Some studies
5
Center of Rproductive Medicine “Clinic of Professor Feskov”, Genetic laboratory, have indicated that damaged placenta may leak high levels of PP13.
Kharkiv, Ukraine Study design, size, duration: Cohort of 115 healthy IVF patients with a
singleton viable embryo in early first trimester.
Participants/materials, setting, methods: Calculation of gestational age
Study question: Does the intrauterus application of platelet-rich plasma (PRP)
(GA); ultrasound evaluation of crown-rump length (CRL), mean gestational sac
allow to improve the condition of the endometrium and IVF outcome in repeated
diameter (GSD) and volume (GSV), mean yolk sac diameter (YSD) and volume
implantation failure patients?
(YSV); fetal heart rate (FHR), mean uterine arteries pulsatility index (UtA-PI);
Summary answer: Application of PRP in patients with refractory emdome-
and maternal blood placental protein 13 (PP13) levels. Patients were divided
trium resulted in satisfactory endometrial thickness. PRP was effective in IVF
into three groups by GA; and early miscarriage versus ongoing pregnancy after
patients with repeated implantation failure (P< 0.05).
GA 10 weeks.
What is known already: The endometrium plays an important role in achiev-
Main results and the role of chance: Early fetal loss occurred in 14.8% of
ing optimal outcomes of assisted reproductive technologies. PRP is a novel
patients; miscarriage group had higher discrepancy between calculated and
method that is used in reproductive medicine to improve the IVF outcome. The
measured GA (P<0.001), lower GSD and GSV (P=0.005 and P=0.02, respec-
mechanisms of PRP have not been completely elucidated, but laboratory studies
tively), significantly different YSD and YSV, lower mean GSD and volume ratios
have shown that the high concentration of growth factors in PRP can potentially
(P=0.001 and P=0.003, respectively), and higher UtA-PI/CRL ratio with mis-
speed up the healing process. Recently, the intrauterine infusion of PRP has been
carriage at GA of 46-48 days and GA ≥49 days (P=0.034 and P=0.026, respec-
described as a way to promote endometrial growth and receptivity.
tively). PP13/CRL ratio was higher in patients with miscarriage at GA ≥49 days
Study design, size, duration: The effect of the intrauterus application of
(P<0.041).
PRP to improve the endometrium thickness in patients with repeated implanta-
Limitations, reasons for caution: Small cohort, only IVF pregnancies.
tion failure (RIF) was studied. The implantation rates were evaluated in RIF
Wider implications of the findings: Possible very early prediction of
patients in IVF programs after PRP procedure. The study’s protocol was
impaired placentation prior to early pregnancy loss. A larger cohort is needed
approved by the Center’s IRB.
to further verify the clinical utility of these miscarriage predictions in IVF
Participants/materials, setting, methods: In total, PRP group included 32
pregnancies.
cycles of 20 patients with the mean age 34.2±3.6 y.o. The control group included
38 cycles of 23 patients with the mean age 35.7±4.2 y.o The studies period was
January - December 2019. PRP was infused per intrauterine catheter on the 9th
day of hormone replacement therapy cycle and progesterone administration P-363 The Association between Serum Estradiol and
day. All patients were transferred two good-quality blastocysts. T-test and Chi- Progesterone on the Same Day of FET and the Pregnancy
squared test were used for data analysis. Outcome
Main results and the role of chance: After PRP infusion, the average endo- K. Rageh1, A. Barakat2, N. Barakat3
metrium thickness on day of progesterone administration in PRP group was 1
Al-Azhar university, Ob/Gyn., Cairo, Egypt ;
8.31±1.18mm, which was significantly thicker than control group (5.87±0.81mm) 2
Al-Baraka Fertility Hospital, Ivf, adliya, Bahrain ;
(Student t-test t=2.13, P = 0.04). The implantation rate and clinical pregnancy 3
Al- Baraka fertility hospital, lab, adliya, Bahrain
rate in PRP group were significantly higher than control group (28.13% vs.
15.79%, P<0.05; 25.0% vs. 10.53%, P<0.05, respectively). There was no differ- Study question: To investigate the impact of serum E2 and P levels on the
ence in cycle cancellation rate in both groups. same day of embryo transfer on pregnancy outcomes for FET cycles.

Summary answer: The association between serum estradiol and progester- and IGF1. Demographic data and pregnancy outcomes were collected. Serum
one levels on the same day of FET and the pregnancy outcome is still not proven levels of PLGF, VEGF, and IGF1 were compared between patients who conceived
What is known already: So far, most studies about FET cycles have focused (n=21) and who did not conceive subsequently (n=19).
on embryological factors and the thickness of endometrium, while little attention Main results and the role of chance: There was no significant difference
has been paid to the serum steroid hormone levels on the day of embryo in maternal age BMI, transferred embryo numbers, endometrial thickness
transfer. between the two groups. The serum concentration of PLGF in non-pregnant
Besides endometrial thickness, E2 and P levels are also regularly monitored group (median 7.24, range 0-14.82pg/ml) was significantly (P=0.009) higher
for endometrial receptivity. However, whether serum E2 and P levels on the day than it in the pregnant group (median 1.92, range 0-9.17pg/ml). IGF1 level was
of embryo transfer can serve as an indicator for clinical pregnancy is doubtful. comparable between pregnant and non-pregnant patients (86.9±23.1 ng/ml
The effect of serum E2 and P levels on the day of embryo transfer is a matter vs. 87.2±26.5 ng/ml, P=0.971). There was no significant difference (P=0.088)
of controversy in the literature and needs further evaluation. in the serum VEGF level between non-pregnant group (median 233.3, range
Study design, size, duration: This was a retrospective cohort study (regis- 78.89- 514.79pg/ml) and the pregnant group (median 139.0, range 50.47-
tered at clinical trials.gov with ID NCT04114500), for 402 FET cycles which 379.34pg/ml).
conducted in Al- Baraka Fertility Hospital, Manama, Bahrain, between April 2018 Limitations, reasons for caution: The sample size is relatively small in this
and May 2019 study and a large-scale study is needed to verify the results.
Participants/materials, setting, methods: Endometrial preparation for Wider implications of the findings: The altered expression of PLGF in
402 patients received estradiol tab. When end.thickness reached 8 mm or serum may provide information on the prognostic value of serum angiogenic
greater, patients were initiated on both vaginal and oral Progest. A depot GnRH factor profile for pregnancy outcome.
agonist was administered in midluteal phase of the preceding cycle. On the same Trial registration number: Nil
day of FET, serum levels of E2 and P were assessed, Then, transfer of PGT-
euploid embryos was performed. 12 days later pregnancy test was assessed,
P-365 To compare effect of two protocol in hormone-replacement
and then ultrasound was scheduled to check the viability
cycle for frozen-thawed embryo transfer - A single center,
Main results and the role of chance: Serum P and estradiol levels the same
retrospective study
day of FET were measured. A multivariable analysis to assess the relationship
between serum E2 & P level and pregnancy outcomes was performed, adjusted J. Gao1, C. Ming-Hui1, X. Yanwen1, Z. Canquan1
1
for confounding variables. Mean E2 value was 931.41 ± 438.65 pg/ml, while First Affilated Hospital Of Sun Yat-sen University, Reproductive Center,
mean P value was 8.47 ± 9.4 ng/ml. Progesterone levels were split in terciles: Guangzhou, China
T1: <15 ng/ml; T2: 15–30 ng/ml; T3: > 30 ng/ml. Out of 402 cases, 240 cases
had positive pregnancy test (59.7%) while the clinical pregnancy rate was 53.9% Study question: What is the optimal dose for dydrogesterone combined used
(217 cases out of 402) with no correlation between serum (E2, P & E/P ratio) with progesterone vaginal gel for luteal-phase support(LPS) in hormone-replace-
and the pregnancy rate ment cycle for frozen-thawed embryo transfer.
Limitations, reasons for caution: its retrospective design, which precludes Summary answer: 90 mg Crinone plus 20mg dydrogesterone BID has higher
drawing conclusions regarding how to improve pregnancy outcomes in FET clinical pregnancy rate than 10mg dydrogesterone BID for LPS in hormone-re-
patients using serum estradiol and progesterone levels as predictors; also in our placement cycle for frozen-thawed embryo transfer.
study, we didn’t evaluate live birth rates (LBR). What is known already: Vaginal progesterone gel (Crinone,8%) contains 90
Wider implications of the findings: The implantation process is the most mg of natural progesterone, which acts on the endometrium and myometrium
vital and the least understood part of reproduction.trying to understand this and has some effect on inhibiting uterine contractions. The literature shows that
inigma of implantation in the future we are planning to go for a prospective the combination of Crinone and oral luteal-phase support drugs(dydrogester-
study using only vaginal progesterone, with evaluation of secondary outcomes one) can improve the pregnancy rate of hormone-replacement cycle for fro-
e.g. live birth rates (LBR), and spontaneous abortions/biochemical zen-thawed embryo transfer. But what is the optimal dose for dydrogesterone
pregnancies. is controversial problem.
Trial registration number: NCT04114500 Study design, size, duration: This retrospective study was conducted on
749 cycles who underwent hormone-replacement cycle for frozen-thawed
P-364 Expression of vascular endothelial growth factor (VEGF), embryo transfer from 1 Jan 2018 to 30 Nov 2019 at the First Affiliated Hospital
placental growth factor (PLGF) and insulin-growth like factor 1 of Sun Yat-sen University.
(IGF-1) in serum from women undergoing frozen embryo transfer. Participants/materials, setting, methods: 749 cycles underwent hor-
X. Guo1, T.C. Li1, X. Chen1 mone-replacement cycle for frozen-thawed embryo transfer were enrolled.
1
The Chinese University of Hong Kong, Department of Obstetrics & Gynaecology, Patients used 4~9mg estradiol for 10~14 days in each treatment cycle. When
Hong Kong, China the thickness of endometrium ≥8mm, progesterone was administered for 17
days. Among them, 427 cycles received 90 mg Crinone plus 10mg dydrogester-
Study question: Is there a difference in serum angiogenic profile on day LH+7 one BID(Group A), and 320 cycles received 90 mg Crinone plus 20mg dydro-
between pregnant and non-pregnant women undergoing frozen embryo transfer? gesterone BID(Group B). Both of them receive single blastocyst embryo transfer
Summary answer: Serum PLGF level on day LH+7 in women who conceived at the sixth day of progesterone conversion.
was significantly lower than those who did not conceive. Main results and the role of chance: There were no significant differences
What is known already: Angiogenesis is essential for successful pregnancy with regard to mean age, embryo transfer cycles, the thickness of endometrium
establishment. VEGF, PLGF and IGF1 are three main angiogenic factors which on the day of progesterone conversion and embryo score between the two
play significant role around the time of embryo implantation. Previous studies groups. The clinical pregnancy rate in group A was significant lower than group
have shown that alternation in expression of angiogenic factors was associated B (57.61% vs 66.56%, P=0.015).
with infertility and they may serve as predictive indicators for pregnancy out- Limitations, reasons for caution: This is a retrospective study. It suggest
come. However, the relationship between serum PLGF, VEGF and IGF1 and that 90 mg progesterone vaginal gel plus 20mg dydrogesterone BID may have
pregnancy outcome has not been fully illustrated. better clinical pregnancy rate than 10mg dydrogesterone BID. Addition random-
Study design, size, duration: It is a prospective pilot study in a tertiary ized prospective study are needed to confirm our finding, which is our future
teaching hospital. A total of 40 infertile patients who underwent frozen research direction.
embryo transfer treatment from Jan 2018 to Dec 2018 were recruited in Wider implications of the findings: We found that compare with 10mg
our study. dydrogesterone BID, 20mg dydrogesterone BID plus 90 mg progesterone vaginal
Participants/materials, setting, methods: All the serum specimens were gel would increase the clinical pregnancy rate in hormone-replacement cycle for
collected precisely on day 7 after LH surge in a natural non-conception cycle. frozen-thawed embryo transfer.
Commercial Elisa kits were used to identify the concentrations of PLGF, VEGF, Trial registration number: no

P-366 The role of chromosomal aberrations of the embryo in the Summary answer: Dydrogesterone addition to vaginal P and 1-day FET post-
genesis of recurrent and sporadic miscarriage ponement provide similar outcome in patients with low P levels compared to
E. Kudryavtseva1, V. Kovalev1, I. Baranov2 normal P “in phase” transfers.
1
Ural state medical university, Obstetrics and gynecology, Ekaterinburg, Russia
What is known already: In artificial cycles for FET, low serum P levels on the
C.I.S. ;
2 day of transfer have been associated to decreased pregnancy and live birth rates.
National research center for obstetrics- gynecology and perinatology,
In our hands, measurement of serum P levels prior to transfer and if required,
organizational and methodological department, Moscow, Russia C.I.S.
adjustment of vaginal P doses associated to 1-day FET postponement significantly
Study question: Does the frequency and structure of chromosomal abnor- improved outcomes. Despite this therapeutic strategy, more than 10% of cycles
malities in the embryo (fetus) differ with sporadic and recurrent pregnancy loss were cancelled for persistent inadequate serum P levels. Therefore, we modified
Summary answer: Chromosomal abnormalities in the embryo are a significant our protocol by adding dydrogesterone to vaginal P in patients with low P levels
cause of a miscarriage in both sporadic and recurrent prior to FET and similarly postponed embryo transfer by one day.
What is known already: About 50% of miscarriages are associated with the Study design, size, duration: This is a retrospective analysis of 249 prospec-
presence of chromosomal abnormalities in the embryo. It is assumed that if the tively collected FET cycles from November 2018 to October 2019. Endometrial
patient suffers from recurrent miscarriage, that is, has a history of at least 2 preparation was achieved by sequential administration of vaginal estradiol until
consecutive cases of pregnancy loss, chromosomal abnormalities in the embryo endometrial thickness >7 mm, followed by transdermal estradiol combined with
are less common, and other reasons for the unsuccessful outcome of pregnancy 800 mg/day vaginal micronized P (400 mg twice a day). Our previous ROC
should be sought. But in recent years there have been reports that, in a com- analysis of serum P levels on FET day showed that optimal P level was 11 ng/
parative analysis of the chromosome set in an embryo with sporadic and recur- mL to maximize sensitivity and specificity for live birth.
rent miscarriage, there was no significant difference in the frequency of Participants/materials, setting, methods: This study was conducted in a
occurrence of chromosomal pathology university hospital. Serum P was measured on D1 following exogenous vaginal
Study design, size, duration: A retrospective cohort study was conducted P introduction in the evening (referred as D0). When P levels were >11 ng/mL,
by the method of continuous sampling, which included 1000 patients from Russia embryo transfer was performed “in phase” on D2, D3 or D5 depending on
with a pregnancy demise diagnosed in the period of 6-12 weeks of gestation. embryo stage at cryopreservation. When P levels were <11 ng/mL, dydroges-
The study was conducted in 2018-2019. terone (10 mg three times a day orally) was added to vaginal P and FET was
Participants/materials, setting, methods: The first group consisted of postponed by one day.
681 patients whose first sporadic miscarriage was registered. The second group Main results and the role of chance: Mean serum P level on D1 was 10.2
consisted of 319 patients who previously had a miscarriage. The products of + 3.8 ng/mL, range (2.6-25.3 ng/mL). Serum P <11 ng/mL were observed in
conception (fresh samples of native biological material) obtained by vacuum 63% of cycles (mean P level: 7.8 + 1.9 ng/mL vs. 14.1 + 2.9 ng/mL in patients
aspiration were delivered to the laboratory where chromosomal microarray with P >11 ng/mL, p<0.0001). On D1, serum estradiol levels were also signifi-
analysis was performed. cantly lower in patients with P <11 ng/mL (271 + 202 vs. 354 + 320 pg/mL,
Main results and the role of chance: In group 1, various CAs in the embryo / p<0.01). Therefore, 158 FET were performed with dydrogesterone addition
fetus were detected in 378 samples (55.5%), in group 2 - in 203 samples (63.5%). from D1 onwards and postponed by one day while 91 FET were performed “in
The frequency of occurrence of chromosomal abnormalities in patients who phase” following introduction of vaginal P. Characteristics of patients in both
already have a history of miscarriage was higher than with sporadic miscarriage groups were similar for age (34 + 5 vs. 34 + 6 years), endometrial thickness
- the differences are statistically significant (p = 0.015). There were no significant prior to P introduction (9.6 + 2.0 vs. 9.7 + 2.2 mm), number of transferred
differences in the structure of chromosomal abnormalities. Autosome trisomy embryos (1.4 + 0.5 vs. 1.5 + 0.5), embryo transfer stage (D2/D3/blastocyst:
and numerical abnormalities of sex chromosomes were most often detected. 7/32/61% vs. 5/38/57%). This strategy led to similar positive pregnancy test
So chromosomal abnormalities in the embryo are a significant cause of a mis- (37.3% vs. 40.6%, NS), heartbeat activity at 8 weeks (29.1% vs. 31.8%, NS) and
carriage in both sporadic and recurrent. Structural rearrangements were ongoing pregnancy rates at 12 weeks (27.2% vs. 31.8 %, NS) between “dydro-
detected in 52 samples (5.2%), and 30 (57.7%) of them were submicroscopic, gesterone addition and 1-day postponement of FET” and embryo transfers
and would not have been detected using standard cytogenetic studies. Genetic performed “in phase”.
analysis of abortive material is an important component of the examination for Limitations, reasons for caution: The number of cycles has to be extended
the selection of further management tactics for patients. CMA is an effective to confirm these preliminary data.
research method when conducting genetic analysis of conception products. Wider implications of the findings: These results suggest that serum P
Limitations, reasons for caution: In different ethnic groups, the frequency measurement prior to ET followed by further addition of dydrogesterone to
and structure of chromosomal abnormalities may vary vaginal P and postponement of transfer might optimise the outcome of patients
Wider implications of the findings: In many cases, a genetic study of abor- with low P levels in hormonally substituted FET cycles and avoid cancellation of
tive material makes it possible to clearly establish the cause of miscarriage, a large number of cycles.
reduce the number of tests and diagnostic procedures assigned to patients, and Trial registration number: not applicable
optimize the algorithm of examination and preconception preparation.
P-368 Endometrial thickness on the day of ovulation trigger is an
effective predictor of pregnancy outcomes after frozen blastocyst
P-367 Dydrogesterone addition to vaginal progesterone and transfer in spontaneous natural cycles
transfer postponement improve outcome in patients with low P S. Onogi1, K. Ezoe2, S. Nishihara1, J. Fukuda1, T. Kobayashi1,
levels following hormonally substituted cycles for frozen-thawed K. Kato1
embryo transfer 1
Kato Ladies Clinic, Gynaecology, Tokyo, Japan ;
I. Cedrin - Durnerin1, C. Vinolas1, F. Krief1, B. Dagher-Hayeck1, 2
Kato Ladies Clinic, R&D division, Tokyo, Japan
M. Peigne1, C. Sifer2, S. Sarandi2, B. Bennani Smires2,
M. Grynberg1 Study question: Can the endometrial thickness (EMT) on the day of ovulation
1
Hôpital Jean Verdier, Service de médecine de la reproduction et préservation de la trigger predict the pregnancy outcomes after single vitrified-warmed blastocyst
fertilité, Bondy, France ; transfers (SVBTs) in spontaneous natural cycles?
2
Hôpital Jean Verdier, Laboratoire de biologie de la reproduction, Bondy, France Summary answer: The increased EMT on the day of the trigger was associ-
ated with improved ongoing pregnancy rate and decreased miscarriage rate in
Study question: Does dydrogesterone addition to vaginal progesterone (P) the first trimester.
and postponement of frozen thawed embryo transfer (FET) improve outcome What is known already: The relationship between the EMT on the day of
in patients with low P levels following hormonal replacement therapy? embryo transfer (ET) and pregnancy outcomes has been controversial for

decades. Numerous studies reported that the decreased EMT on the day of ET abnormalities and preeclampsia. It is a well-known fact that several factors like
was associated with a reduced likelihood of pregnancy; therefore, the EMT has cigarette smoking, ethnicity and type of conception affect the levels of these
been used as a predictor of pregnancy outcomes. However, recent studies two markers. The levels of PAPP-A are reduced and the levels of free β-hCG
demonstrated that the EMT on the day of ET is a poor predictor and has only are increased in women who conceived through ART when compared with
a small independent prognostic value. The correlation of the EMT on the day naturally conceived pregnancies. Moreover IVF/ICSI procedure, type of embryo
of ovulation trigger with pregnancy outcomes after SVBT in spontaneous natural transfer as well as freezing and thawing of embryos, might have an impact on
cycles is currently undetermined. the levels of first trimester biochemical markers.
Study design, size, duration: A total of 901 SVBTs in spontaneous natural Study design, size, duration: A cross-sectional, observational, collaborative,
cycles, from November 2018 to October 2019, were analysed. Associations of retrospective study was conducted between June 2009 and December 2019 in
EMT on the days of the trigger (EMT-Trigger) and SVBT (EMT-SVBT) with clinical women with singleton pregnancies after either spontaneous conception, or IVF
and ongoing pregnancy rates were statistically evaluated. The factors possibly with PGT-A and frozen embryo transfer (FET) or after IVF without PGT-A and
associated with EMT on the day of the trigger, such as patient and cycle charac- fresh transfer (ET) or FET and who had first trimester combined screening test
teristics, were also investigated. between 10 and 14 weeks of gestation.
Participants/materials, setting, methods: After monitoring follicular devel- Participants/materials, setting, methods: In IVF/PGT-A-cycles, trophec-
opment and serum hormone levels, ovulation trigger was performed using a toderm biopsy was performed. Serum PAPP-A and free β-hCG were measured
nasal spray containing buserelin, a gonadotropin-releasing hormone agonist. After at 10 to 14 weeks of gestation and analysed by Cobas E and Kryptor, certified
ovulation was confirmed, SVBTs were performed on day 5. The EMT-Trigger by Fetal Medicine Foundation(FMF). Levels for both parameters were converted
and EMT-SVBT were evaluated using ultrasound on the day of the trigger and into Multiples of the Median (MoM) for corresponding gestational age by soft-
immediately before the SVBT procedure. ware of FMF. Both parameters were compared between the following groups:
Main results and the role of chance: The patients were allocated according IVF/PGT-A/FET versus IVF/noPGT-A/FET; IVF/noPGT-A/ET versus IVF/
to the quartiles of EMT-Trigger as follows: EMT <8.1 mm, 8.1 mm ≤ EMT < 9.1 noPGT-A/FET, spontaneous pregnancy versus IVF/PGT-A/FET or IVF/noP-
mm, 9.1 mm ≤ EMT < 10.6 mm, and EMT ≥10.6 mm. The increase in EMT- GT-A/ET and IVF/noPGT-A/FET.
Trigger was significantly associated with the improvement of clinical and ongoing Main results and the role of chance: The Kolmogorov-Smirnov test was
pregnancy rates (P = 0.0003 and P < 0.0001, respectively) and decreased mis- used to analyse the normality of the distribution of variables. Difference of
carriage rates in the first trimester (P = 0.0487). The patients were also allocated median of PAPP-A and free β-hCG were analysed by the Wilcoxon rank-sum
according to the quartiles of EMT-SVBT: EMT <9.1 mm, 9.1 mm ≤ EMT < 10.1 test with nonparametric ANOVA. SAS studio™ software was used for statistical
mm, 10.1 mm ≤ EMT < 12.1 mm, and EMT ≥ 12.1 mm. EMT-SVBT was asso- analysis.
ciated with clinical and ongoing pregnancy rates (P = 0.0053 and P = 0.0010, A total number of 728 patients were included. Patients were divided into four
respectively), but not with the miscarriage rate (P = 0.0848). Additionally, mul- groups; (i) IVF/PGT-A/FET (n=143); (iia) IVF/noPGT-A/ET (n=346) and (iib)
tivariate logistic regression analysis demonstrated a significant correlation IVF/noPGT-A/FET (n=100); (ii) naturally conceived pregnancies (n=139).
between the EMT-Trigger and ongoing pregnancy (adjusted odds ratio [AOR], The median levels of PAPP-A were 0.97MoM; 1.12MoM; 1.20MoM and
1.154; 95% confidential interval [CI], 1.046-1.274; P = 0.0042). However, no 1.03MoM respectively.
correlation was observed between EMT-SVBT and ongoing pregnancy (AOR, The median PAPP-A was the lowest in the group with IVF/PGT-A/FET and
1.043; 95% CI, 0.958-1.136; P = 0.3251). The decreased EMT-Trigger was sig- the highest in the group with IVF/noPGT-A/FET.
nificantly associated with an increase in female age (P < 0.0001) and shortened A statistically significant difference was observed in the median PAPP-A
follicular/proliferation period (P < 0.0001). between IVF/PGT-A/FET group versus IVF/noPGT-A/FET group (p=0.01),
Limitations, reasons for caution: The data used in this study were obtained spontaneous pregnancies versus IVF/noPGT-A/FET (p=0.01), and spontaneous
from a single-centre cohort; therefore, multi-site studies will be required to pregnancies versus IVF/noPGT-A/ET (p=0.06).
ascertain whether these findings can be generalised to other clinics with different No difference was observed when the group with naturally conceived preg-
protocols and/or patient demographics. nancies was compared with IVF/PGT-A/FET group (p=0.5) and in the group
Wider implications of the findings: This is the first report demonstrating with IVF/noPGT-A/FET versus IVF/no PGT-A/ET (p=0.3).
a correlation between the EMT on the day of the trigger and pregnancy out- The median levels of free b-HCG are 1.12MoM; 1.00MoM; 1.01MoM and
comes after frozen blastocyst transfer in spontaneous natural cycles. Our results 0.98MoM respectively.
suggest that EMT on the day of the trigger could be an effective predictor of Median free b-HCG was the lowest in the group with natural conception.
pregnancy outcomes. No statistically significant difference was observed in the concentrations of
Trial registration number: not applicable median level of free b-HCG amongst the compared groups (p>0.05).
Limitations, reasons for caution: The limitations of the study are the ret-
P-369 Impact of Preimplantation Genetic Testing for Aneuploidies rospective design and the ethnic diversity in the groups due to the multicentric
(PGT-A) on first trimester biochemical markers - PAPP-A study design.
(placenta associated plasma protein) and free β-hCG (human Wider implications of the findings: PGT-A and ET-type have an implication
chorionic gonadotropin) on PAPP-A levels. The impact and pathophysiology of IVF related procedures
D. Markova1, K.O. Kagan2, M. Hoopmann2, H. Abele2, E. Abecia1, (freezing and thawing of the embryo, trophoectoderm biopsy) on levels PAPP-A
H. Fatemi3, B. Lawrenz3 require further evaluation in order to assess the need for adjustments on the
1
IVI RMA Middle East Fertility Clinic, Fetal medicine, Abu Dhabi, United Arab first trimester biochemical markers for pregnancies after IVF/PGT-A.
Emirates ; Trial registration number: Not applicable
2
Women’s University Hospital- Tuebingen- Germany, Feto-Maternal Medicine,
Tuibingen, Germany ; P-370 Blastocyst morphology is not associated with maternal first
3
IVI RMA Middle East Fertility Clinic, Reproductive medicine, Abu Dhabi, United trimester serum markers after fresh single embryo transfer (SET)
Arab Emirates A. Pérennec1, A. Reignier1, T. Goronflot2, P. Barrière1, T. Fréour1,
T. Lefebvre1
Study question: Is there a difference in concentrations of first trimester bio-
1
chemical markers amongst patients with IVF/PGT-A, IVF/noPGT-A (fresh / Department of Biology and Reproductive Medicine, Nantes University Hospital,
frozen embryo transfer) and spontaneous conception? Nantes, France ;
2
Summary answer: There is a statistically significant difference in the concen- Data Clinic- INSERM, Nantes University Hospital, Nantes, France
trations of PAPP-A, but not in the levels of free β-hCG amongst the anal-
ysed groups. Study question: Is there an association between blastocyst morphology and
What is known already: PAPP-A and free β-hCG are the two serum bio- maternal first trimester serum markers in In Vitro Fertilization pregnancies
markers used in calculation of the first trimester risk for chromosomal obtained after fresh single embryo transfer ?

Summary answer: Blastocyst morphology is not associated with serum levels Study question: Is it possible to define a simple, sensitive and reproducible
of PAPP-A and b-hCG in first trimester screening after fresh single embryo low-invasive extracellular vesicles (EVs) -based method to allow the quick iden-
transfer. tification of an implantative endometrium?
What is known already: Blastocele expansion, inner cell mass (ICM) and Summary answer: It is possible to obtain and analyze EVs and EVs-associated
trophectoderm (TE) morphology are used to evaluate blastocyst implantation miRNAs from a small volume of endometrial fluid samples.
potential. Serum levels PAPP-A and b-hCG are used for first trimester combined What is known already: Increasing embryo implantation rates has become
screening, as they are associated with the risk of fetal aneuploidy and placental one of the greatest challenges in ARTs. Usually an endometrial-biopsy is done
disorders. Several studies analyzed first trimester serum markers in IVF (In Vitro to identify a receptive endometrium, which prevents embryo transfer in the
Fertilization) pregnancies and found lower PAPP-A level than in spontaneous same cycle, as it is detrimental for the implantation. Implantation is a complex
pregnancies, while the results remained unclear regarding b-hCG levels. Several process, which requires a synchrony between the development of the embryo
hypotheses have been raised to explain this apparent decrease, but the patho- and the endometrium, but also, an adequate embryo-endometrial cross-talk.
physiology remains unclear, as well as the potential association between embryo The presence of EVs as mediators of this communication has been describe in
morphology and early placentation. the endometrial fluid (EF). The molecular analysis of the content of the EVs from
Study design, size, duration: This bi-centric retrospective study was con- EF could be a non-invasive method to improve the implantation rates.
ducted between January 2012 and August 2018. We included 122 women aged Study design, size, duration: The population under study consisted of a
from 18 to 43 years-old, whose pregnancy progressed at least beyond 13 weeks group of 45 women who assisted at the Human Reproduction Unit from January
after a single blastocyst transfer and who participated in the first trimester com- 2018 to June 2019.
bined screening test: 43 in a public IVF Center and 79 in a private IVF center. The study was approved by our Institutional Ethical and Investigation Board
Participants/materials, setting, methods: Day 5 and day 6 blastocysts (CEIC 09/54 and CEIC 11/45).
were evaluated according to Gardner and Schoolcraft classification. Blastocysts Participants/materials, setting, methods: The main inclusion criteria were;
were divided into three groups : excellent (≥ 3AA), good (3-6AB, 3-6BA, B2), cycle duration between 27 and 29 days, normal uterine and ovarian ultrasound,
medium to poor (3-6BB, 3-6AC, 3-6CA, B1, 3-6CB, 3-6BC). First trimester no previous gynecological pathologies and age between 18 and 37 years.
serum markers were measured in maternal blood between 11 and 13 weeks of A pool of EF was used for the establishment of a robust methodology for
amenorrhea + 6 days with Roche method and were expressed in MoM (multiple analyzing EVs from endometrial fluid in clinical settings, where the sample is
of median). Univariate and multivariate analyses were performed. limited and no sophisticated equipment is available. For that, five different meth-
Main results and the role of chance: Body mass Index, smoking status, odologies were compared in triplicate.
type of infertility, geographical origin, anti-mullerian hormone (AMH) level, Main results and the role of chance: From the five methods compared,
ovarian stimulation characteristics, pregnancy outcomes and obstetrical compli- two of them consisted in the direct extraction of RNA while in the other three;
cations were comparable between the three groups. Female age was statistically an enrichment of EVs was done before the RNA extraction. SmallRNAseq was
lower in the « medium to poor » group than in other groups. There were no performed to determinate the most efficient method and showed that the pro-
significant differences in mean first trimester serum markers between the three tocols with a previous enrichment step of EVs obtained a higher miRNA
groups (PAPP-A : p = 0,20 ;b-hCG : p = 0,12). No significant difference was expression.
found either between mean first trimester serum markers and ICM morphology Once the best method was selected, it was applied in a set of real samples
(PAPP-A : p = 0,67 ; b-hCG : p = 0,60), TE morphology (PAPP-A : p = 0,66 ; with different implantation outcome. The content of miRNAs (mainly associated
b-HCG : p = 0,25), or blastocele expansion (PAPP-A : p = 0,22 ; b-hCG : p = with EVs) of endometrial fluid samples from women in whom the implantation
0,48). After adjustement on potential confounding factors (female age, type of was successful (n=15) and unsuccessful (n=15) were analyzed. The results
gonadotropin, parity, number of oocytes retrieved and occurrence of ovarian obtained from the differential analysis of the set of samples with different implan-
hyperstimulation syndrom), the multivariate analyses did not report any signifi- tation outcome are being analyzed and it is expected that the results will be
cant association between PAPP-A or bHCG levels and blastocyst morphology. available by the time this communication is presented.
Limitations, reasons for caution: “Poor” (≥B3CB and ≥B3BC) and Limitations, reasons for caution: The main problem when working with
“medium” (≥B3BB, ≥B3AC, ≥B3CA and B1) quality blastocysts were grouped biological replicates is the impossibility to quantify the sample of origin before
together and represented only 27 blastocysts. starting with the analysis. Therefore, finding an adequate way to normalize the
Wider implications of the findings: Our study concluded that first trimester samples is crucial for analyzing the results.
serum markers were not statistically different according to blastocyst morpho- Wider implications of the findings: This work demonstrate that it is pos-
logical characteristics. Although this needs further confirmation, this suggests sible to obtain and analyze EVs and EVs-associated miRNAs from a small volume
that blastocyst morphology could have an impact on implantation, but not on of endometrial fluid samples, which allows the use of EV-miRNAs as a non-in-
placentation. Therefore, these findings are reassuring for couples undergoing vasive biomarkers for the detection of an implantative endometrium.
IVF and blastocyst transfer. Trial registration number: not applicable
P-372 An increased % of CD56+CD16+ NK cells in the

P-371 Endometrial fluid derived extracellular vesicles as low- endometrial biopsy is not related with recurrent miscarriage
invasive diagnostic biomarkers of implantative endometrium when maternal Killer cell Immunoglobulin-like Receptors are
J. Ibañez Perez1, M. Diaz-Nuñez2, L. Lainz2, M. Diez-Zapirain2, considered
M. Iglesias2, A. Expósito2, M. Gonzalez3, J.J. Lozano4, D. Alecsandru1, A. Pacheco2, N. Garrido3, A. Barrio4, P. Aparicio5,
A.M. Aranzay3, E. Gonzalez5, F. Royo6, N. Subiran7, R. Matorras2, A. Pellicer6, J.A. Garcia -Velasco4
J.M. Falcon-Perez8 1
IVIRMA MADRID, Department of Immunology- IVI RMA- Madrid- Spain,
1
CIC biogune- Cruces University Hospital, Human Reproduction Unit - Biocruces, MADRID, Spain ;
2
Barakaldo, Spain ; IVI RMA Madrid, Department of Andrology- IVI RMA- Madrid- Spain, Madrid,
2
Cruces University Hospital, Human Reproduction Unit - Biocruces, Barakaldo, Spain ;
3
Spain ; IVI Foundation- Health Research Institute La Fe- Valencia- Spain., IVI Foundation-
3
CIC bioGUNE, Genome Analysis Platform, Derio, Spain ; Health Research Institute La Fe- Valencia- Spain., Valencia, Spain ;
4 4
Bioinformatics Unit- Centre Esther Koplovitz CEK, CIBERehd, Barcelona, Spain ; IVI RMA Madrid, Department of Reproductive Endocrinology and Infertility- IVI
5
CIC bioGUNE, Exosomes Laboratory, Derio, Spain ; RMA- Madrid- Spain, Madrid, Spain ;
6 5
Ciberehd- CIC bioGUNE-, Exosomes Laboratory-, Derio, Spain ; IVI RMA Madrid, Department of Immunology- IVI RMA Madrid- Spain, Madrid,
7
Department of Physiology- Faculty of Medicine and Dentistry- University of the Spain ;
6
Basque Country UPV/EHU, Biocruces, Leioa, Spain ; IVI RMA- Roma- Italy, Department of Reproductive Endocrinology and Infertility-
8
Ikerbasque- CIC bioGUNE, Exosomes Laboratory, Derio, Spain IVI RMA- Roma- Italy, Roma, Italy

Study question: Could the increased % of CD56+CD16+ NK cells in the Summary answer: Endometrial Scratching in stimulated IUI cycle is a cost-ef-
endometrial biopsy in oocyte donation recipients be considered as a risk factor fective and easy technique which may improve clinical pregnancy rate in women
for recurrent miscarriage? with previous failed IUI cycles
Summary answer: An increased % of CD56+CD16+ NK cells in the endo- What is known already: Endometrial Scratching(ES) has been suggested as
metrial biopsy is not related with recurrent miscarriage when maternal KIR an intervention to increase the probability of pregnancy in women undergoing
genotype is considered IVF. Majority of studies reported that ES improves outcome in IVF, IUI and natural
What is known already: There is still a current controversy regarding the conceptions: however, the size and quality of studies are poor which really
relationship between an increased % of uNK cells in mid-secretory endometrium questions the presence of any beneficial effect. Endometrial injury is often per-
and its association with recurrent miscarriage. Increased risk of recurrent mis- formed by pipelle biopsy that has an established safety profile. However, it is
carriage (RM) has described in KIR AA mothers when the fetus has more HLA- also known to be associated with a moderate degree of discomfort/pain and
C2 genes than the mother, and this HLA-C2 are paternally or egg donor bleeding and requires an additional pelvic examination. Neither of these studies
inherited. In ART oocyte donor cycles, oocyte HLA-C behaves as the paternal reported the pain/discomfort and bleeding secondary to the procedure.
HLA-C and KIR-HLA-C combination is not currently taken into consideration Study design, size, duration: A prospective randomized controlled trial.
on donors’ selection. KIRAA women have lower live birth rates (LBR) after One hundred sixty-two women ( 81 in each group) with primary/secondary
double embryo transfer (DET). infertility were randomized into intervention and control group. Sample size was
Study design, size, duration: Between January 2017 and June 2019 we estimated using the statistical formula for comparing two proportions. The min-
performed a prospective study of 139 women undergoing one cycle of oocyte imum expected difference in the pregnancy rate between the group is considered
donation-ART. They had recurrent miscarriage (RM) of unknown etiology. Thirty as 20% and the sample size is estimated at 5% level of confidence and 80% power.
nine out of 139 patients underwent an endometrial biopsy under sterile condi- The study was conducted from June 2017 to June 2019
tions with a Pipelle catheter in the month prior of the embryo transfer (ET). Participants/materials, setting, methods: Couples aged between 25 to
Twenty-one healthy oocyte donors were included as a control group and they 35years attending the Infertility outpatient clinic in OBG department, JIPMER,
had an endometrial biopsies too. Pondicherry, India. Unexplained or mild male factor infertility with previous up
Participants/materials, setting, methods: All the patients were selected to 3 failed IUIs were randomised according to random blocks. Three cycles of
from IVI RMA Clinics. KIR and HLA-C typing was performed for patients and ovulation induction with Clomiphene citrate and Gonadotrophins followed by
HLA-C for partners and oocyte donors. They had a normal thrombophilia tests. IUI was done. ES was done using pipelle on day8/ 9 of the stimulated cycle in
The miscarriage and LBR has analyzed. The immune cell populations on biopsy the intervention group. Pain and bleeding were noted after the procedure.
were analyzed by 3 techniques: flow cytometry, immunohistochemistry and gene Main results and the role of chance: The mean age of women was 29.1
expression. A HLA-C tetramer has used to investigate the mother KIR biding ± 3.4 years. There was a significant difference in the mean duration of infertility
to the embryo HLA-C. between two groups with a P-value of 0.041. There was no statistical difference
Main results and the role of chance: The median age of our patients in the distribution of type of infertility or unexplained and mild male factor
was 40 years, and 25 years for oocyte donors. In our cohort the pregnancy infertility (p=0.17, p=0.807 respectively). There was no statistically significant
rate was 71.2%, the miscarriage rate was 31.6% (20.8% were clinical mis- difference with respect to the number of prior failed IUI (p=0.279). The clinical
carriages), and the live birth rate was 39.5%. A higher miscarriage rate after pregnancy rate in the ES group was 22.2% in comparison 9.8% in the control
DET in KIR AA (47.8%) compared to KIR AB (10.5%) and KIR BB (6.7%) (p group. In the intent to treat analysis, with a P-value of 0.03 calculated from Chi-
< 0.003) was observed. A significantly decreased LBR has observed after square test(p<0.05) there was statistically significant difference in the pregnancy
DET in KIR AA (4.3%) compared to AB (26.3%) or BB (46.7%) (p=0.009). rate between Intervention and Control group. There is less than a 3% chance
We analyzed the % and gene expression of CD56bright, CD56+CD16+, that the differences are not real. Efficacy of intervention was found to be
TregCD25+CD4+FoxP3+ cells. No differences were observed on Fourteen Percent (14%). Two patients in ES group had abortions and either of
CD56bright or Tregs when compared to controls. A higher CD56+CD16+ the group had no multiple pregnancies. There was an increased pregnancy rate
% was observed in patients (median 10.2), when compared to controls (6.3) (38.8%) in the third IUI cycle in the Intervention group compared to the Control
p<0.01, but no differences were observed by KIR genotype among patients. group (12.5%)(p=0.43). Fifty-one women (63%) had marked a VAS pain score
A lower gene expression (1/FC) of IL-10 was observed in the patients group of 4-5 with Mean VAS score of 3.42 ± 1.35. Twelve women (12.2%) experienced
compared with the control group (p<0.02), but no differences were observed by mild spotting post-procedure.
KIR genotype among patients. No differences have observed on TNFalpha gene Limitations, reasons for caution: Small Sample size. Older age group might
expression between patients and control nor either by KIR genotype among patients. have led to a low pregnancy rate. Many unknown factors besides poor endo-
There weren´t any differences on HLA-C tetramer binding to the uNK cells metrial receptivity might have led to repeated failure cycles and reduced CPR.
between groups or by maternal KIRs. ES was done using the gentle movement of Pipelle which might have led to less
Limitations, reasons for caution: Our sample was small and this is the first inflammatory response for implantation.
report analyzing the reproductive outcome in oocyte donation by the endome- Wider implications of the findings: Endometrial Scratching in IUI will be
trial % of CD56+CD16+ NK cells and maternal KIR. However, apart from an inexpensive alternative to IVF for couples after IUI failures especially in devel-
statistical significance, the association strength was noticeably high, which confers oping countries, with an acceptable pregnancy rate and does not demand any
our findings more confidence. special qualification. Larger and adequately powered studies are needed to elu-
Wider implications of the findings: A higher miscarriage rate was observed cidate the beneficial effects of endometrial scratching on Implantation.
in KIR AA patients but there weren´t differences in the % of endometrial Trial registration number: Trial REF/2017/10/015540
CD56+CD16+ NK cells between KIR AA, AB or BB. Their increased % observed
in the patients when compared to control group cannot be related with an
increased risk of miscarriage. P-374 When is a pregnancy of unknown location considered to be
Trial registration number: Not applicable non-viable and ready to treat?
K. Barnhart1, B. Nolan2, T. Thomas3, E. Steider4, W. Youssef5,
P-373 Efficacy of Endometrial Scratching on Pregnancy rate in K. Hansen6, A. Steiner7, M. Stephenson8, M. Alston9, M. Cedars10,
women with Unexplained and Mild Male Factor Infertility after Failed E. Jungheim11, M. Diamond12, H. Zhang13
Intrauterine Insemination Cycles A Randomised Controlled Cycle 1
University of Pennsylvania Medical Centre, Dept of Ob/Gyn, Philadelphia
C. Thyagaraju1, M.S. Madhuri1, P. Dasari1 Pennsylvania, U.S.A. ;
2
1
JIPMER, Obstetrics and Gynecology, Pondicherry, India University of Pennsylvania, Department of Ob/Gyn, Philadepphia, U.S.A. ;
3
Univiersity of Pennsylvania, Dept of Ob/Gyn, Philadelphia, U.S.A. ;
4
Study question: Does Endometrial Scratching(ES) in proliferative phase University of Pennsylvania, Dept of Ob/Gyn, Philadpephia, U.S.A. ;
5
increase the probability of pregnancy in women undergoing Ovulation Induction University of Pennsylvania, Dept of Ob/Gyn, Philadelphia, U.S.A. ;
6
and Intrauterine Insemination(IUI) after previous failed IUI cycles? University of Oklahoma, Dept of Ob/Gyn, Oklahoma City, U.S.A. ;

7
Duke University, Dept of Ob/Gyn, Durham, U.S.A. ; What is known already: Threshold values for predicting a clinical pregnancy
8
University of Illinois at Chicago, Dept of Ob/Gyn, Chicago, U.S.A. ; for a fresh blastocyst were 111 IU/l and for a vitrified-warmed blastocyst
9
Denver Health, Dept of Ob/Gyn, Denver, U.S.A. ; 137 IU/l.
10
University of California San Francisco, Dept of Ob/Gyn, San Francisco, U.S.A. ; Study design, size, duration: A cryopreserved thawing embryo transfer
11
Northwestern University, Dept of Ob/Gyn, Chicago, U.S.A. ; (T-ET) cycles were retrospectively analyzed a database of clinical and laboratory
12
Augusta University, Dept of Ob/Gyn, Augusta, U.S.A. ; information between 1 January 2015 and 30 February 2018 at a single fertil-
13
Yale University, Dept Biostatistics, New Haven, U.S.A. ity center.
Participants/materials, setting, methods: In the present study, a total of
Study question: Are criteria that determine when an early pregnancy is 166 patients with T-ET were included and only serum HCG levels drawn 7 days
nonviable conservative enough to enroll women in a clinical trial regarding after the T-ET were included. The percentage for the area under the curve and
treatment? the 95% confidence interval were generated for each ROC curve. The AUC
Summary answer: Conservative criteria adopted to diagnose a nonviable measures the diagnostic accuracy of HCG on pregnancy outcome; an AUC
pregnancy of unknown location as part of a RCT did not eliminate enrollment closer to 1.0 denotes a perfect test.
of a viable gestation. Main results and the role of chance: Among 166 patients who underwent
What is known already: Current clinical criteria to consider a pregnancy T-ET, 89 resulted in a positive maternal serum HCG; 31 resulted in a biochemical
non-viable vary in clinical guidelines. When an early gestation is not visualized pregnancy, and 14 in clinical abortion, and 44 patients achieved an ongoing
with ultrasound, serial hCG values are used to determine viability prior to poten- pregnancy and 43 of them finally achieved live births. Of the 44 ongoing preg-
tial intervention. NICE 2019 defines a change of 63% or greater over 48 hours nancies, 43 live births were successful in the T-ET cycle of which difference were
as potentially viable. ACOG 2018 defines a minimal rate consistent with a viable very small, however they were shown separately because of the significant
gestation to be 49% or greater over 48 hours (for initial values under 1500 mIU/ meaning of live births. The cut off values predicting a clinical pregnancy was
mL). A viable intrauterine gestation has been noted with a rate of as low as 35% 22.8 IU/L with sensitivity of 91.4% and a specificity of 91.7%. The cut off values
over 48 hours (Seeber et al. Fert and Sert,2006;86:454-459.). predicting an ongoing pregnancy for a T-ET was 26.45 IU/L, with a sensitivity
Study design, size, duration: ACT or NOT is an RCT that evaluated the of 93.2%, a specificity of 84.4%, and live birth was 27.95 IU/L, with a sensitivity
treatment of women with a persistent pregnancy of unknown location. Inclusion of 90.7 %, a specificity of 84.6 %.
criteria included no definitive ultrasound evidence of intrauterine or extrauterine Limitations, reasons for caution: This is a retrospective study with small
gestation and a plateau in hCG, defined as a < 30% in 2 days, < 50% in 3 days, sample size and it is a single centered study.
< 75% at 4 days, < 100% at 5 days, < 130% at 6 days or >166% at 7 days. Two Wider implications of the findings: Clinicians could inform patients who
clinicians confirmed eligibility. have undergone thawing single embryo(blastocyst) transfer whether their pos-
Participants/materials, setting, methods: After determination of eligibility itive serum hCG would present better outcomes or not based on these cut
subjects were randomized 1:1:1 to expectant management and to two different off values.
active treatment arms. Trial registration number: GBIRB2019-310
Main results and the role of chance: A participant was enrolled, random-
ized to expectant management, and was later noted to have a viable gestation. P-376 4D ultrasonographic evaluation of uterine peristalsis
A 33-year-old, G2 P1001, who conceived with use of clomiphene citrate and correlates with progesterone levels in patients with repetitive
IUI was enrolled at 4 5/7 weeks gestation. She presented with abnormal serial implantation failure.
hCG values of 7% in 2 days and 24% over 4 days: 86 at GA 4 0/7, 92 at 4 2/7 B. Moliner, M.D.1, J. Llacer1, A. Bernabeu1, J.C. Castillo1,
and 107 at 4 5/7. Ultrasound at GA of 4 5/7 demonstrated no evidence of an A. Fuentes1, F. Sellers1, R. Bernabeu1
intrauterine or extrauterine gestation. Subsequently hCG values rose normally: 1
Instituto Bernabeu, Reproductive Medicine Department, Alicante, Spain
348 at 5 0/7, 803 at 5 2/7, and 2477 at 6 0/7. A viable IUP was diagnosed at 6
5/7 with a CRL 7.7mm and cardiac activity. She elected to proceed with prenatal
Study question: Does 4D ultrasonographic evaluation of uterine peristalsis
care and delivered at term without complication. This event was reported to
on the day of embryo transfer correlate with progesterone levels in patients
the DSMB and IRB and judged to be unanticipated based on current clinical
with repetitive implantation failure (RIF)?
standards. However, a memo was circulated to all participating centers to use
Summary answer: Uterine peristalsis evaluated by 4D ultrasound shows
caution against declaring non-viability when; a) using early hCG values prior to correlation with progesterone levels when assessed on the day of embryo trans-
6 wks gestation when the date of conception is known, b) the hCG is continu- fer in patients with RIF.
ously rising, and c) in those who may be at greater risk for multiple pregnancy. What is known already: The role of progesterone in the inhibition of uterine
Limitations, reasons for caution: While this case represents a single event peristalsis is widely known. Uterine peristalsis and progesterone levels on the
from a larger RCT, it highlights the limitations of clinical guidelines based solely day of embryo transfer are described as potential factors influencing pregnancy
on laboratory tests. Criteria to classify an early gestation as non-viable, based outcomes in IVF cycles. However, reliable methods demonstrating a correlation
on only hCG values, may not be 100% predictive and should include all aspects between these two entities specifically in patients with RIF are still under
of the clinical setting. research.
Wider implications of the findings: Despite conservative criteria, a viable Study design, size, duration: Our retrospective cohort study (November
gestation was unintentionally enrolled in trial designed to manage a non-viable 2017-January 2020) included all consecutive IVF cycles of RIF patients (n=153)
gestation. If this participant was actively treated her pregnancy may have been who underwent uterine contractility/progesterone levels assessment on the day
interrupted. Current clinical standards may need to be amended to ensure a of embryo transfer. RIF was defined as an unsuccessful implantation of a total
pregnancy of unknown location is nonviable prior to treatment. number of ≥3 blastocysts originating from oocyte donation or autologous PGT-A
Trial registration number: NCT01800162 cycles. Participants underwent frozen embryo transfer cycles (FET) and were
evaluated for uterine contractions and serum progesterone levels 30 minutes
P-375 Predictive value of serum HCG concentrations in before their embryo transfer procedure.
pregnancies achieved after thawing embryo transfer Participants/materials, setting, methods: Uterine contractions (UC) were
S.J. Chon1 assessed by recording a 6-minute-long video using 2D vs 4D mode (Voluson
1
Gil Hospital, Obstetrics and Gynecology, Incheon-si, Korea- South E10) by a single operator (BM). Contractions were counted visually on a 15x
accelerated recording. In order to define low progesterone levels, we separated
Study question: What would be cut off values of serum HCG level in maternal progesterone levels in quartiles, counting last quartile as Low progesterone level
blood measured on seventh day of thawing blastocyst transfer to confirm stable group (LP=< 9,2 ng/ml; N=38), considering the rest as Normal Progesterone
clinical pregnancy and live birth rates? level (NP>9,2 ng/ml; N=115).
Summary answer: The cut off values predicting a clinical pregnancy was Main results and the role of chance: Overall, patients mean age was 41
22.8 IU/L and for ongoing pregnancy was 26.45 IU/L. years, with an average of 4.1 embryo transfers performed previously and with

an average of 5.1 embryos transferred. The average of contractions measured higher than the proportion of patients with RIF and low expression of AMHRII
in 2D was 0.62 contractions per minute (cpm) while the average of contractions and the proportion of normal individuals.The ER, IGFBP1, and PRL expression
measured in 4D was 0.99 cpm. There were no differences between the groups in the endometrium of patients with RIF and high expression of AMHRII was
of uterine factor (adenomyosis, myomas, adhesions or polyp) or endometriosis, significantly lower than that in patients with RIF and normal expression of
as well as of the variables previously mentioned such as age, the number of AMHRII. A negative correlation was found between the expression of AMHRII
embryo transfers made previously or the number of embryos transferred pre- and ER, IGFBP1 and PRL in the endometrium during the luteal phase in patients.
viously. Differences were observed in uterine contractility measured by 4D Limitations, reasons for caution: Clinical samples are more difficult to
ultrasound, observing 1.3 cpm in LP group while in HP group an average of 0.9 obtain.Further studies with more samples are required to tested. Interfering
cpm was observed. This difference was statistically significant (p = 0.012) giving gene expression at the cellular level to elucidate the exact underlying molecular
a RR of 1.6 (95% CI 0.6 to 4). However, uterine contractility assessment using mechanisms about AMHRII in RIF.
2D didn’t show statistically significant differences (p = 0.315), observing a mean Wider implications of the findings: This study was novel in reporting the
of 0.8 cpm in the LP group and a mean of 0.6 cpm in the NP group. expression of AMH and AMHRII at the normal maternal–fetal interface. The
Limitations, reasons for caution: The inherent risk of bias associated to abnormally increased expression of AMHRII might participate in the occurrence
retrospective studies. Additionally, the small sample size -especially for the sub- of RIF via a mechanism associated with the interference of endometrial decid-
group analysis (normal vs low progesterone levels)- precludes robust conclu- ualization and apoptosis, ultimately resulting in impaired endometrial receptivity.
sions. Caution is warranted in extrapolating these results to patients without Trial registration number: not applicable
previous implantation failure.
Wider implications of the findings: Our findings suggest that low proges- P-378 Human endometrial glandular organoids: a powerful
terone levels constitute a significant factor for increased uterine contractility. 3D-cell model to mimic morphological and molecular changes of
Furthermore, 4D ultrasound appears to be a reliable method for assessment of the receptive endometrium at the implantation window
uterine peristalsis, thus should be considered for routine evaluation in RIF patients A. Luddi1, V. Pavone1, G. Morgante1, L. Governini1, B. Semplici1,
undergoing cryotransfer in artificial cycles. L. Benincasa1, V. De Leo1, P. Piomboni1
Trial registration number: None 1
University of Siena, Molecular and Developmental Medicine, Siena, Italy
P-377 Anti-Müllerian hormone receptor type II causing repeated Study question: Does a 3D-culture model of human endometrial glandular
implantation failures through the endometrium apoptosis of the organoids mimic the morphology and the gene expression profile of a receptive
luteal phase endometrium at the implantation window?
F. Yunxing1, H. Rong2 Summary answer: Endometrial organoids are surrogate as regard morphol-
1
18309609178, Ningxia Medical University Shuang Yi Campus, Yinchuan, China ; ogy, creation efficiency, expandability, gene expression profile; pinopodes, reli-
2
Ningxia Medical University / Ningxia Medical University General Hospital, able marker of endometrial receptivity, are detectable after in vitro
Reproductive Medicine Center of Ningxia Medical University General Hospital, decidualization.
Yinchuan, China What is known already: Many evidences support the existence in the human
of a narrow window of uterine receptivity, which opens during the mid-luteal
Study question: To detect the expression of AMH and anti-Müllerian hormone phase, during which endometrium becomes receptive toward the embryo. At
receptor (AMHR)II at the maternal–fetal interface and explore whether AMH this time, formation of pinopodes on the apical membranes of the endometrial
and AMHRII participated in the occurrence of repeated implantation failure (RIF). epithelial cells occurs, a phenomenon that is considered as a reliable morphologic
Summary answer: AMHRII had an effect on RIF via the AMH/AMHRII sig- marker of endometrial receptivity. Due to ethical concerns to study human
naling pathway. It participated in the occurrence of RIF by interfering with endo- implantation in vivo, this in vitro three-dimensional culture system offers the
metrial decidualization and apoptosis. opportunity to evaluate the endometrial microenvironment and to investigate
What is known already: The abnormal expression of key proteins and hor- the molecular mechanisms underlying the implantation process.
mones at the maternal–fetal interface affected the maternal–fetal communication Study design, size, duration: Endometrial biopsies have been collected
and led to adverse pregnancy outcomes. The expression of AMHRII in endo- from healthy women, who underwent ovarian surgery from January to October
metrial tissues was closely related to the reproductive ability of mice. 2019. The epithelial cells were isolated and 3D culture was applied to obtain
Study design, size, duration: From January 2015 to May 2016. The 154 organoids. Morphology, ultrastructure as well as gene expression profile of key
endometrial tissues in the early follicular, follicular, and luteal phases were col- genes known to be involved in the implantation process have been evaluated,
lected from 41 women with infertility due to tubal or male factors alone; 41 before and after in vitro hormonal treatments to induce morphological and
samples were also collected from patients with RIF. The 20 placental tissues were molecular modifications mimicking the proliferative and secretive phase of the
collected in early, mid and late pregnancy respectively. 12 blastocysts were cul- uterine cycle.
tured from nontransplanted fourth-order embryos with poor quality on the third Participants/materials, setting, methods: Endometrial specimens have
day.All patients age were less than 35. been collected from women of proven fertility (n=5; 22-39 years old). Organoids
Participants/materials, setting, methods: 1. Relative expression of AMH of epithelial cells have been obtained, and then exposed for 4 days to estrogen
and AMHRII in Endometrium and placenta were analyzed using immunohisto- alone or in combination with progesterone and cAMP, mimicking the proliferative
chemistry,WB and qRT-PCR. 2. Expressions of AMH and AMHRII mRNA in and the secretory phase, respectively. Then, organoids were fixed and processed
blastocysts detected using a single-cell real-time polymerase chain reaction. 3. for TEM and SEM analysis or processed for RNA extraction and gene expression
Apoptosis in the endometrium during the luteal phase detected by TUNEL and analysis by digital droplet PCR (ddPCR).
WB. 4. The expression of estrogen receptor (ER), IGFBP1, and PRL were tested Main results and the role of chance: Our data highlight that organoids
via immunohistochemistry. 5. The relationship between AMHRII and ER, IGFBP1, obtained from human endometrium represent a good model to study the phys-
and PRL. iological function of the endometrial epithelium. TEM analysis showed that
Main results and the role of chance: AMH and AMHRII were less expressed organoids preserve glandular organization as well as ultrastructural characteris-
in the normal endometrium during the menstrual cycle. AMHRII was slightly tics. Moreover, a scanning electron microscopic (SEM) survey of endometrial
highly expressed in the endometrium during the menstrual cycle, especially in organoids, definitively demonstrates the dramatic changes occurring in the lumi-
the luteal phase. AMHRII was highly expressed in the placental tissues during nal surface according to hormonal treatment. In organoids treated with E2,
early pregnancy and in the second and third trimesters. In addition, the expres- mimicking the proliferative phase, the luminal surface is relatively smooth or
sion of AMHRII mRNA was detected in the blastocysts, but no expression of exhibit irregular micro-extensions; indeed, it appears overlaid with a rich net of
AMH mRNA was observed. Positive expression of AMHRII in patients with RIF microvilli and cilia clusters of different length. The organoids mimicking the secre-
was 29.3%, which was higher than that in patients of normal childbearing age, tory phase are characterized by the presence of large cytoplasmic apical pro-
especially during the luteal phase. TUNNEL and WB results indicated the protrusions traditionally called “pinopodes”, that bulge out of the entire luminal cell
portion of patients with RIF and high expression of AMHRII was significantly surface. These protrusions have regular contours and are characterized by a

significant loss of free surface micro-extensions. These data are also confirmed Limitations, reasons for caution: This is a retrospective case-control study,
by ddPCR analysis, that revealed significant modulation of genes involved in the which theoretically has selection bias, information bias and confounding bias.
endometrial receptivity depending on hormone treatment: when 3D-cultured Although it has been controlled in the design and implementation, relevant bias
organoids were supplemented with E2, the expression of PGR, MMP26 and PAEP interference cannot be completely excluded. In addition, the sample size is rel-
significantly increased if compared to control. More interestingly, E2+MPA+cAMP atively small, although literature with larger samples never have been published.
treatment increased the expression of HOXA 10, IGF1, VEGF, MMP26 and PAEP, Wider implications of the findings: Surgical management is a safe and
while ESR1 significantly decreased. effective approach for IP and even for IHP. Compared with cornual resection,
Limitations, reasons for caution: Larger study needs in order to confirm cornuostomy by laparoscopy showed no difference in the removal of ectopic
these data, above all for supporting conclusions from gene expression profiles. pregnancy and the preservation of intrauterine pregnancy, but the risk of per-
Moreover, further development of this approach based on the incorporation of sistent ectopic pregnancy was worthy of attention.
other endometrial cell types is looked-for a better understanding of mechanisms Trial registration number: Not applicable
underlining the embryo implantation establishment.
Wider implications of the findings: 3D-culture model of glandular endo- P-380 Female lifestyle factors and risk of recurrent pregnancy
metrium not only represents a valuable research tool to study molecular mech- loss: systematic review and meta-analysis
anisms underlying human implantation in vitro, but also will provide a valuable K.Y.B. Ng1,2, G. Cherian1, A. Kermack1,2, S. Bailey1,2, N. Macklon3,4,
model to carry out comparative studies on specific mechanisms that may drive S. Sunkara5, Y. Cheong1,2
endometrial dysfunction, paving the way to the setup of personalized treatments. 1
Princess Anne Hospital, Obstetrics and Gynaecology, Southampton, United
Trial registration number: Not applicable Kingdom ;
2
University of Southampton, School of Human Development and Health,
P-379 management and outcome of interstitial pregnancy after Southampton, United Kingdom ;
in vitro fertilization/embryo transfer: a retrospective consecutive 3
University of Copenhagen, Zealand University Hospital, Copenhagen, Denmark ;
series of 185 cases 4
London Women’s Clinic, London Women’s Clinic, London, United Kingdom ;
C. Wang1, S. Yang1, R. Li1, J. Qiao1 5
Kings College London, Department of Women and Children’s Health, London,
1
Peking University Third Hospital, Obstetrics and Gynecology, Beijing, China United Kingdom
Study question: Surgical management of interstitial pregnancy (IP) with or Study question: Do female lifestyle factors influence the risk of recurrent
without intrauterine pregnancy (IUP): cornual resection or cornuostomy? pregnancy loss (RPL)?
Summary answer: Compared with cornual resection, cornuostomy by lap- Summary answer: Overweight and obese women had a significantly higher
aroscopy showed no difference in the removal of ectopic pregnancy and the risk of RPL; the impact of smoking, alcohol consumption and caffeine intake on
preservation of intrauterine pregnancy. RPL remains uncertain.
What is known already: IP is a rare type of ectopic pregnancy, but its inci- What is known already: Recurrent pregnancy loss, defined as the occurrence
dence rises among patients undergoing in Vitro Fertilization/Embryo Transfer of two or more consecutive miscarriages occurs in 1-2% of couples, and is a
(IVF/ET) treatment. Most patients show no specific clinical manifestations, but diagnosis that remains unexplained in a large proportion of cases. Whilst the
once the interstitial portion ruptured, patients may face serious life threats. specific mechanisms leading to RPL are still relatively unknown, poor lifestyle is
Study design, size, duration: A total of 185 cases who were diagnosed with associated with a hostile reproductive environment, which may compromise
IP followed IVF/ET and treated in our hospital between Jan 2010 and Oct 2019 optimal embryo implantation and securement of a pregnancy. Although associ-
were included, of which 66 (35.68%) were interstitial heterotopic pregnancy ations between certain lifestyle factors and the risk of sporadic miscarriage have
(IHP). The data of each patient, including age, history of gynecologic surgeries, been reported, whether lifestyle factors impact the risk RPL is less well known.
details of IVT/ET process, therapeutic interventions and reproductive outcome, It is important to understand any probable associations to improve patient
were collected through electronic medical record database and analyzed management.
retrospectively. Study design, size, duration: This is a systematic review and meta-analysis
Participants/materials, setting, methods: Measurement data were pre- addressing the impact of female lifestyle factors on the risk of RPL. Electronic
sented as mean±SD or median (minimum-maximum), and statistical significance databases including Medline, Embase, Cochrane Library, CINHL and Pubmed
of differences among the groups was analyzed by Student’s t-test or Mann– were searched until August 2019. The lifestyle search terms included ‘diet’,
Whitney U test. Categorical variables were reported as n%, and statistical sig- ‘smoking’, ‘alcohol’, ‘caffeine’, ‘exercise’ and ‘BMI’. The search term for miscar-
nificance of differences among the groups was analyzed by chi-square test or riage history included ‘miscarriage’, ‘recurrent pregnancy loss’, ‘recurrent mis-
Fisher’s exact test. A two-sided p value< 0.05 was considered statistically sig- carriage’ and ‘spontaneous abortion’.
nificant. All statistical analyses were performed with the use of SPSS statistics Participants/materials, setting, methods: Full text manuscripts were
for Windows version 22.0 software. reviewed for relevancy by two independent authors. Studies were included if
Main results and the role of chance: The average number of embryos they explored women of reproductive age, who had been exposed to an aspect
transferred was 2.08±0.416. 171 patients received Day 3 embryos and 14 of female lifestyle, such as obesity, being overweight, underweight, smoking and
patients received Day 5 blastocysts. 118 patients underwent fresh ET and alcohol intake. The outcome assessed was the risk of having RPL in this popu-
67 patients underwent frozen ET. There were no differences between IP lation. Where possible, individual study estimates were pooled using either fixed
without IUP patients and IHP patients on number of embryos transferred or random effects meta-analysis.
(2.034±0.43 vs 2.136±0.35, p=0.098) or type of ET (fresh ET 68.91% vs Main results and the role of chance: A total of 24,705 records were
54.55%, p=0.052), but IP without IUP patients received more Day 3 embryos identified from the electronic search and an additional 192 records identified
transferred (94.96% vs 86.36%, p=0.040), and showed earlier gestational through hand searching of references. Fifteen studies were included in the sys-
age (days) at diagnosis (43.84±6.08 vs 50.68±10.16, p<0.001). Patients are tematic review and meta-analysis; 8 case control studies, 4 cohort studies, 2
treated mainly by surgery, including 9 laparotomies, 113 laparoscopic corn- survey based studies and 1 observational study. Meta-analyses of 4 studies
uostomies (LC), 30 laparoscopic cornual resections (LCR) and 7 laparoscopic showed that the odds of RPL in the obese compared to women with normal
loop ligatures. LC and LCR shared the similar duration of surgery (minutes) BMI is 1.79 (95% CI 1.34, 2.41). The odds of RPL in women with BMI>25
[51 (9-189) vs 61(23-155), p=0.317] and intraoperative blood loss [5 (1-300) compared to women with normal BMI is also significantly increased (OR 1.33,
vs 10 (2-200), p=0.317]. There were no significant differences between LC 95% CI 1.13, 1.56). Meta-analysis of 5 studies showed that being underweight
and LCR on live birth rate (91.4% vs 77.8%, p=0.574) among IHP patients does not increase the risk of RPL (OR 1.00, 95% CI 0.59, 1.70). Meta-analysis
and persistent ectopic pregnancy rate (6.3% vs 0.0%, p=0.560) among IP of 3 studies shows no increase in risk of RPL in women who smoked cigarettes
without IUP patients. No surgical complications (including infection, uncon- compared to non smokers (OR 1.62, 95% CI 0.90, 2.93), and no increased risk
trolled bleeding, uterine rupture during subsequent pregnancy or delivery) of RPL in women who consumed alcohol compared to those who do not (OR
were observed in our cohort. 1.12, 95% CI 0.88, 1.44). Meta-analyses of 2 studies showed no increased risk

of RPL in women who have higher caffeine intake (>99mg/day) than women are comparing the cytokine and hormone profile in the two patient groups and
who have lower caffeine intake (≤99mg/day) OR 1.35 (95% CI 0.97, 1.89). will correlate it to the level of HLA-G. Cell assays analyzing the effect of cytokine
Limitations, reasons for caution: This review is limited by heterogeneity and hormone cocktails will be studied to evaluate the direct effect on HLA-G
in the definition of RPL and methods of quantifying smoking, alcohol and caffeine regulation.
intake between studies. Some studies did not restrict on whether the miscar- Limitations, reasons for caution: As the JEG-3 cell line only represents an
riages were consecutive. This review was limited to studies in English and we in vitro model for human trophoblast cells, it might not reflect in vivo settings.
did not assess impact of male lifestyle. Also, cytokines co-exist within the local milieu and could have synergistic, antag-
Wider implications of the findings: Being overweight and obese contrib- onistic or indirect effects mediated through regulation of other immune factors
uted significantly to increased risk of RPL by over 1.3- and 1.8-fold respectively. on HLA-G expression in vivo.
Whether lifestyle interventions including weight loss programmes are beneficial Wider implications of the findings: This study provides a possible link
require further investigation. Smoking, alcohol and high caffeine intake did not between the increased levels of anti-inflammatory cytokines and reduced
increase the risk of RPL, but evidence is limited given very few studies. levels of sHLA-G. This could prove to be important for both early establish-
Trial registration number: not applicable ment of pregnancy and pregnancy complications such as PE. Also, systemic
rather than placental inflammation might be involved in the manifesta-
P-381 Regulation of the immune tolerance-inducing Human tion of PE.
Leukocyte Antigen-G (HLA-G) for induction of peripheral and Trial registration number: not applicable
placental maternal tolerance of pregnancy.
G. Persson1, C.S. Stæhr1, F.S. Klok1, J.B.S. Bork1, C. Isgaard1, P-382 Evaluation of endometrial expression of cell adhesion genes
M. Lebech2, T.V.F. Hviid1 in recurrent implantation failure (RIF) patients
1
Zealand University Hospital, Department of Clinical Biochemistry, Roskilde, A. Pathare1, I. Hinduja2
1
Denmark ; P. D. Hinduja Hospital and Medical Research centre, Department of IVF and
2
Zealand University Hospital, Department of Gynaecology and Obstetrics, Research, Mumbai, India ;
2
Roskilde, Denmark P. D. Hinduja Hospital and Medical Research centre, Department of IVF,
Mumbai, India
Study question: How is the expression and the isoform profile of HLA-G
regulated at the feto-maternal interface and systemically in the mother in uncom- Study question: Does the endometrium of recurrent implantation failure (RIF)
plicated pregnancy versus pre-eclampsia? patients reveal differential expression of cell adhesion genes than fertile women
Summary answer: Levels of peripheral and placental soluble HLA-G in controlled ovarian stimulation (COS) cycle.
(sHLA-G) are not correlated and may represent different compartments. HLA-G Summary answer: Out of seven cell adhesion genes evaluated in the study,
expression profile can be regulated directly by cytokines. five showed significant differential expression in endometrium of RIF patients
What is known already: The expression of paternally derived antigens by than fertile women under COS.
the fetal cells during establishment of pregnancy must involve induction of mater- What is known already: The cell adhesion molecules expressed by endo-
nal tolerance. HLA-G expression by the endometrium and placental trophoblast metrium during window of implantation are proven to be important for endo-
cells is an important factor that allows generation of tolerance. HLA-G expres- metrial receptivity. Based on our previously published whole genome microarray
sion may be modulated by local factors such as hormones and cytokines (e.g. study, genes like CD300A, Cadherin-3 (CDH3), Cartilage Oligomeric Matrix
progesterone and IFN-γ). Implantation failure, recurrent abortion and pregnancy Protein (COMP), Hyaluronan Binding Protein 2 (HABP2), Collagen Type XXII
complications such as pre-eclampsia (PE) are often characterized by an altered Alpha 1 Chain (COL22A1), Thrombospondin 1 (THBS1), Microfibril Associated
HLA-G, cytokine and hormone profile. However, these measurements are often Protein 4 (MFAP4) were associated with cell adhesion as one of the most sig-
based on systemic serum levels. No clear picture of the local HLA-G regulation nificant biological functions among RIF patients during COS. Thus, it is worth-
has yet been established. while to study their expression in larger cohort of RIF patients under COS.
Study design, size, duration: The direct effect of cytokines on HLA-G Study design, size, duration: Case-control study included RIF patients (n=25,
expression was studied in vitro by stimulating the chorioncarcinoma cell line cases) and healthy fertile oocyte donors (n=25, controls), both undergoing COS.
JEG-3 with regulatory, anti- and pro-inflammatory cytokines. From October The endometrial tissues were collected in the period of 2016-2019.
2017 until present 30 women with uncomplicated pregnancies and 20 cases of Participants/materials, setting, methods: Endometrial tissue was collected
PE diagnosed according to the Danish guidelines have been recruited at Zealand from cases and controls on Human chorionic gonadotropin (HCG)+7th day
University Hospital, Denmark. Peripheral blood was collected pre-partum. during COS. RNA extracted from tissue was converted to cDNA which was
Cord blood, placental biopsies and blood were collected within 30 minutes further subjected to real time PCR to study ‘gene expression’ by SYBR green
from birth. chemistry. The differential expression of genes ‘CD300A, CDH3, COMP,
Participants/materials, setting, methods: JEG-3 cells were stimulated HABP2, COL22A1, THBS1, MFAP4’ in cases were calculated by delta-delta-CT
for 24 or 72 hours with IL-2, IL-4, IL-6, IL-10, IL-12, IL-15, IL-17A, TGF-β1, method, using fold change (FC) as parameter. Level of significance was deter-
TNFα and IFN-γ1b. RNA was isolated from the placental biopsies, and serum mined using Mann-Whitney T test.
and EDTA plasma were obtained from placental and maternal blood samples. Main results and the role of chance: CDH3 (P=<0.0001, FC:-2.81), COMP
Cord blood was used for HLA-G genotyping. HLA-G expression level was (P=<0.0001, FC:-6.46), HABP2 (P=0.026, FC:-2.00), THBS1 (P=0.0238, FC:-
analyzed by flow cytometry, ELISA and digital droplet PCR. The mRNA iso- 1.70), MFP4 (P=0.0350, FC:-1.88) genes majorly involved in functions of cell
form profile was analyzed using reverse transcription PCR and fragment adhesion like extracellular matrix organization, adhesion junctions in endome-
analysis. trium epithelium were observed to be significantly down-regulated with fold
Main results and the role of chance: Peripheral and placental sHLA-G levels change > 2 in ‘cases’ compared to ‘controls’
were not correlated. Cases with PE showed lower levels of peripheral sHLA-G, Limitations, reasons for caution: The study was performed in COS cycle,
while no difference between groups was observed in placental sHLA-G levels. to rule out the hormonal bias. Therefore, the data cannot be generalized to
The expression pattern of the HLA-G mRNA isoforms were overall cohort of RIF patients. Also, due to modest sample size of the study, it
G1>G3=G2/4>G5=G6 for both the JEG-3 cell line and the placental biopsies. is necessary to be substantiated in larger population of RIF patients.
In vitro analysis showed that IL-2, IL-6 and TNF-α decreased expression of HLA- Wider implications of the findings: Down-regulation of cell adhesion
G, while IFN-γ upregulated its expression in JEG-3 cells. Interestingly, stimulation panels in RIF patients indicate the impairment in endometrial receptivity which
of JEG-3 with IL-10 changed the isoform profile, leading to an almost two-fold possibly contributes to repeated failure of implantation. On validating these
increase of the level of HLA-G1 over HLA-G2/4 and -G3 transcripts compared markers in larger cohort of RIF patients, the panel can be implemented for
to the control. No differences in the isoform profile were observed between assessment of endometrial receptivity to evaluate cell adhesion function in
placental biopsies from uncomplicated pregnancies and PE placental biopsies, endometrium.
although cases of PE did have lower levels of HLA-G1. In ongoing analyses we Trial registration number: Not applicable

P-383 Circulating miRNome during the implantation window as a 3

Arnaud de Villeneuve Hospital- CHU Montpellier, ART-PGD department,
non-invasive diagnostic tool to predict successful IVF/ICSI cycle Montpellier, France
C. Baron1, D. Haouzi1, F. Entezami2, S. Brouillet3, S. Hamamah3
1
Study question: What outcome benefits are expected in RIF patients after
Inserm U1203, IRMB- Hôpital St-Eloi- CHU Montpellier, Montpellier, France ;
2
customized embryo transfer based upon identification of endometrial receptivity
American Hospital of Paris, IVF department, Neuilly-Sur-Seine, France ;
3
window by transcriptomic approach.
Arnaud de Villeneuve Hospital- CHU Montpellier, ART/PGD Department,
Summary answer: Customized embryo transfer according to window of
Montpellier, France
implantation testing improves pregnancy and birth outcomes for RIF patients.
What is known already: A receptive endometrium is mandatory for success-
Study question: Do circulating miRNAs profile during the implantation window
ful implantation of a competent embryo. For years, endometrial receptivity
under the same HRT treatment for frozen embryo replacement cycle, predict
assessment was established by indirect methods and several non-invasive
successful pregnancy ?
approaches have been reported with controversial results and poor predictive
Summary answer: We identified a miRNA signature in serum during the
value. Transcriptomic and proteomic approaches have also been used to define
implantation window that can predict successful pregnancy.
molecular signatures and identify specific biomarkers of human endometrial
What is known already: The development of non-invasive tests predicting
receptivity. We previously identified a set of genes as biomarkers of endometrial
the pregnancy outcomes remains a challenge in assisted reproductive technology
receptivity and developed an innovative test based on quantification by RT-qPCR
(ART). However, to date, very few studies analyzed the miRNome in blood-
of these biomarkers in endometrium biopsies and tested its relevance in fertile
stream during the implantation window to identify potential circulating biomark-
patient.
ers related to the pregnancy outcomes.
Study design, size, duration: Endometrial biopsies were performed during
Study design, size, duration: Serum (n=14) were collected during the
the implantation window 7-9 days after the LH surgein natural cycle or 5-9 days
implantation window from RIF patients (mean ± SD, age: 38.6 ± 5.0 years) under
after progesterone administration under HRTrespectively. According to tran-
hormone replacement therapy (HRT) treatment for subsequent frozen embryo
scriptomic testing result in the subsequent frozen embryo transfer cycle (FRET),
transfer. RNAs were extracted to perform the miRNA expression profile. Then,
blastocysts were transferred at the specific day where endometrium was iden-
miRNA expression was analyzed according to the following pregnancy outcome:
tified as receptive and cleavage stageD2/D3 embryos were transferred72/48
successful implantation (n=7) or implantation failure (n=7). Successful implan-
hours before the specific cycle day where endometrium was identified as
tation was defined as both positive b-hCG and clinical pregnancy and implanta-
receptive.
tion failure as negative b-hCG.
Participants/materials, setting, methods: During 2015-2018 a total of
Participants/materials, setting, methods: Serum were obtained during a
217 RIF patients (4.4±1.9 failed fresh/frozen transfers of 6.4±3.6 embryos)
mock HRT treatment cycle from 14 repeated implantation failure (RIF) patients.
were enrolled. Genomic testing of endometrial biopsies was performed under
Then, miRNA expression profile between groups, successful vs. implantation
natural/HRTcycles and mRNA expression of genes predictive of receptivity
failure after frozen embryo replacement was evaluated by NGS using the HTG
were established by RT-qPCR. Customized embryo transfer (n=157 patients)
EdgeSeq miRNA Whole Transcriptome Assay (Illumina, Firalis). miRNA sequenc-
was performed in a subsequent FRET cycle based upon these results. Clinical
ing data were normalized using the method recommended by HTG molecular
pregnancy and live birth rates (LBR) were compared to control RIF patients with
diagnostic. Then, statistical analysis and receiver operating characteristic (ROC)
standard FRET in natural/HRT cycles (n=60 patients).
analysis were applied to miRNA sequencing data.
Main results and the role of chance: Customized embryo transfer using
Main results and the role of chance: We identified 44 miRNAs differentially
the genomic testing strategy yielded spectacular increase in outcome for 157
expressed between groups with a fold change > 2 and a P-value < 0.05. All
RIF patients (age 37.2±4.3 years). Comparison of the results between the study
miRNAs were overexpressed in serum from patients with successful implanta-
group and the control group showed significantly higher rates for implantation
tion. Supervised hierarchical clustering of these 44 miRNAs showed a good segrega-
(22.7% vs 7.2%, p=0.0001), clinical pregnancy (38.8% vs 15.0%, p=0.0006),
tion of serum samples from patients with and without a successful implantation.
ongoing pregnancy (36.3% vs 8.3%, p=0.00002) and live birth rates (31.8% vs
related to these 44 miRNAs, we note four of them which are members of the let-7
8.3%, p=0.0002). Analyses of endometrial receptivity status revealed mostly a
family [miR-1 (x2.4, p = 0.017, AUC = 0.88), miR-2 (x2.2, p = 0.004, AUC =
0.94), miR-3 (x2.1, p = 0.001, AUC = 0.90), miR-4 (x2.1, p = 0.001, AUC = delay both in natural and HRT cycles between 1 to 3 days. Most patients achieved
pregnancy after the first customized FRET (70%). Better clinical pregnancy and
0.98)]. The microRNA target filter function from Ingenuity software predicted
live birth rates were obtained with blastocyst transfer compared to cleavage
that 1375 mRNAs were targeted by the let-7 family that are involved in cell
stage transfers (respectively 36.3% vs 22.5% and 29.5% vs 14.0%). Whatever
invasion, proliferation, growth and survival via integrin subunit beta 3 (ITGB3),
the day of transfer, results were significantly higher in the study group compared
vimentin (VIM), B-cell CLL/lymphoma 2 like 1 (BCL2L1) that play a central role
to the control group, with ongoing pregnancy rate for transfer of cleavage-stage
in endometrial receptivity acquisition and implantation process. These results
embryos of 21.1% compared to 4.5% in the control group (p=0.03). In blastocyst
might have potential clinical applications to develop a non-invasive diagnostic
transfer group ongoing pregnancy rate of 33.9% was significantly greater than
tool to predict successful implantation, to avoid endometrium biopsy and con-
8.6% in the control group (p=0.003).
sequently, to increase IVF/ICSI success.
Limitations, reasons for caution: The benefit of this innovative strategy
Limitations, reasons for caution: Further investigations with a larger num-
should be analysed with respect to the genetic status of the transferred embryos
ber of patients are in progress to validate theses results.
after PGT for aneuploidy.
Wider implications of the findings: Circulating miRNAs profile of the
Wider implications of the findings: In RIF patients customized embryo
implantation windows during a cycle preceding frozen embryo transfer cycle can
transfer according to endometrial transcriptomic testing improves dramat-
predict the successful implantation. This information is crucial and can lead to
ically the clinical outcome by increasing almost four-fold the LBR. In our
the development a prognostic tool of the attempt, opening new perspectives
study, the majority of RIF patients displayed a delay in their receptivity
in the patient care management.
window of 1- 3 days, revealing a potential cause for their previous implan-
Trial registration number: ID: NCT04192396
tation failures.
P-384 Four-fold increase in live birth rate: proof of the benefits of
window of implantation testing in RIF patients by a prospective
multicenter trial P-385 Hysteroscopic endometrial peeling: a different approach to
F. Entezami1,2, D. Haouzi2,3, A. Ferrières-Hoa2,3, C. Vincens3, endometrial scratching
S. Bringer-Deutsch3, E. Vintejoux3, S. Hamamah2,3 T. Alkon1, M. Luna-Rojas1, C. Hernandez-Nieto1, D. Cassis-
1
American Hospital of Paris, IVF, Neuilly sur Seine, France ; Bendeck1, B. Sandler1
1
2
Université Montpellier- CHU Montpellier- Hôpital Saint Eloi, Inserm U1203-, Reproductive Medicine Associates of New York, Reproductive endocrinology and
Montpellier, France ; Infertility, Mexico City, Mexico

Study question: Does hysteroscopic endometrial peeling improve reproduc- What is known already: Implantation is considered the “black box” in ART.
tive outcomes in women with implantation failure? Implantation is a highly regulated process that requires: receptive endometrium,
Summary answer: Women with implantation failure may benefit from intra- functional blastocyst, and interaction between them. Human chorionic gonad-
operative endometrial peeling prior to a frozen embryo transfer (FET). otropin (hCG) is a hormone synthesized and released by the syncytiotrophoblast.
What is known already: Trials on endometrial scratching with pipelle and It stimulates the ovarian production of progesterone during the first trimester
diagnostic hysteroscopies in women with implantation failure have demon- of pregnancy. Several studies suggest that intrauterine hCG increases the preg-
strated methodological limitations and high risk of bias, making it unclear nancy rate. The hCG may promote peritrophoblastic immune tolerance, which
whether these procedures improve reproductive outcomes. To date, no study facilitates trophoblast invasion by inducing an increase in endometrial T-cell
has described the effects of intentional endometrial lesion through apoptosis. It may also support trophoblast apposition and adhesion by regulating
hysteroscopy. proteins involved in implantation as well as altering endometrial secretory
Study design, size, duration: A retrospective, cohort study included infertile parameters.
patients who failed transfers of at least two good quality blastocysts. Thereafter, Study design, size, duration: We retrospectively reviewed all single euploid
underwent an endometrial peeling through hysteroscopy and subsequently a frozen embryo transfers (FET) of euploid embryos into GCs at the CReATe
blastocyst FET from January 2018 to December 2019. A sample size of 70 Fertility Center between Jan 2017 and Mar 2019. Mosaic embryos were excluded.
patients per group was calculated to be necessary to detect a 15% difference in We compared the cycles in which 50 µL of hCG was introduced into the uterine
clinical pregnancy rates with 80% power. Alpha=0.05. cavity 10-15 min prior to embryo transfer vs. cycles where no hCG was admin-
Participants/materials, setting, methods: Women <40 years, with implan- istered. We then compared the first transfer and all transfers after a failed transfer
tation failure, normal saline sonogram and, no previous uterine surgeries were in both groups.
segregated into two groups: A) 70 patients underwent hysteroscopic endome- Participants/materials, setting, methods: We analyzed FET cycles of
trial peeling, which consists on removing the superficial endometrial layer of the Euploid embryos in GCs. The GC’s underwent a pelvic US and sonohysterogram
whole uterine cavity with hysteroscopic biopsy forceps and B) 85 controls who before ET. The patients received HRT for uterine preparation. hCG insertion
did not undergo surgical endometrial peeling. All patients underwent a subse- was used in the transfer cycle at the discretion of the fertility physician.
quent blastocyst FET. Trophectoderm biopsy cases were excluded from the Main results and the role of chance: First transfer
analysis.
Main results and the role of chance: In total, 155 women were included
in the cohort. No differences were found in age, body mass index, baseline FSH,
With hCG Without hCG
AMH, baseline antral follicle count, previous number of stimulation/IVF cycles, P Value
n(%) n(%)
number of embryos transferred and, embryonic quality among cohorts.
Evaluating the endometrium, no differences were observed in the endometrial Age 26.44±4.78 27.62±4.47 .289
pattern, however, a significant difference in the endometrial thickness
(8.5±1.4mm vs 9.0±1.6mm, p=0.03) was noted among groups. When analyzing Endometrial thickness 10.8±2.02 10.43±2.02 .52
the subsequent FET cycle, statistical significance was observed in implantation Morphology .350
rates (38.8% vs 16.6%, p= 0.01) and clinical pregnancy rates (65.7% vs 30.3%, 1 6/18 (33.3%) 29/196 (14.8%)
p=0.0009). No difference was found in clinical loss rates (10.4% vs 17.1%,
p=0.08) among Group A vs Group B respectively. 2 9/18 (50%) 128/196 (65.3%)
Of the patients who underwent surgical endometrial peeling, 15.7% (11/70) 3 2/18 (11.1%) 28/196 (14.3%)
had the following incidental intraoperative findings: mild intrauterine and/or Egg donation 15/18 (83.3%) 152/196 (77.6%) .57
cervical adhesions (4/70, 0.05%), endometrial polyps (8/70, 11.4%), polypoid
endometrium (6/70, 0.8%). 63.6% of the patients with incidental findings became Adjunct therapy 18/18 (100%) 153/196 (78.1%) <.001
pregnant. Pregnancy 13/18 (72%) 132/195 (67%) .69
Limitations, reasons for caution: This study is limited by its retrospective
Clinical Pregnancy 13/18 (72%) 119/195(61%) .87
nature. Additionally, in patients with noted uterine pathology, appropriate surgical
management was administered in the same setting, this could have biased the
results. Future randomized controlled trials are needed with euploid frozen
embryo transfers. After at least 1 failed transfer
Wider implications of the findings: This study describes for the first time
the clinical utility of mechanically peeling the superficial endometrial layer through
hysteroscopy in women with implantation failure. Our findings suggest that sur-
With hCG Without hCG
gically removing superficial endometrial tissue, promotes the generation of new P Value
n(%) n(%)
receptive tissue, favoring embryo implantation.
Trial registration number: Not applicable Age 26±3.67 27.9±4.61 .035
Endometrial thickness 10.81±2.27 9.92±2.27 .52
P-386 HCG improves pregnancy chances in embryo transfers
Morphology .51
using euploid embryos and Gestational Carriers after one
implantation failure 1 4/34 (11.7%) 16/111 (14.4%)
J. Mashiach1, A. Roumia1, M. Dviri2, N. Weizman1, S. Baram1, 2 23/34 (67.6%) 57/111 (51.3%)
C. Librach3 3 4/34 (11.7%) 25/111 (22.5%)
1
CreATe Fertility Center, CreATe Fertility Center, toronto, Canada ;
2 Egg donation 30/34 (88.2%) 82/111 (73.8%) .21
CreATe Fertility Center, CrATe Fertility center, Toronto, Canada ;
3
CreATe Fertiity Center- Unviversity of Toronto, Obstetrics and Gynecology, Toronto, Adjunct therapy 31/34 (91.1%) 60/111 (54%) <.0001
Canada Pregnancy 21/34(61.7%) 59/111 (53%) .02
Clinical pregnancy 20/34(58.8%) 44/111(40%) .04
Study question: Does intrauterine hCG improve pregnancy rate in frozen
single embryo transfer of euploid embryos in Gestational Carriers (GC)?
Summary answer: After a first failed euploid single embryo transfer into a
GC, intrauterine administration of hCG appears to improve the chances of Using a logistic regression model, accounting for age at OPU, adjunct therapy
subsequent implantation and pregnancy. and intrauterine hCG administration, in those with 1 or more previous failed

transfers, we observed that intrauterine hCG administration was significantly Gr 1 -0; Gr 2 - 2 (10,53%) ; Gr 3 - 7 ( 25,00%) , Gr 4 – 9 (56,25%) ( P 1-3 <0.03;
associated with the chance of achieving pregnancy (CI 1.048-6.57 p=0.039). P 1-4 <0.001).After correction of the revealed disorders in women with RIF– ET
Limitations, reasons for caution: The retrospective nature of the study according to the implantation window was performed pregnancy rate - 48 (62,34 %)
and the relatively small study size are the main limitations of this study. Limitations, reasons for caution: Gr1 the main reason of RIF is the pecu-
Wider implications of the findings: In our model of euploid embryos in liarity of IW, expected to have a high clinical effect using genetic methods. Gr 2,
“proven” uteri of GCs, we can investigate implantation in the most optimal Gr3 clinical efficacy depends not only on IW, but also deviations in the structure
clinical setting possible. When implantation of a euploid embryo fails (i.e failed of the endometrium. Gr1 -dyschronosis is the most complex and needs further
first transfer ), intrauterine administration of hCG appears to increase the chance research.
for pregnancy. Wider implications of the findings: The proposed examining technology
Trial registration number: n/a of endometrium receptivity helps to obtain pregnancy in complex category of
patients with RIF. The proposed examining technology remains an informative,
P-387 Endometrial receptivity - Is it possible to make the “old” reproducible, accessible method. The technology help determine the indications
research methods more informative? for molecular genetic studies of the endometrium and interpret the results
I. Gyulmamedova1, T. Zadorozhna2, T. Tatarchuk3, O. Girina4, Trial registration number: not applicable
N. Vladykina4, M. Sokolova5, O. Gyulmamedova6, V. Sosnina7,
P-388 Significance of invasion of decidualized endometrial
O. Nosenko8
stromal cells by human embryonic stem cells derived
1
Adonis- Medical Group of Companies- Kyiv- Ukraine, Iivf, Кyiv, Ukraine ; trophoblastic spheroids (BAP-EB) in pregnancy outcomes of in
2
Institute of Pediatrics Obstetrics and Gynecology- National Academy of Ukraine, vitro fertilization (IVF)
Laboratory of Morphology, Kyiv-, Ukraine ;
3 Y.L. Lee1,2, S.W. Fong1, K.C. Lee1, A.C.H. Chen1,2, K.F. Lee1,2,
Institute of Pediatrics Obstetrics and Gynecology- National Academy of Ukraine,
W.S.B. Yeung1,2, E.H.Y. Ng1,2
Department of Endocrine Gynecology-, Kyiv-, Ukraine ;
1
4
Adonis- Medical Group of Companies- Kyiv- Ukraine, ivf, Kyiv, Ukraine ; The University of Hong Kong, Obstetrics and Gynaecology, Hong Kong, Hong
5
Adonis- Medical Group of Companies-, ivf, Kyiv-, Ukraine ; Kong ;
2
6
Medical Center IPF-, ivf, Kyiv-, Ukraine ; The University of Hong Kong Shenzhen Hospital, Shenzhen Key Laboratory of
7
Medical Center IPF, -ivf-, Kyiv-, Ukraine ; Fertility Regulation, Shenzhen, China
8
Odessa National Medical University, Department of Obstetrics and Gynecology,
Study question: Does invasion of decidualized endometrial stromal cells
Odessa, Ukraine
(hEnSC) towards trophoblast-like BAP-EB relate to IVF pregnancy outcome?
Study question: Is the proposed examining technology of endometrium recep- Summary answer: The invasion of decidualized hEnSC from patients with a
tivity informative in women with Recurrent Implantation Failure (RIF)? live birth towards BAP-EB was significantly higher than those with negative preg-
Summary answer: The complex application of the morphological, immuno- nancy outcome.
histochemical methods, scanning electron microscopy is informative and effective What is known already: The success rate of IVF remains low even after the
for the diagnosis of endometrial receptivity in women with RIF. transfer of good quality embryos. Implantation-competent blastocysts can induce
What is known already: Single competent blastocyst transfer in maximum migration of decidualizing hEnSC. The encapsulation of the invading blastocyst
endometrium receptivity is the key goal for reproductive medicine. Knowledge by the decidua supports subsequent trophoblast invasion. The study of the
of the implantation window (IW, is especially important in patients with recurrent dynamic fetal–maternal interaction between the decidua and the implanting
implantation failure (RIF). Significant efforts are focused on the search and imple- blastocyst requires a proper implantation model. We have derived trophoblastic
mentation of non-invasive endometrial receptivity biomarkers - proteomics and spheroids (BAP-EB) from human embryonic stem cells that resemble human
lipidomics in intrauterine fluid in the cycle of embryo transfer (ET). Gene-array trophectoderm and trophoblast during early implantation process. They attach
methods are used to analyze the global gene profile of endometrial cells. specifically onto receptive endometrial epithelial cells. BAP-EB can be used as
However, in the daily work, morphological research and endometrium dating, human embryo surrogate for studying the early implantation process.
as well as scanning electron microscopy (SEM), continue to be of clinical interest Study design, size, duration: hEnSC were isolated from endometrial biopsies
due to their availability. obtained from IVF patients in their natural cycle 7 days after luteinizing hormone
Study design, size, duration: During 2010-2019 In 77 patients with RIF on surge (LH+7). Human embryonic stem cells were differentiated into early tro-
the 7-10th day of the menstrual cycle, an endometrial biopsy / hysteroscopy phectoderm-like BAP-EB-48h or trophoblast-like BAP-EB-96h. An invasion assay
was performed. A morphological , immunohistochemical (CD-138) study was using Matrigel Invasion Chamber was established with decidualized hEnSC and
performed; After correction of the revealed disorders, in the following cycles, BAP-EB-48h or -96h. hEnSC isolated from patients with live births (n=5) and
the endometrial biopsy was obtained at P+6; P+8 or P + 7; P + 9 day of expo- negative pregnancy outcomes (n=5) in their subsequent stimulated IVF cycle
sure to progesterone. A morphological study (MS) and SEM were carried out. were compared.
Descriptive statistics methods used. Participants/materials, setting, methods: Human endometrial stromal
Participants/materials, setting, methods: The age of the patients - 34.8± cell line (T-HESC) was induced to decidualize in vitro. Purity of primary hEnSC
0.59,, the duration of infertility 8.48 ± 0.64 , the number of IVF attempts 3,11 was confirmed by immunohistochemical staining of stromal (Vimentin) and epi-
± 0.19. According to the results of MS, patients were divided: Gr1 - 13 (16,88%) thelial (cytokeratin) markers. The 24h-invasion of decidualized hEnSC through
- the endometrium without morphological abnormalities.Gr2 - Chronic endo- Matrigel towards trophectoderm-like BAP-EB-48h or trophoblast-like BAP-EB-
metritis with micro- macropolyposis, confirmed by CD 138 – 19 (24, 68 %); 96h was compared to medium alone control. The expression levels of decidu-
Gr3 - Stromal fibrosis, endometrial hypoplasia - 29 (37,66%).Gr4- dyschronosis alization markers (PRL, IGFBP1) and the invasion ability towards BAP-EB were
of glands and stroma - 16 (20,78%) compared between hEnSC isolated from patients with live births and negative
Main results and the role of chance:Not informative results (NIR), repeated pregnancy outcomes.
biopsy (RB) at a later date after additional treatment : Gr 2 - 6 (31,58 %) , Gr3 Main results and the role of chance: After treatment with cAMP for 3, 6
- 3 (10,34%); Gr 4 - 2 (11.1%)The reason NIR: Dense film, an abundance of and 9 days, the decidualization markers PRL and IGFBP1 were significantly
red blood cells, absence pinopodia - P + 6; P + 8; Gr1 - no indication for a RB induced from day 6 onwards. The isolated primary hEnSC were of high purity
(P1-2<0.02). In all 11 patients with RB, temporary trends in the formation of as demonstrated by positive vimentin staining (>95%) but not cytokeratin stain-
pinopodia persisted Few pinopodia: Cr1 - 1 (7,69%) , Gr 2 - 12 (63,16%); Gr ing. The invasion of hEnSC through Matrigel towards medium alone without
3- 16 (59%), Gr 4 – 6 (37,5%) patients (P <0.04;1-2,1-3,1-4) Dyschronosis BAP-EB, BAP-EB-48h or BAP-EB-96h was quantified by the absorbance of crystal
of the development of pinopodia: 1 (7,69 %), 2 (10,53%), 3 (11,11%), 14 violet stained cells. It was found that significantly more hEnSC invaded towards
(87,5 %) respectively (P 1-4 <0.001) The lag of the formation of pinopodia BAP-EB-96h than BAP-EB-48h when compared to medium alone control, sug-
for 2 or more days: 8 (61,54%) , 13 (68,42%) , 19 (65,52%), 5 (31,25%) respec- gesting the specific invasion of hEnSC towards the trophoblast-like but not the
tively (P> 0.05; 1-2,1-3.1-4). Lack of regression, polymorphism of microvilli; trophectoderm-like BAP-EB. The decidualization and invasion abilities of hEnSC

isolated from patients with live births and negative pregnancy outcomes were with pregnancy loss of aneuploid origin. The AmiGO gene ontology database
compared. The expression levels of PRL and IGFBP1 were similar in the two was used as a reference to establish the functional role currently attributed to
groups of patients 6 days after in vitro induction of decidualization (p>0.05). the genes reported.
The invasion of hEnSC through Matrigel in the presence or absence of BAP-EB- Structural factors inherited from mothers directly by embryos were the major-
96h was measured. Interestingly, the percentage of BAP-EB-96h-induced invasion ity of the cases reported and included: Robertsonian translocations, deletions
was significantly higher in hEnSC from patients with live births (54.3% ±11.1%) and insertions. Cases of maternal mosaicism leading to aneuploidy in the embryo
than those with negative pregnancy outcomes (20.8%±9.1%, p<0.05). The data included: trisomy 9 and duplication of chromosome 4p. Several of these cases
indicated a correlation of responsiveness of hEnSC towards BAP-EB with preg- were identified using FISH in addition to traditional karyotyping techniques.
nancy outcome. Germline factors with a plausible role in aneuploid pregnancy loss of maternal
Limitations, reasons for caution: BAP-EB and the isolated endometrial origin included: skewed X-inactivation and FMR1 repeats.
stromal cells may not fully represent the in vivo developed human blastocysts Studies that reported the association of single gene mutations with aneuploid
and endometrial cells, respectively. The in vitro nature of the hEnSC experiments pregnancy loss were conflicting. Single gene mutations with an uncertain role in
and the limited sample size in this study may also limit the interpretation of aneuploid pregnancy loss included: SYCP3 and MTHFR. Both genes have an
the data. established role in cell division in animal models however the research in humans
Wider implications of the findings: The study of early implantation failure is inconclusive.
is limited by the number and the ethical concerns of human embryos donated One study reported an association of a single gene mutation of NLRP7 in
for research. The current data demonstrated the potential use of BAP-EB as women with recurrent molar pregnancies.
human embryo surrogate for assessing the responsiveness of hEnSC towards Limitations, reasons for caution: Whilst this review was based on a broad
implanting embryos and as predictive tool for pregnancy outcome. systematic search of the literature most studies identified were case reports:
Trial registration number: nil only a fraction were cohort studies of moderate quality. The role of paternal
germline factors in aneuploid pregnancy loss was not addressed in this systematic
review and warrants future research.
P-389 Maternal germline factors associated with aneuploid
Wider implications of the findings: Identifying maternal genetic factors
pregnancy loss: a systematic review.
associated with an increased risk of aneuploidy will expand our understanding
U. Blyth1, L. Craciunas2, M. Choudhary3 of cell division, nondisjunction and miscarriage secondary to embryo
1
Newcastle upon Tyne Hospitals NHS Foundation Trust, Academic Foundation aneuploidy.
Programme, Newcastle upon Tyne, United Kingdom ; The candidate germline factors identified may be incorporated in a screening
2
Newcastle upon Tyne Hospitals NHS Foundation Trust, Obstetrics and panel for women suffering miscarriage of aneuploidy aetiology to facilitate coun-
Gynaecology, Newcastle upon Tyne, United Kingdom ; selling for subsequent pregnancies.
3
Fertility Centre at Life- Newcastle upon Tyne Hospitals NHS Trust- Biomedicine Trial registration number: PROSPERO registration number
West Wing- Newcastle upon Tyne- NE1 4EP- UK, Reproductive Medicine, CRD42019153653
Newcastule upon Tyne, United Kingdom
P-390 Pattern of early beta human chorionic gonadotropin
Study question: Which maternal germline factors have been associated with (β-hCG) in intracytoplasmic sperm injection (ICSI) for
pregnancy loss secondary to numerical chromosome abnormalities of pregestational testing for monogenic disorders (ICSI/ PGT-M)
the embryo? versus ICSI only cycles
Summary answer: A range of maternal germline factors such as insertions,
N. Samara1, E. Haikin-Herzberger1, A. Dick1, H. Amir1, A.
deletions, translocations, copy number variants or single gene mutations have
Reches1, M. Malcov1, D. Ben Yosef1, F. Azem1
been associated with aneuploid pregnancy loss.
1
What is known already: Miscarriage describes the spontaneous loss of preg- lis maternirty hospital- Sourasky medical center.affiliated with the Sackler Fac,
nancy before the threshold of viability; the vast majority occur before 12 weeks Wolf PGD Stem Cell Lab- Racine IVF Unit, Tel Aviv, Israel
of gestation. Miscarriage affects 1 in 4 couples and is the most common com-
plication of pregnancy. Chromosomal abnormalities of the embryo are identified Study question: Is there a difference in β-hCG pattern in pregnancies achieved
in approximately 50% of first trimester miscarriages; aneuploidy accounts for following ICSI/PGT-M versus ICSI? 
86% of these cases. Summary answer: A comparable pattern of early β-hCG rise were observed
The majority of trisomic miscarriages are of maternal origin with errors during in ICSI/PGT-M cycles compared with ICSI only.
meiotic division of the oocytes. Chromosome segregation errors in oocytes What is known already: Initial serum β-hCG level is used to confirm preg-
may be sporadic events secondary to advancing maternal age; however, there nancy following IVF, scheduled follow-up testing is accepted to reassure normal
is increasing evidence to suggest possible maternal germline contributions to this. progression and prediction of outcome. β-hCG levels and pattern of increase
Study design, size, duration: A systematic review of the literature was are well correlated with pregnancy outcome. In our unit, fertilization in PGT-M
conducted to identify maternal germline factors associated with pregnancy loss cycles is routinely achieved by ICSI, to avoid sperm contamination of analysis
secondary to numerical chromosome abnormalities of the embryo. The litera- result. On day 3 or 5, embryonic biopsy is performed for genetic analysis, which
ture search was run in September 2019 using the electronic databases: OVID might affect syncytiotrophoblast secretion of β-hCG. Our aim was to assess
MEDLINE, EMBASE and the Cochrane Library. No time or language restrictions early β-hCG rise in ICSI/PGT-M compared to ICSI cycles in order to evaluate
were applied to the searches, only primary research was included. The review possible effect of biopsy on early β-hCG rise
protocol was registered with PROSPERO (CRD42019153653) following PRISMA Study design, size, duration: This retrospective cohort study was performed
guidelines. at a tertiary university-affiliated medical center. It included patients with single
Participants/materials, setting, methods: Participants were women who embryo transfer, from ICS/PGT-M and ICSI only cycles, performed in our IVF
had suffered pregnancy loss secondary to numerical abnormalities of the embryo. unit between June 2017 and September 2018. Both fresh and frozen embryo
Study identification and subsequent data extraction was performed by two transfer (FET) cycles were included. A total of 163 cycles with a positive β-hCG
authors independently and in duplicate. The Newcastle-Ottawa Scale was used result were available for analysis, of which 85 of ICSI/PGT-M and 78 of ICSI
to judge included studies on three broad perspectives: the selection of study Participants/materials, setting, methods: In ICSI/PGT-M cycles embryo
groups; the comparability of the groups; and the ascertainment of either the biopsy took place on day 3 or 5 according to genetic analysis method, control
exposure or outcome of interest for case-control or cohort studies respectively. group included patients undergoing ICSI during same period. First β-hCG was
The results were synthesised narratively. timed on day 10-12 according to transfer day and second test was usually taken
Main results and the role of chance: The literature search identified 2639 two days later. In order to adjust for different period of time between the
titles, out of which 54 were eligible for inclusion in this systematic review. They two tests and for the none-linear elevation of Beta-HCG we used a calculated
reported on maternal germline factors having variable degrees of association parameter - ln (Delta-Beta-HCG/period).

Main results and the role of chance: The two groups were comparable Main results and the role of chance: From adjusted odds ratios (aOR) of
with respect to age, BMI and parity. Gravidity was significantly higher in the ICSI/ demographic factors, older (aOR 1.2, 95%CI [1.1; 1.3], p < 0.01) or obese
PGT-M than in ICSI group (2.0±1.8 vs.0.8±1.1, p<0.01). In the majority of (>35kg/m2) women (aOR 3.4, 95%CI [1.1; 10], p = 0.03) had increased risks
PGT-M cycles embryos were transferred on day 5 and none were transferred of miscarriage. Risk was reduced after having previously delivered >2 babies
on day 3, compared to ICSI cycles in which the majority of embryos were (aOR 0.1, 95% CI [0.01; 0.6], p = 0.02). Regarding the serially collected data,
transferred on day 3, in ICSI/PGT-M cycles some embryo were transferred on maternal age remained a risk factor for miscarriage especially in women aged
day 4 shortly after receiving genetic analysis results (61.9% vs 33.8% for day 5-6, ≥35 years (HR 2.6, 95%CI [1.2–5.6]). Ultrasonical alterations in crown-rumP-
38.1% vs 1.3% for day 4 and 0% vs. 64/9% for day 2-3, p<0.01). Once β-hCG length, mean gestational sac diameter or presence of bleeding were also import-
was positive, the clinical pregnancy rates were 98.2% in the ICSI/PGT-M group ant (HR 0.76, 95%CI [0.69–0.82], HR 0.72, 95%CI [0.64–0.79] and HR 1.13,
compared to 92.2% in the ICSI group. Similarly, live birth rates (65.8% in the 95%CI [1.03–1.27], respectively). Biochemically, serum hCG, progesterone and
PGT-M vs. 72%% in ICSI) were not significantly different. Differences between estradiol, respectively showed the most distinct effect with hazard ratios of 0.36,
serum β-hCG levels were not statistically significant between two groups in both 95%CI [0.26–0.48], 0.76, 95%CI [0.75–0.77]and 0.80, 95%CI [0.75–0.84],
first and second tests, neither when comparing the ln(delta-β-hCG/period). respectively. From the tested 56 different combinations, a model including mater-
After adjustments for age, number of previous pregnancies, fresh vs. frozen and nal age (>35 years or not), bleeding (yes/no), CRL (mm) and hCG (U/L) data
embryo day of transfer, ln(delta-Beta-hCG/period) was still comparable was the best to predict miscarriage (WAIC score 3118, differences were eval-
between ICSI/ PGT-M group and ICSI group. uated by the statistical Watanabe-Akaike information criterion (WAIC)). The
Limitations, reasons for caution: Our study is a retrospective one, with best combination without sonographic data (WAIC 3424) was the combination
potential biases inherent to its design. In some cases, lag time between first and of maternal age, bleeding, hCG and estradiol
second β-hCG levels was not constant due to scheduling constraints. Limitations, reasons for caution: Fetal karyotype was unavailable due to
Wider implications of the findings: Our study shows no difference in the logistics and heart rate frequency was not measured due to risk of doppler
early pattern of serum β-hCG levels between ICSI/PGT-M cycles and ICSI cycles. heat-induced damage. Because of the demands of serial data collection, the
It reassures that embryo biopsy for ICSI/PGT-M, taken at day 3 or 5 in both sample size was too small for other estimates than the WAIC for the quality of
fresh and frozen cycles, does not affect early β-hCG rise, correlated with preg- prediction in each model
nancy outcome. Wider implications of the findings: Reassuring, the widely applied approach
Trial registration number: not applicable of combining hCG, CRL and bleeding data for early pregnancy outcome turned
out to be the best. However, without sonography the best model had nearly
P-391 Pregnancy viability in relation to demographic and repeated the same ability. Future trials might explore this combination in women with
sonographic and serum biomarkers pregnancies <7 weeks’ gestation or unknown location
J.F. Petersen1, L. Friis-Hansen2, T. Bryndorf3, A.K. Jensen4, Trial registration number: NCT02761772
A. Nyboe Andersen5, E. Løkkegaard1
1 P-392 Extracellular vesicles: an important biomarker in recurrent
North Zealand Hospital, Obstetrics and gynecology, HIllerød, Denmark ;
2 pregnancy loss?
North Zealand Hospital, Clinical Biochemistry, Hillerød, Denmark ;
3
Private Practice, Gynecological clinic, Copenhagen, Denmark ; N. Rajaratnam1, N.E. Ditlevsen2, M.M. Jørgensen3, O.B.
4
Institute of Public Health- University of Copenhagen, Section of Biostatistics, Christiansen4
1
Copenhagen, Denmark ; Aalborg University Hospital, Emergency- and Trauma Center, Aalborg, Denmark ;
5 2
Rigshospitalet, The Fertility Clinic, Copenhagen, Denmark Region Hospital North, Emergency Department, Hjoerring, Denmark ;
3
Aalborg University Hospital- The Faculty of Medicine, Clinical Immunology and
Study question: Which individual and combined variables of demography, Clinical Diagnostics, Aalborg, Denmark ;
transvaginal sonography and serum markers are the best dynamic predictors of 4
Aalborg University Hospital- The Faculty of Medicine, Obstetrics and Gynaecology,
viability in the first trimester? Aalborg, Denmark
Summary answer: Combined maternal age, hCG, crown-rumP-length and
bleeding data was the best model to dynamically predict miscarriage, closely Study question: Can different types of extracellular vesicles (EVs) markers be
followed by maternal age, hCG, estradiol and bleeding used as predictive markers of pregnancy outcome in recurrent pregnancy loss
What is known already: Early pregnancy losses are common and about 25% (RPL) patients?
of recognized pregnancies end in a miscarriage. More precise prediction of Summary answer: Although tendencies were found, there were no significant
miscarriage could potentially alleviate the anxiety of couples experiencing symp- differences in the progression of EV levels throughout pregnancy between RPL
toms but remains challenging. The diagnosis often depends on the development patients with live birth or miscarriage.
of ultrasonic or biochemical evidence. Maternal age is the principal risk factor What is known already: RPL defined as ≥ 3 miscarriages has an estimated
and the current tendency towards delayed childbearing leads to an increased incidence of 1-3% in all couples. The etiology of RPL is considered to be
incidence of clinical visits for viability reassurance. Early pregnancy diagnostics multifactorial. EVs are small particles secreted into plasma that take part in
is usually based on retrospective demographic, ultrasonic or biochemical data. numerous physiological processes and their contents provide information
Potentially, all three sources prospectively combined carries an increased accu- about the originating cell and pathophysiological states in different diseases.
racy compared with any one-parameter diagnostics In pregnancy disorders, changes can be seen in the composition, bioactivity
Study design, size, duration: A prospective cohort of pregnant women with and concentration of placental and non-placental EVs. So far no laboratory
asymptomatic and assumed healthy pregnancies of no more than 55 days’ ges- test has a good diagnostic value in predicting new pregnancy loss in RPL
tation was recruited and followed from June 2016 to March 2017 at two patients.
University Hospitals in Copenhagen, Denmark. Participants had repeated visits Study design, size, duration: Prospective cohort study performed at a ter-
every second week from 4–14 weeks’ gestation until either miscarriage or con- tiary university-affiliated centre for RPL from the period July 2018-June 2019.
firmed viability at the end of the first trimester. The study population included The study includes 26 pregnant RPL patients, n=20 with live birth (LB) and n=6
203 women with 715 visits with a new miscarriage (MI). Within these groups 4 patients in the MI group
Participants/materials, setting, methods: Before inclusion, participants received intravenous immunoglobulin (IVIG) as treatment, and 8 in the LB group.
and male partners completed a validated questionnaire for baseline demographic The decision to provide IVIG was mainly made on the basis of a high number
data. At each visit, a panel of 29 commonly used blood-samples, transvaginal of previous miscarriages.
sonography and symptoms of miscarriage was recorded. Based on updated Participants/materials, setting, methods: Up to 5 blood samples were
values, unadjusted hazard ratios (HR) identified the eight most influential bio- obtained from each patient: 1 baseline before pregnancy and 1-4 during preg-
markers on the risk of miscarriage. Testing these in adjusted groups of three, nancy in gestational week 6, 8, 10 and 16. The samples were analyzed for EV
56 different combinations were evaluated for their ability to individually and type and concentration with EV Array, a method able to capture EVs by using
dynamically predict viability at each visit an antibody panel targeting membrane proteins. 32 different markers were

analyzed specific for immunology/inflammation, coagulation, hypoxia, placenta In contrast, the abundance of p16+ stromal cells showed low, but a significant
and hormones. correlation with the percentage of CD56+ stromal cells (R=0.28; p<0.01).
Main results and the role of chance: The analysis showed that within the Limitations, reasons for caution: The study was limited in sample size.
LB group (n=20, IVIG n=8, no IVIG n=12) there was a statistically significant Wider implications of the findings: Increased cell senescence in endome-
(from p < 0.01 to p < 0.0001) increase in 47 % of the EV subtypes in the group trial stroma during the mid-luteal phase is associated with rise in uNK cells in
receiving IVIG, between the sample taken before pregnancy and IVIG treatment women with recurrent pregnancy failure. These results confirm the role of uNK
and the sample taken at gestational week 6 after initiation of IVIG. Regarding cells in the selective clearance of senescent cells in the endometrium during the
pregnancy outcome, several graphs were made with median values for the LB period of embryo implantation.
group compared with graphs for each of the 6 miscarried pregnancies, These Trial registration number: NA
showed clear tendencies towards different progressions of the EVs positive for
markers CD9, CD81, Annexin V (general EV markers), CAXII (hypoxia marker),
P-394 Aspirin and low-molecular-weight heparin combination
FSHR, LHR (hormone receptors), TRAIL (immunological marker) and PLAP
therapy provides favorable immunomcompetent milieu to protect
(placental marker). One patient with miscarriage of a normal but severely
pregnancy in hyperhomocysteinemic women with recurrent
growth-restricted fetus in gestational week 21 already exhibited a marked decline
pregnancy loss
of almost all EV subtypes from week 12.
Limitations, reasons for caution: A limitation is the small study size. It is P. Chakraborty1, S. Chatterjee2, D. Patra2, S. Parvin1,
unclear which changes in EV levels are induced by the growing, and in some cases S. Kalapahar1, R. Chattopadhyay1, S. Ghosh1, M. Chakraborty1,
probably ischemic placenta and which are exclusively due to the IVIG S.N. Kabir1, B. Chakravarty1
1
administration. Institute of Reproductive Medicine- Kolkata., Department of Assisted
Wider implications of the findings: This is the first study of EVs in preg- Reproduction, Kolkata, India ;
2
nancies of RPL patients.In a larger study it will be possible to investigate whether Vidyasagar University, Tissue culture and molecular biology, Kolkata, India
the clear tendencies for different progressions of EV subtypes during pregnancies
with successful and unsuccessful outcomes still applies and whether it can be Study question: Does aspirin and low-molecular-weight heparin (LMWH)
used as a prognostic biomarker. combination therapy offer a favorable immunological milieu to salvage pregnancy
Trial registration number: Ethics committee approval number: N-20180025 in hyperhomocysteinemic women with recurrent pregnancy loss (RPL)?
Summary answer: Aspirin-LMWH combination therapy attenuates Th17 cell
P-393 Relationship between the percentage of endometrial response and stimulates Th2-associated cytokine and chemokine production to
senescent cells and uterine natural killers (uNK) cells during the confer immunocompetency and protect pregnancy in hyperhomocysteinemic
mid-luteal phase in women with recurrent pregnancy failure women with RPL.
D. Parvanov1, R. Ganeva1, N. Vidolova1, G. Stamenov2 What is known already: The fetal immune tolerance is effected by a balance
1
Nadezhda Women’s Health Hospital, Research, Sofia, Bulgaria ; between T-helper (Th)-1, Th2, Th17, and regulatory responses involving both
2
Nadezhda Women’s Health Hospital, Obstetrics and Gynecology, Sofia, Bulgaria innate and adaptive immune cells orchestrated by signaling molecules (chemo-
kines and cytokines). Women with RPL and/or recurrent implantation failure
Study question: Is there an association between the presence of p16INK4a-pos- are reported with an elevated Th1/Th2 cell ratio and increased immune inflam-
itive cells in different endometrial compartments and CD56+ uNK cells during matory responses at maternal-fetal interface. We have previously demonstrated
the mid-luteal phase in RIF patients? that combined aspirin-LMWH anticoagulant therapy improves rate of pregnancy
Summary answer: The percentage of endometrial stromal p16INK4a-positive salvage in hyperhomocysteinemic women with RPL (PLOS One 2013). We
cells positively correlates with the abundance of CD56+ uNK cells in RIF women hypothesized that LMWH might persuade an immune-modulatory milieu, which
during the mid-luteal phase. could balance the pro-inflammatory response implicated in pathogenesis of RPL.
What is known already: P16INK4a is commonly measured in order to assess Study design, size, duration: This prospective cohort study in 53 hyperho-
the level of senescence in the human endometrial tissue. Recent studies have mocysteinemic (homocysteine>12mmol/L) women with RPL is conducted
shown that p16-positive senescent cells are involved in endometrial receptivity between August 2017 and December 2019 at Institute of Reproductive Medicine,
and in the acute cellular remodelling during the embryo implantation period. Kolkata. Patients (n=24) were supplemented with aspirin (5mg/day) alone or
Recognition and elimination of these senescent cells by immune cells, especially in combination with LMWH (2500IU/day) (n=29) for 12 weeks. They are ana-
CD56+ uNK cells, play an essential role in tissue repair and homeostasis. lyzed for cytokines related to Th1 (interferon (IFN)-g, tumor-necrosis-factor
However, data about the association between the abundance of senescent cells (TNF)-a), Th2 (interleukin (IL)-10, IL-13) and Th-17 (IL-6, IL-23, transform-
and uNK cells in human endometrium is still scarce. ing-growth-factor (TGF)-b) with their respective chemokines (CXCL10,
Study design, size, duration: This is an observational study of 92 women CXCL11), (CCL17, CCL22) and (CCL20,CXCL1,CXCL8).
with recurrent implantation failure (RIF) who had an endometrial biopsy during Participants/materials, setting, methods: Plasma samples obtained at
the mid-lutheal phase. We used immunohistochemical (IHC) markers, p16 and gestational weeks (g.w.) 6 and 12 are analyzed by multiplex fluorochrome and
CD56, in order to analyze the association between senescent cells and uNK in Luminex performance assay for levels of Th1, Th17, and Th2-related cytokines
human endometrium. and chemokines respectively. Percentage of Th17 and T-regulatory (Treg) cells
Participants/materials, setting, methods: Our study was performed at in peripheral-blood-mononuclear-cells (PBMC) is detected by flow-cytometry.
Nadezhda Women’s Health Hospital, Bulgaria. Patient biopsies with known Differences in chemokine levels in between the groups were assessed by linear
pregnancy outcome were retrieved from our tissue bank. Tissue sections were mixed models. Statistical significance was set at p < 0.05 as evaluated by stu-
stained immunohistochemically for CD56 (a uNK-specific cell surface antigen) dent’s T-test.
using CD56 antibody (E-AB-62136, Elabscience, USA), or p16 senescent bio- Main results and the role of chance: No differences are observed in
marker using p16 ink4a antibody (Master Diagnostica, Granada, Spain). The per- cytokine and/or chemokine level/s at g.w. 6 between the groups. The
centage of positively stained cells was calculated after enumeration by two percentage of Th17 cells (IL-17+/CD4+) significantly decreased (2.98 ± 0.68%
independent investigators in multiple endometrial sections. vs. 1.74 ± 0.29%; p<0.01) with a simultaneous increase (4.94 ± 1.38% vs.
Main results and the role of chance: The percentage of p16-positive cells 3.03 ±0.91%; p<0.02) in percentage of Tregs (CD4+CD25brightFoxp3+ T cells)
in the endometrial stroma during the mid-luteal phase in the studied group of in aspirin-LMWH-treated cohort. The combination group, although
patients ranged between 0.03% and 6.73%, while it varied between 0.06% and documented a decreased concentration/s (pg/ml) of Th17-type cytokines
42.79% in the glands, and between 1.05% and 88.90% in the luminal epithelium. (IL-6 (1.88 ± 0.65 vs. 0.91 ± 0.43; p<0.04 and IL-23(17.94 ± 9.76 vs. 7.78 ±
CD56-positive uNK cells were found only in the stroma and their abundance 3.43; p<0.01)), at g.w. 12, concentration of TGF-β was significantly higher
ranged between 0.01% and 34.17%. (3909.05 ± 1248.35 vs. 2469.83 ± 1058.71; p<0.01) which cue to probable
No correlation was found between the frequencies of occurrence of senes- maintenance of balance of inflammatory response in first trimester. Improvement
cent p16+ glandular and luminal epithelial cells and CD56+ cells (p>0.05). in IL-10 concentration (6.32 ± 2.46 vs. 8.87 ± 4.32; p<0.02) was documented

after combination treatment. No differences were observed in Th1-type cytokine P-396 Multidisciplinary approach in repeated implantation failure
level/s (IFN-γ (1.97 ± 0.69 vs. 1.76 ± 0.34) and TNF-α (1.03 ± 0.37 vs. 0.98)) patients’ evaluation
providing a favorable shift in the combination cohort indicating a Th2 bias. Mixed S. Bordoli1, M. Scarrone1, F. Pasi2, V. Canti3, L. Quaranta3,
linear model test showed an increase in CCL22 (p<0.05); and a decrease (p < R. De Lorenzo1, S. Cella1, V.S. Vanni4, E. Papaleo3, P. Giardina3,
0.04) in CCL20 during g.w.12 while CXCL10, CXCL11, CXCL1, CXCL8, P. Rovere Querini1
CCL17, CCL22 do not differ during the treatment. 1
Vita-Salute San Raffaele University, Internal and Specialistic Medicine, Milan, Italy ;
Limitations, reasons for caution: Future studies with higher sample size 2
IRCCS San Raffaele Hospital, Obstetrics and Gynecology, Milan, Italy ;
should corroborate the current findings. Moreover, this is an observational study 3
IRCCS San Raffaele Hospital, IVF unit, Milan, Italy ;
not supported by appropriate/untreated controls because of stringent regula- 4
Vita-Salute San Raffaele University, IVF Unit, Milan, Italy
tion/s from local ethics board. LMWH treatment was not continued beyond
12 weeks of gestation for absence of systematic network monitoring in Study question: Do repeated implantation failure (RIF) patients benefit from
our set-up. medical multidisciplinary evaluation?
Wider implications of the findings: Aspirin-LMWH supplementation aug- Summary answer: An internist evaluation from an immunological, endocri-
ments the number of Treg cells and increases IL-10 that perhaps provide a nological and hematologic point of view, associated with the gynaecological one
favorable immunocompetent milieu to protect pregnancy in hyperhomocyste- in RIF patients can improve their reproductive prognosis.
inemic women with RPL. Further ongoing studies will look into the duration of What is known already: No consensus regarding the definition of RIF has
LMWH regimen for better immunosupression. been reached. A widely accepted definition is the failure of implantation of at
Trial registration number: NA least three good quality embryos, in at least three fresh or frozen cycles trans-
ferred in women aged <40 years. These women are usually evaluated only in
P-395 Evaluating the clinical utility of endometrial receptivity the context of IVF units. Pathogenesis is still unclear and multifactorial (embryo
analysis test in women with recurrent pregnancy loss factors, maternal/endometrial factors, gamete factors and ART techniques asso-
ciated factors). Recurrent pregnancy loss (RPL) is instead well known to benefit
M. Luna1, T. Alkon1, C. Hernandez-Nieto1, D. Cassis-Bendeck1,
from thorough medical investigations since many causes other than gynaecolog-
B. Sandler1
1
ical ones have been proved to be associated to RPL.
Reproductive Medicine Associates of New York, Reproductive Endocrinology and Study design, size, duration: We performed a prospective observational
Infertility, New York- NY, U.S.A. study of patients referred to recurrent pregnancy loss and implantation failure
outpatient clinic at San Raffaele Hospital (Milan, Italy) between March 2019 and
Study question: Does endometrial receptivity analysis (ERA) test offer clinical March 2020 (still ongoing). The patients reproductive complaints were RIF
utility in patients with recurrent pregnancy loss (RPL)? (n=43) or RPL (n=110).
Summary answer: Women with RPL may benefit from a personalized adjust- Participants/materials, setting, methods: We studied patients according
ment in timing of frozen embryo transfer. to demographic data, past medical history (genetic diseases, hypertension,
What is known already: Successful embryo implantation requires an appro- abnormal thyroid function, congenital or acquired thrombophilia, history of DVT
priate embryonic development, coincident with a receptive endometrium. or pulmonary embolism, autoimmune diseases), past gynecological and obstetric
Defects in endometrial receptivity contributing to implantation failure, remain history. Laboratory test have been prescribed to investigate autoantibodies,
not only the major rate-limiting step in vitro fertilization (IVF) success, but also inflammatory indexes or dysmetabolic conditions. In order to assess the rele-
a major culprit in unexplained infertility and spontaneous abortions. Because of vance of the evaluation and management, we have longitudinally followed
this, the ERA test was developed. ERA test enables the determination of a patients during fertility procedures after the first visit and collected pregnancy
personalized window of implantation (WOI). However, clinical evidence is still rates and outcomes.
scarce regarding the value of performing an endometrial receptivity test in Main results and the role of chance: When we compared the prevalence
women with RPL. of abnormal findings in RIF and RPL we found no statistically significant (p>0.5)
Study design, size, duration: Monocentric prospective study from June 2018 difference in the distribution, suggesting that the factors contributing to the poor
to April 2019. 10 women with at least 2 consecutive miscarriages under 10 reproductive outcomes in these two dramatically different groups of patients
weeks of gestation of unexplained origin after IVF conceptions, underwent an may be similar. In a significant proportion of RIF patients we detected positive
ERA test. Subsequently, a single frozen euploid embryo transfer (SET) with antinuclear antibodies (32.5%) antithyroid antibodies (18.6%), prothrombin
appropriate adjustment in timing according to the ERA test was performed. mutation (11.5%), hyperhomocysteinemia and positive antiphospholipid anti-
Participants/materials, setting, methods: Women with RPL of unex- bodies (6.9%). Moreover, after appropriate treatment and proper support, 15
plained origin, <40 years, history of a full-term pregnancy after a euploid embryo patients underwent a further IVF cycle and n=8 (implantation rate 0.53) had a
transfer, normal karyotypes, uterine cavity assessed by hysteroscopy and no positive pregnancy test, compared to an implantation rate of 0.15 normally
identified thrombophilia’s were included. Participants were administrated oral/ reported in RIF patients undergoing a subsequent embryo transfer.
transdermal estradiol on cycle day two for endometrial preparation. When the Limitations, reasons for caution: The small number of patients assessed
endometrial thickness was ≥ 7 mm/ triple line pattern, luteal support was begun so far in our multidisciplinary clinic is the main limitation of this work. An
using vaginal and intramuscular progesterone. Endometrial biopsy was performed increment of data collection and pregnancy rate and outcomes will be of
upon completion of 5 days of progesterone. interest.
Main results and the role of chance: 100% of the patients were found to Wider implications of the findings: The establishment of a multidisciplinary
have a non-receptive endometrium. Five were pre-receptive (24 hours), three team comprising an infertility specialist flanked by an internist could improve
early receptive (12 hours), and two late receptive (12 hours). After a personal- assisted reproductive technology (ART) outcomes by detecting immunological/
ized SET was performed, 70% (7/10) became pregnant. Of the 70% of the endocrinological problems that may play a role in both implantation failure patho-
women that achieved a pregnancy, 1 patient had a miscarriage at week 10 and genesis and physiological pregnancy development.
the remaining had a live birth. Trial registration number: non applicable
Limitations, reasons for caution: This a pilot study performed to test the
utility of ERA test in women with RPL. Due to the size of the study, the effects
P-397 Low iron stores are associated with Recurrent Pregnancy
of more subtle covariates would not have been detected. Future randomized
Loss but does not predict subsequent loss or ability to conceive
controlled trials are needed.
Wider implications of the findings: A displaced implantation window was M. Georgsen1, M.C. Krog2, A.S. Korsholm3, H.W. Hvidman3,
found in all the patients analyzed with RPL. These findings suggest for the first A.M. Kolte1, A.S. Rigas4, H. Ullum4, S. Ziebe3, A.N. Andersen3,
time, that the WOI may change after a full-term pregnancy or surgical uterine H.S. Nielsen5, M.B. Hansen6
1
manipulation. These results enable appropriate study design of future studies. Copenhagen University Hospital- Rigshospitalet, The Recurrent Pregnancy Loss
Trial registration number: Not Applicable Unit- The Fertility Clinic, Copenhagen, Denmark ;

2 2
Copenhagen University Hospital- Rigshospitalet, The Recurrent Pregnancy Loss University of Liverpool, Institute of translational medicine, Liverpool, United
Unit- The Fertility Clinic and Department of Clinical Immunology, Copenhagen, Kingdom ;
3
Denmark ; Homerton University Hospital, Homerton fertility centre, London, United Kingdom
3
Copenhagen University Hospital- Rigshospitalet, The Fertility Clinic, Copenhagen,
Denmark ; Study question: Does the pregnancy implantation site and trophoblastic ring
4
Copenhagen University Hospital- Rigshospitalet, Department of Clinical thickness in an early scan predict outcomes and complication rates of intrauterine
Immunology, Copenhagen, Denmark ; pregnancies?
5
University of Copenhagen- Rigshospitalet- and Amager-Hvidovre Hospital, The Summary answer: Pregnancies located in the lower half of the uterine cavity
Recurrent Pregnancy Loss Unit and Department of Gynaecology-and-obstetrics, with a thinner trophoblastic ring were more likely to miscarry.
Copenhagen, Denmark ; What is known already: Women routinely have early gestation scans to
6
Copenhagen University Hospital- Rigshospitalet, Department of Clinical assess the location and to confirm viability of pregnancy. This reassures most
Immunology, Copenhagen, Denmark women who continue to have an uneventful pregnancy, although unfortunately
some will later have a miscarriage or later pregnancy complications. Trophoblastic
Study question: Is low iron stores associated with Recurrent Pregnancy Loss ring may be related to placentation and no previous studies have examined the
(RPL) and does low iron stores impair the ability to conceive? predictive role of trophoblastic ring thickness at time of early gestation scanning
Summary answer: S-ferritin levels were inversely related to number of pre- in pregnancy outcomes and late pregnancy complications. Implantation sites after
vious losses in women with RPL. Low iron stores did not affect the ability to an embryo transfer had not been associated with pregnancy outcomes.
conceive. Study design, size, duration: This was a prospective observational study in
What is known already: Iron requirements are increasing during pregnancy a tertiary referral publically funded IVF unit in UK. 300 patients who attended
and iron deficiency is common among women of reproductive age. Iron is essen- for early pregnancy scan following single embryo transfer over a 14-month period
tial for hemoglobin production, cellular metabolism and the immune system. were included after obtaining Informed consent.
Iron deficiency has been suggested to affect fertility due to the iron requirements Participants/materials, setting, methods: Patient typically underwent their
of the developing oocytes. Low maternal iron stores result in lower birth weight early pregnancy scan between 6-8 weeks gestation. Inclusion criteria included
and increase the risk of preterm birth. Studies of iron stores and sporadic preg- any live ongoing singleton pregnancy following single embryo transfer and a 3D
nancy loss are sparse and contradictory. Furthermore, an association between image was obtained of the uterus. Exclusion criteria were Women with uterine
iron deficiency and RPL has not previously been studied. cavity abnormalities, multiple pregnancies or double embryo transfers were
Study design, size, duration: A cohort study comparing s-ferritin levels in excluded. Patients were followed up to until the end of pregnancy, either mis-
women with RPL (n=84) referred between 2013-2016 with a control group of carriage, term or preterm birth. Any adverse pregnancy outcomes (pre-eclamp-
women of reproductive age with no known fertility problem (n=153). The study sia, IUGR etc) were also recorded.
investigated whether s-ferritin correlated to the ability to conceive and time to Main results and the role of chance: Of the 300 patients recruited, at this
conception during the first two years after sampling and whether s-ferritin were interval review 286 pregnancies were ongoing with the remaining 14 ending up
associated with outcome of the first pregnancy after referral for RPL women. as miscarriages. Women who miscarried were older than those with ongoing
Participants/materials, setting, methods: The study included women with pregnancies (mean ages 36.3 vs 33.5 years; p=0.002) but there were no other
RPL (≥3 pregnancy losses) referred to the Danish RPL unit and women who demographic differences were identified (e.g. BMI). Mean trophoblastic ring was
consulted the Fertility Assessment and Counselling Clinic both located at thicker in the ongoing pregnancy group when compared with women who mis-
Copenhagen University Hospital, Denmark. Pregnancy history were recorded carried (7.14mm vs. 5.4mm; p=0.008).
and serum samples were collected at first consultation and stored at -20°C until Of the 14 miscarriages, 4 were located in the lower half of the uterine cavity.
analysed. Follow-up on verified pregnancies was done after two years for both 18 pregnancies were located in the lower half of the cavity in the remaining
groups. Women who did not try to conceive or underwent fertility treatment ongoing pregnancies. This indicated a significant increase in miscarriage rate when
were excluded. implantation occurred in the lower half of the cavity (p=0.01).
Main results and the role of chance: Women with RPL had lower s-ferritin Limitations, reasons for caution: This is an observational study without
compared with the control group, 39.9 µg/L versus 62.2 µg/L (p=0.001) and intervention. The observed difference may be attributable to the differences in
had a higher prevalence of low iron stores (s-ferritin <30 μg/L), 35.7 % versus patient age, but the slight difference of <3 years is unlikely to be the sole reason.
13.7 % (p<0.001). There was an inverse relationship between s-ferritin levels Wider implications of the findings: Investigation of trophoblastic ring thick-
and number of pregnancy losses before referral for the RPL women. When ness as a marker of pregnancy outcome/complications is warranted. Identifying
divided into groups of 1) 2-3 losses, 2) 4 losses, and 3) ≥5 losses, median s-fer- non-invasive biomarkers facilitating studies of trophoblast invasion could lead
ritin levels differed significantly between the groups (p=0.010). There was no to development of therapeutic strategies targeting complications of pregnancy.
association between s-ferritin levels and the ability to conceive or time to preg- Avoiding embryo deposition in the lower cavity and correlating implantation site
nancy in neither the RPL nor the control group. S-ferritin did not predict the risk following embryo transfer also requires further investigation.
of losing the first pregnancy after referral in the RPL group. Trial registration number: not applicable
Limitations, reasons for caution: This study is limited by the relatively small
sample size and the low prevalence of iron deficiency in the studied population P-399 Proteomic signatures in Assisted Reproduction Technology
which may explain why we did not find a correlation between iron deficiency (ART) cycles
and subsequent pregnancy loss or ability to conceive. V. Kanaka1, A. Papadopoulou2, S. Proikakis2, P. Drakakis1,
Wider implications of the findings: The findings indicate that low s-ferritin T.G. Tsangaris1, D. Loutradis1
could be a relevant risk factor in women with RPL. It should be investigated 1
Department of Obstetrics and Gynecology, Alexandra Hospital, School of Medicine ;
whether iron status should be assessed routinely before pregnancy in both 2
Proteomics Research Unit, Biomedical Research Foundation of the Academy of
healthy women and women with RPL and whether iron supplementation may Athens, Athens, Greece
increase the chance of live birth. Study question: The identification of biomarkers for successful embryo
Trial registration number: Not applicable implantation and pregnancy onset in In Vitro Fertilization (IVF) by proteomic
analysis
Summary answer: Twenty four proteins were differentially expressed among
P-398 Does pregnancy implantation site and trophoblastic
women achieving and not achieving pregnancy after IVF and embryo implantation
ring thickness at early gestational scan predict outcomes of an
What is known already: More than 400,000 babies from 1,6 million Assisted
intrauterine pregnancy? An observational study
Reproduction Technology (ART) cycles are born around the world every year.
L. Nancarrow1, S. Vinayagam1, M. Swanson1, K. Ford1, The efficacy of in vitro fertilization cycles (IVF) has improved significantly over
D. Hapangama2, R. Hombury3, R. Russell1 the past 40 years. However better understanding of the reproductive process
1
Liverpool Womens Hospital, Hewitt fertility centre, Liverpool, United Kingdom ; may help increase birth rate. Proteomic analysis of the embryo secretome that

has already been done has shown a positive correlation between high levels of stimulated cycles (endometrium in STIM ET and EP = 10.7 mm versus STIM ET
leptin and human leukocyte antigen (HLA) with the implantation potential of and no EP = 11.8 mm; p = 0.029). In contrast, with Fr ET cycles, the size of the
the embryo. Moreover previous studies have shown that protein AMBP and lining was not important for the occurrence of EP (Fr ET and EP = 10.6 mm
Leucine-rich alpha-2-glycoprotein play an important role in the implantation versus Fr ET and No EP = 11.14 mm; p = 0.2). This comparison emphasizes the
process. observation that EP is more often detected in stimulated than in frozen cycles
Study design, size, duration: Proteomic analysis was performed for the (61.98 % versus 39.02%; p = 0.04). Something more, most of the EP are resulted
analysis of peripheral blood serum of women undergoing IVF treatment age after a transfer of blastocyst stage embryos but only in stimulated cycles (EP
30-43. Blood was collected prior to the IVF program, at the day of embryo rate 52% versus 21%, p = 0.0009). Also, we have established that high level of
transfer as well as 6 and 12 days post embryo transfer. In order to compare Progesterone on the day of hCG trigger could contribute for an ectopic embryo
successful and unsuccessful cycles, twenty women is the size of the study by implantation (progesterone in STIM ET and EP = 1.05 ng/ml versus STIM ET
which ten has successful implantation and ten unsuccessful in a three years dura- and no EP = 0.59 ng/ml; p = 0.0014).
tion of the study. Limitations, reasons for caution: The study was a retrospective cohort
Participants/materials, setting, methods: After serum was obtained, two study at a single center, larger trials with an increased number of patients needed
dimensional gel electrophoresis (2DE) followed by MALDI-TOF–MS/MS Mass to confirm our finings.
Spectrometry were performed to analyze the differential expression of proteins Wider implications of the findings: This study shows an increased risk of
in the serum samples among women having successful and unsuccessful IVF an ectopic pregnancy in stimulated ET cycles than in FrET. These findings suggest
cycles. elevated progesterone and hormonal milieu of ovarian stimulation was detri-
Main results and the role of chance: Twenty four proteins were differen- mental for ectopic pregnancy. Freeze all strategy followed by Fr ET would
tially expressed among women achieving and not achieving pregnancy. Specifically, decrease chance of ectopic pregnancies in ART.
protein AMBP, Apolipoprotein A-IV, Growth/differentiation factor 2, Trial registration number: Not applicable
Glucosamine-6-phosphate isomerase 2, G patch domain and ankyrin repeat-con-
taining protein 1, Hemoglobin subunit beta, Kelch-like protein 23, Zinc-alpha-2- P-401 Characterization of bHCG trends in Euploid Frozen-
glycoprotein, Leucine-rich alpha-2-glycoprotein, Vitronectin, protein NATD1 Thawed Embryo Transfer Cycles by Initial Value
and Plasminogen were expressed only in successful cycles. Proteins that were J. Chervenak1, R. Setton2, A. Petrini2, K. Hancock2, P. Chung2,
present only in unsuccessful resulting cycles are proteins that belong mainly to Z. Rosenwaks2
the immune system and include Complement C1r subcomponent, Complement 1
New York Presbyterian/Weill Cornell, Obstetrics and Gynecology, New York,
C1s subcomponent, Complement C1q subcomponent subunit B, Ig mu heavy U.S.A. ;
chain disease protein, Ig mu chain C region as well as Hemopexin, Serum para- 2
Center for Reproductive Medicine, Obstetrics and Gynecology, New York, U.S.A.
oxonase/arylesterase 1, Alpha-2-macroglobulin, Tyrosine-protein kinase STYK1,
Angiotensinogen, Keratin, type I cytoskeletal 9 and Keratin, type II cytoskeletal 1 Study question: What is the predictive value of initial bHCG change stratified
Limitations, reasons for caution: by initial bHCG value in frozen-thawed embryo transfer (FET) cycles with pre-
–No Donor cycles implantation genetic testing (PGT)?
–Age limited to 30-43 years old Summary answer: This study affirms that the initial bHCG level and its initial
–The results must be evaluated in a larger cohort and longer duration of time. rate of change are strong independent predictors of live birth in euploid FET.
Wider implications of the findings: Predicting pregnancy and improving What is known already: Existing studies have established initial bHCG level
drugs for IVF treatment. Improve IVF pregnancy rates and success. and subsequent rate of rise after IVF as one of the earliest means of predicting
Trial registration number: non applicable pregnancy outcome. Over the past decade, the proportion of FET and PGT
have increased. There is limited data to inform prognostic counseling in this
population. Further, the effect of the initial value of bHCG on the expected rate
P-400 Ovarian hyperstimulation and high levels of progesterone
of rise of bHCG has not been well characterized.
are detrimental for ectopic pregnancy in ART
Study design, size, duration: This is a retrospective cohort study in which
I. Oprova1, N. Magunska2, T. Milachich3, A. Shterev2, P. Andreeva1 all FET cycles that had a transfer of a single PGT-euploid embryo performed in
1
Shterev Hospital, Assisted Reproduction Department, Sofia, Bulgaria ; an academic medical center from 2013 to 2019 were screened for inclusion.
2
Shterev Hospital, Department of Obstetrics and Gynecology, Sofia, Bulgaria ; Only cycles that resulted in a positive initial serum bHCG on CD 28 or 29,
3
Shterev Hospital, IVF and Andrology Laboratory, Sofia, Bulgaria followed by a rise in the serum bHCG value 48 hours later, and a subsequent
maximum of one gestational sac were included in the study.
Study question: Could ovarian hyperstimulation and high levels of Participants/materials, setting, methods: There were 596 FETs during
Progesterone be a risk factor for an ectopic pregnancy in ART? the study time period that met the inclusion criteria. Groups were stratified by
Summary answer: Endometrium thickness over 12 mm and progesterone initial bHCG level (group 1: <50 mIU/mL; group 2: ≥50 mIU/mL and ≤100;
level under 1 ng/ml and FrET could be a preventive measure for EP development. and group 3: >100 mIU/mL) and compared across the rate of change of this
What is known already: The ectopic pregnancy (EP) rate has been reported value over two days (≤50%, 51-99%, and ≥100%). Statistical analysis was per-
higher after assisted reproductive technology (ART) than after a spontaneous formed using Chi-square and Fisher’s exact t-test.
conception. The factors associated with abnormal implantation are widely dis- Main results and the role of chance: Baseline age and BMI were similar for
cussed and not completely clear. all groups. For group 1, the live birth rate (LBR) was the lowest of the cohorts
Study design, size, duration: The present study was an observational ret- (LBR 19.57%) and there was no significant difference in the chance of live birth
rospective trial, performed in a private IVF clinic. It encompassed 366 women regardless of whether bHCG rose <50% (LBR 10%), between 51-99% (LBR
who have undergone in vitro fertilization treatment: stimulated cycles with fresh 10%), or ≥100% (LBR 26.9%, p=0.16). For group 2, the LBR significantly
embryo transfers (STIM ET) and hormone replacement therapy with frozen increased across the three categories of rate of bHCG rise (LBR 25% for ≤50%
embryo transfers (Fr ET). rise, vs. 54.5% for 51-99% rise, vs. 70.5% for ≥100% rise, p<0.05). Similarly for
Participants/materials, setting, methods: Four features were evaluated: group 3, the LBR significantly increased across the three categories (LBR 10%
endometrial thickness at the day of ET, stage of the embryos transferred – for ≤50% rise, vs. 84.1% for 51-99% rise, vs. 90.1% for ≥100% rise, p<0.05 for
cleavage stage (day 3) or blastocyst stage (day 5), the progesterone level on the all). For groups 2 and 3, an increase of bHCG of more than 50% is strongly
day of hCG in stimulated cycles, ovarian hyperstimulation (OHS). The primary associated with an increased LBR (p=0.02 and p=<0.0001, respectively). In the
outcome measure was a development of EP. Logistic regression was performed absence of a bHCG rise of >50% over 2 days, there was no significant difference
to utilize the significance of the numbers. noted between the 3 groups and prognosis was poor (LBR 14.29%).
Main results and the role of chance: At total of 366 Fresh ET or Fr ET Limitations, reasons for caution: This study examines bHCG values taken
cycles were compared, 41 with EP and 325 with an intrauterine pregnancy. We during a narrow interval of days after FET. The results of this study may not be
detected that thin endometrium could be risky for developing EP, but only in generalizable to values taken outside of this timeframe.

Wider implications of the findings: The findings of this study elucidate the 1
University Women’s Hospital and University of Bern, Gynaecological
relative impact of initial serum bHCG values in FET with PGT cycles for predicting Endocrinology and Reproductive Medicine, Bern, Switzerland ;
2
live birth and offer a counseling tool for the clinician. University Women’s Hospital, Gynaecological Endocrinology and Reproductive
Trial registration number: not applicable Medicine, Bern, Switzerland ;
3
Bern University Hospital and University of Bern, Institute of Pathology, Bern,
Switzerland ;
P-402 The impact of chronic endometritis on pregnancy 4
University of Bern, Institute of Social and Preventive Medicine, Bern, Switzerland
outcomes in women with recurrent implantation failure
Y. Liu1, X. Chen1, W.C. Cheung1, Y. Zhao1, R. Qi1, J.P.W. Chung1, Study question: What is the impact of diagnostic endometrial biopsy and
C.C. Wang1, T.C. Li1 treatment of chronic endometritis (CE) on the time-to-pregnancy and live birth
1
The Chinese University of Hong Kong, Department of Obstetrics and in women with recurrent pregnancy loss or recurrent implantation failure?
Gynaecology, Shatin- Hong Kong, China Summary answer: Diagnostic endometrial biopsy and treatment of chronic
endometritis reduces time-to-pregnancy an time to live birth in women with
Study question: Does the presence of chronic endometritis (CE) affect preg- recurrent pregnancy loss or recurrent implantation failure.
nancy outcomes in women with recurrent implantation failure (RIF)? What is known already: Up to 57% of women with recurrent pregnancy
Summary answer: Chronic endometritis was associated with a lower suc- loss (RPL) and up to 66% of women with recurrent implantation failure (RIF)
cessful implantation rate in women with RIF. suffer from CE. Recently, a meta-analysis showed higher pregnancy and live birth
What is known already: Chronic endometritis is a state of persistent inflam- rates after antibiotic treatment for CE in women with RIF. This was also found
mation in the endometrial lining, which has been reported to be associated with in a meta-analysis of trials on endometrial scratch injury in women who had two
reproductive failure, including RIF. Plasma cell density in the entire endometrial or more implantation failures. However, there remains a need to investigate the
biopsied tissue has been put forward as a more reliable and accurate method effect of diagnosis and treatment of CE and endometrial scratch injury specifically
of plasma cell assessment for the diagnosis of CE, and that the prevalence of in women with RPL and RIF.
CE in women with reproductive failure was only 10%. However, the proof of Study design, size, duration: This is a retrospective analysis of a cohort of
this measurement requires clinical studies to confirm that it is of useful prognostic women at the University Women’s Hospital of Bern, Switzerland diagnosed for
value and leads to effective treatment on CE. recurrent pregnancy loss or recurrent implantation failure between January 2014
Study design, size, duration: This is a case-control observational study on and April 2019 (n=127). 108 fulfilled inclusion criteria, 61 with RPL and 47 with
pregnancy outcomes in 64 reproductive-age women with RIF undergoing endo- RIF. 41 served as historical controls as we only performed hysteroscopic assess-
metrial scratch in a natural cycle preceding frozen-thawed embryo transfer with ment without biopsy and detection of plasma cells until 2016.
the use of nondonor oocytes. All the women are classified with CE status using Participants/materials, setting, methods: We defined RPL as ≥ three con-
a stringent plasma cell density threshold, > 5.15 CD138+ plasma cells/ secutive miscarriages and RIF as ≥ six toP-quality cleavage-stage embryos unsuccess-
10 mm2, which was the 95th percentile of a fertile control population and served fully transferred. We performed diagnostic hysteroscopy and endometrial biopsy.
to establish a normal reference range. The pathologist stained endometrial tissue with hemalaun-eosin and immunohisto-
Participants/materials, setting, methods: Women failing to achieve a chemically for CD138. In cases of chronic endometritis, defined as presence of ≥
clinical pregnancy after transfer of at least four good-quality embryos in three one plasma cell, we treated with antibiotics. We used Cox regression to adjust for
or more transfer cycles and seeking treatment in our unit were included. confounding factors such as age of mother and parity and stratified by RPL and RIF.
Endometrial specimens were collected precisely 7 days after LH surge (LH+7) Main results and the role of chance: Of 108 women, forty-one (38%) had
and plasma cell density was determined based on Syndecan-1 (CD138) positive standard hysteroscopic assessment and served as control (control group). The
cells in the entire biopsy section. The clinical information (e.g. successful implan- remaining 67 (62%) women had a diagnostic endometrial biopsy and histopatho-
tation rate and clinical pregnancy rate) for these patients was reviewed and logical assessment. In 25 (37%) women, no plasma cells were detected (CEneg).
analyzed. Forty-two (63%) women were positive for chronic endometritis and subsequently
Main results and the role of chance: Among 64 patients recruited, 24 of treated with doxycycline 100mg twice a day orally for two weeks (CEpos). The
them were excluded because of the loss of follow-up and high uterine killer cells women in CEpos group (hazard ratio = 2.86; 95% CI 1.45 – 5.63; p= 0.002) had
count. Thus, a total of 40 patients were studied. 10% (4/40) of them were found a higher, and women in CEneg group (HR =2.11; 95% CI 0.87 – 5.08; p=0.095) a
to have CE, and the other 90% (36/40) did not have CE. There was no difference possibly higher chance for a clinical pregnancy than women did in the control group.
in maternal age, body mass index, history of pregnancy or duration of infertility We observed similar results for the probability of a live birth: for women of CEpos
between the CE and non-CE groups. group, the hazard ratio is 4.00 (95% CI 1.65 – 9.71; p=0.002) and for women of
We observed a lower successful implantation rate in the CE group when com- CEneg the hazard ratio is 2.02 (95% CI 0.71 – 5.70; p=0.184), both compared to
pared with the non-CE group [50.0% (2/4) vs. 52.8% (19/36), respectively], the control group. In all women with biopsy (groups CEng and CEpos), the hazard
while the unsuccessful implantation rate is higher in the CE group [47.2% ratio was 2.62 (CI 95% 1.39 – 4.94; p=0.003) for a clinical pregnancy and 3.00 (CI
(17/36)] than the non-CE group [50.0% (2/4)], but without a significant statistical 95% 1.38 – 6.52; p= 0.005) for a live birth compared to the control group.
difference (p =1.000). Limitations, reasons for caution: This is a retrospective analysis of data
However, the clinical pregnancy rate is higher in CE group than non-CE group from our center. We did not perform test-of-cure after Treatment unless a
[50% (2/4) vs. 36.1% (13/36), respectively]. Further, the non-pregnancy rate is woman had a subsequent miscarriage. However, our interest lied in the general
lower in CE group [50.0% (2/4) vs. 63.9% (23/36), respectively, p =0.622]. impact on the time-to-pregnancy of an endometrial diagnostic biopsy and sub-
Limitations, reasons for caution: The limitation of this study is that it only sequent Treatment of CE.
involved four women with CE due to its prevalence of 10% in our unit using the Wider implications of the findings: In women suffering from RPL or RIF,
novel diagnostic method, which precludes the analysis of the relationship we suggest performing endometrial biopsy, diagnosis and treatment of CE to
between CE and pregnancy outcomes in women with RIF. shorten the time-to-pregnancy and live birth. Endometrial biopsy itself might
Wider implications of the findings: We reported the presence of CE may have an effect on time-to-pregnancy and live birth. Possible rehabilitative healing
affect the successful implantation rate in women with RIF. We reason that CE processes may need some time.
may provide a role in predicting the prognostic value in other subgroups of Trial registration number: not applicable
reproductive failure, such as recurrent miscarriage and infertility.
Trial registration number: ChiCTR-IOC-16007882

P-403 Shortened time to live birth in subfertile women
MALE AND FEMALE FERTILITY PRESERVATION
undergoing endometrial diagnostic biopsy
V. Mitter1, S. Meier2, T.T. Rau3, M. Von Wolff1, M. Zwahlen4,
A.S. Kohl Schwartz3

P-404 The impact of the BRCA gene mutations on the P-405 Low in vitro maturation rates of oocytes recovered during
reproductive potential in patients with active breast cancer ovarian tissue cryopreservation of very young girls compared to
undergoing fertility preservation post-menarche patients undergoing fertility preservation before
S. Arab1, A. Badeghiesh2, W. Buckett3 oncotherapy
1
McGill University, Department of Obstetrics and Gynecology-Reproductive Y. Fouks1, E. Hamilton2, Y. Cohen1, Y. Hasson1, Y. Kalma1, F. Azem1
1
Endocrinology and Infertility Centre, Montreal, Canada ; Tel-Aviv Medical Center affiliated to the Sackler Faculty of Medicine- Tel-Aviv
2
McGill University, Department of Obstetrics and Gynecology, Montreal, Canada ; University, Lis Maternity Hospital- Department of Obstetrics and Gynecology-
3
McGill University, Department of Obstetrics and Gynecology-Reproductive Fertility Division-, Tel Aviv, Israel ;
2
Endocrinology and Infertility Center, Montreal, Canada Tel Aviv University, Sackler Faculty of Medicine, Tel Aviv, Israel
Study question: Do breast cancer patients with the BRCA gene mutations Study question: What are the outcomes of IVM of oocytes in very young
have lower ovarian reserve and poorer ovarian response than non-BRCA breast patients scheduled for onco-therapy?
cancer patients? Summary answer: Oocytes recovered from girls younger than 5 years of age
Summary answer: Breast cancer patients with the BRCA gene mutations who undergo fertility preservation have a lower maturation rate in IVM com-
have similar ovarian reserve and ovarian stimulation respond compared to pared to older females.
non-BRCA breast cancer patients. What is known already: In vitro maturation (IVM) of oocytes recovered
What is known already: Controversy exists about fertility and ovarian reserve during ovarian tissue cryopreservation is often practiced although it is still con-
in BRCA mutation carriers. Some studies suggest that these patients may have sidered experimental. To date, only a few studies have examined the success of
low ovarian reserve and poor ovarian stimulation response. However, most data this maturation process in pre-menarchal girls.
exists in carriers without breast cancer, rather than women with breast cancer Study design, size, duration: A total of 93 patients aged 0–25 years who
undergoing fertility preservation prior to gonadotoxic treatment. What data underwent ovarian tissue cryopreservation as part of onco-fertility preservation
does exist in breast cancer patients is from very small studies with conflicting between 2007 and 2019 were included in the study.
results. Concerns about poorer response to stimulation in BRCA carriers may Participants/materials, setting, methods: Oocytes were recovered from
lead to women electing not to pursue fertility preservation or to being the medium following ovarian tissue cryopreservation protocol and matured
overstimulated. over 48 hours. Their development and maturation rate were recorded and
Study design, size, duration: A retrospective cohort study of 131 breast compared between the different age groups.
cancer patients with known BRCA status who had had undergone fertility pres- Main results and the role of chance: The patient’s age was positively and
ervation at a university teaching hospital ART center from 2005-2019 inversely correlated with the total number of mature oocytes. The absolute mat-
Participants/materials, setting, methods: Breast cancer patients < 40 uration rate in post-menarche and pre-menarche patients differed significantly
years old who had undergone fertility preservation with either IVF or IVM (32.7% vs 11.3%, P > 0.001, respectively), while the degeneration rate of the
before starting gonadotoxic therapy; known BRCA status; both ovaries were oocytes did not (38.9% vs 33.4%, P = 0.167). The pre-menarche group had signifi-
present and no ovarian disease or surgery. Before starting the treatment, antral cantly lower rates of total mature oocytes compared to the post-menarche group
follicle count (AFC) was determined. Total of oocytes retrieved, number of (99 vs 9, P = 0.004). A subanalysis of the oocytes recovered from patients aged
MII oocytes , oocyte fertilization rate (where appropriate) and the number of 1-5 years demonstrated that very few mature oocytes completed the IVM protocol.
oocytes and/or embryos cryopreserved were compared according to Limitations, reasons for caution: This study was limited by its small sample
BRCA status. size and retrospective nature.
Main results and the role of chance: Of 244 breast cancer patients Wider implications of the findings: Oocytes recovered from girls younger
<40 years who underwent fertility preservation between 2005 and 2019, than 5 years of age who undergo fertility preservation have a lower maturation
131 had known BRCA status and 125 fulfilled the above criteria. A total of rate in IVM compared to older females. This may indicate a need for alternative
94 (75%) patients were BRCA negative, of whom 52 (55%) underwent methods for preserving the oocytes of very young patients.
fertility preservation with stimulation (IVF) and 42 (45%) without stimulation Trial registration number: N/A
(IVM), and 31 (25%) patients were BRCA positive, of whom 16 (52%) under-
went IVF and 14 (45%) underwent IVM .The patient mean ages (30+/-4.1 P-406 Pregnancy outcomes and affecting factors in the
vs 32+/-4.2)(P=0.98), AFC (17+/-10 vs 17+/-9) (P=0.83) and, where patients undergoing fertility treatments after high-dose
appropriate, total gonadotrophin dose (2468 IU vs 1961 IU )(P=0.46) and medroxyprogesterone acetate therapy for endometrial cancer
days of stimulation (8.7+/-62 days vs 7.2+/-5.4 days)(P=0.26) were similar A. Kawasaki1, Y. Mori1, H. Itagaki1, H. Ijiri1, T. Sato1
in BRCA positive compared to BRCA negative patients. In terms of repro- 1
University of Tsukuba, Department of Obstetrics and Gycecology, Tsukuba, Japan
ductive response, there were no significant different between BRCA positive
and BRCA negative patients in the total number of eggs retrieved overall Study question: Are fertility treatments beneficial for the patients after con-
(12.1+/-9 vs 11.0+/-8)(P=0.65), the total number eggs retrieved following servative management of endometrial cancer? What are affecting factors for the
IVF (14.1+/-7.2 vs 12.7+/-6.1)(P=0.47) or IVM (11.9+/-10.6 vs 10.4+/- success of fertility treatments?
9.2)(P=0.60), the number of MII oocytes collected (5.5+/-6.5 vs 5.2+/-5.1) Summary answer: Fertility treatments bring good pregnancy outcomes for
(P=0.76), the number of fertilized oocytes (where appropriate) (4.9+/-6.0 the patients after high-dose medroxyprogesterone acetate (MPA) therapy for
vs 3.2+/-4.9)(P=0.18), the number of cryopreserved oocytes (3.1+/- 3.6 endometrial cancer. Several factors may reduce pregnancy outcomes.
vs 2.0+/-2.9)(P=0.16 )and the number of cryopreserved embryos (4.2+/- What is known already: MPA therapy is an option for young patients in an
7.1 vs 4.8+/-6.5)(P=0.37). There was no difference between BRCA1 early stage of endometrial cancer who wish to preserve their fertility. While
and BRCA2 fertility treatments can lead to early pregnancy and bring relatively good preg-
Limitations, reasons for caution: The main limitations are the retrospective nancy outcome for patients after MPA therapy, we must pay attention to disease
cohort study design which could introduce unidentified biases (although the recurrence. Some affecting factors for pregnancy outcomes such as endometrium
BRCA status was not known beforehand in most cases) and the relatively small damage, tumor characters etc. are indicated but enough evidence has not
sample size which would miss small but real differences. been shown.
Wider implications of the findings: Comparable ovarian reserve and repro- Study design, size, duration: In this study, 40 patients of endometrial cancer
ductive performance when undergoing fertility preservation should be reassuring after MPA therapy are recruited from the database of our institute from 1998
for BRCA mutation carriers with breast cancer that outcomes are similar to to 2018. In those patients, 22 patients underwent fertility treatments just after
non-BRCA carriers. BRCA carriers with breast cancer should be offered fertility getting remission of endometrial cancer (FT group). Age-matched control groups
preservation and protocols should not be changed solely on the basis of were made of 213 general infertility patients who underwent reproductive treat-
BRCA status. ments (C group) and 127 patients who underwent IVF-ET in our insti-
Trial registration number: MUHC (ARSU6002/2020-6219) tute(C-IVF group).

Participants/materials, setting, methods: Clinical pregnancy rate and live lymphomas (52%), and testicular cancer (20%) and others (28%). At the time
birth rate were compared between FT group and C group or C-IVF group. We of cryopreservation patients were in childbearing age (median age: 35 years,
also calculated recurrence rate and compared between FT group and a group range 21-53) and at the time of IVF treatments the median age was 38 years
of patients who underwent no fertility treatments after MPA therapy (non-FT (range: 26-54). As regard as outcomes, 33 ICSI cycles were performed with the
group). Next, we investigated the affecting factors on pregnancy outcomes of frozen sperms, fertilization rate was 68%, implantation rate was 25%, cumulative
patients in FT group. The data were analyzed using t-test and chi-square test. birth rate was 30%, with a total of 15 healthy babies born from 2009 to 2019.
The level of significance was set at P<0.05. Limitations, reasons for caution: This was a single-center study reporting
Main results and the role of chance: The pregnancy rate in FT group was data from a limited number of patients. Since our center is the regional referent
59.1% while 56.8% in C group. The pregnancy rate was 44.9% in C-IVF group semen bank, we will continue on banking semen of oncological patients and on
and 35.7% in patients who underwent IVF-ET in FT group. No statistically sig- collecting follow-up data. Thus, clinical value and limitations will be deepened in
nificant difference was observed in the pregnancy rate between FT group and a larger cohort.
control groups. The disease recurrence rate was 31.6% in FT group and 21.4% Wider implications of the findings: Sperm cryostorage is simple and feasible
in non-FT group. Regarding patients who failed to get pregnancy in FT group, for male adults and adolescents and it should be part of fertility management
thin endometrium, low ovarian reserve, changing institute for fertility treatments for patients candidate for receiving gonadotoxic therapies. The usage rate may
and disease recurrence were common factors related to the failure. increase during the years and a regular follow-up may prevent storage of unnec-
Limitations, reasons for caution: To confirm the results of this study, the essary samples.
sampling numbers are too small. A multi-center study is desirable to collecting Trial registration number: not applicable
more data. The large scale analysis is necessary to rule out cofounding factors
and prove true variables affecting pregnancy outcomes.
Wider implications of the findings: Patients undergoing infertility treat- P-408 Clinical evidence of follicle activation and loss in human
ments after MPA therapy should pay attention to the possibility of increasing ovaries post chemotherapy
risk of disease recurrence. The adverse factors are considered in performing D. Shai1, H. Roness1, I. Spector1, H. Raanani1, S. Aviel-Ronen2,
fertility treatments. A close relationship between specialists of oncology and M. Shapira1, M. Baum1, I. Gat1, D. Meirow1
reproduction is important not to miss disease recurrence. 1
Sheba Medical Center, Morris Kahn Fertility Preservation Center, TEL AVIV, Israel ;
Sheba Medical Center, Department of Pathology and Talpiot Medical Leadership
Program, Tel aviv, Israel
P-407 Fifteen year single center experience in sperm banking for
cancer patients: use and reproductive outcomes in survivors Study question: What are the immediate pathophysiological findings related
S. Stigliani1, C. De Leo2, E. Maccarini1, P. Scaruffi1, I. Casciano1, to follicle survival, dynamics and stromal injury in ovaries of patients treated with
P. Anserini1 alkylating and non-alkylating agent chemotherapy.
1
IRCCS Ospedale Policlinico San Martino, UOS Physiopathology of Human Summary answer: Follicle activation and not primordial follicle apoptosis or
Reproduction, Genoa, Italy ; stromal fibrosis may be an underlying mechanism of chemotherapy-induced
2
University of Genoa, DINOGMi, Genoa, Italy follicle loss in patients treated with alkylating agents.
What is known already: Alkylating agents are the most ovotoxic of the
Study question: Which are usage rates of frozen semen and results of in vitro chemotherapy agents, causing significant loss of ovarian follicle reserve, however
fertilization (IVF) cycles in cancer patients that cryopreserved their semen before very little data exists to explain the mechanisms underlying this loss in the ovaries
gonadotoxic treatments? of patients exposed to chemotherapy treatments Some mechanisms have been
Summary answer: The percentage of patients who used their frozen semen suggested, such as apoptosis of primordial follicles, stromal and blood supply
was 6% and the outcomes of the IVF cycles confirmed the validity of the changes and accelerated activation of primordial follicles. Examining the acute
procedure. changes in human ovaries recently exposed in vivo to alkylating and non-alkylating
What is known already: Some cancer diagnoses and treatments can place chemotherapy will increase our understanding of the mechanisms of chemo-
patients at risk for infertility, since gonadotoxic treatments may damage sper- therapy--induced follicle loss and enable development of targeted Fertility
matogenesis. Sperm cryopreservation is an established and safe technique rec- Preservation strategies.
ommended by international guidelines for cancer-related fertility issues, to Study design, size, duration: Histological analysis of ovarian cortical tissue
preserve fertility of cancer patients who are treated during their reproductive removed from 96 women aged 15-39 with cancer diagnoses who were under-
years. Literature data report not uniform rates - ranging from 5% to 29% - of going ovarian tissue cryopreservation for fertility preservation. Forty-eight had
cancer survivors’ use of their cryopreserved semen. Collection of follow-up received chemotherapy treatment within six months of tissue retrieval; 24 with
data about reproductive outcomes of cancer survivors who use their frozen alkylating agents and 24 with non-alkylating agent chemotherapy. Ovarian cortical
semen in IVF cycles is necessary to evaluate the clinical impact of this procedure. tissue from 48 age-matched patients who had not received chemotherapy were
Study design, size, duration: This was a retrospective study which reported used as controls.
data of 683 cancer patients who referred to our center from 2004 to 2019 for Participants/materials, setting, methods: Fresh ovarian tissue samples
fertility counseling and preservation before gonadotoxic therapies. were paraffin embedded, serially sectioned and histologically evaluated for follicle
Participants/materials, setting, methods: A total of 632 oncological numbers by three independent researchers. Immunohistochemical staining was
patients cryopreserved their own semen following international guidelines. conducted for apoptosis (cleaved Caspase-3), FOXO3a, collagen (Sirius red)
Semen was evaluated according to the World Health Organization 2010 manual and neovascularization (CD34).
and it was cryopreserved using standard slow freezing method with nitrogen Main results and the role of chance: Ovaries of women treated with
steam. The ICSI cycles were performed according to conventional procedures alkylating chemotherapy contained significantly more growing follicles compared
and outcomes were collected. with age-matched controls (mean of 4.2 ± 0.6 growing follicles per section
Main results and the role of chance: Among the 683 cancer patients who compared to 2.2 ± 0.3, p=0.004). The number of growing follicles was signifi-
referred to our laboratory, the most frequent pathologies were leukemia and cantly inversely correlated with time from treatment (r = -0.52, p=0.009), such
lymphoma (44%), followed by testicular cancer (35%). Patients’ median age was that the highest number of growing follicles was seen in ovaries removed within
32 years (range 13-65). A total of 632 patients cryopreserved their semen. Over a few days of chemotherapy exposure. Despite a significant drop in primordial
the years, 46% of patients continued to renew maintenance of their cryopre- follicle population in ovaries treated with alkylating chemotherapy, no increased
served samples; 32% of patients were lost at follow-up; 10% were discarded expression of apoptosis marker cleaved Caspase-3 was seen in primordial fol-
according to patient’s will; 6% of banked samples were discarded because licles even within a week post chemotherapy, while increased staining of cleaved
patients died; 2% of patients moved their sperm samples to another fertility Caspase-3 was observed in granulosa cells of growing follicles. Decreased
center for IVF treatments; 4% used their frozen semen for IVF at our center. nuclear staining of FOXO3A was seen in oocytes of PMFs in ovaries treated
Men who used their frozen semen for IVF cycles were survivors of leukemia and with alkylating agents compared with untreated ovaries. Marked fibrosis and

neovascularization was observed in ovaries exposed to alkylating agent chemo- Wider implications of the findings: Fertility outcomes in oncology patients
therapy 4-6 months earlier, but was not seen as acute changes in ovaries exposed have not been adequately studied due to the small number of patients with
more recently. cancer undergoing FP. Few studies investigated the pattern of follicular growth
Limitations, reasons for caution: We were unable to perform molecular to better characterize the outcome of ovarian stimulation not only in oocyte
analysis on human ovarian samples as we had access to paraffin embedded not number and maturity, but also in oocyte quality.
fresh-frozen samples. Therefore, the study was restricted to histological Trial registration number: not applicable
evaluations.
Wider implications of the findings: This study provides evidence that che- P-410 Infertility and access to parenthood after an adolescent and
motherapy induces follicle activation and loss in vivo in ovaries of patients treated young adult (AYA) cancer: a second ‘obstacle course’?
with alkylating agents. Prevention of this increased follicle activation may there- M. Vialle1, J. Saïas-Magnan2, P. Bottin2, A. Dezamis3, C. Metzler-
fore be a potentially successful and highly targeted method of fertility preserva- Guillemain2, A.D. Bouhnik4, J. Mancini4, B. Courbiere2,5
tion, which would not interfere with the effectiveness of chemotherapy. 1
Centre Norbert Elias UMR 8562, Ecole des Hautes Etudes en Sciences Sociales
Trial registration number: Helsinki #8065-10-SMC EHESS- Centre National de la Recherche Scientifique CNRS, Marseille, France ;
2
Hôpital de la Conception - AP-HM, Department of Reproductive Medicine /
P-409 Fertility preservation and oocyte quality in women with CECOS, Marseille, France ;
breast cancer and ovarian tumors: a prospective analytical study. 3
Institut Paoli Calmettes, Department of Clinical Psychology, Marseille, France ;
C. Fabiani1, A. Guarino1, E. Licata1, G. Paciotti1, C. Meneghini1, 4
Sciences Economiques et Sociales de la Santé & Traitement de l’Information
R. Rago1 Médicale SESSTIM, Institut National de la Santé et de la Recherche Médicale
1
ASL ROMA 2 - Sandro Pertini Hospital, REPRODUCTION PHYSIOPATOLOGY and INSERM U125- Aix-Marseille Université- Institut de Recherche pour le
ANDROLOGY, roma, Italy Développement IRD UMR 259, Marse ;
5
Aix Marseille Univ, Avignon Univ- CNRS- IRD- IMBE, Marseille, France
Study question: Does the type of cancer disease influence the oocyte quality
and ovarian response to stimulation for fertility preservation (FP) in female oncol- Study question: What are the experiences of the medical care of fertility and
ogy patients? access to parenthood for French adults in age to procreate who have had an
Summary answer: Fewer mature oocytes are retrieved in ovarian and breast AYA cancer?
cancer patients than infertile patient, the number of dysmorphic oocytes is higher Summary answer: Become parent after an AYA cancer is still a difficult path-
with a recurrent abnormality. way: patients report information and medical support gaps, feel guilty or are
What is known already: Cancer survival has improved significantly and main- sometimes blamed.
taining fertility is an important factor for the quality of life and for the manage- What is known already: The medical literature has so far mainly shown the
ment of cancer patient. Breast cancer is the most frequent cancer in women, important effects of cancer treatments on fertility, the positive results of fertility
and ovarian cancer, specially border-line one, is increasing in young patients. preservation methods, or the uncertain conditions for maintaining or regaining
Currently mature oocyte cryopreservation is a standard technique for fertility fertility after cancer. Research in the humanities and social sciences has more
preservation. The negative impact of cancer therapy on fertility is well known. specifically analysed the difficulties experienced when cancer is diagnosed and
However the data on the pattern of ovarian response after ovarian stimulation during treatment. Few have studied the post-cancer experiences of access to
in oncology patients are limited. Few studies analyzed the effect of the type of parenthood according to fertility conditions. The few existing surveys are quan-
malignancy on oocyte quality, in cancer patients. titative, but none are based on in-depth qualitative interviews.
Study design, size, duration: This is a prospective analytical study performed Study design, size, duration: This study was conducted between December
in the IVF unit at the Sandro Pertini Hospital in Rome between 2016 and 2019. 2018 and November 2019 and was carried out by a multidisciplinary team of
The aim of this study is to investigate the effect of cancer diseases on number, sociologists and doctors. A qualitative survey was performed through in-depth
above all on quality of oocyte and on dysmorphic oocyte ratio, in women with interviews with 22 women and 14 men, who were adolescents or young adults
breast cancer and ovarian tumors compared to women age-matched who under- at the time of cancer diagnosis and in remission for at least one year. Three
gone ICSI treatment for tubal or male factor infertility. types of cancer were studied: breast, testicles and malignant hematological
Participants/materials, setting, methods: A total of 64 patients diseases.
recently diagnosed with breast cancer (Group A) and 14 patients (Group B) Participants/materials, setting, methods: Respondents were recruited
with ovarian tumors were referred for counseling on FP. A total of 168 patients from different locations and by different means: oncology or reproductive
under 38 age underwent ICSI treatment for tubal or male infertility is the control departments, snowball sampling, testimony call. Interviews were conducted by
group (Group C). Baseline characteristics are age, body mass index (BMI) and a sociologist face to face, by visioconference or telephone. The interviews cov-
antimulleran hormone (AMH) value. The primary outcome was number and ered, in a retrospective and chronological way: the personal, conjugal and pro-
quality of retrievd oocyte from ovarian pick-up (OPU). fessional history, the occurrence of cancer and its impact, especially in relation
Main results and the role of chance: Baseline characteristic are comparable. to treatment. The effects on fertility, desire for children and access to parenthood
Despite similar ovarian stimulation protocol, similar mean stimulation duration were discussed throughout the narrative.
(Group A: 10.72; Group B: 10.4; Group C: 11.05), the recombinant FSH cumu- Main results and the role of chance: A total of 36 interviews ranging in
lative dose was significantly higher in breast patients (Group A: 2103.61 U.I.; length from one to three hours were conducted with 22 women and 14 men,
Group C: 1514.77 U.I.) and the mean oestradiol peak at triggering was signifi- aged between 22 and 45 years. Among them, 8 women had breast cancer and
cantly lower due to the administration of letrozole in this patients (Group A: 14 had malignant hematological disease; 6 men had testicular cancer and 8 had
597.16; Group C: 1131.49). Also the number of MII was significantly lower in malignant hematological disease. Among women, 9 underwent a fertility pres-
cancer patients (Group A: 68.7%; Group B: 72.1%; Group C: 78.8%), the number ervation before cancer treatment, 2 after and 9 did not have preservation.
of immature oocytes dose was significantly higher in breast cancer (Group A: Among the men, 13 did sperm cryopreservation and 1 did not. Among the 22
21.7 %; Group C: 13.5%), while the ratio of dysmorphic oocyte was significantly women interviewed, 8 had a child since the end of their cancer treatment,
higher in cancer patients, expecially in ovarian cancer ones (Group A: 9.6% ; conceived naturally or by assisted reproductive technologies (ART) with their
Group B 16.5%; Group C: 7.7%). In Group A and B dysmorphic analyzed cryopreserved or donated oocytes; 6 of 14 men had or were expecting a child,
oocyte have a particular abnormality: a perivitelline space (SPV) with conceived naturally or by ART with their cryopreserved or donated sperm. A
granularity. few knew if they were fertile or infertile at the time of interview, but most were
Limitations, reasons for caution: Limitations concern the paucity of specific unaware of their fertility condition. This diversity of situations makes it possible
cancer group, especially ovarian tumors and BRCA mutated breast cancer to analyse a variety of experiences of medical care after cancer. However, despite
patients. We still have not a follow up data to evaluate the competence of vit- this diversity, difficulties linked information or medical support gasps are system-
rified MII oocytes for oncology patients and we cannot report information on atically reported, appearing at different stages of the pathway towards parent-
spontaneous conceptions and births. hood according to the respondents.

Limitations, reasons for caution: This qualitative study is based on a limited Wider implications of the findings: boECM2 combined with alginate may
number of interviews and a sample of people with limited characteristics. A prove to be a promising new scaffold material for creation of an artificial ovary
study with a larger sample of people with broader characteristics would allow for women in remission from cancer, but at risk of malignant spread from reim-
to reinforce or discuss the results. A selection bias is present, related to the plantation of their tissue.
volunteering process to participate. Trial registration number: not applicable
Wider implications of the findings: The results showed the importance of
a broader diffusion to the medical community and patients of the post-cancer P-412 In depth follow up of childhood development in two
existing ‘self-reconstruction’ difficulties, related to the treatment consequences generations of spermatogonial stem cell transplantation derived
on fertility, sexuality and couple life and the lack of information and medical offspring in a mouse model
support towards parenthood. J. Serrano1, R. Van Eekelen1, C.M. De Winter-Korver1,
Trial registration number: Not applicable S.K.M. Van Daalen1, C.L. Mulder1, A.M.M. Van Pelt1
1
Amsterdam UMC- University of Amsterdam, Centre for Reproductive Medicine,
P-411 Hydrogel derived from decellularized bovine ovarian Amsterdam, The Netherlands
extracellular matrix supports human follicle survival in vitro
M.C. Chiti1, L. White2, J. Vanacker1, E. Ouni1, A. Des Rieux3, Study question: What is the effect of transplantation of in vitro propagated
M.M. Dolmans1,4, C. Amorim1 spermatogonial stem cell (SSCT) on neonatal health and development in the
1
Institut de Recherche Expérimentale et Clinique, Pôle de Recherche en derived offspring?
Gynécologie- Universitè Catholique de Louvain, Brussels, Belgium ; Summary answer: Systematic preclinical safety testing of SSCT derived off-
2
University of Nottingham, Wolfon Centre for Stem cells- Tissue Engineering and spring reveals no major differences in congenital abnormalities, birthweight,
Modeling, Nottingham, United Kingdom ; physical development and reflex ontogeny of the derived offspring.
3
Louvain Drug Research Institute, Advanced Drug Delivery and Biomaterials What is known already: Pediatric cancer treatments improved greatly in
Unit-Université Catholique de Louvain, Brussel, Belgium ; recent years leading to increased life expectancy, but infertility is a late effect.
4
Cliniques Universitaries St. Luc, Départemnt de Gynecologie, Brussel, Belgium Spermatogonial stem cells (SSCs) enable long-term male fertility through a tight
balance between self-renewal and differentiation in the testis. To preserve fer-
Study question: Is a thermosensitive hydrogel derived from decellularized tility in prepubertal male cancer patients, a testicular biopsy is offered prior to
bovine ovarian extracellular matrix (boECM) able to support human follicle gonadotoxic treatment. SSC in vitro propagation followed by auto-transplanta-
survival and development after 7 days of in vitro culture? tion is proposed to restore spermatogenesis allowing natural conception. As
Summary answer: When combined with alginate, this thermosensitive hydro- several assisted reproductive techniques are linked to increased congenital
gel constitutes what appears to be an efficient three-dimensional (3D) matrix abnormalities, low birthweight or developmental deviations, detailed pre-clinical
for in vitro culture of human ovarian follicles. follow-up of development of SSCT offspring is warranted before clinical
What is known already: To successfully assemble a working artificial ovary, application.
we need to create a 3D matrix able to accommodate isolated follicles and cells. Study design, size, duration: Mouse neonatal SSCs were propagated in vitro
While encouraging results have been achieved with fibrin matrices, the next step and subsequently transplanted into sterile males (n=30). Control and fertile
is designing an ECM-derived scaffold more closely resembling human ovarian transplanted males (n=3 per group) were bred with control females (n=27) for
ECM in its biochemical composition. This new generation of biomimetic scaffolds one (control) and two generations (SSCT) of offspring, powered for congenital
might well prove to be a breakthrough for ovarian follicle survival in an artificial abnormalities (n=144 control, n=124 F1 SSCT, n=117 F2 SSCT). To assess
ovary prototype. childhood development, the pups were checked (double blinded) for congenital
Study design, size, duration: Two hydrogels (boECM1, boECM2) were abnormalities, physical and behavioral development during the first 28 days
analyzed for dsDNA, collagen and glycosaminoglycans (GAGs), and compared of life.
to bovine ovarian tissue. Human ovarian tissue (n=3 patients) was then digested Participants/materials, setting, methods: SSCs were isolated from neo-
to isolate ovarian follicles and encapsulate them inside 4 selected hydrogel com- natal DBA/2J mice and propagated in vitro. Sterile males (W/W-v) received
binations: 1) 100% boECM2; (2) 90% boECM2 + 10% alginate; (3) 75% boECM2 SSCT to recover fertility. Control (DBA/2J) and transplanted (W/W-v) males
+ 25% alginate; (4) 100% alginate (control). Follicle count, viability and growth were placed in breeding with control females (DBA/2J). At birth, the pups were
were evaluated on day 0 (D0) and D7 after in vitro culture. checked for congenital abnormalities, birthweight and length. During the first 28
Participants/materials, setting, methods: Ovarian biopsies were obtained days, birth defects, ear opening, eye opening, complete fur growth, incisor erup-
from fertile patients (n=3) with no ovarian pathology, and live/dead assays were tion were checked, along with behavioral reflex testing of negative geotaxis,
performed to evaluate follicle survival before in vitro culture. Follicle recovery grasp and righting reflexes.
rates (n follicles encapsulated on D0/n follicles recovered on D7) were assessed Main results and the role of chance: Overall, the health and development
after one week. Follicle growth was measured by comparing mean follicle diam- of naturally conceived SSCT derived pups during the first 28 days of life was
eters between D0 and D7 of in vitro culture. On D7, follicle viability was deter- similar to control pups, both in the first and second generations. However,
mined using a live/dead assay kit. statistically significant physical differences were found in the birthweight of
Main results and the role of chance: While dsDNA was significantly second-generation pups, with a 0.068g decrease from 1.18 grams in control to
reduced in both boECM1 and boECM2 (p<0.0001) compared to native tissue, 1.11 grams in SSCT derived offspring (95%CI: [-0.134; -0.003]). Furthermore,
the greatest reduction (p<0.01) was observed in boECM2. There was no dif- the eruption of the upper incisor occurred 1.16 days earlier in SSCT derived
ference in collagen content between groups. GAG content was significantly pups of the second generation (95%CI: [-2.161; -0.167]). No statistically sig-
higher (p<0.01) in native tissue than in boECM1, but not boECM2. Indeed, a nificant differences were found between control and SSCT derived offspring,
significantly higher concentration of GAGs was found in boECM2 (P-value in both generations, for the remaining physical and behavioral assessments:
<0.05). A total of 114 isolated human follicles were encapsulated in boECM2-de- birth length, eye and ear opening, lower incisor eruption, fur growth, righting
rived hydrogels and in vitro cultured for 7 days. On D7, overall follicle recovery reflex, grasping reflex and negative geotaxis. The study was powered for con-
rates were: 0%, 23%, 65% and 85% in groups 1 to 4 respectively, rising propor- genital abnormalities, which was a rare event (n=8), with no statistically signif-
tionately with increased alginate content. There was no difference in follicle icant differences found between groups in either generation (control n=1/142,
viability between group 2 and 3 and the control group (group 4) (87%, 97% and SSCT F1 n=3/121, SSCT F2 n=4/113). However, the estimated odds ratios
87% respectively) but follicle viability was not applicable in group 1 as no follicles were relatively high at 3.57 (95%CI: [0.13; 293.33]) for F1 and 4.19 (95%CI:
were recovered. On D7, statistically significant follicle growth was accordingly [0.60-82.95]) for F2.
detected in all groups except group 1 (P-value <0.05, P-value <0.01, P-value Limitations, reasons for caution: Although powered for congenital abnor-
<0.001 respectively). Moreover, similar growth rates were found compared to malities, the sample size of this study was insufficient to determine the impact
the control group on D7. of SSCT in the incidence in the offspring, given the rarity of congenital
Limitations, reasons for caution: This was a pilot study. abnormalities.

Wider implications of the findings: Given the overall normal development P-414 impact of cancer treatment on ART outcomes: study of
of SSCT derived offspring, and after improvement of in vitro propagation of male cancer survivors
human SSCs and development as an advanced therapeutic medical product S. Shibasaki1,2, Y. Takeshige1,2,3,4, H. Hattori1,2, Y. Nakamura1,2,
(ATMP), ethical approval should be requested for introduction of SSCT in a N. Aono1,2,3,4, Y. Nakajo1, N. Okuyama2,3, T. Tai1,3, M. Toya1,
phase 1 clinical trial, alongside follow up of the children. T. Hashimoto2,3, H. Igarashi1, K. Kyono1,2,3,4
Trial registration number: Not applicable. 1
Kyono ART Clinic Sendai, Department of Gynecology, Sendai, Japan ;
2
Human Ovariantissue Preservation Enterprise, Department of Gynecology, Tokyo,
P-413 Modified RNA Encoding for Anti-Müllerian Hormone Japan ;
Has Fertoprotective Potential in Human Ovary Exposed to 3
Kyono ART Clinic Takanawa, Department of Gynecology, Tokyo, Japan ;
Cyclophosphamide 4
Kyono ART Clinic Morioka, Department of Gynecology, Morioka, Japan
L. Man, M.D.- M.Sc.1, E. Kallinos1, Z. Rosenwaks1, D. James1
1
Weill Cornell Medical College, Center for Reproductive Medicine, New York City, Study question: Does chemotherapy and/or radiation therapy in male cancer
U.S.A. patients affect the fertility potential of sperm, embryonic development, or clinical
outcomes?
Study question: Does the administration of modified RNA encoding AMH Summary answer: There was no significant difference in embryonic devel-
confer a fertoprotective effect on the primordial follicle pool in the context of opment or clinical outcomes between the pre-cancer treatment group and the
cyclophosphamide? post-cancer treatment group.
Summary answer: Administration of modified RNA AMH conferred a fer- What is known already: It is well known that cancer treatment, particularly
toprotective effect from cyclophosphamide, preserving the primordial follicle chemotherapy and radiation therapy, for male cancer patients have negatively
pool in both mouse ovary and human ovarian cortical xenografts. affected spermatogenesis. For this reason, sperm cryopreservation is recom-
What is known already: The risk of ovarian failure post-chemotherapy is mended before cancer treatment to preserve fertility. However, it is not clear
determined largely by the type, amount of chemotherapy, and the age of the whether chemotherapy and radiation therapy affect embryonic development
patient at treatment (Meirow, et al., 2010). Several studies have investigated and clinical outcomes of ART.
the fertoprotective potential of a wide range of compounds in rodents, Study design, size, duration: This study included the patients who used
non-human primates, and patients (Spears, et al., 2019). In mice, recombinant ART from January 1997 to June 2019. We defined 25 patients who underwent
or adenovirus-encoded AMH has been shown to protect the ovary from 45 ICSI cycles with cryopreserved ejaculated sperm before cancer treatment
cyclophosphamide (Cp)-induced follicle loss (Kano, et al., 2017, Sonigo, as group A. Of 109 patients who visited our clinic after cancer treatment, 31
et al., 2019). cancer survivors who underwent 70 ICSI cycles with ejaculated sperm were
Study design, size, duration: Cross-sectional study. defined as group B. A total of 63 frozen embryo transfers were performed
Murine model: intraovarian injection (saline/murine Mod-RNA-AMH) fol- (group A: 27, group B: 36 cycles).
lowed by an intraperitoneal (IP) injection 24 hours later (saline/Cp). A total of Participants/materials, setting, methods: Of the cancer survivors,
4 arms, 6 ovaries in each. sperm could not be collected in 42 patients and ICSI was performed in 31
Xenotransplantation model: human ovarian tissue transplanted into patients. Only semen analysis was performed for the rest. The cancer treat-
immunocompromised mice, intra-graft injection (buffer/human recombi- ment was performed by chemotherapy and/or radiation therapy. Semen
nant-AMH/human Mod-RNA-AMH) followed by an IP injection 24 hours later parameter, embryonic development, clinical outcomes were compared
(saline/Cp). A total of 4 arms, 4 grafts in each. between groups A and B. Chi-squared test, Mann-Whitney U test, and Fisher’s
Ovaries/grafts were harvested 2 weeks after the first chemotherapy injection. exact test were used for statistical analysis. P<0.05 was considered statistically
Participants/materials, setting, methods: In the murine model, we used significant.
6-7-week-old C57/B6 females. Main results and the role of chance: The major cancer types in group
In the xenograft model, we co-transplanted human ovarian cortical tissue from A and B were testicular tumor (40.0%, 25.8%) and hematopoietic malignancy
a 12-year-old organ donor, with endothelial cells, into NOD scid gamma (28.0%, 51.6%). The mean age of females at the ovum pick-up in group B
(NSG) mice. (37.1±4.5) was significantly higher than in group A (35.3±4.0) (P<0.05).
Cp was administered at the dose of 60mg/Kg, and the protocol was repeated There were no significant differences in the anti-Mullerian hormone levels
a week later. or antral follicle counts, or baseline FSH levels. The mean age of males at
After harvest of the ovaries/xenografts, the ratio of follicles of all growth the sperm collection in group B (39.9±7.9) was significantly higher than in
stages was measured in histologic sections. group A (35.9±6.4) (P<0.05). In group B, the mean period from the end of
Main results and the role of chance: In the murine model, we found the cancer treatment to the start of ART was 10.8±9.7 years. The median of
retention of primordial follicles similar to the controls: saline-intraovarian/ sperm concentration in group A [37.7×106 (20.5-78.9)] was significantly
saline-IP, 49±12.10, Mod-RNA-AMH/saline, 53.33±7.81, compared to Mod- higher than in group B [5.5×106 (0.9-26.4)] (P<0.05). Two cases in group A
RNA-AMH/Cp, 47.17±13.61. A 50% decrease was noted with saline/Cp Tx: showed azoospermia after cancer treatment. There were no significant dif-
26.5±9.01. In the human xenografts, the retention of early primordial follicles ferences in fertilization rates [63.5% (167/263), 65.7% (229/326)], and
was markedly improved with Mod-RNA-AMH pre-treatment. Buffer intra- blastocyst formation rates [48.7% (73/150), 49.3% (75/152)] between
graft/Cp IP 9.41±10.91% primordial follicles versus 37.48±17.12% with groups A and B. Similarly, there were no significant differences in the preg-
Recombinant-AMH, and 56.24±39.44% with Mod-RNA-AMH. Notably, the nancy rates of frozen-thawed embryo transfer [59.3% (16/27), 50.0%
percentage of primordial follicles was similar to grafts treated with buffer/ (18/36)] and live-birth rates [44.4% (12/27), 41.7% (15/36)] between
saline: 51.67± 4.04%. groups A and B.
Limitations, reasons for caution: Human ovarian tissue available for Limitations, reasons for caution: In this study, the ART outcome was com-
research is restricted. Repeating the experiment with more replicates will pared between the patients with sperm frozen before cancer treatment, and the
strengthen the results. Moreover, demonstrating the benefit to human xenografts only patients with sperm obtained after the treatment, which might cause a
at later time points will provide further support for the positive effect con- certain bias in grouping. We could not have enough information about cancer
ferred by AMH. treatment, which should be shared by oncologist.
Wider implications of the findings: Due to its high efficiency, transient Wider implications of the findings: Considering the effects of gonad toxicity
protein expression, and the fact that it does not elicit a substantial innate immune from chemotherapy treatment, sperm cryopreservation before cancer treatment
response, Mod-RNA is an optimal mode of delivery. Administration before is basically recommended. However, if sperm can be collected after a withdrawal
chemotherapy may confer a pronounced fertoprotective effect with minimal period, ART treatment outcomes might be comparable. Since the number of
expense and intervention. target patients was still small, further study would be necessary.
Trial registration number: na Trial registration number: not applicable

P-415 No sign of actin polymerization or Hippo pathway inhibition P-416 Ovarian stimulation for fertility preservation in young
in fragmented human ovarian tissue women: is there a right time to start?
S. Aagaard Lunding1, S.G. Kristensen2, L. Hardardottir2, R. Quintana Berto1, A. Santolaria Baig1, N. Garcia Camuñas1,
H.Ø. Olesen2, A. Nyboe Andersen1, C.Y. Andersen2, S.E. Pors2 J. Subirá Nadal1,2, P. Polo Sanchez1, J.M. Rubio Rubio1
1 1
Rigshospitalet, The Fertility Clinic, Copenhagen, Denmark ; La Fe University Hospital, Women’s Health Area, Valencia, Spain ;
2 2
Rigshospitalet, Laboratory of Reproductive Biology, Copenhagen, Denmark IVI-RMA Castellon-Valencia, Reproductive Endocrinology, Castellon, Spain
Study question: Does tissue fragmentation of human ovarian cortex result in Study question: Is there an optimal time to start an ovarian stimulation in
polymerization of the actin cytoskeleton and subsequent activation of follicle young women who initiate an oocyte cryopreservation (OC) cycle for fertility
growth through Hippo pathway inhibition? preservation (FP)?
Summary answer: Tissue fragmentation of human ovarian cortex did not Summary answer: Starting OC cycles out of the early follicular phase
result in actin polymerization nor did it result in follicle activation by inhibition (EFP) – or random-start protocol – may provide better ovarian response and
of the Hippo pathway. oocyte yield.
What is known already: The Hippo pathway has been associated with What is known already: OC is the preferred method for FP compared to
regulation of early ovarian follicle growth in mammals including humans. Studies other treatments. Traditionally, stimulation protocols start during the EFP but
of murine ovaries suggest that changes in the actin cytoskeleton, caused by this methodology sometimes requires more time depending on patient’s ovarian
fragmentation, result in inhibition of the Hippo pathway by dephosphorylation cycle. However, in certain settings – cancer, for instance – we do not have enough
of YAP and in turn may activate follicle growth by transcription of growth time for delaying other therapies in order to preserve women’s fertility. Some
factors (eg. CCN’s) and apoptosis inhibitors. In humans, in vitro and in vivo authors have argued that is possible to achieve a synchronized follicular devel-
studies of fragmented ovarian tissue demonstrated upregulation of growth opment and an optimal oocyte retrieval despite starting the stimulation out the
factors and growth of preantral follicles. However, the connections between EFP based on the different follicles waves. Random-start ovarian stimulation
fragmentation, the actin cytoskeleton and follicle activation in humans are yet protocols are a feasible alternative to conventional cycles in such scenery.
to be confirmed. Study design, size, duration: This is a retrospective study performed from
Study design, size, duration: Donated frozen ovarian cortex from six January 2013 to January 2020 at a tertiary university hospital. Our research was
women (34-37 years), undergoing ovarian tissue cryopreservation for fertility focused in the first OC cycle of young women (< 25 years old) which are
preservation between 2001 and 2011 prior to gonadotoxic therapy, were expected to have a better cycle response due to their clinical characteristics.
thawed for in vitro and xenotransplantation study. From each woman one Data was retrieved from patients’ electronic medical record.
cortex piece of 5x4x1mm was fragmented into 20 cubes of 1x1x1mm and Participants/materials, setting, methods: We collected data from epide-
another intact piece served as control tissue. Both fragmented and control miology, stimulation and oocyte retrieval from a total number of 78 OC cycles.
tissue were incubated for either 0, 10, 30, 60, 120 or 240 minutes prior to Two groups were stablished depending on the phase in which the treatment
examination. began: EFP n=63 and random-start n=15. Continuous variables were summa-
Participants/materials, setting, methods: Actin polymerization was rized by the median and standard deviation for a descriptive analysis. After
assessed with western blot of the ratio of F-actin to G-actin in fragmented and evaluating the normal distribution of both groups, we performed a nonpara-
control ovarian tissue. Inhibition of the Hippo pathway was assessed with west- metric statistical analysis to compare them.
ern blot of the ratio of phosphorylated YAP (pYAP/YAP) and gene expression Main results and the role of chance: Both groups were comparable in
analysis of the downstream growth factors CCN2, CCN3 and CCN5 in frag- terms of Body Mass Index (BMI), Anti-Müllerian Hormone (AMH) or Antral
mented and control ovarian tissue. Fragmented and control tissue were xeno- Follicular Count (AFC).
grafted to immunodeficient mice for six weeks before histological assessment Total dosing of FSH was similar in both groups with a mean of 2401,441 ±
of follicle growth. 1044,6 IU in the EFP group and 2295 +/- 636,90 IU in the random-start
Main results and the role of chance: Both F-actin and G-actin were detect- grouP- (p=0.941).
able in all samples, but no significant difference was found in the ratio in frag- The total number of follicles at triggering day was comparable in both groups.
mented tissue compared with control tissue for each timepoint (p=0.8). Both However, there was a higher number of follicles >16 mm in the random-start
YAP and phosphorylated YAP (pYAP) were expressed in all samples. At the group (12 ± 5) compared with the EFP group (7,47 ± 5) (p=0,011). Estradiol
three selected timepoints (0, 60 and 120 minutes) there were no difference in values at the trigger day were similar, as well as Follicular to Oocyte Index (FOI)
the ratio of YAP/pYAP in fragmented and control tissue (p=0.6). Moreover, and Follicular Output Rate index (FORT).
CCNs were expressed in all tissue samples. Only CCN5 was on average 0.56- Nevertheless, the oocyte retrieval was more successful in the random-start
fold lower in control tissue compared with fragmented tissue (p=0.0026). group with a mean number of 11,72 (±7,25) oocytes in the EFP patients and
However, when assessing CCN5 in the fragmented tissue at the different time- 17,43 (±9,84) in the random-start patients (p=0.018). Even though the maturity
points, expression of CCN5 did neither increase nor decrease over time (p=0.8). rate was comparable in both groups, the number of metaphase II oocytes was
Thus, fragmentation did not induce upregulation of CCN5 gene expression. also higher in the random-start group (p= 0,017).
Histological assessment of the xenografted ovarian tissue revealed fewer follicles Limitations, reasons for caution: The data from this study was retrospective
in the fragmented tissue compared with control tissue, 239 versus 975 preantral and from a single center. Further studies including more patients would ensure
follicles, respectively. A logistic regression model accounting for paired samples a stronger statistically significant difference.
showed that the proportions of growing follicles, defined as all non-primordial Wider implications of the findings: Random-start protocol for OC cycles
follicles, in the exposed tissue (34.7%) and the control tissue (27.4%) were not provides an advantage in terms of timing. In settings in which an urgent ovarian
different (p = 0.56). stimulation is needed, starting it without delays not only allows to attempt fertility
Limitations, reasons for caution: Donated ovarian tissue were only preservation for patients who have important time constraints, but also, does
available from six women, which limits the statistical power. Initially, ovarian not compromise oocyte retrieval.
cortex is cut into pieces prior to cryopreservation and some activation could Trial registration number: Not applicable
have been induced in the control tissue. Furthermore, the unequal follicle
distribution throughout the cortex warrants caution when interpreting the
P-417 Effect of BRCA genetic status on ovarian stimulation
findings.
response for fertility preservation in breast cancer patients.
Wider implications of the findings: We were not able to confirm that
A retrospective cohort study
fragmentation of human cortical tissue results in actin polymerization and sub-
sequent follicle activation through disturbance of the Hippo pathway. Thus, L. Perez Martin1, C. Alonso Mayo1, S. Caballero Sanz1, F.J. Amor
these in vitro and in vivo results may indicate that ovarian tissue fragmentation Valera1, D. Trobo Marina1, F.M. Pérez Milán1
1
is an ineffective method to activate follicle growth in humans. Hospital General Universitario Gregorio Marañón, Reproduction Unit, Madrid,
Trial registration number: Not applicable Spain

Study question: Do BRCA1 and BRCA2 mutation carriers diagnosed with 2

Department of Obstetrics and Gynecology, Seoul National University College of
breast cancer have an increased risk of poor ovarian response to ovarian stim- Medicine,
ulation for fertility preservation?
Summary answer: No association was observed between the presence of a Study question: Is there any difference in ovarian reserve after chemotherapy
BRCA gene mutation and ovarian response to ovarian stimulation in terms of between triple negative breast cancer (TNBC) patients and other breast cancer
number of oocytes retrieved. patients?
What is known already: Breast cancer is the second most common malig- Summary answer: Decreased ovarian reserve after chemotherapy is more
nancy diagnosed worldwide. 5-year survival rate extents to more than 80% due obvious in TNBC group than other group. Early referral before chemotherapy
to early detection with screening programmes and the recent advances in its is crucial especially in TNBC patients.
treatment. Successful treatment often requires the use of gonadotoxic chemo- What is known already: Triple Negative Breast Cancer (TNBC) is a cancer
therapeutic agents, so fertility preservation is now advised to most premeno- that is estrogen receptor (ER) negative, progesterone receptor (PR) negative
pausal patients. Predisposing mutations are present in around 10-15% of cases, and human epidermal growth factor receptor 2 (HER2) negative, based on
being BRCA genes the most commonly involved. There have been reports immunohistochemistry (IHC). Some studies suggest that women with BRCA1
suggesting a poorer response to ovarian stimulation of BRCA mutation carriers, or BRCA2 mutation have fewer oocytes in their ovaries and large portion of
and recent studies showed lower serum AMH levels compared to non-carriers. TNBC patients were proven to have BRCA mutation. But there is no report
Study design, size, duration: This is a retrospective cohort study. All breast about association between TNBC and ovarian reserve.
cancer patients who underwent fertility preservation in our unit from January Study design, size, duration: In this retrospective study, data from 138 breast
2010 to December 2019 were eligible to be included. Our aim was to compare cancer patients who referred for fertility preservation at a single tertiary center
the number of oocytes retrieved between BRCA mutation carriers and were examined from January, 2017 to December, 2018. All options were
tested-negative controls. Secondary outcomes were number of metaphase II explained by specialist of endocrinology, and patients selected the method of
oocytes retrieved, rate of low or suboptimal ovarian response (<6 and < 9 fertility preservation. Most patients (n=122) chose GnRH agonist injections only,
oocytes, respectively) and cancelation rate due to insufficient response. and other patients (n=16) wanted to perform oocyte or embryo
Participants/materials, setting, methods: All breast cancer patients tested cryopreservation.
for BRCA mutations who underwent a fertility preservation treatment were Participants/materials, setting, methods: Subjects were classified into
included in the study. We compared the number of oocytes retrieved in exposed two groups, TNBC group (n=33) and other group (n=105) in which at least
and non-exposed patients using appropriate statistical tests. Effect size was esti- one of ER, PR, HER2 is positive. Clinical characteristics and markers of ovarian
mated by relative risk, which was adjusted considering potential confounders by reserve (AMH, FSH, E2) of the two groups were compared before, 6 months,
multivariate analysis (logistic regression). The study was conducted in Gregorio and 1 year after chemotherapy.
Marañón General University Hospital (Madrid, Spain), and was approved by the Main results and the role of chance: Baseline characteristics including age
internal institutional committee. were not significantly different between two groups. In TNBC group, initial AMH
Main results and the role of chance: We included 113 patients. BRCA levels were significantly lower than other group (3.25 ± 0.61 vs. 3.85 ± 1.04,
mutations were present in 29 (25.7%). No differences were observed between p<0.001). After 1 year of chemotherapy, FSH levels were higher in the TNBC
groups regarding age, smoking habit, previous infertility and/or pregnancy, antral group (22.98 ± 13.08 vs. 11.15 ± 5.38, p=0.051).
follicle count (AFC), initial and total dose of FSH consumed during ovarian Limitations, reasons for caution: Limitations of this study is that difference
stimulation and cotreatment with LH and/or letrozole. in drug regimen and duration of chemotherapy between two groups was not
No association was detected between BRCA status and number of oocytes considered thoroughly. Further prospective studies with larger, homogenous
retrieved (mean difference: 0.65; 95% CI: -2.29—3.59; p>0.05; log rank test cohorts are needed.
p value >0.05). Multivariate analysis did not show association after adjusting Wider implications of the findings: In this study, TNBC patients showed
by age, AFC, smoking habit and total FSH dose (regression coefficient: -0.99; trend to have lower ovarian function before chemotherapy and slower recovery
CI 95%: -3.62—1.64). of ovarian function after chemotherapy. Therefore, urgent referral for fertility
BRCA status showed no effect on number of metaphase II oocytes retrieved preservation should be considered especially in TNBC patients.
(mean difference: 1.71; 95% CI -0.70—4.12; log rank test p value >0.05). Trial registration number: not applicable
Cancelation rate due to insufficient response was similar in both groups
(6.1% vs 9.3%; p >0.05). Risk of low and suboptimal response was also similar P-419 A streamlined andrology workflow for elective sperm
(RR: 0.70 CI 95%: 0.28—1.74; and RR: 0.96 CI 95%: 0.57—1.60; respectively). freezing before chemotherapy is pertinent to timely preservation
The impact of BRCA genetic status on low and suboptimal response was not of male fertility
affected by cofactors such as age, smoking habit, AFC and total FSH dose Z. Huang1, J. Mathew2, C. Ho2, S. Chew2
(adjusted OR: 0.99 CI: 0.87—1.83; and OR: 0.39 CI 95%: 0.08—1.83, 1
Institute of Molecular and Cell Biology- National University Health Systems,
respectively).
Obstetrics and Gynaecology, Singapore, Singapore Rep. of ;
Limitations, reasons for caution: BRCA status was unavailable in many 2
National University Health Systems, Obstetrics and Gynaecology, Singapore,
cases. The low number of patients included rendered a low power to conclude
Singapore Rep. of
about the main objective, despite observed frequency of BRCA mutations being
higher than expected. Pre-treatment AMH levels were unknown in most patients, Study question: Does a streamlined andrology workflow help improve time-
so ovarian reserve could only be assessed by AFC. to-treatment for men who require elective sperm freezing before
Wider implications of the findings: As more BRCA mutation carriers are chemotherapy?
aware of their genetic status, more couples are seeking preimplantation genetic Summary answer: A standardized workflow with instructions and checklist
testing, which ideally requires a large number of oocytes and embryos. According for all clinicians to refer for elective sperm freezing before chemotherapy is
to our findings, BRCA mutation carriers do not require oocyte cryopreservation efficient and reduce time to treatment.
to maximize chances before they decide to have children. What is known already: Men who require elective sperm freezing to preserve
Trial registration number: Not applicable their fertility prior to chemotherapy to treat their medical conditions (e.g. cancer
or debilitating medical conditions) are often referred to multiple clinics and spe-
cialists (referring physicians, fertility specialist and andrologist), undergo a mul-
titude of investigations and go through many consent forms before receiving the
P-418 Ovarian reserve before and after chemotherapy in triple
appropriate treatment. This causes additional financial costs and further delays
negative breast cancer patients
as well as precious time lost before these men can receive life-saving medical
J. Han1,2, K. Hye In1, K. Sung Woo1,2, K. Hoon1,2, K. Seung-Yup1,2, treatments. Many men become confused by conflicting instructions, frustrated
S. Chang Suk1,2, K. Seok Hyun1,2, M.C. Young1,2 by numerous appointments and lack accurate information about elective sperm
1
Department of Obstetrics and Gynecology, Seoul National University Hospital, ; freezing.

Study design, size, duration: Retrospective analyses on all the elective sperm including predictive factors for success is lacking, and limited data exists as to
freezing cases performed in 2018 was done to assess, 1. the duration required which ovarian stimulation and embryo transfer regimens are the most optimal
from time of referral to time of sperm freezing, 2. costs bore by these men and for this group of patients.
3. types of cases referred. An institution-approved checklist and counselling form Importantly, negative outcomes are generally underreported.
designed by the andrology service for elective sperm freezing is implemented Study design, size, duration: Retrospective study in a cohort of 28 patients,
from January 2019 and a prospective analysis of the referral process and out- undergoing OTT at Aarhus University hospital in Denmark followed by IVF
comes was performed. treatment during the period 2012 to 2017.
Participants/materials, setting, methods: Participants: Men referred from Participants/materials, setting, methods: A cohort of 28 women under-
all disciplines for elective sperm freezing going IVF-treatment in Danish fertility clinics after OTC and OTT. The study
Setting: The andrology service attends to men with a wide range of psychosexual included evaluation and analysis of patient characteristics, including diagnosis
and fertility issues; limited appointment slots available. prior to OTC, details of the OTT surgery, ovarian stimulation regimens and
Method: An approved standardized checklist deployed in 2019 with indications, reproductive outcomes, including number of follicles, oocytes, embryo transfers,
requirements and counselling form which a clinician (from any discipline) can pregnancy, and live birth. The possible impact of time to fertility treatment was
access on the hospital internal web-portal to reference, download and employ also evaluated.
in their clinics for direct onward referral to do elective sperm freezing for Main results and the role of chance: In 19 patients responding to ovarian
these men. stimulation, a median of 3.0 cycles per patient (range: 1-14 cycles) was per-
Main results and the role of chance: Elective sperm freezing requests formed and 2.0 mature oocytes were retrieved per cycle. Eleven women (39%)
comprises 20% the andrology service’s workload. With the limited appointment achieved 15 pregnancies of which 60% were lost during first or second trimester,
slots and usually lengthy consults for psychosexual issues and complex andro- resulting in 5 patients (17,9%) having one or more live births, and 7 healthy
logical issues, it can result in long waiting times, increased number of clinic visits, children being born. In breast cancer patients (mean age at OTC=33 years),
increased costs and delays to life-saving treatment for these men just to access the pregnancy (PR) and live birth rates (LBR) were 35.0% and 5.0% per embryo
the service. Typically it takes up to 3 clinic visits before the men can schedule transfer, respectively. For all other diagnoses (mean age at OTC=26.6 years),
their elective sperm freezing prior to January 2019. Additionally, relevant inves- PR and LBR were 50.0% and 37.5% per embryo transfer, respectively. None
tigations before sperm freezing such as sexually transmitted infections screen of 12 women aged ≥35 years at OTT achieved a live birth. A total of 1.42
must be done as mandated by the Ministry of Health, Singapore - however prior fertilized oocytes per cycle, was obtained using the long GnRH agonist protocol
to 2019, this was haphazard and not all men have their screen performed rou- compared to 0.71 fertilized oocyte per cycle in GnRH antagonist cycles
tinely - resulting in further delay to elective sperm freezing and initiating their (p=0.004, Mann-Whitney). Frozen-thaw embryo transfer resulted in more
medical treatment (range 4 -14 days). The referring physicians from multiple pregnancies than fresh embryo transfer (P<0.05). There were no significant
disciplines (medical-oncology, haematology, endocrinology, urology) had to delay differences in age at OTC, diagnosis or transplantation site between patients
commencing vital medical treatments for these men until their fertility preser- who had oocytes retrieved and non-responders to stimulation, but in non-re-
vation plans were completed. With the implementation of the streamlined sponders, the mean time from OTT was 15.7 months compared to 9.9 months
workflow and checklist in January 2019, the referring physician can follow the in responders (p<0.05).
checklist, counsel patients and refer directly for elective sperm freezing with no Limitations, reasons for caution: Although this study includes the largest
delay, waiting time or need to see the andrologist anymore. single center Scandinavian cohort of OTC-OTT patients undergoing IVF treat-
Limitations, reasons for caution: The analysis is largely descriptive and ment, the sample is still low, limiting the statistical power. Moreover, since live
applicable only in a tertiary referral hospital with a broad range of disciplines birth was rare, parameters related to this outcome may be masked.
and high number of complex cases. Wider implications of the findings: Based on the present results, the long
Wider implications of the findings: This resolves the unnecessary delay GnRH agonist down-regulation protocol, followed by freeze-all and frozen-thaw
and extra costs for men to have elective sperm freezing done. The standardized embryo transfer should be recommended and multiple OTT procedures and
checklist and counselling form will create a model of care for other clinical repeated IVF-treatment may be necessary to achieve a live birth. Patients may
services to adopt. This is to avoid delay in commencing life-saving medical-treat- benefit from being referred to fertility treatment soon after OTT.
ment due to the need for fertility preservation. Trial registration number: 3-3013-2790/1
P-421 Spermatogenesis enhancement in neonatal mouse frozen -
thawed testis after 3- dimensional culture with growth factors.
P-420 Reproductive outcomes after IVF treatment in a cohort of A. Alrahel1, M. Movahedin2, Z. Mazaheri3, F. Amid4
Danish women transplanted with cryopreserved ovarian tissue 1
Reproductive Biotechnology Research Center- Avicenna Research Institute- ACECR-
I.M. Dueholm Hjorth1, S.G. Kristensen2, M. Dueholm3, Tehran- Iran., Reproductive Biotechnology Research Cente, tehran, Iran ;
P. Humaidan4 2
Anatomical Sciences Department- Faculty of Medical Sciences- Tarbiat Modares
1
The Regional Hospital in Horsens and Aarhus University Hospital, Department of University- Tehran 14115331- Iran, Anatomical Sciences Department, Tehran,
Obstetrics and Gynecology, Aarhus C, Denmark ; Iran ;
2
The Juliane Marie Centre for Women- Children and Reproduction- University 3
Basic Medical Science Research Center- Histogenotech Company, Basic Medical
Hospital of Copenhagen- Blegdamsvej 9- 2100 Copenhagen- Denmark., Science Research Center- Histogenotech Company, Tehran, Iran ;
Laboratory of Reproductive Biology, Copenhagen, Denmark ; 4
Anatomical Sciences Department- Faculty of Medicine- Tehran University of
3
Aarhus University Hospital, Department of Obstetrics and Gynecology, Aarhus, Medical Sciences-, Anatomical Sciences Department, Tehran, Iran
Denmark ;
4
Skive Regional Hospital and Faculty of Health- Aarhus University, The Fertility Study question: Are growth factors (GF) provide profound impact on sper-
Clinic and Department of Clinical Medicine, Skive and Aarhus, Denmark matogenesis after 8 weeks of in-vitro 3- dimensional culture?
Summary answer: The level of TnP1 gene expression and ACRBP were
Study question: How were the reproductive outcomes, ovarian stimulation significantly higher in the fresh culture group with growth factors compared to
regimens and predictive factors for successful fertility treatment in women under- control groups.
going ovarian tissue cryopreservation (OTC) and transplantation (OTT)? What is known already: Aggressive chemotherapy may lead to permanent
Summary answer: Although low, reproductive outcomes after OTT are more male infertility. Three-dimensional(3D) in-vitro culture can operate as an effective
favorable in younger patients swiftly referred to IVF treatment, followed by strategy for studies on spermatogenesis and male infertility treatment. Sperm-like
frozen-thaw embryo transfer. cells derived from the human and mice spermatogonial stem cells after fro-
What is known already: The reproductive outcomes after OTT are diverse zen-thawed has already been carried out by researchers. In this study, 3D culture
as some women conceive spontaneously, whereas others fail to conceive even of testicular tissue by GF were studied in 4 groups of immature mice testes after
after repeated IVF treatment. Knowledge on the causes of this variability, 8 weeks to evaluate the progress of the spermatogenesis process.

Study design, size, duration: The testes were removed from 5 NMRI neo- under 50 was undertaken to evaluate fertility preservation discussion and overall
natal male mice (six days old) and repeated 3 times for further accuracy in each referral rates were measured.
group. The testis tissue fragments were transferred to the hexahedrons, incu- Main results and the role of chance: The median age of patient respondents
bated in a culture incubator and cultured for 8 weeks was 30 years and 69% were in a relationship. The majority (89%) of patients
Participants/materials, setting, methods: The fragments of testes tissue perceived oncology HCPs as the most trusted source to learn about fertility
are placed in the freezing culture. The tissue was examined from the aspect of preservation and 11% preferred to learn from another channel of information
morphology.PLZF, SCP3, ACRBP antibodies were assessed to identify sper- (videos, brochures, websites). Following the survey and in order to fulfill the
matogonia, spermatocyte and sperm-like cell respectively. Four studied groups need expressed by respondents, a series of seminars were organized to bring
are as follow: Culture of fresh testicular tissue fragments on agarose with and awareness and knowledge to healthcare providers about fertility preservation
without growth factors for 8 weeks ,the culture of frozen-thawed neonatal and how to access the service. The mean pre-session knowledge score was 2.08,
mouse testicular tissue fragments on the agarose with and without growth factors 2.46 and 2.06 for nurses, physicians and allied HCPs respectively, whereas the
for 8 weeks. mean post-session knowledge score was 3.73 (p<0.05) for all. Documented
Main results and the role of chance: The findings showed that the size and evidence of discussion was 15% and referral rate for fertility preservation was
diameter of seminiferous tubules were increased compared to control groups. 7.7% (104/1345) pre-intervention. The discussion rate was 35% and the referral
Different types of germ cells including spermatogonia and spermatocyte cells rate 21% (379/1807) post-intervention (p<0.001). The reasons for non-discus-
were observed in Fresh and frozen-thawed testicular tissue. sion and non-referral were assessed.
The expression level of ACRBP was significantly increased in the fresh culture Limitations, reasons for caution: The study samples size are small, and the
group with growth factors and in the frozen-thawed culture group with growth fact that the seminars were performed in university-based hospitals limit the
factors (P≤0.05). generalization of these findings. Furthermore, the evaluation of discussion and
Limitations, reasons for caution: Further studies are needed, where the referral rates overlap the period of the educational intervention.
culture medium enriched with several supplements or growth factors, for better Wider implications of the findings: Oncology HCPs are perceived as
results. the most trusted source of health information and advice among young male
Wider implications of the findings: The slow programmed freeze of the adults with cancer. Therefore, increasing knowledge of healthcare providers
3D agarose testicular tissue culture appears to be a useful method to maintain in fertility preservation could enable proactive and open discussions with
long-term preservation, especially for children with cancer before initiating treat- patients, thus enhancing the quality of counselling and improving access
ment with chemotherapy or radiotherapy.These results will encourage research- to care.
ers to set up a culture system for in vitro spermatogenesis. Trial registration number: Not applicable
P-423 Oncofertility cares provided to patients: results from a
P-422 Translating male cancer patients’ needs into a professional systematic review of literature revealed a low adhesion to fertility
development intervention that improved fertility preservation preservation services due to accessibility barriers
knowledge and referral among oncology healthcare providers J. Daolio1, L. De Panfilis2, M. Perin2, M.C. Bassi3, L. Mangone4,
W.M. Buckett, M.B.B.Ch.-M.D.1, B. Herrero2, G. Raskovic3, M.T. Villani1, A. Nicoli1
M.C. Brabant4, P. Chan5 1
Azienda Unità Sanitaria Locale - IRCCS di Reggio Emilia, Departement of
1
MUHC Reproductive Centre, Obstetrics and Gynecology- McGill University, Obstetrics & Gynecology, Reggio Emilia, Italy ;
2
Montreal, Canada ; Azienda Unità Sanitaria Locale - IRCCS di Reggio Emilia, Unit of Bioethics, Reggio
2
MUHC, Obstetrics and Gynaecology, Montreal, Canada ; Emilia, Italy ;
3 3
MUHC, Oncology, Montreal, Canada ; Azienda Unità Sanitaria Locale - IRCCS di Reggio Emilia, Medical Library, Reggio
4
MUHC, Obstetrics and Gynecology, Montreal, Canada ; Emilia, Italy ;
5 4
MUHC, Urology, Montreal, Canada Azienda Unità Sanitaria Locale - IRCCS di Reggio Emilia, Unit of Epidemiology,
Reggio Emilia, Italy
Study question: How do male cancer patients want to learn about fertility
preservation and how can this need be addressed improving care and refer- Study question: What kind of barriers could hinder the use of fertility pres-
ral rates? ervation services?
Summary answer: Cancer patients perceived their oncology healthcare pro- Summary answer: Barriers in knowledge, attitude and organization of onco-
viders as the most trusted source from fertility preservation. Targeted seminars fertility cares could be present at patient, health providers and organizational
for oncology healthcare professionals improved knowledge and referral. levels limiting the adherence to service.
What is known already: Clinical practice guidelines for fertility preservation What is known already: International guidelines on oncofertility cares rec-
recommend clinicians to discuss the potential impact of cancer treatments on ommend to counsel patients of childbearing age about the risk of infertility
future fertility with reproductive-aged cancer patients, and be prepared to refer following cancer treatment and options available to preserve fertility. Patients
them to a reproductive specialist, if appropriate. However, multiple publications without fertility preservation choices before treatments have negative psycho-
consistently show that pre-treatment fertility counselling is disseminated to only logical consequences during the course of treatments and, in case of survival,
a minority of newly diagnosed cancer patients. Effective and co-ordinated strat- on quality of life. Poor adherence to guidelines is acknowledged due to the
egies to overcome this and improve quality of life after cancer are urgently presence of barriers in daily clinical practice regarding healthcare providers and
needed. One barrier is the lack of understanding of cancer-related infertility and information provided to patients hindering the access to fertility preservation
available resources amongst both patients and their oncology practitioners. services (FPS). In our realty, only 16,7% of patients were counseled for FPS
Study design, size, duration: Co-ordinated and targeted regional strategy between 2015-2017.
comprising: (1) Cancer patient surveys (March-July 2016) to determine fertility Study design, size, duration: This study was conducted from August 2019
preservation needs; (2) Assessment of efficacy of educational seminars to oncol- to November 2019 as part of a major healthcare pathway about oncofertility
ogy health care professionals (January 2017-December 2018); and (3) Evaluation services. In the first phase, we conducted a comprehensive systematic review
of discussion of the effect of cancer care on subsequent fertility in men aged of literature with the keywords “fertility preservation”, “cancer”, “education”,
under 50 years and, where appropriate, referral rates (January 2018-July 2019) “engagement”; in the second phase, results will be used by an expert team of
Participants/materials, setting, methods: Male cancer patients (n=45) Embryologists and Bioethicists to define educational interventions to overcome
completed surveys about cancer and fertility and to choose most trusted sources barriers in our network.
for information - oncology healthcare professional (HCP); other HCP; peers; Participants/materials, setting, methods: The electronic search was con-
educational material; websites. Seminars were given at 7 different regional sites. ducted in Medline, Embase and Cinahl, and led to identify a set of 1540 items.
Oncology HCPs (n=109) completed a pre- and post-surveys and differences The selection was confined to articles in English language only. Three authors
were analyzed. Targeted chart review of men with a new diagnosis of cancer screened individually all the items by title and abstract, and, after removing

duplicates and discussing about discrepancies, a selection of 320 articles was apoptosis was 6.93%±2.8 SSC and 38.2%±13.2 EL4 (P≤0.05), and miR 206 was
finally considered for the evaluation of contents. (14.6%±5.5 SSC and 38.3%±23.7 EL4) (P≤0.05).
Main results and the role of chance: The analysis by text revealed 4 key Limitations, reasons for caution: The biggest limitation of this study was
items on which the educational interventions could be focused: healthcare pro- the failure to perform it on human specimens
viders, patients, patient’s engagement and implementation of educational FPS Wider implications of the findings: Taken together, this study suggests that
programs aimed at identifying criticisms on oncofertility decision-making coun- MiR -therapy may lead to the development of novel therapeutic strategies for
seling before treatments. Both healthcare providers and patients challenge with cancer, and apoptotic MiRs may be a potential therapeutic agent for human
FPS barriers concerning communication, inappropriate support for a conscious tumors and is worthy of further investigation.
decision-making process or lack of knowledge about fertility preservation strat- Trial registration number: 96-01-30-29861
egies, etc. These represent core items on which building educational programs
aimed to increase the discussion rate on FPS between clinicians and patients, by
P-425 Does letrozole supplementation during ovarian stimulation
ensuring a personalized care path to patients in terms of time, use of available
for fertility preservation impact early luteal progesterone levels
resources, quality and continuity of care, and a good quality of life. Suggested
following GnRH agonist trigger
tools are the development of a multidisciplinary team, a script-based approach,
the partners or relatives involvement at the time of counseling, the release of I. Lalami1, M. Peigné1, C. Vinolas2, F. Krief2, I. Cédrin1,
informative material and the training of professional figures working between M. Grynberg1
1
oncologic and reproductive care services (nurses or midwives). Hôpital Jean Verdier, Department of Reproductive Medicine and Fertility
Limitations, reasons for caution: It is important to keep on sensitizing all Preservation, Bondy, France ;
2
multidisciplinary healthcare professionals involved in cancer care to avoid the Hôpital Jean Verdier, Department of Reproductive Medicine and Fertility
underestimation of FPS barriers. Preservation-, Bondy, France
Wider implications of the findings: The evaluation of effectiveness of FPS
awareness-raising interventions in terms of patient engagement can increase the Study question: To investigate early luteal progesterone levels following GnRH
number of FPS accesses, improve the knowledge of cancer therapies and the agonist (GnRHa) trigger after controlled ovarian stimulation (COS) in cancer
quality of the clinician/patient relationship. The implementation of multidisci- patients with or without letrozole supplementation (COSTLES).
plinary FPS programs can also increase the adherence to national guidelines on Summary answer: Following GnRHa trigger, early luteal phase progesterone
oncofertility cares. levels are significantly lower in COSTLES when compared with stimulation per-
Trial registration number: not applicable formed without aromatase inhibitor.
What is known already: Oocyte vitrification after COS is the first line and
most effective method for fertility preservation (FP) prior to cancer treatment.
P-424 In vitro nanoparticle-mediated delivery of MicroRNA 143
However, COS leads to supraphysiologic serum estradiol levels that could be
and MicroRNA 206 on spermatogonia and cancer cell apoptosis
potentially harmful in estrogen-sensitive diseases such as breast cancer. To pro-
A. Shams1, M. Koruji2, R. Shabani3, M. Asghari4, M. Karimi5
tect patients from the potential adverse effects of elevated estradiol levels during
1
Iran univesity of medical sciences, reproductive biology, tehran, Iran ; COS, specific FP protocols using aromatase inhibitors (letrozole) have been
2
Iran univesity of medical sciences, anatomy, tehran, Iran ; developed recently. However, data on luteal progesterone levels following COS
3
iran university of medical sciences, anatomy, tehran, Iran ; are lacking despite rising evidence of the role of progesterone in breast
4
Tabriz university of medical sciences, statistics, tabriz, Iran ; tumorigenesis.
5
Iran univesity of medical sciences, nanotechnology, tehran, Iran Study design, size, duration: This retrospective study was conducted from
July 2014 to December 2019. Serum progesterone levels following GnRHa trig-
Study question: to assess the efficiency of PLGA-miR 143/206 transfection ger measured in 84 breast cancer patients undergoing COSTLES were compared
on apoptosis in mouse leukemia cancer cells (El4) and spermatogonial stem to those obtained in 162 FP patients stimulated without letrozole
cells (SSCs) administration.
Summary answer: we were able to induce apoptosis on cancer cells without Participants/materials, setting, methods: This study was performed in a
any significant damage to spermatogonial stem cell by using smart transfection. public tertiary hospital. All women underwent COS with GnRH antagonist pro-
What is known already: A promising method to preserve fertility in children tocol. In COSTLES, the aromatase inhibitor was started 2 days before recom-
with cancer is via testicular biopsy before the onset of cancer treatment, fol- binant FSH administration and continued until GnRHa trigger. Final oocyte
lowed by isolation, proliferation, maintenance and transplantation. One of the maturation was induced by injection of 0.2 mg of triptorelin, 36 hours before
methods for the decontamination of cancerous cells from testicular cell sus- oocyte pick-up in both groups of patients. Serum hormonal levels of estradiol,
pensions can be smart nanoparticles. Smart gene delivery via Nanoparticles LH and progesterone were measured in the morning following the injection of
can be targeted and concurrently minimize damage to healthy cells in cancer triptorelin.
treatments.Many microRNAs (MiRs) are used to induce apoptosis in cancer Main results and the role of chance: Patients in COS without letrozole and
cells.Many studies have shown the effect of MiRs 143 and 206 on induction of COSTLES groups were comparable in terms of age (31.7±0.4 vs. 32.7±0.5,
apoptosis in many cancer cells. ransfection of MiRs via nanoparticles has been respectively), BMI (23.6±0.5 vs. 23.4±0.4 kg/m²), antral follicle count (22.2±1.0
very promising. vs. 22±1.7 follicles) and serum anti-Müllerian hormone levels (3.1±0.2 vs.
Study design, size, duration: This study was in vitro animal experiment and 2.6±0.8 ng/mL). They received comparable total amount of exogenous FSH
it was done on 50 neonate mouse. This study was conducted over a period of (3167.3±251.9 vs. 2981.2±237.2 IU). As expected, on the day following GnRHa
9 months administration serum estradiol levels were lower in COSTLES cycles (650.3±57.7
Participants/materials, setting, methods: mouse spermatogonia and can- vs. 2451.4.0±144.0 pg/mL, p<0.01). However, GnRHa-induced LH surge was
cer cell were used. To obtain a suitable miR dose that can induce apoptosis in significantly higher in patients having received aromatase inhibitors (71.9±4.6
cancer cells, while not harming SSCs, several doses were evaluated. Cells were vs. 51.2±2.6 UI/L, p<0.01). In addition, serum progesterone levels were signifi-
treated separately at 3 doses of each miR (for miR 143, doses of 25, 50 and 75 cantly lower in the COSTLES group (8.6 ± 0.7 vs. 10.5 ± 0.5 ng/mL, p< 0.03).
nmol and for miR 206, doses of 50, 100 and 150 nmol) at 24, 48 and 72 hours. Otherwise, the mean number of oocytes recovered (14.2±0.7 vs. 14.1±0.9
Viability and apoptosis were investigated by MTT and Annexin Kits oocytes, respectively) and vitrified at metaphase 2 stage (10.1±0.6 vs. 10.0±0.7
Main results and the role of chance: Based on MTT assay results, the oocytes) did not differ significantly between COS without letrozole and
optimal dose of miR 143 was 75 nmol (61.24%±2.85 SSC and 45.57%±0.78 COSTLES.
EL4) (P≤0.05), and for miR 206, the optimal dose was 150 nmol (53.82%±6.7 Limitations, reasons for caution: Serum progesterone levels were available
SSC and 28.14%±3.01 EL4) (P≤0.05). The optimal time was 48h. these doses, only once, during the very early luteal phase.
the survival rate of the EL4 cells was below IC50 and SSC survival was above Wider implications of the findings: The higher LH surge following GnRHa
50%. Annexin V staining also confirmed the selected doses (for miR 143 total administration in COSTLES protocol may be in relation with a weaker negative

1
feed-back of estradiol on the hypothalamic-pituitary axis. However, the lower Oslo University Hospital, Department of Reproductive Medicine, Oslo, Norway ;
2
serum progesterone levels might suggest a suppressive effect of letrozole in its Oslo University Hospital, Department of Pathology, Oslo, Norway
production. Further analysis is required to confirm the mecanisms.
Trial registration number: not applicable Study question: What is the spatial distribution of primordial follicles the
human ovarian cortex during the reproductive lifespan?
Summary answer: Follicles lie clustered in the human ovarian cortex; average
P-426 Gonadotrophin stimulation and risk of relapse in breast
cluster radius is ~270μm. At older age, degree of clustering increases and the
cancer
cluster size decreases.
A. Fredriksson1, E. Rosenberg2, Z. Einbeigi3, C. Bergh1, What is known already: In mice, the distribution of follicles is clustered, and
A. Strandell1 the degree of clustering is increased in aged mice. In human, primordial follicles
1
Sahlgrenska University Hospital, Reproductive Medicine, Gothenburg, Sweden ; distribute unevenly in the ovarian cortex, but the pattern of distribution has
2
Sahlgrenska University Hospital, Gynaecology, Gothenburg, Sweden ; been unknown.
3
Southern Älvsborg Hospital, Department of Medicine- Oncology, Borås, Sweden Study design, size, duration: Observational study using ovarian tissue sam-
ples derived from patients undergoing fertility preservation treatment at an
Study question: Is gonadotrophin stimulation as part of IVF associated with academic hospital.
an increased risk of relapse in breast cancer? Participants/materials, setting, methods: Ovarian tissue samples derived
Summary answer: Controlled ovarian hyperstimulation (COH) in connection from 14 patients (aged 20.8 – 35.2, mean 28.1 years) were obtained after uni-
with IVF in women with previous breast cancer was not associated with an lateral oophorectomy. Cortical fragments were processed for quantitative his-
increased risk of breast cancer relapse. tological assessment, including recording of two-dimensional coordinates of
What is known already: Breast cancer is the most common malignancy primordial follicles and stage of follicle development. The spatial distribution of
among women in the world and the leading cause of cancer death among females. follicles was tested and cluster radius was calculated. Loss of follicles was mod-
The use of COH with gonadotrophins in order to rescue the fertility by cryo- eled with computer simulation, comparing alternative models of follicle loss
preservation of oocytes or embryos prior to cancer treatment is currently the dynamics.
most established fertility preservation method for women with breast cancer. Main results and the role of chance: Primordial follicles form clusters with
To date, there are only a few small retrospective hospital-based controlled stud- a radius of ~270 µm in the ovarian cortex, which is increasingly apparent with
ies evaluating the risk of breast cancer relapse in patients undergoing fertility ageing. At older age, follicle density declines and the distance to the nearest
preservation with or without COH showing no evident risk of relapse in breast neighbouring follicle increases, the cluster radius decreases, but the degree of
cancer after the use of chemotoxic agents. clustering increases. Computer simulation of follicle loss dynamics indicate the
Study design, size, duration: This was a retrospective, population-based, close-range follicle-to-follicle signalling may contribute to the emergence of clus-
matched cohort study comprising a total of 5857 women, previously treated ters during reproductive ageing.
for breast cancer of whom 337 were exposed to COH. Exposure for COH and Limitations, reasons for caution: The tissue was obtained for diagnostic
outcomes as relapse and death were identified for all patients from 2003 to 2014 purposes. The small size of the fragments may limit the validity of assumptions
by assessing national registries holding data on IVF, births, prescription of drugs, of the spatial statistical analysis. All tissue samples were sectioned in the same
causes of death and cancer diagnoses. Relapse, when non-registered, was cal- orientation, limiting comprehensive three-dimensional analysis.
culated based on typical diagnoses and procedures, indicating relapse. Wider implications of the findings: Clustering may impede precise estima-
Participants/materials, setting, methods: Women aged 20-44 years, pre- tion of total follicle count, the imprecision increasing with age. Excessive dissec-
viously diagnosed with breast cancer and exposed to COH, were matched for tion of ovarian tissue fragments on ~ 0.5 mm scale may disrupt clusters and
age at breast cancer diagnosis +/- five years, tumour size and lymph node interfere with close-range follicle-to-follicle signalling, causing rapid graft
involvement to a non-exposed control group. In a subsequent secondary analysis exhaustion.
the entire cohort was assessed with adjustments for N- and T-stages. Trial registration number: not applicable
The risk of r2elapse in breast cancer was estimated as crude hazard ratios
(HRs) and 95% confidence intervals using Cox proportional hazards models. P-428 Semen quality and cryopreservation in patients with brain
Main results and the role of chance: In the matched cohort relapse tumors.
occurred in 27 of 337 (8.0%) women having undergone COH compared with D. Rivet-danon1, C. Jean1, A.S. Gille1, C. Chalas1, C. Patrat1,
71 of 334 (21.3%) among the non-exposed (HR=0.60; 95% CI 0.37-1.00; V. Drouineaud1
p=0.051). In the secondary analysis with adjustments for T- and N stages the 1
Assistance Publique – Hôpitaux de Paris AP- HP Centre- Université de Paris,
risk remained low after COH exposure; 27/337 (8%) in the exposed cohort
Service de Biologie de la Reproduction - CECOS, Paris, France
compared with 1176/5520 (21.3%) in the control group (HR=0.54; 95% CI
0.34-0.84; p=0.006). In the exposed cohort nine breast cancer related deaths
Study question: Do initial sperm parameters of a patient with brain tumor
(9/27; 8%) occurred compared with 555 in the non-exposed control group
make cryopreservation possible?
(555/5220; 10.1%). The coding template for assessing relapse from diagnoses
Summary answer: Overall, initial mean sperm parameters are normal except
and procedures in regions where relapse was not a registered variable, compared
for progressive motility in pineal region tumors and make possible cryopreser-
with a reference region, resulted in a sensitivity of 95% and specificity of 87%.
vation in the vast majority of cases.
Limitations, reasons for caution: A substantial degree of missing data on
What is known already: Baseline data on semen quality and cryopreservation
important prognostic variables was a limitation. Further, data on confounding
in patients presenting brain tumors are mainly based on small sample sizes and
factors were not completely covered. Another limitation was that the variable
results of various studies are controversial. Melatonin is a neurohormone
‘relapse’ had been introduced only in one region and had to be calculated from
secreted by the pineal gland. Literature data show that melatonin affects secretion
other variables.
of both gonadotropins and testosterone, and can influence sperm quality.
Wider implications of the findings: In this large, retrospective, matched
Moreover, men having reduced sperm motility present lower levels of melatonin
cohort study we found no risk of relapse in breast cancer among women who
in semen than patients with normal sperm parameters. Furthermore, in vitro
had been exposed to gonadotrophins as part of IVF. This is reassuring but prob-
addition of melatonin in semen shows an increase of sperm progressive motility.
ably confounded by selection to IVF of a group of women with a more favourable
Study design, size, duration: This was an 8 years retrospective and obser-
prognosis.
vational study including patients with brain tumors, referred for sperm cryopres-
ervation before a potentially gonadotoxic treatment, to the Paris Cochin
University Hospital sperm bank (Centre d’Etude et de Conservation des Œufs
P-427 Clustering of primordial follicles during reproductive ageing et du Sperme humains, CECOS). The location of brain tumors plus clinical and
A. Schenck1, G. Greggains1, M.V. Rodriguez1, B. Davidson2, histological data were collected as well as sperm parameters before freezing and
P. Fedorcsak1 after thawing. Fertile sperm donors were recruited between 2014 and 2016.

Participants/materials, setting, methods: Two hundred and fourteen brain Participants/materials, setting, methods: Eleven transgender boys (mean
tumor patients aged 14-58 years old and 92 fertile sperm donors were included. age 16.4y) and 39 cisgender girls (mean age 15.5y) were referred to FP at IVF
Sperm parameters were analyzed before freezing and after thawing, according units of two tertiary university-affiliated medical centers. The transgender boys
to the World Health Organization’s (WHO) 2010 guidelines. Sperm parameters (study group) were referred before initiating testosterone treatment and the
and the quality of sperm straws were compared on one part between patients cisgender girls (control group) were referred due to cancer diagnosis before
with brain tumors, in particular according to the location of the tumor, and on starting anticancer treatment. Control ovarian stimulation was carried out by
other part between patients and fertile sperm donors. the GnRH antagonist protocol. Statistical analyses compared ART data and FP
Main results and the role of chance: Among the 214 patients, brain tumor outcomes between two groups.
location was recorded in 183 patients: pineal gland (26), hypothalamic-pituitary Main results and the role of chance: The values of the accepted ovarian
axis (7) and other periphery brain regions (150). reserve markers, follicle-stimulating hormone (FSH: 5.7±1.9 mIU/mL) and antral
In all patients, each mean initial sperm parameter was normal according to follicle count (AFC: 20.4±5.5) of all the adolescent transgender boys were within
the WHO 2010 criteria (total sperm count: 326.5 ± 433.2 x 106, sperm con- the normal range. The transgender boys were slightly older compared to the
centration: 95.9 ± 108.16 x 106/ml, progressive motility: 32.7 ± 14.8%), except cisgender girls (16.4±1 vs 15.5±1.3 years, respectively, P = 0.03). The amount
sperm vitality which was low (53.2 ± 19.2%). of FSH used to stimulate the ovaries was significantly lower among the trans-
However, mean progressive motility (p<0.001), vitality (p<0.001), morphol- gender boys compared to the cisgender girls (2308±945 IU vs 4372±1877 IU,
ogy (p=0.03) and number of motile spermatozoa per straw (NMSPS) (p=0.008) respectively, P < 0.001), but the duration of ovarian stimulation was similar for
were significantly lower in brain tumor patients than in donors, respectively. both groups (12.2±3.6 and 10.1±2.8 days, respectively, P = 0.092). There were
In pineal brain tumor patients, mean sperm progressive motility was low no significant differences between the transgender boys and cisgender girls in
according to the WHO 2010 criteria (27.8 ± 13.4%) and significantly lower than peak estradiol levels (2660±2532 pg/mL vs 1269±975 pg/mL, respectively,
in the case of hypothalamic-pituitary tumors (27.8% vs 45.0%; p=0.009). All P = 0.053), the number of retrieved oocytes (30.5±11.4 vs 22±13.2, respec-
mean sperm parameters of patients with pineal tumor were significantly reduced tively, P = 0.062), the number of MII oocytes (24.9±11.7 vs 18.8±11.2, respec-
compared to those of fertile donors (total sperm count, p=0.02; sperm con- tively, P = 0.122), and the maturity rates (80±10.5% vs 85.4±14.6%, respectively,
centration, p=0.005; progressive motility, p<0.001; vitality, p<0.001; morphol- P = 0.118).
ogy, p=0.01; NMSPS, p=0.01, respectively). Limitations, reasons for caution: Our control group consisted of oncology
Severe oligozoospermia (sperm concentration <1 x 106/ml) was significantly patients in whom the ovarian reserve and the ovarian stimulation response may
more frequent in patients with pineal tumor than in periphery locations be affected by the underlying disease and it may, therefore, not be representative
(p=0.007). of the general healthy population.
More than 90% of patients could bank sperm regardless of tumor location. Wider implications of the findings: Adolescent transgender boys have a
Limitations, reasons for caution: The abstinence period was high in normal ovarian reserve and an excellent response to ovulation stimulation before
patients with brain tumors and could explain the low sperm vitality and motility initiating testosterone treatment. Oocyte cryopreservation is, therefore, a fea-
observed. The disease itself and/or brain surgery and/or the associated sible and effective way for them to preserve their fertility for future biological
altered general state in those patients probably influenced the long absti- parenting.
nence delay. Trial registration number: Not required
Wider implications of the findings: Patients with brain tumors showed
suitable sperm parameter allowing cryopreservation in most cases, even in pineal P-430 Passive slow freezing is an easy, cost-effective and
gland tumor patients. Importantly, sperm straws could be used for Assisted efficacious alternative freezing method for slow controlled
Reproductive Technologies (ART) if necessary. Those results confirm the impor- cryopreservation of ovarian tissue.
tance of sperm cryopreservation in those patients before a potentially high S. Lierman1, A. Bus2, S. Andries2, P.E.J. Bols2, K. Tilleman1
gonadotoxic treatment. 1
University Hospital Ghent- Fertility and Stem cell Team, Department for
Trial registration number: not applicable Reproductive Medicine, Ghent, Belgium ;
2
Laboratory for Veterinary Physiology and Biochemistry- Gamete Research Centre,
P-429 Fertility preservation outcomes among adolescent Faculty of Pharmaceutical Biomedical and Vetrinary Science- University of
transgender boys before testosterone treatment compared with Antwerp, Antwerp, Belgium
adolescent cisgender girls
H. Amir1, A. Oren2, E. Klochendler Frishman3, O. Sapir3, Study question: Is there a difference in follicle activation after xenotransplan-
Y. Shufaro3, F. Azem1, A. Ben-Haroush3 tation of passive slow frozen ovarian tissue compared to standard controlled
1
Tel Aviv Sourasky Medical Center, Racine IVF Unit- Lis Maternity Hospital, Tel frozen tissue?
Aviv, Israel ; Summary answer: Human primordial follicles have similar developmental
2
Tel Aviv Sourasky Medical Center, Pediatric Endocrinology and Diabetes capacity in a xenotransplantation model after passive slow rate freezing (PSF)
Unit- Dana-Dwek Children’s Hospital, Tel Aviv, Israel ; compared to the control slow rate freezing (CSF).
3
Rabin Medical Center, IVF and Infertility Unit, Petah Tikva, Israel What is known already: For the moment, CSF is the established method for
the freezing of cortical fragments of ovarian tissue (OT). CSF requires expensive
Study question: What are the fertility preservation (FP) outcomes among computerized equipment and the process is time-consuming. There is a risk for
adolescent transgender boys before testosterone treatment compared with technical failures with the potential loss of precious material. Other techniques
adolescent cisgender girls? like tissue vitrification are promising. However, the technique is expensive and
Summary answer: FP outcomes from assisted reproductive technology (ART) laborious. PSF is a technique that is successfully used in freezing of in vitro cell
among adolescent transgender boys are excellent compared to adolescent cis- cultures and testicular tissue of different animal species including human
gender girls. (Goossens et al., 2013). Until today no studies have described PSF for human
What is known already: ART enables young transgender men to have genet- ovarian tissue.
ically related children and overcome gender-affiliation hormone (i.e. testoster- Study design, size, duration: Xenotransplantation was performed in the
one) therapy-related fertility impairment. Although medical FP outcomes in pouch between the musculus obliquus and peritoneum of Balbc/nude mice
young cancer patients who preserve fertility before exposure to anticancer (n=24). On each side of the incision at the linea alba, a PSF and a CSF fro-
therapy are well known, there are no data on FP outcomes among transgender zen-thawed OT-piece of the same patient was introduced. After 2,4 and 6 weeks
boys before initiating testosterone therapy. transplantation, mice were euthanized by cervical dislocation and grafts were
Study design, size, duration: This retrospective cohort study included 11 collected, fixed in 4% buffered formalin and embedded in paraffin for HE staining.
adolescent transgender boys and 39 adolescent cisgender girls who underwent All xenotransplantation experiments were performed in duplo.
FP between January 2017 and April 2019 and September 2013 and April 2019, Participants/materials, setting, methods: In total, 48 OT strips from 4
respectively. transgender patients were frozen using L-15+0.45% HSA+DMSO (1.5M). For

CSF a programmable freezer was used (2°C/min:4°C to –9°C; seeding at group, grafts were retrieved on days 3 (n=5) and 10 (n=5). One fragment was
–9°C; –0.3°C/min to –40°C and to –140°C by 10° C/min). PSF was per- taken from each patient to serve as non-grafted controls.
formed using an isopropyl alcohol container (Mr. Frosty, Nalgene) and stored Participants/materials, setting, methods: Prospective experimental study
at –80 °C overnight at ≈ -1°C/min. Follicle activation and the fibrotic surface conducted in an academic gynecology research laboratory.
area by Masson Trichrome staining were analyzed. Fisher’s exact-test was Retrieved xenografts were analyzed by histology (follicle count and classifica-
performed. tion), immunohistochemistry (caspase-3 for apoptosis and LC3B for autophagy)
Main results and the role of chance: The grafting recovery rate was 95.8% and immunofluorescence (analysis of follicle activation pathways): (i) FOX01
(46/48 OT-strips). Both freezing procedures, PSF versus CSF, showed similar cytoplasm translocation for PI3K/pAkt pathway activation; and (ii) YAP nuclear
amounts of primordial follicles after 2 hours of warming before transplantation translocation for Hippo pathway activation. Immunostained cellular localizations
(53.0% (1690/3188) versus 54.7% (1309/2393) (P=0.220)). were determined in primordial follicles by image acquisition using the Axio Imager
After 2 weeks of transplantation, a clear and significant activation of primordial with ApoTome at 63x magnification.
follicles in both cryopreservation methods was seen compared to the non-trans- Main results and the role of chance: The ASCs+OT group showed
planted tissue. For PSF (85.3% (290/340) versus 47.0% (1498/3188) and CSF significantly higher follicle density and lower follicle atresia than the OT group
(72.7% (288/396) versus 45.3% (1084/2393), (P<.0001).This activation shift over time. Nevertheless, ASCs did not appear to modulate apoptosis, as the
was significantly higher in PSF compared to CSF (85.3% (290/340) versus 72.7% rate of caspase-3+ follicles was higher in both groups after 3 days compared to
(288/396) (P<.0001). After 4 weeks, the PSF technique showed a higher per- non-grafted controls (2-step/ASCs+OT group: 24.3 %; OT group: 25%; non-
centage of secondary follicles in the growing follicle pool in comparison to CSF grafted controls: 3.75%; p<0.05). Regarding autophagy, average LC3B staining
37.6% (65/173) versus 6.8% (4/59) (P<0.001). concentrations were similar in the ASCs+OT group and non-grafted controls,
After 6 weeks, the proportion of secondary follicles was not significantly but significantly lower in primordial follicles in the OT group, indicating less active
different between the two methods (CSF 47.5% (19/40) versus PSF 38.5% cell repair.
(15/39)). FOX01 localization was mainly nuclear in oocytes of primordial follicles in
The percentage of fibrosis in the xenograft was comparable for PSF versus non-grafted controls. A significant shift to cytoplasmic localization (follicle acti-
CSF for the 2 and 4 weeks (10.25±0.37 versus 10.50±0.37) and (9.42±0.47 vation marker) was observed in the OT group on day 3, while the ASC+OT
versus 10.02±0.37). After 6 weeks transplantation, graft tissue from the PSF group showed no difference from non-grafted controls.
showed a higher percentage fibrotic area compared to the tissue of the CSF YAP localization was largely cytoplasmic in granulosa cells of primordial folli-
(13.56±0.48 versus 11.55±0.46, (P=0.003)). cles in non-grafted controls, but shifted significantly to nuclear (follicle activation
Limitations, reasons for caution: Although the follicle activation and growth marker) in both grafted groups 3 days after OTT. ASCs appeared to confer a
in xeno transplanted human OT after PSF is similar to CSF, PSF has not yet been protective effect on primordial follicles by suppressing the PI3K/pAKT pathway.
used in transplantation in human. The true competence of the PSF frozen tissue However, there was no evidence of Hippo pathway modulation by ASCs.
still needs proof of concept in human. Limitations, reasons for caution: These findings warrant long-term studies
Wider implications of the findings: Passive slow freezing (PSF) shows a investigating maintenance of the primordial follicle pool over time, with a view
significant higher follicle activation after xenotransplantation than the conven- to clinical application in patients undergoing OTT.
tional method of freezing (CSF). PSF could be an easy and cost-effective alter- Wider implications of the findings: The present study provides further
native to CSF for fertility preservation of human ovarian tissue. insights into the mechanisms of action involved in follicle activation and cell repair,
Trial registration number: This research is conducted with the approval of underlining the beneficial impact of ASCs in grafted ovarian tissue. We also
the local ethics committee 2015/0124 – B670201523543), Ethical Committee demonstrated that ASCs exert positive effects on the ovarian reserve by pro-
on Animal Experimentation (ECD18/12). tecting primordial follicles.
P-431 Early follicle activation, repair and death after human
ovarian tissue transplantation using adipose tissue-derived stem P-432 TurnerFertility study: preliminary data on fertility
cells preservation by ovarian tissue cryopreservation in young girls with
L. Cacciottola1, G. Courtoy2, C. Hossay1, J. Donnez3, Turner syndrome
M.M. Dolmans1 S. Nadesapillai1, M. Schleedoorn1, R. Peek1, D. Braat1,
1
Institut de Recherche Expérimentale et Clinique- Université Catholique de Louvain, J. Van der Velden2, K. Fleischer1
1
Gynecology Research Unit, Brussels, Belgium ; Radboud University Medical Center, Obstetrics and Gynaecology, Nijmegen, The
2
Institut de Recherche Expérimentale et Clinique- Université Catholique de Louvain, Netherlands ;
2
Imaging Platform, Brussels, Belgium ; Radboud University Medical Center, Pediatric Endocrinology, Nijmegen, The
3
Society for Research into Infertility, Av. Grandchamp 143, Brussels, Belgium Netherlands
Study question: Does ovarian tissue transplantation (OTT) using adipose Study question: Which girls with Turner syndrome (TS) could benefit from
tissue-derived stem cells (ASCs) yield better follicle outcomes shortly after trans- fertility preservation by ovarian tissue cryopreservation (OTC), based on the
plantation than the standard procedure? presence of follicles in relation to karyotype, clinical and hormonal data?
Summary answer: Improved survival, reduced activation and better mainte- Summary answer: Girls with TS who have favourable predictive parameters
nance of repair mechanisms were observed in primordial follicles 3 days after (e.g 46,XX cell line, a measurable AMH or spontaneous puberty) could benefit
grafting using ASCs compared to standard OTT. from fertility preservation by OTC.
What is known already: OTT has gained ground as a valid fertility restoration What is known already: Infertility due to premature ovarian insufficiency is
approach thanks to its established effectiveness. Nevertheless, more than 50% a major concern for girls with TS and their parents. Physicians are often asked
of follicles are lost after transplantation due to two main mechanisms: prolonged about possible options to preserve their fertility. However, evidence for suc-
hypoxia and massive follicle activation, the latter known as the ‘burn out’ effect. cessful fertility preservation by OTC/autotransplantation in these girls is lacking
ASCs have been shown to shorten the hypoxic/ischemic period by boosting and many questions remain. Without evidence on the effectiveness of OTC it
revascularization in human xenografts, thereby increasing follicle survival. It is cannot routinely be offered to girls with TS.
also known that hypoxia-related signaling is able to modulate several pathways Study design, size, duration: A national prospective exploratory intervention
involved in cell survival and follicle activation, like the PI3K/pAKT and Hippo study. Ovarian cortex will be obtained after unilateral ovariectomy from 100
pathways. girls with TS aged 2-18 years. Patients will be included between 2017 and 2020.
Study design, size, duration: Twenty nude mice (Swiss Nu/Nu) were grafted Participants/materials, setting, methods: All girls with TS who have com-
with frozen-thawed ovarian fragments from 5 patients. Ten of them underwent pleted the diagnostic work up of TS were included. After unilateral ovariectomy,
OTT in 2 steps using ASCs, as previously described by Manavella et al. (ASCs+OT one fragment of the ovarian cortex was used to determine the number of follicles
group), and the remaining ten using the standard procedure (OT group). In each by serial sectioning and staining. Karyotyping of ovarian cells, lymphocytes, buccal

cells and urine cells was performed by Fluorescence in situ hybridization (FISH). centre, ovarian activity resumed in 90% of grafts, and in 64% when cryopreserved
Blood samples obtained before ovariectomy and during the yearly clinical visit elsewhere. This was after a mean period of 5.3 months. Follicle density and
after ovariectomy will provide information on hormonal parameters. tissue volume grafted appear to be the strongest predictors of ovarian activity
Main results and the role of chance: Currently, we have received duration over 12 months, however this does not reach statistical significance.
98 informed consent forms and 66 girls (age 3-19) had an unilateral ovariectomy. Stimulated cycles in 25 patients with low-dose modified recombinant FSH pro-
Oocytes were found in 27% of the girls (n=18; age 5-17). In this group, 5 girls tocol from 2012, yielded an oocyte from 68% of follicles (32% rate of empty
were prepubertal, 12 girls had a spontaneous thelarche (age 10-17) and 4 girls follicles) and 75% of these were retrieved from non-ovarian grafts. Mature
had a spontaneous menarche (age 13-16). Hormone values and the chromosome oocytes were retrieved from mean follicular size 13.5mm (range 6-20 mm). The
pattern were found to be predictive parameters for the presence of follicles. In overall 2PN fertilisation rate was 64%. A total of 25 fresh and frozen embryos
this cohort, the chance of finding follicles in girls with a 46,XX cell line is three were transferred in 15 women, yielding a clinical pregnancy rate 28% (7/25) and
times higher than in girls without a 46,XX cell line. Until now, there was only 1 live birth rate of 20% (5/25). There was additionally a single, ongoing sponta-
girl with 45,X cells in lymphocytes and buccal cells (n=25) who had follicles and neous pregnancy. All pregnancies occurred in patients aged under 32 years at
a measurable AMH. FISH was used to karyotype the ovarian cortex cells of 5 cryopreservation and only one of the successful patients had prior chemotherapy
patients with a mosaicism and revealed that 42 of the 46 oocytes that were exposure.
analysed had a normal X-chromosomal content. Granulosa cells were largely Limitations, reasons for caution: Trends alone can be observed from this
45,X, but showed different levels of X chromosome mosaicism, not only data as the numbers are small.
between patients but also between individual follicles of the same patient. Wider implications of the findings: This case series adds to international
Despite the presence of a low percentage (10–45%) of 46,XX ovarian stromal understanding of this technique’s results. It encourages detailed and consistent
cells, normal macroscopic ovarian morphology was observed. reporting, as ongoing research is needed to improve efficacy. These results sup-
Limitations, reasons for caution: The final analysis will be performed when port offering ovarian cryopreservation to young women who are undergoing
the dataset of 100 TS girls is completed. Further research is necessary to deter- fertility threatening treatment, with optimism for future pregnancy.
mine the functionality of the follicles in the ovaries of TS girls and to elucidate Trial registration number: not applicable
if OTC is an effective method for fertility preservation in TS.
Wider implications of the findings: A combination of clinical, hormonal P-434 Dynamic in vitro culture of human ovarian cortical tissue
and karyotypic data could provide predictive parameters to define which girls improves follicle growth and the expression of genes involved in
with TS might benefit from fertility preservation. These parameters could help follicle activation and progression.
physicians during fertility preservation counselling to determine if OTC is an R. Talevi1, M.M. Pallotta1, V. Barbato1, M. Di Nardo1, V. Costanzo1,
option for a certain girl with TS. R. Ferraro2, G. Mondrone3, K.S. Adiga4, T. Capriglioine1,
Trial registration number: NCT03381300 G. Catapano5, R. Gualtieri1
1
Università degli studi di Napoli Federico II, Dipartimento di Biologia, Napoli, Italy ;
P-433 Experience with 54 cryopreserved ovarian tissue grafts in a 2
Genesis Day Surgery, IVF center, Caserta, Italy ;
single centre: the devil in reporting details! 3
IVF RED srl, research, Caserta, Italy ;
G. Rozen1, F. Agresta2, D. Gook1, S. Siii3, K. Stern1 4
Kasturba Medical College Manipal Academy of Higher Education, Clinical
1
Royal Women’s Hospital and Melbourne IVF, Reproductive Services, Melbourne, Embryology, Manipal, India ;
5
Australia ; Università della Calabria, Ingegneria Meccanica- Energetica e Gestionale, Rende
2
Melbourne IVF, Reproductive clinic, Melbourne, Australia ; CS, Italy
3
South West Healthcare- 25 Ryot Street- Warrnambool- 3280, Obstetrics and
Gynaecology, Victoria, Australia Study question: Does dynamic (D) versus static culture of human ovarian
cortical tissue (HOCT) better support follicle growth and expression of genes
Study question: Following transplantation of cryopreserved ovarian tissue, involved in follicle activation and progression?
what are IVF outcomes per cycle and are there factors related to this technique Summary answer: D vs static culture of HOCT improves follicle progression
which impact on success. to the secondary stage and the expression of genes involved in follicle activation
Summary answer: Mature oocytes which resulted in pregnancies were largely and progression
harvested from non-ovarian grafts. Longer duration of ovarian activity correlated What is known already: Oxygen availability, disruption of solutes gradients
with increased tissue volume and follicle density. and application of physiological fluid mechanical stress, through dynamic culture
What is known already: Transplantation of cryopreserved ovarian tissue has in vitro of HOCT have been shown to enhance follicle growth and health. Several
a unique role for fertility preservation and over 140 live births have now been pathways, e.g. Hippo, Wnt, and P3K/AKT, are modulated by fragmentation and
published. Nevertheless, uncertainties remain regarding its efficacy, with success subsequent culture of HOCT in vitro.
of this technique reported inconsistently and mostly reported per patient, rather Study design, size, duration: Biopsies of HOCT were fragmented into
than per cycle started or embryo transferred as is used in conventional IVF. This 1×1×0.5mm strips and cultured, in groups of ten, in gas-permeable dishes (PD)
hampers identification and optimisation of factors which may predict success, and in D condition for 6 days. At the end of culture, the following endpoints
such as grafting sites and tissue volumes. were assessed: 1) follicle quality and stage; 2) follicle viability; 3) expression of
Study design, size, duration: We analysed a case series of reproductive genes involved in follicle activation and growth. This study was approved by our
outcomes throughout the IVF treatment process following ovarian tissue trans- local ethical committee.
plantations at our centre (2006-2019), with tissue frozen in the years 1996-2018. Participants/materials, setting, methods: Biopsies were obtained from
This included 54 grafting procedures in 40 patients, following a diagnosis of five consenting patients (aged 27.2±5.8) during laparoscopic surgery for benign
premature ovarian insufficiency. Results of modified low-dose stimulation and gynecological conditions, fragmented using a tissue chopper and cultured 6 days
oocyte retrieval, including both oocyte cryopreservation and IVF cycles were in PD and D conditions. Follicle health and stage were assessed through histology,
reviewed. and live-dead far-red/Hoechst 33342 labeling. Total RNA was extracted to
Participants/materials, setting, methods: Ovarian tissue was harvested analyze the expression of BMP15, BMP7 (Wnt pathway), INHA (follicle growth),
from 40 patients at risk from gonadotoxic treatment at median age of 26 years FOXO3, FGF2 (P3K/AKT pathway), MST2, YAP1 (Hippo signaling).
(range 18-39). Almost 30% of patients had their tissue frozen elsewhere. Nine Main results and the role of chance: Overall, 2300 follicles were analyzed.
of 31 patients (29%) had prior cytotoxic treatment. Following a diagnosis of At day 0 most follicles were primordial (primordial, 78.7%; primary, 18.3%;
premature ovarian insufficiency, ovarian tissue was grafted to pelvic sites including secondary, 3%), with high viability (78.8%) and good quality (grade 1, 43.6%;
the ovary and adjacent pelvic side walls, as well as the anterior abdominal wall, grade 2, 29.2%; grade 3, 27.2%). At day 6, D was superior to PD culture in terms
on average five years following extraction. of follicle progression (primordial, 27.8 vs 40%; primary, 50 vs 45.3%; secondary,
Main results and the role of chance: Fifty-four grafting procedures were 22.2 vs 14.7%; p<0.01) and quality (grade 1, 43.6 vs 33.6%; grade 2, 27.4 vs
performed in 40 women. For patients whose tissue was cryopreserved in our 24.6%; grade 3, 29 vs 41.8%; p<0.01). Viability was not significantly different

between samples (day 0: 78.8%; PD: 67.9%; D: 65.3%). At day 6 of in vitro D the day of fertilization showed no significant differences but progesterone levels
versus PD culture, the expression of genes involved in follicle activation (FGF2; were significantly higher in recipients of ganirelix protocol (8.19 ± 3.08 vs. 10.15
p<0.001) and progression (BMP7, p<0.05; BMP15, P<0.01) increased, whereas ± 7.05, p=0.03).
the expression of the inhibitors of cell proliferation (INHA, FOXO3; p<0.001) Mean number of transferred embryos showed no significant differences (1.29
decreased. The expression of MST2 and YAP1, components of the Hippo path- ± 0.62 in desogestrel vs 1.13 ± 0.6 in ganirelix, p=0.09). No statistically significant
way, was different between the two in vitro culture systems. In particular, MST2 differences were found in clinical pregnancy rates (48.2% in desogestrel vs. 53%
was uP-regulated and YAP1 was down-regulated (p<0.05) after D versus PD in ganirelix) and live birth rates (37.3% in desogestrel vs. 43.4% in ganirelix).
culture. Limitations, reasons for caution: Retrospective study carried out in oocyte
Limitations, reasons for caution: This is an In vitro study carried on a limited donors that vitrified oocytes, which is a suitable model to suggest outcomes in
number of patients non-medical fertility preservation, but the study did not analyze fertility preser-
Wider implications of the findings: Dynamic vs static culture, which vation cycles specifically. Further well conducted clinical trials and long-term
enhances the number and health of secondary follicles, and expression of genes safety studies are needed.
involved in cell proliferation and follicle growth, could represent a new tool for Wider implications of the findings: According to our findings, no significant
in vitro folliculogenesis differences were observed in oocyte competence, embryo development and
Trial registration number: not applicable live birth rates from vitrified oocytes coming from desogestrel or ganirelix sup-
pressed cycles. Therefore, non-medical fertility preservation cycles could be
P-435 Can progesterone primed ovarian stimulation (PPOS) be suppressed with progesterone instead of GnRH-antagonists, maintaining the
introduced in non-medical fertility preservation? Results from efficacy, increasing patient convenience and decreasing costs.
vitrified oocytes from the oocyte donation program. Trial registration number: not applicable
M. Devesa Rodriguez De La Rua1, N.P. Polyzos1, E. Clua1,
D. Rodríguez1, S. García1, L. Coll1, F. Martínez1 P-436 Choosing a triggering agent in oocyte preservations
1 cycles - A comparison between gonadotropin-releasing
Dexeus Mujer, Obstetrics- Gynecologý and Reproductive Medicine, Barcelona,
hormone agonist (GnRH-a) and recombinant human chorionic
Spain
gonadotropin (r-hCG).
Study question: Does the type of LH peak suppression (PPOS vs. GnRH E. Haikin Herazberger1, S. Knaneh1, D.B.Y. Ben Yosef1, H. Amir1,
antagonist) have any effect on oocyte competence, embryo development and A. Reches1, Y. Kalma1, F. Azem1, N. Samara1
live birth rates in recipients of vitrified donated oocytes? 1
Tel-Aviv university, IVF Unit- Department of Obstetrics and Gynecology- Tel-Aviv
Summary answer: PPOS protocol results in equally competent oocytes / sourasky Medical centere- Tel-Aviv- Israel, Tel aviv, Israel
embryos and comparable live birth rates as compared with GnRH antagonist
(ganirelix) in recipients of vitrified oocytes. Study question: Is there a difference in outcomes of oocyte preservation
What is known already: It has been reported that progesterone is effective cycles according to the type of triggering agent?
in LH peak suppression in oocyte donation and IVF cycles, with no differences Summary answer: GnRH-a significantly increases the mature oocyte rate in
in the number of retrieved oocytes or embryo quality. Although some studies medically indicated cycles, with no similar effect in social fertility preservation
have found lower pregnancy rates in recipients of oocytes coming from a PPOS cycles.
cycle suppressed with medroxyprogesterone acetate, results with desogestrel What is known already: Fertility preservation is no longer reserved for
did not reveal significant differences. However, no study has analyzed results patients with malignancy before medical intervention. More and more women
coming specifically from vitrified oocytes. Assessing outcomes from vitrified seek to extend their fertility options due to personal choices. One main advan-
oocytes coming from donation cycles suppressed with progesterone in compar- tage of an antagonist protocol is that it provides the option to trigger ovulation
ison to GnRH antagonist, would allow applying the same protocol to non-medical with GnRH-a, which lowers the risk of ovarian hyperstimulation syndrome
fertility preservation. (OHSS). However, it also lowers pregnancy and live birth rates in fresh embryo
Study design, size, duration: Retrospective cohort study carried out from transfer cycles due to an inadequate luteal phase. In fertility preservation luteal
2016 to 2018 in a University affiliated private fertility clinic. phase is irrelevant. We set out to compare other relevant outcomes according
Donors: age between 18-35 years, regular menstrual cycles, BMI 18-28 Kg/ to triggering type in this subgroup of patients.
m2 and no relevant medical history. LH suppression with desogestrel (PPOS): Study design, size, duration: This retrospective cohort study was performed
n=83. LH suppression with ganirelix: n=83. at a tertiary university-affiliated medical center. Overall, 226 fertility preservation
Recipients: age <50 years, reception of vitrified oocytes coming from oocyte cycles between May 2013 and September 2018 were included in the analysis, of
donation cycles suppressed either with desogestrel (n=83) or with ganirelix which 126 cycles were triggered with GnRH-a and 100 cycles with r-hCG.
(n=83). Medical indications accounted for 130 cycles and social indications accounted
Participants/materials, setting, methods: Donors: pretreatment with oral for 96 cycles.
contraceptives, stimulation carried out with gonadotrophins and downregulated Participants/materials, setting, methods: Fertility preservation indications
with oral desogestrel (75mg daily, from stimulation day 1 until day of trigger) or were extension of fertility or fertility protection preceding chemotherapy, radio-
subcutaneous ganirelix (0.25 mg/day, flexible protocol). therapy, or endometriosis surgery and fragile X carriers at risk for premature
Recipients: endometrial preparation with oral estradiol valerate (2mg/8 hours) ovarian insufficiency. Triggering with GnRH-a (Decapeptyl 0.2 mg) or r-hCG
and vaginal progesterone (200 mg/8 hours). Embryo transfer was performed (Ovidrelle 250 microgram) was administrated when the leading follicle was ≥17
on third or fifth day of embryo development. Treatment was continued until the mm with adequate estradiol levels. Mature oocyte rates were calculated as the
day of βhCG and, if positive, until 10th week of pregnancy. number of MII oocytes/total retrieved oocytes per cycle. A cut-off of 80%
Main results and the role of chance: Donors: There were no significant indicated good oocyte maturation rate.
differences concerning mean age, BMI, AMH, AFC, number of mature (21.05 Main results and the role of chance: No significant differences were
± 7.52 vs. 23.05 ± 9.28) and vitrified oocytes (12.93 ± 6.66 vs. 15.17 ± 9.92) observed between patients triggered by GnRH-a compared to r-hCG in terms
between desogestrel and ganirelix cycles, respectively. of gonadotropin dosage and BMI (p=0.95 and 0.62, respectively). The majority
Recipients: Age and BMI did not differ significantly between both groups of of patients with social indication were triggered with GnRH-a (63.5%). Patients
recipients. The number of thawed oocytes was significantly lower in the desoge- with medical indications were divided equally between GnRH-a and r-hCG
strel group compared to the ganirelix group (9.89 ± 4.05 vs. 10.69 ± 3, p=0.02). (p=0.043). The average age of the GnRH-a patients was significantly lower than
Warming survival rates (80.1% vs. 81.1%), fertilization rates (71.3% vs. 75%) and that of the r-hCG patients (p<0.01), and the estradiol blood levels of the former
embryo development rate (53.1% vs. 52.1%) didn’t differ significantly between were higher than those of the latter (p=0.04). There was no significant difference
desogestrel and ganirelix groups, respectively. in oocyte maturation rate between triggering by GnRH-a or r-hCG (p= 0.48).
Embryo transfer rate did not differ significantly between the groups (91.6% Comparison of the percentage of GnRH-a- and r-hCG-triggered cycles with an
in desogestrel vs. 88% in ganirelix). Endometrial thickness and estradiol levels MII rate >0.8 revealed a slightly higher percentage for the GnRH cycles, however,

difference was not statistically significant. (59.6% versus 47.1%, p=0.08). Data (FG = 1.13 ± 0.19 vs Non-FG = 0.58 ± 0.22; mean ± SD). Contrastingly,
were then analyzed according to both indication for fertilization therapy and the genetic and epigenetic controls comprehended with form of programmed
triggering agent. GnRH-a significantly increased the likelihood of a high matura- cell death; Bcl-2, FOXO1 and microRNA-218 were markedly lower or
tion rate by 3.55 (1.30-9.66) among treatment cycles for medical indications, down-regulated in the ovarian tissue-fibrin encapsulated group (Bcl-2;
while there was no significant effect of the triggering agent in treatment cycles Non-FG = 7.31 ± 1.52 and FG = 5.51 ± 1.16; FOXO1; Non-FG = 9.42 ± 3.01
for social indication. and FG = 7.98 ± 0.43; microRNA-218; Non-FG = 2.39 ± 0.99 and FG =
Limitations, reasons for caution: This cohort analysis is based on retro- 0.63 ± 0.25; mean ± SD).
spective data collection, with potential biases inherent to that design. Pregnancy Limitations, reasons for caution: The present research findings preliminary
rates and live birth rates were not evaluated, and future studies on these aspects document outcomes from a small number of participants following oophorec-
are warranted. tomy procedure. Additionally, the limitation of retrieved ovarian tissue size and
Wider implications of the findings: The results affirm that triggering with number theoretically leads to the restriction of research or study design expan-
GnRH-a and r-hCG have similar ART outcomes in women planned to no fresh sion and recapitulation in all aspects.
embryo transfer. GnRH-a triggering administration should be considered first Wider implications of the findings: Our preliminary research outcomes
choice in fertility preservation for reducing the risk of OHSS and for greater speculate that the novel technology focused on tissue engineering with biode-
likelihood of good maturation rates in medically indicated cycles. gradable gel matrix may be applicable for ovarian tissue cryopreservation and
Trial registration number: not applicable fertility preservation. Furthermore, the cortical striP-fibrin encapsulation may
subsequently lead to the restriction of cell apoptosis during ovarian tissue cryo-
P-437 Epigenetic factor regulation; microRNA-9 and preservation and transplantation.
microRNA-218 related to cell proliferation and apoptosis, by Trial registration number: 384/62
tissue engineering-fibrin encapsulation: Preliminary study on
ovarian tissue cryopreservation for fertility preservation P-438 The use of LH for preserving reproductive health in
P. Thuwanut1, W. Sereepapong1, K. Pruksananonda1, cancer patients
C. Suebthawinkul1, P. Tuntiviriyapun1, C. Taweepolcharoen1, S. Marcozzi1, R. Vicenti2, G. Orlando3, C. Centonze3,
P. Sirayapiwat1 M. Macciocca2, F. Di Rella4, M. De Felici1, R. Fabbri2, F.G. Klinger1
1 1
Department of Obstetrics and Gynecology, Faculty of Medicine- Chulalongkorn University of Rome Tor Vergata, Dep. of Biomedicine and Prevention, Rome, Italy ;
2
University, Bangkok, Thailand S.Orsola-Malpighi Hospital- University of Bologna, Department of Medical and
Surgical Sciences DIMEC, Bologna, Italy ;
Study question: Could fibrin encapsulation regulate alteration of epigenetic 3
Merck Serono spa, Medical Affairs Department, Rome, Italy ;
factors related to cell proliferation and apoptosis; microRNA-9 and 4
National Cancer Institute- IRCCS Foundation G. Pascale, Medical Oncology-
microRNA-218, during ovarian tissue cryopreservation procedure? Department of Senology, Naples, Italy
Summary answer: The tissue engineering technology by fibrin encapsulation
tended to down-regulate microRNA-218 (cell apoptosis) while uP-regulate Study question: Is luteinizing hormone (LH) able to protect ovarian reserve
microRNA-9 (cell proliferation) during ovarian tissue cryopreservation. against cyclophosphamide (CPM)-induced damage in women?
What is known already: Application of biodegradable matrix i.e. hydrogel Summary answer: LH appears able to significantly reduce primordial follicle
or fibrin for mammalian cell encapsulation has been extensively considered one (PMF) loss in ovarian strips treated in vitro with phosphoramide mustard (PM),
of potential strategies which offer utility of cell-based system regulation under the active metabolite of CPM.
complex microenvironment. Recently, this fascinating technology was introduced What is known already: Cancer therapies cause a severe reduction of PMFs,
to reproductive medicine research i.e. immature testicular tissue fragment encap- often inducing premature ovarian insufficiency (POI) in female patients. For this
sulation or mesenchymal stem cell encapsulation in 3D ovarian cell construction. reason, several compounds have been analyzed as adjuvant therapies to protect
This novel approach allows molecule cocktails; angiogenic, growth factors, hor- the ovarian reserve without interfering with cancer treatment on tumor cells.
mones or essential nutrients delivered to cell or tissue leading to cell/graft sur- We recently demonstrated, in vitro, the protective effect of LH against ovotox-
vival during culture, cryopreservation and transplantation. This technology, icity induced by cisplatin on the ovary of prepuberal mice. These results suggest
however, mainly presents in animal research whereas in terms of human ovarian a possibility for to use of LH prior or in concomitance with an anticancer treat-
studies are limited. ment in order to preserve oocytes in human patients, therefore preventing the
Study design, size, duration: The cortical pieces of ovarian tissues collected early onset of menopause and/or infertility.
from two patients during July-August, 2019 (there cortical pieces per patient) Study design, size, duration: The ovarian tissues were collected from three
were allocated into three sub-groups; I) fresh (F), II) ovarian tissue without fibrin patients (age-pathology : 25 yrs-Ewing’s sarcoma, 15 yrs-medulloblastoma and
gel encapsulation (Non-FG) and III) ovarian tissue encapsulated with 10% fibrin 14 yrs-Hodgkin lymphoma) who have cryopreserved their ovarian tissue before
gel (FG). The cortical pieces from each designed group (Non-FG and FG) were receiving anticancer treatment. For each patient, eight ovarian strips were thawed
processed for slow freezing procedure and evaluated for relative microRNA and randomly assigned to the experimental conditions (two strips each): Control
(microRNA-9 and microRNA-218) and mRNA (Bcl-2 and FOXO1) expres- (CTRL), PM, PM+LH-200mIU and PM+LH-500mIU. LH was added 1 hour
sion levels. before the treatment with PM. Samples were analyzed after 24 and 48 hrs of
Participants/materials, setting, methods: Total of six cortical strips treatment.
(0.5 x 0.5 x 1.0 mm; width x length x thickness) from premenopausal patients Participants/materials, setting, methods: Ovarian strips were cultured
(age 39 and 48 years old; diagnosed with breast cancer and uterine fibroid) were for 24-48 hours at 37°C and 5% CO2, in α-MEM supplemented with 40% human
allocated as mentioned in study design. Ovarian tissues were cryopreserved by serum with/out 10mM PM and/or 200mIU/ml LH or 500mIU/ml LH. The
-1°C/minute slow freezing (4°C to -80°C and plunged into -196°C). Fresh and samples were processed for:
post-thawed samples were analysed for relative microRNA (reference to U6) –Histology, for PMFs and primary follicles (PFs) analysis;
and mRNA (reference to GAPDH) expression levels using qRT-PCR. –Western blotting, for the expression of protein involved in DNA damage and
Main results and the role of chance: The present findings preliminary apoptosis;
indicated that the fresh ovarian cortical strips present the highest relative –Real-Time PCR, for the expression of genes involved in apoptosis and
microRNA-9 expression level (2.97 ± 1.43; mean ± SD). Nonetheless, the lower inflammation.
relative microRNA-218 (2.06 ± 2.27; mean ± SD), Bcl-2 (4.42 ± 0.38; mean ± Main results and the role of chance: Histological analysis showed that in
SD) and FOXO1 (7.23 ± 5.43; mean ± SD) expression levels compared the the untreated group, the follicle density varied from 450.45 to 921.38 PMFs/
post-thawed samples were observed. Regarding to the recent advance in tissue mm3 and from 94.45 to 430.30 PFs/mm3 between the three patients analysed.
engineering technology, the uP-regulation of microRNA related to cell prolifer- Therefore, relative follicular density (%) was performed to analyse statistical
ation (microRNA-9) was notably revealed in the post-thawed ovarian cortex differences between the control and treated groups. Notably, whereas relative
encapsulated with fibrin gel which was 1.95 times higher than non-FG samples PMFs density (%) was significantly reduced in PM vs CTRL either after 24 and

48 hrs, this reduction was partially counteracted by both LH dosages tested (24 any significant increase in antrum differentiation upon PMSG treatment
hrs: CTRL=74.47±5.12; PM=42.31±5.66; PM+LH-200mIU=70.77±3.72; (4, 0.4, 40 IU) was observed. The chorionic gonadotropin was more effective
PM+LH-500mIU=59.19±9.49 - 48 hrs: CTRL=68.15±1.75; PM=25.96±16.91; than oFSH in promoting ivF outcomes. Indeed, preantral follicles stimulated
PM+LH-200mIU=42.62±11.79; PM+LH-500mIU=56.35±12.96). No significant with 4 IU PMSG displayed a significantly greater growth than those exposed to
differences were observed in relative PFs density (%). Real-Time PCR and oFSH (Δ%: 63.7±28.9 vs 52±31; p=0,038). Moreover, PMSG promoted a
Western blotting revealed no significant differences, between groups, in the complete meiotic competence in the majority of collected oocytes (58.3% MII,
expression of molecules involved in apoptosis (NOXA, PUMA, Bax, Bak and 2.8% GVBD, 38.8% GV). Of note, the MII oocytes were all isolated by early
Bcl2), DNA damage (γH2AX), probably due to the already occurred loss of the antral follicles with Δ%>40 (p<0.01). On the contrary, only a very small fraction
follicles, and in the expression of IL6, a pro-inflammatory cytokine. of the oocytes isolated from oFSH-treated follicles was able to resume meiosis
On the contrary, treatment with PM is able to significantly increase IL1β gene and no one reached the MII stage (83,30% GV, 16,70% GVBD; p<0.001).
expression, another pro-inflammatory cytokine; interestingly, pre-treatment with Limitations, reasons for caution: -
LH, at both tested dosages, counteracted this increase (24 hrs treatment: Wider implications of the findings: These proof-of-concept PMSG exper-
CTRL=1.00±0.01; PM=2.03±0.04; PM+LH-200mIU=1.37±0.04; PM+LH- iments establish a way for new protocols optimizing ivF and allow to identify a
500mIU=1.50±0.03 and 48 hrs treatment: CTRL=1.00±0.12; PM=1.75±0.09; trans-species hormone to functionalize biomaterials. Both strategies might sup-
PM+LH-200mIU=1.30±0.10; PM+LH-500mIU=1.63±0.20).Limitations, port follicle development in large animal models and humans with the hope of
reasons for caution: Main limitation of this preliminary study includes its small translating this technology for fertility preservation purposes.
sample size (only ovarian cortex from three subjects were used) that needs to Trial registration number: not applicable
be improved. The present study did not elucidate the molecular mechanisms
involved in LH ovoprotection against PM P-440 Fertility preservation in adolescent males: Setting up a
Wider implications of the findings: These preliminary results showed that national service.
LH is able to preserve PMFs in human ovarian cortex exposed to PM in vitro. M. Horan1, L. Hartigan1, H. Groarke1, L. Glover1, M. Wingfield1
These findings encourage the possibility to use the hormone as a treatment to 1
Merrion Fertility Clinic- Dublin and The National Maternity Hospital Dublin,
prevent the premature onset of menopause and/or infertility in women under- Obstetrics & Gynaecology, Dublin, Ireland
going anticancer treatments.
Trial registration number: Not applicable Study question: Does a multi-disciplinary approach to sperm cryopreservation
in adolescent males enable quality care?
P-439 Production of ovine fertilizable oocytes using innovative in Summary answer: Within 18 months of inception, an efficient national care
vitro (iv) technologies applied to preantral follicles growth. pathway has been established with successful cryopreservation and positive
C. Di Berardino1, G. Capacchietti1, N. Bernabò1, M. Crociati2, patient feedback.
M. Monaci2, L. Sylla2, B. Barboni1 What is known already: The incidence of both childhood and adolescent
1
Faculty of Bioscience and Agro-Food and Environmental Technology- University of cancer is increasing throughout Europe. Around 200 children and young adoles-
Teramo, Unit of Basic and Applied Biosciences, Teramo, Italy ; cents are diagnosed with cancer in the Republic of Ireland every year. Thanks
2
University of Perugia, Department of Veterinary Medicine, Perugia, Italy to advances in medicine and surgery, survival rates in this group are now greater
than 80%. As a result, there is increased emphasis on the long-term effects of
Study question: Considering the early pool of gametes potentially able to cancer treatment. Studies from other units outside of Ireland have shown that
generate offspring, what actions should be taken to optimize ivF thus increasing semen cryopreservation was possible in most adolescent cancer cases regardless
fertilizable oocytes availability? of age or disease diagnosis, however to date, no formal service for CAYA male
Summary answer: The standardization of long-term gonadotropin stimulation sperm cryopreservation exists in Ireland.
is a key step to implement ivF in order to promote both reproductive targets, Study design, size, duration: We developed a National CAYA Fertility
oogenesis and follicle steroidogenesis activation. Preservation Consortium with fertility specialists at our clinic and paediatric
What is known already: Increasing in childhood cancer survivors points up oncology specialists at the National Children’s Hospital. The infrastructure and
an urgent need in fertility preservation (FP) strategies to overcome the negative combined expertise through this collaborative consortium allowed us to provide
impact of cancer treatments on reproductive cell cycle. Because of a concern a sperm cryopreservation service for adolescent boys, with the option of pro-
exists on the possible re-introduction of malignant cells following ovarian trans- ducing samples at either site. We performed a retrospective review of all ado-
plantation, in vitro (iv) culture of early stage follicles still remains a potential lescent male patients referred for consideration of sperm cryopreservation in a
alternative to obtain fertilizable oocytes that contribute to biological offspring. 16-month period.
Since technologies advances are required to transfer iv Follicologenesis (ivF) from Participants/materials, setting, methods: Patients were identified through
bench to bedside, monovulatory large mammal becomes ideal translational the National Paediatric Haematology/Oncology Programme. Clinical and demo-
models to validate cultural conditions enhabling synergic follicles-oocyte iv graphic data was recorded on patients. Disease diagnosis was provided by the
development. primary oncologist or rheumatologist in the setting of benign disease. We eval-
Study design, size, duration: The present research aimed to study the influ- uated patient demographics and semen parameters. Sperm parameters analyzed
ence of 2 different gonadotropins (oFSH vs Pregnant Mare’s Serum were volume, sperm concentration and motility. Number of failed samples,
Gonadotropin/PMSG) on ivF cultures of single ovine preantral follicle (PA). The number of straws stored per patient, and number of visits required for each
ivF outcomes obtained by using the specie-specific (oFSH) or the trans-species patient to produce a sufficient sample was also recorded.
chorionic gonadotropin (PMSG) were analyzed after 14 days of culture by com- Main results and the role of chance: This project addressed deficiencies
paring follicle/oocyte growth, timing and percentage (%) of antrum differentia- and current barriers in the provision of fertility preservation services to children
tion as well as % of Methaphase II (MII) oocytes after iv Maturation (IVM). in Ireland.
Participants/materials, setting, methods: Preantral follicles, mechanically Fifteen patients, aged between 12 and 17 years old, were referred to our
isolated from slaugterhoused prepubertal ovaries, were incubated as 3D single service for consideration of sperm cryopreservation between August 2018 and
follicle culture and exposed to 25 ug oFSH or to different PMSG doses (from December 2019. Of these, 93% (14/15) presented with malignant disease. One
0,4 to 40 IU/ml). In order to compare the degree of meiotic competence, MII patient was referred for sperm cryopreservation prior to commencing gonad-
oocytes obtained adopting an advanced IVM, consisting of cumulus oocytes otoxic treatment for non-malignant disease. 93% of the male patients elected
complexes co-cultured with walls antral follicle-derived somatic cells, were to come to our fertility clinic rather than provide their samples at the National
analyzed. Children’s Hospital.
Main results and the role of chance: The PMSG influence on ivF perfor- Of the 15 patients referred, 12 attempted sperm production. Eight of these
mances was strictly dose-dependent. More in detail, 4 IU was the concentration patients (67%) had sperm successfully cryopreserved. The youngest patient in
of PMSG able to enhance follicular diameter increment (Δ%: 63.7±28.9 vs our study was 12 years old and successfully cryopreserved four straws. Four
43.4±19.3 and 52.7±33.4, respectively for 4, 0.40, 40 IU; p<0.05). Conversely, patients (36%) did not achieve sperm cryopreservation: 1 patient failed to collect

any sample, 2 patients had no sperm in the sample produced. Unfortunately 1 hypothesized a uniform distribution model for the referrals, assuming therefore
patient had very poor semen quality that was not suitable for freezing, this was that the frequency of cancer is constant over the years.
following one cycle of chemotherapy. Five patients attended the clinic twice to Wider implications of the findings: The frequency of adolescents who
attempt specimen production. Of the 8 patients who successfully had sperm bank their semen prior to initiate gonadotoxic therapies is lower than it should
cryopreserved, only 1 patient had semen analysis results entirely within WHO be. Future research is needed to delineate the barriers and to develop interven-
‘normal’ limits. tions that could overcome them. Meanwhile, physicians engaged in oncology
Limitations, reasons for caution: The study cohort was limited in size. We should dedicate more attention on fertility preservation in young boys.
currently have no long term data on the rate of straw utilisation in our cohort Trial registration number: not applicable
of patients, given that this is a very new service.
Wider implications of the findings: Prior to this study, the number of P-442 Females’ fertility preservation for cancer patients: lessons
adolescent males availing of cryopreservation in Ireland was unknown. This study learned of ten years’ experience in a French center
will enable collection of data and the establishment of a national database. A. LE BRAS1, V. Windal1, V. Puy1, L. Hesters1, V. Gallot2,
Success thus far will improve awareness of this service among oncologists in B. Alexandra2, M. Grynberg2, C. Sonigo2, N. Frydman1
Ireland. 1
Hospital A. Béclère, Reproductive Biology Unit CECOS, Clamart Cedex, France ;
Trial registration number: N/A 2
Hospital A. Béclère, Department of Reproductive Medicine and Fertility
Preservation, Clamart Cedex, France
P-441 Sperm cryopreservation in adolescents with cancer.
F. Filippi1, E. Somigliana1,2, M. Reschini1, C. Meazza3, M. Podda3, Study question: For female cancer patients who underwent fertility preser-
C. Guarneri1, D. Giacchetta4, M. Massimino3, L. Restelli1, vation, what are the outcomes of oocyte- or embryo-thawing after Controlled
M. Terenziani3 Ovarian Stimulation or in vitro maturation ?
1
IRCCS Fondazione Ca’ Granda Policlinico di Milano, IVF Unit, Milano, Italy ; Summary answer: Regardless of FP technique, live births were obtained for
2
Università degli Studi di Milano, Dept. Clinical sciences and Community Health, all of the emergency in vitro fertilization procedures in former cancer patients.
Milano, Italy ; What is known already: Cryopreservation of embryos was considered the
3
IRCCS Istituto Tumori di Milano, Pediatric Unit, Milano, Italy ; best FP method before the development of vitrification. Whereas vitrification
4
Ospedale Santi Paolo e Carlo, Assisted Reproduction Techniques Unit, Milano, Italy has improved the success rate for cryopreserved oocytes in female cancer
patients, no live births have ever been reported using in vitro matured oocytes.
Study question: Is sperm banking underused in adolescents with cancer? Study design, size, duration: This was a single-center, retrospective study
Summary answer: Semen cryopreservation prior to initiate gonadotoxic from January 2009 to December 2018 in France. We reviewed the biological
therapies is insufficiently used in adolescents with cancer. and clinical outcomes of female patients treated for cancer who returned to use
What is known already: Sperm cryopreservation was the first validated their oocytes or embryos that were cryopreserved using Controlled Ovarian
technique for fertility preservation. It is simple, promptly available, cheap and Stimulation (COS) or In Vitro Maturation (IVM).
safe. Even if only a minority of subjects who stored their semen will ever thawed Participants/materials, setting, methods: After a cancer diagnosis, four
them, there is a general consensus that sperm cryopreservation could be possible cryopreservation procedures were used: either frozen oocytes or frozen
cost-beneficial and that it should be systematically offered to men with cancer embryos, after either COS or IVM treatment cycles. Among the 667 patients
prior to initiate gonadotoxic treatments. However, evidences on sperm cryo- who underwent emergency FP before gonadotoxic cancer treatments, 40
preservation in adolescents and young men are sparse. They generally show that returned to thaw their oocytes or embryos. We compared the clinical and
sperm banking is feasible but also indirectly suggest that the proportion of those laboratory data of freeze/thaw cycles for these four groups in these 40 patients.
actually storing their semen may be hindered. Main results and the role of chance: Following emergency FP, 40 patients
Study design, size, duration: Men undergoing semen cryopreservation at (6%) returned to attempt a pregnancy with a mean time lapse of 3.3 ± 1.5 years.
the Infertility Unit of the Fondazione IRCCS Ca’ Granda Ospedale Maggiore Most were cured of breast cancer (n = 31). One hundred and thirty-four oocytes
Policlinico of Milan, Italy between January 1986 and December 2018 were ret- were used in 25 thawing cycles (7 after IVM-FP and 18 after COS-FP). Eighty-
rospectively reviewed. Information were obtained from patients’ charts. seven embryos were used in 24 thawing cycles (14 after COS-FP and 10 after
Participants/materials, setting, methods: Inclusion criteria were as follows IVM-FP). Eight pregnancies (3 from frozen oocytes, 5 from frozen embryos)
1) Age 14-21 years, 2) diagnosis of malignancy. Subjects those performing surgical resulted, three of them from fertilized, mature oocytes after IVM. All babies
sperm collection were excluded. The analysis focussed on the following infor- were delivered at term with normal perinatal characteristics. We report the first
mation: age at semen preservation, semen parameters at the time of preserva- births in previous cancer patients using vitrification of in vitro matured
tion and oncological diagnosis. oocytes for FP.
Main results and the role of chance: Overall, 317 subjects satisfied our Limitations, reasons for caution: Some confounding factors such as smok-
selection criteria. Cancer diagnoses were as follows: lymphomas (n=159, 50%), ing, alcohol consumption, and toxic environmental exposure at the time of
leukemia (n=21, 7%), testicular cancer (n=60, 19%) and others (n=77, 24%). cryopreservation are missing. Only a few patients have returned to attempt
The frequencies of referred cases according to age was not consistent pregnancy after being cured from cancer. We have no information about natural
(p<0.001) with the theoretical uniform distribution of referrals (ie with a similar pregnancies in patients who have had FP.
number of cases per group of age): a clear sharp and constant increase with Wider implications of the findings: The present study is one of only a few
age emerged. This unbalanced distribution remained also when restricting the published series conducted in patients treated for cancer comparing the out-
analysis to boys older than 16 years (p<0.001) or to those older than 18 years comes of thaw cycles according to the FP procedure chosen at the time of
(p=0.01), suggesting that discrimination for age persisted also in older boys. diagnosis. We report the first pregnancy obtained from matured oocytes
The total number of cases referring before 1999, between 1999 and 2008 and after IVM.
between 2009 and 2018 were 88, 98 and 131, respectively. An improvement Trial registration number: NA
of the distribution of cases per age group emerged over time (Chi square for
trend, p=0.002). However, within each historical period, distribution per age
P-443 Effects of high concentrations of Resveratrol on human
group remained inconsistent with the theoretical uniform distribution (p<0.001
sperm cryopreservation
for all). No statistically significant differences were identified in semen quality
between the different age groups. The proportion of boys who could not E. Porcu1, L. Notarangelo1, L. Cipriani1, G. Damiano1, P.M. Ciotti1,
freeze their specimen because of the absence of viable spermatozoa was also N. Calza1, S. Zuffa1, M. Nadalini2, E. Nardi3, E. Borini2, A. Borini2
1
similar. Infertility and IVF Unit, University of Bologna-Sant’Orsola Hospital, Bologna, Italy ;
Limitations, reasons for caution: Number of boys who failed to make a 2
9.baby, Family and Fertility Center, Bologna, Italy ;
3
collection was not recorded. Secondly, we cannot exclude that our results may University of Bologna, Department of Medical and Surgical Sciences, Bologna,
reflect a local low sensitivity towards fertility preservation. Thirdly, we Italy

Study question: Can high concentrations of resveratrol as antioxidants in Study design, size, duration: Ovarian tissue was collected from 7 prepu-
sperm cryopreservation be useful for post thawing viability and morphology? bertal girls (age 1-10 years) and 6 adult women (age 20-35 years). Primordial
Summary answer: High concentrations of resveratrol do not improve vitality and primary follicles were isolated from frozen-thawed prepubertal and adult
and post-thaw morphology in sperm cryopreservation. ovarian tissue. Frozen-thawed ovarian tissue from the same patients was also
What is known already: It was demonstrated that the cryopreservation of evaluated by immunohistochemistry and transmission electron microscopy.
human semen produces reactive oxygen species (ROS), which cause important Participants/materials, setting, methods: Mitochondrial activity in isolated
sperm damages such as lipid peroxidation of the cell membrane and DNA dam- follicles was explored by MitoTracker CM-H2XRos, which is a mitochondria-spe-
age. Several studies have examined the role of in vitro supplementation of cific fluorescent and cell-permeant probe, selectively sequestered only in mito-
antioxidants, such as resveratrol, to protect sperm DNA from ROS oxidative chondria with active ΔΨm, depending on their oxidative activity. Mitochondrial
damage and improve post-thawing sperm quality. content was investigated in prepubertal and adult ovarian tissue by TOMM20
Study design, size, duration: The effects of Resveratrol will be examined immunostaining, a peptide receptor located on the surface of the outer mito-
based on the assessment of sperm motility and morphology. Prospective study: chondrial membrane. Transmission electron microscopy was performed to
100 waste seminal samples of patients undergoing a IVF cycle at the University evaluate mitochondrial morphology.
of Bologna Infertility and IVF Center and 9.baby Center, Bologna from March Main results and the role of chance: Prepubertal follicles showed higher
to December 2019. Three groups were considered: Group A = post thawing fluorescence intensity by MitoTracker (p=0.03), compared to adult follicles. This
control group; Group B = post thawing 30 µM/L Resveratrol; Group C = post result suggests that mitochondrial activity is enhanced in prepubertal patients
thawing 50 µM/L Resveratrol. compared to adult patients. Quantification of TOMM20 immunostaining, based
Participants/materials, setting, methods: One hundred patients selected on the intensity and extent of the signal, revealed significantly stronger TOMM20
for normozoospermia and mild or moderate oligoastenoteratozoospermia. staining in prepubertal follicles than adult follicles (p=0.01), indicating the pres-
Patients age ranged from 23 to 53 years with a mean value of 38.18±6.00. ence of more mitochondria in prepubertal follicles. Last, TEM allowed to identify
Diagnosis of normozoospermia and mild or moderate oligoastenoteratozo- ultrastructural differences in the morphology of mitochondria: in prepubertal
ospermia was made considering not only the cellularity of seminal fluid samples follicles, mitochondria were heterogeneously dispersed throughout the cyto-
but also the total and progressive motility and morphology of spermatozoa. plasm in small clusters and were mostly elongated in shape, rather than round
Sperm cryopreservation was performed using Test Yolk Buffer, Resveratrol and as in adult mitochondria. Moreover, cristae in prepubertal mitochondria were
Sherman freezing Protocol. mostly longitudinal rather than archiform and, in some cases, shaped in the form
Main results and the role of chance: Semen standard analysis showed that of a fingerprint.
in the pre-freezing group the parameters related to total motility, progressive Limitations, reasons for caution: The present study evaluates follicle mat-
motility and morphology were 48,70 ± 8,63, 24,45 ± 14,70 and 19,73 ± 9,32 uration and competence in prepubertal patients, by assessing mitochondria
respectively. In the control group, total motility, progressive motility and mor- function and activity. Nevertheless, these parameters only provide indirect infor-
phology were respectively 26,30 ± 7,09, 12,30 ± 8,39 and 13,60 ± 6,59. In the mation and additional cellular organelles should be studied to further corroborate
30 µM/L Resveratrol group the total motility, progressive motility and morphol- these results.
ogy were respectively 22,77 ± 6,72, 9,47 ± 6,48 and 13,05 ± 6,41. In the Wider implications of the findings: With this study, we provided for the
50 µM/L Resveratrol group total motility, progressive motility and morphology first time information on the function, content and morphology of mitochon-
were respectively 19,05 ± 6,54, 6,87 ± 5,71 and 11,83 ± 6,09. Thawing results dria from prepubertal patients. We revealed that in prepubertal follicles mito-
showed that, 30 µM/L and 50 µM/L of resveratrol produce pejorative effects chondria are more abundant and that mitochondrial activity is enhanced,
on motility and morphology (P < 0.0001) of sperm after thawing, compared to suggesting that intrisic maturation changes will occur in ovarian follicles
the control group. with aging.
Limitations, reasons for caution: The scientific community opinion is still Trial registration number: /
controversial about the use, in vivo and in vitro, of antioxidants in IVF cycles.
Wider implications of the findings: In IVF cycles, the occurrence of oxida- P-445 Elective fertility preservation: do women come back as
tive events during in vitro culture and in cryopreservation processes is well much as they freeze?
demonstrated; the use of antioxidants during these phases must be further M. Miguens1, A.M. Quinteiro Retamar2, A. Coscia2, D. Acosta2,
investigated. G. Vega Balbuena1, N. Allende2, S. Papier2
Trial registration number: None 1
cegyr, fertility, buenos aires, Argentina ;
2
cegyr, clinical, buenos aires, Argentina
P-444 Mitochondrial activity in prepubertal and adult ovaries
R. Masciangelo1, M.C. Chiti1, A. Camboni1, C.A. Amorim1, Study question: What is the return rate for counseling in patients who had
J. Donnez2, M.M. Dolmans3 chosen to do elective fertility preservation and the proportion of women who
1
Université Catholique de Louvain, Pôle de Recherche en Gynécologie- Institut de used their stored eggs?
Recherche Expérimentale et Clinique, Bruxelles, Belgium ; Summary answer: A quarter of patients who preserved their fertility return
2
Société de Recherche pour l’Infertilité, Avenue Grandchamp 143, Bruxelles, around two years after for counseling and only 12,58% of the total performed
Belgium ; an IVF cycle.
3
Cliniques Universitaires Dt. Luc, Départment de Gynécologie, Bruxelles, Belgium What is known already: Increasing numbers of women are seeking elective
oocyte cryopreservation (EOC). This is due to the improving success rates of
Study question: Are mitochondrial content and activity in prepubertal ovarian the oocyte vitrification technique over the years and also social freezing is pro-
follicles different from adult ovarian follicles? moted to single women as a way of preserving their fertility potential. Recent
Summary answer: Differences in the number, activity and morphology of studies have found usage rates are less than ten percent. Studies have proven
mitochondria of prepubertal patients were evidenced. that vitrified oocytes can be fertilized, implant and achieve normal pregnancies
What is known already: Mitochondria generate most of the energy of cells after a short period but the chance of having desired outcomes in long term
through oxidative phosphorylation, possible thanks to generation of mitochon- cannot be predicted by physicians based on real-life experience.
drial membrane potential. Reduced efficiency of oxidative phosphorylation and Study design, size, duration: A cross-sectional retrospective study of 469
energy production in oocytes is related to poor embryo development. Moreover, women who did an EOC between 2009 and 2018 in a single-center at Buenos
it is known that mitochondria play a central role in ovarian aging via their major Aires, Argentina.
contribution to cell survival and apoptosis and that maternal age is associated Participants/materials, setting, methods: Women who did an EOC
with increased oxidative stress in oocytes, resulting in mitochondrial dysfunction. between 2009 and 2018 were included. The primary outcomes were the return
The role of mitochondria in determining oocytes’ developmental capacity is rate to the clinic for counseling and the proportion of women who used their
increasingly evident, nevertheless, literature on mitochondrial function in pre- storage eggs. Other analyzed variables were mean age at the moment of the
pubertal follicles at early stages of growth is scarce. freezing, mean number of oocytes vitrified per patient, the average time it took

patients to return and ongoing pregnancy rate in the patients who used their Supplement of ascorbic acid might be a potential therapy for MEHP-induced
oocytes. All analyses are descriptive. ovarian injury.
Main results and the role of chance: The total of cryopreserved oocytes Trial registration number: not applicable
over the years was 3376 being the average 7,2 per patient (from 1 to 32). The
mean age at the moment of the EOC was 37,17 years (19 to 45). The 24,95% P-447 Fertilisation outcomes after cryopreservation of human
(n: 117) of the total of patients returned for counseling with a mean age of immature and in vitro matured oocytes
39,57. The average time it took women to return was 2 years. From this group,
M. Petrushko1,2, V. Piniaiev3, T. Yurchuk2
27,35% (n:32) have decided to perform a second oocyte retrieval cycle (main
1
reason was a low number of eggs vitrified in the cycle before -median: 4-) and ART Clinic of Reproductive Medicine, Embryology, Kharkiv, Ukraine ;
2
around a half-used their stored eggs for an IVF treatment (50,43% n:59) which Institute for Problems of Cryobiology and Cryomedicine of the NAS of Ukraine,
is a 12,58% of the total of patients included. The thawed oocyte viability rate Cryobiology of reproductive system, Kharkiv, Ukraine ;
3
was 72%, the fertilization rate was 66% and the ongoing pregnancy rate was ART Clinic of Reproductive Medicine, Gynecology, Kharkiv, Ukraine
32.2% (n:19).
Limitations, reasons for caution: The main limitation of this study was its Study question: What are maturation and fertilization outcomes of GV
retrospective design based on data from a single center which may be subject oocytes in vitro matured prior and after vitrification?
of bias. Summary answer: Oocytes in vitro matured prior vitrification have higher
Wider implications of the findings: The cycles of oocyte cryopreservation, maturation, fertilization and blastocyst formation rate than oocyte matured after
in our clinic, have increased at least 800% since 2009. Even though, our rate is vitrification.
a little above compared with the literature, is still a low number. Prospective What is known already: Emergency of fertility preservation for patients who
cost-effectiveness and long-term success analysis are needed to determine if it’s are in danger of losing their ovarian function or donation programs apply oocyte
worth it or not. preservation for further usage in assisted reproductive technologies in spite of
Trial registration number: Not applicable being at different maturation stage (germinal vesicle stage (GV), MI, MII). There
is lack of data concerning oocyte cryopreservation efficiency and timing of in
P-446 Ascorbic acid protects against ovarian dysfunction induced vitro maturation regards to cryopreservation procedure.
by mono (2-ethylhexyl) phthalate in fetal mouse ovaries in vitro Study design, size, duration: GV oocytes (n=108) were cryopreserved prior
via reducing the oxidative stress and after in vitro maturation oocytes and their survival, fertilisation and blastocyst
formation rates were analyzed. All manipulations have been carried out between
C. Sui1, C. Liu1
January 2017 and November 2019.
1
Tongji Hospital- Tongji Medical College- Huazhong University of Science and Participants/materials, setting, methods: This single-center cohort study
Technology, Reproductive Medicine Center, Wuhan, China included 31 patients diagnosed with polycystic ovary syndrome (PCOS) and
poor responders (mean age of women was 34.3±4.2 years). GV oocytes were
Study question: Whether ascorbic acid protects fetal ovaries from the injury randomly selected for in vitro matured (IVM) prior (group 1) and after vitrification
effect induced by mono (2-ethylhexyl) phthalate (MEHP) and what is the (group 2). After thawing MII oocytes were fertilized by ICSI and cultured up to
mechanism. day 5. All manipulations with gametes were performed according with written
Summary answer: Supplement of ascorbic acid can rescue the injury effect consent of patients.
induced by MEHP in fetal mouse ovaries in vitro via reducing the oxidative stress Main results and the role of chance: It has been shown maturation rate
What is known already: Di-(2-ethylhexyl) phthalate is considered to be was 73.2 vs 52.0% in group 1 and 2 respectively. Overall no differences were
toxicant to female reproduction, potentially by its active metabolite MEHP. found for survival rate of MII oocyte matured prior cryopreservation and cryo-
Toxicological studies affirmed oxidative stress is one possible pathway for MEHP- preserved GV oocyte (97.6 % vs 96.2 %; P>0.05). Fertilisation and blastocyst
adjuvant effect. Ascorbic acid is known to be an antioxidant and a powerful formation rate were higher in group 1 (77.5 % and 38.7%) than in group 2 (38.5%
reducing agent and plays an important role in attenuating oxidative damage to and 20.0%). These differences are significant between the groups (P<0.05).
the reproductive system. Limitations, reasons for caution: Limited number of GV oocytes in the
Study design, size, duration: Fetal mouse ovaries exposed to different con- study group. Further studies are recommended and should include also group
centration of MEHP (0 M, 10-4 M, 10-5 M, 10-6 M) were treated with or without advanced age patients.
ascorbic acid (100nM) in vitro for 6 days. Then the ovarian follicle constitution Wider implications of the findings: These results revealed that using imma-
and the oxidative stress parameters of ovaries in each treatment were detected ture oocytes at GV stage and long-term preservation can increase the number
respectively to evaluate the influence of MEHP and ascorbic acid. of embryos available for transfer into the uterus of patient with PCOS and poor
Participants/materials, setting, methods: Ovaries from post-natal day 4 responders.
Kunming mice were recruited and assigned to different groups randomly. After Trial registration number: not applicable
treated with MEHP alone or a combination of MEHP and ascorbic acid for 6
days, ovaries were collected to histologically qualify the follicle numbers, and the
ovarian malonaldehyde (MDA) levels were tested. Besides, ovarian mRNA P-448 Fertility preservation in adolescent women: should we
related to Redox (Sod-1, GSS, CAT) were examined using quantitative reverse expect the same response?
transcription polymerase chain reaction. A. Santolaria Baig1, R. Quintana Berto1, N. García Camuñas1,
Main results and the role of chance: MEHP accelerated primordial follicle J. Subirá Nadal1,2, P. Polo Sánchez1, J.M. Rubio Rubio1
activation by increasing the proportion of primary and secondary follicles in 1
La Fe University Hospital, Women’s Health Area, Valencia, Spain ;
ovaries at the concentration of 10-4 M. The expression of SOD was increased 2
IVI-RMA Castellón-Valencia, Reproductive Endocrinology, Castellón, Spain
significantly in ovaries treated with 10-4 M MEHP (p<0.05). Moreover, MEHP
increased the MDA level in the ovaries at 10-4 M (p<0.01) and 10-5 M (p<0.05). Study question: Are oocyte cryopreservation (OC) cycles in adolescent
When ovaries exposed to 10-4 M MEHP, supplement of ascorbic acid rescued women (<20 years old) comparable to OC cycles in ages we expect the best
the follicular constitution, and normalized the MDA level. Ascorbic acid also response (20-25 years)?
reversed the increasing expression of SOD and GSS in the ovaries, which was Summary answer: OC cycles in adolescent women have similar results com-
caused by MEHP. pared with ages we expected the best response.Fertility preservation(FP) is a
Limitations, reasons for caution: This study only demonstrated the pro- feasible option in this age group.
tective effect of ascorbic acid to ovaries in vitro. Whether ascorbic acid func- What is known already: Women from 20 to 25 years old are expected to
tioned as a powerful agent to rescue the MEHP-induced ovarian injury will need have the best response to OC cycles for FP, based on the age-related distribution
in vivo studies to explore. of AMH. Few studies are published in adolescents (<20 years), even though FP
Wider implications of the findings: Ascorbic acid ameliorated the dis- in this group of patients is increasing nowadays. Some authors hypothesized that
ruption of folliculogenesis induced by MEHP via its antioxidant capacity. adolescent have differences in their stimulation cycles regarding the oocyte

maturity or dosing requirements of gonadotropins.The last publications obtained patients (AMH < 1.1 ng/ml by The Bologna criteria) (n=206). These patients
similar results in stimulation in adolescents and in women of other age groups, underwent 1.6 ±0.9 previous ovarian stimulation cycles for oocyte vitrification,
but with a higher likelihood of cancellation. followed by a fresh oocyte retrieval where all oocytes previously vitrified were
Study design, size, duration: This is a retrospective study performed from warmed for ICSI, as a strategy to increase the potential number of embryos
January 2013 to January 2020 at a tertiary university hospital. Our research available.
was focused in OC cycles of adolescent women (between 13 and 20 years of Participants/materials, setting, methods: Oocyte vitrification and warm-
age) and the group we expected the best response according to AMH levels ing were performed with the Cryotop method (Kitazato, Dibimed). Analysis on
(20-25 years). Data were retrieved from patients electronic records. pre- and post-embryonic developmental quality and perinatal evaluation was
Participants/materials, setting, methods: We collected data from epide- performed after ICSI on 580 freshly collected oocytes (FO) and on 614 vitri-
miology, stimulation and oocyte retrieval from 49 cases in adolescent women fied-warmed oocytes (VO) and fresh embryo transfer.
and 104 cases in the 20-25 years group. Numerical variables were summarized Main results and the role of chance: Patients had 36.4 ± 3.7-year-old,
by the median and standard deviation and a comparation of both groups was AMH of 0.7 ± 0.4 and FSH ± 10.3 4.1. The mean number of 2.8 ± 2.1 FO and
performed with T Student test. 3.0 ± 2.2 VO were used for ICSI on the same day per patient. Although fertili-
Main results and the role of chance: The main indication in adolescents sation rates were similar between FO and VO (64% vs 62%), embryonic devel-
for FP was Hodgkin lymphoma(36,73%) followed by gynecological pathol- opmental quality was affected on day 3 (p < 0.01) and day 5 (p < 0.05) of
ogy(24,48%). In the 20-25 years group the gynecological pathology was the main development being lower in vitrified oocytes compared to their fresh counter-
indication(26,92%), followed by Hodgkin lymphoma(22,11%) and breast cancer parts, which resulted in less embryos chosen for transfer in VO. A total of 161
(9,6%). patients underwent embryo transfer: 81 had 1.3 ± 0.5 embryos transferred from
Both groups were comparable in terms of epidemiological variables about FO, while 49 received 1.3 ± 0.4 embryos from VO. Thirty patients received mix
ovarian reserve. In all ages, the most common stimulation protocol was short transfer (one FO and one VO). Implantation rates were 22% and 28% for FO
antagonist protocol. and VO (NS), respectively. All embryos derived from vitrified oocytes that
Results from total days of stimulation, total number of follicles, follicles >16 implanted progressed to delivery (n=14), while 3/17 (18%) and 3/8 (38%)
mm and serum estradiol on the trigger day and FORT(Follicular Output Rate) embryos from freshly collected oocytes and mixed transfer miscarried. Perinatal
index were similar in both groups. Regarding the gonadotropin dosing, adolescent outcomes from this preliminary evaluation showed no difference in gestational
used a mean FSHr dosed of 2219,46 IU(±909,88) compared with 2520,50 age and body weight of children between freshly collected (38.67 weeks and
IU(±934,79) in women 20-25 years(p=0,07). However the initial dosing was 3092 kg) and vitrified oocytes (39.07 weeks and 3298 kg) (NS).
higher in the 20-25 years group: 222,90 IU(±53,66) vs 246,39 IU(±52,53) Limitations, reasons for caution: Retrospective design of the study and
(p<0,05). No significant differences were found between the groups for oocytes limited sample size concerning the data on delivery rates.
retrieval or the maturity rate. Adolescents retrieved a mean of 13,69(±10) Wider implications of the findings: In view of the lower in vitro develop-
oocytes with a maturity rate of 67,76%(±28,32) compared with 11,53(±8,15) mental potential of embryos derived from vitrified oocytes in low ovarian reserve
oocytes and 75,42%(±23,45) in women 20-25 years. The Follicle to Oocyte patients, accumulation of oocytes in patients undergoing ICSI cycles should be
Index (FOI) was also comparable in both groups. further evaluated. However, the preliminary data on perinatal outcomes seems
There were more cancellations in the adolescent group:20,4 % of cycles in to reassure the safety of this technology once implantation occurs.
adolescents vs 6,7% in women 20-25 years(p=0,025). Trial registration number: not applicable
Limitations, reasons for caution: The data from this study were retrospec-
tive and from a single center. P-450 Low value of AMH did not affect oocyte quality and
Wider implications of the findings: This study shows that OC cycles in pregnancy outcome in patients with severe endometriosis
adolescents have similar results to 20-25 years women, the age expected to have A. Pacchiarotti1
a better response.It is plausible to include adolescents in FP programs due to 1
San Filippo Neri Hospital, IVF Unit, ROMA, Italy
their good response. This is very reassuring because these patients are less likely
to have completed childbearing. Study question: The aim of this pilot study was to determine that low
Trial registration number: Not applicable anti-müllerian hormone (AMH) serum levels due to severe endometriosis did
not affect IVF clinical outcomes in young patients (<37)
P-449 Developmental quality post-insemination of vitrified versus Summary answer: young patients with low AMH level due to severe endo-
fresh oocytes from patients with low ovarian reserve metriosis displayed a diminished oocyte yield but not a reduction in embryo
F. Bonald1, M. Montagut1, M. Clouet2, V. Guitard1, A. Chiron1, quality and pregnancy outcomes.
D. Nogueira3 What is known already: diminished ovarian reserve is associated to reduced
1
Labosud Garonne, Reproductive Biology Laboratory, Toulouse, France ; fertility and poor response to ovarian stimulation during in vitro fertilization (IVF)
2
Clinique Croix du Sud, Gynecology, Toulouse, France ; treatments
3
INOVIE Fertilité - Labosud Garonne, Reproductive Biology Laboratory - Clinique Study design, size, duration: A total of 50 IVF cycles of patients younger
Croix du SUD, Toulouse, France than 37 with severe endometriosis were retrospectively analyzed in a single
center between November 2016 and July 2018 and compared to a control group
Study question: Does oocyte developmental quality is affected after vitrifica- of 84 patients with no story of endometriosis and normal AMH value. AMH
tion in women with low ovarian reserve? value has been evaluated within three months before the stimulation. number
Summary answer: Embryonic developmental quality seems affected after and maturation of retrieved oocytes, embryo quality, and pregnancy outcomes
vitrification compared to freshly collected oocytes from the same patient were compared using Student’s t-test and Fisher’s test.
What is known already: Oocyte vitrification in low ovarian reserve patients Participants/materials, setting, methods: A total of 50 IVF cycles of
can be used for fertility preservation, to accumulate oocytes for optimizing the patients younger than 37 with severe endometriosis were retrospectively ana-
use of frozen-thawed-testicular and donor sperm, to augment the number of lyzed in a single center between November 2016 and July 2018 and compared
embryos in PGT cycles. Embryos from vitrified oocytes have similar implantation to a control group of 84 patients with no story of endometriosis and normal
potential than freshly collected oocytes from infertile patients but the number AMH value. AMH value has been evaluated within three months before the
of vitrified oocytes has an affect on delivery rates. Data on donors showed that stimulation. number and maturation of retrieved oocytes, embryo quality, and
in vitro development of embryos derived from vitrified-warmed donor oocytes pregnancy outcomes were compared using Student’s t-test and Fisher’s test.
is altered compared to their fresh sibling counterparts. Data on oocyte devel- Main results and the role of chance: The number of oocytes retrieved per
opmental quality after oocytes vitrification in patients with low ovarian reserve cycle and the percentage of mature oocytes (MII) were significantly (P<0.001)
is lacking. lower in IVF patients with severe endometriosis AMH value ≤ 1.1 ng/ml (Group
Study design, size, duration: A retrospective analysis performed A; 3.8±2.6 retrieved oocytes, 70% MII) compared to patients without endome-
between January 2016 and December 2018 including low ovarian reserve triosis and AMH levels > 1.1 ng/ml (Group B; 6.9±4.6 retrieved oocytes, 83%

MII). On the other hand, embryo morphology, implantation rate (31% vs 33%; “There is a lack of information. We do touch upon it but I think that with the
P=0.833) and pregnancy rate (50% vs 49%; P=1) were comparable in the current climate it’s such a significant part of people’s lives. I don’t think there’s
two groups. enough emphasis on training clinical staff when it comes to fertility…”
Limitations, reasons for caution: none HCP3, Nurse.
Wider implications of the findings: young patients with an impairment of Limitations, reasons for caution: One of the main methodological limita-
the ovarian reserve due to severe endometriosis displayed a diminished oocyte tions of this study is the self-selection process, which has implications for gen-
yield but not a reduction in embryo quality and pregnancy outcomes. These eralisability. The results necessarily reflect the views of those who were willing
results suggest that serum AMH levels should not be adopted as a criterion for to participate. For the lay population, the online recruitment method could result
discouraging those patients from undergoing IVF treatments. in potential bias towards respondents of higher socioeconomic status.
Trial registration number: not applicable Wider implications of the findings: School education remains a con-
sistent source of information but does not adequately cover fertility educa-
tion. In addition to websites based on robust scientific evidence and centres
specialising in sexual education; as an important source, there remains an
important need for additional training for primary HCPs on reproductive
NURSING AND MIDWIFERY health.
P-452 The Gestational Diabetes Mellitus (GDM) and Risk Factors

P-451 I trust the healthcare professional most: Exploring fertility After Assisted Reproductive Techniques (ART)
knowledge among healthcare professional and lay population N.N. Aypar Akbağ1, M. Aluş Tokat2, S. Fata3, E. Ünsal Avdal4,
groups R.E. Okyay3, Ö.E. Doğan5
B. Grace1, J. Shawe2, J. Sarah3, S. Judith1 1
Karamanoğlu MehmetbeyUniversity, Gynecology Nursing, Karaman, Turkey ;
1 2
University College London, Institute for Women’s Health, London, United Kingdom ; Work adres, Dokuz Eylul University Nursing Faculty, Izmir, Turkey ;
2 3
The University of Plymouth, Institute of Health and Community, Plymouth, United Dokuz Eylul University, Gynecology Nursing, Izmir, Turkey ;
4
Kingdom ; Katip Çelebi University, Internal Medicine Nursing, Izmir, Turkey ;
3 5
SPD GmbH, Clinical Research, Bedford, United Kingdom Dokuz Eylul University, Gynecology Medicine, Izmir, Turkey
Study question: What is the level of fertility knowledge amongst healthcare Study question: Do assisted reproductive techniques play a pivotal role in
professionals compared to lay men and women in the UK? gestational diabetes mellitus (GDM) incidence and risk factors for GDM in preg-
Summary answer: Healthcare professionals in this study did not appear to nancies after ART?
have better knowledge than the lay population in this study. Summary answer: The hormones stimulated in ART may cause lipid, carbo-
What is known already: As the average age of first time parents continues hydrate metabolism disturbances. Obesity, age, polycystic ovary syndrome in
to rise, health policies have highlighted the importance of optimising reproductive addition to ART may increase incidence of GDM.
health through better knowledge and awareness. This study therefore aimed to What is known already: External stimulation of hormones in assisted repro-
assess current patterns of fertility knowledge, attitudes and practices; and identify ductive therapies is known to cause disharmony in lipid, carbohydrate and pro-
improvement opportunities. In this study, fertility knowledge of lay men and tein metabolism. However, it is stated in the literature that the increase in the
women and HCPs was assessed using the same test instrument, which provided incidence of GDM may be caused by some confounding factors such as age,
a new and unique perspective compared to previous studies. obesity, polycystic ovary syndrome (PCOS) and multiple pregnancies developing
Study design, size, duration: Mixed method research was conducted using after ART.
a UK wide cross-sectional survey and semi-structed interviews. Results were Study design, size, duration: The retrospective cohort study was performed
obtained from 1,082 survey respondents which included 347 HCPs, 332 men, between December 2018 and February 2019 by using the file information of
and 413 women, 115 of whom were trying to conceive. There was variation in women who had been treated at Dokuz Eylul University InVitroFertilization (IVF)
age, level of education, ethnicity and training. Interviewees were purposively Center and gave birth between 2006-2018. One hundred eigth women were
sampled to include men and women from the reproductive age range reached.
(18-45 years) and of varying ethnic and educational backgrounds. Participants/materials, setting, methods: Women who treated at the IVF
Participants/materials, setting, methods: Survey participants were center between 2006-2018 were included. Giving birth after ART was inclusion
recruited nationwide via online newspaper and social media adverts. Of those criteria; Polycystic Ovarian Syndrome (PCOS), age 40 and over, Body Mass Index
who agreed to a follow-up interview, 35 were included this study. Healthcare (BMI) 30 and over, glucose intolerance, previous GDM history and corticosteroid
professionals (HCPs) were recruited from professional bodies such as RCN, treatment were the exclusion criteria. The women were called from center and
RCGP, RCP, doctors.org.uk. Interview data were transcribed and analysed using data was collected via telephone. A total 621 women registered in center, 108
the framework method. Interviews lasted an hour on average. Favourable ethical women were reached.
opinion was given by University College London Research Ethics Committee. Main results and the role of chance: The GDM was reported by 16.7% of
Main results and the role of chance: Survey questions covered the following women participating in the study. The results was interpreted as high incidence
areas: demographics, basic definitions, male and female reproductive biology, age-re- according to general incidence of GDM reported by American Diabetes
lated fertility decline and conception. The proportion of HCPs correctly answering Association’s incidence of GDM (1-14%). Ovulation induction drugs used by all
the survey knowledge questions was 47.0%(95% CI= 41.6%,52.4%) compared to women were rFSH (recombinant Follicle Stimulating Hormone) (70.4%), Human
47.3% for women(95% CI= 41.0%,52.2%); 50.9%(95% CI= 40.1%, 61.0%) for Menopausal Hormone (HMG) (16.7%), FSH and HMG (13.0%). The mean drug
Women trying to conceive sub group; and 36.3% (95% CI= 30.8%,41.6%) for men. starting dose was 257.98±110.95 and the total dose was 2797.47±1457.20 units.
Chi-squared test confirmed statistically significant difference amongst groups. 98.8% They used drugs for an average of 10.60±1.92 days. Progesterone use rate after
of HCPs stated that they provide fertility advice in their role. transfer was 65.7%. Using rFSH, HMG, together rFSH and HMG (KW=1.162,
Although the internet and school education remained popular sources, p=0.281), drug starting dose (U=668.000, p=0.236), the total dose of drug
HCPs were ranked as the most trusted source for seeking fertility and repro- (U=722.500, p=0.470), the duration of drug use (U=767.000, p=0.718), using
ductive health information, however, they did not appear to have better or not using of progesterone after transfer (KW=0.204, p=0.652) did not
knowledge than the lay population in this study. From interviews, there were increase the incidence of GDM.
inconsistencies among HCPs regarding where responsibility lies for providing Limitations, reasons for caution: Since the data were obtained from the
the right information and support to patients on fertility awareness. sample treated in a wide period of time, problems related to the recall factor
Recommendations were made for additional training on the topic part of were experienced. Also, some women could not be reached from their regis-
general HCP training. tered numbers in the centers.

Wider implications of the findings: This study suggests that women who Older women put a higher monetary value on increases in the livebirth rate
will receive ART should be screened for risk of GDM from the begining of than younger women. Partners tended to place a greater value on reductions in
pregnancy.The incidance can be higher in this group compared with natural treatment associated risks than those intending to get pregnant.
pregnancy. Studies with larger sample should be performed for specific deter- Limitations, reasons for caution: DCEs can elicit intentions which may
mimining the risk factors that can affect this process. not reflect actual behaviour. The external validity of this study is limited by
Trial registration number: not applicable the fact that it was conducted in a single centre with generous public funding
for IVF.
Wider implications of the findings: Couples undergoing IVF may be willing
to forgo higher livebirth rates for reduced maternal and perinatal risks. Along
POSTER VIEWING SESSION with evidence from randomised trials, these preferences should be used to
PSYCHOLOGY AND COUNSELLING inform policy as well as individualised decision making in IVF.
P-454 Pre-implantation genetic testing for aneuploidy;

motivations, concerns and perceptions in a UK population
P-453 Eliciting couples’ preferences for fresh versus elective
frozen embryo transfer in IVF: a discrete choice experiment B. Jones1, T. Bracewell-Milnes1, M. Spearman2, M. Jalmbrant3,
J. Green4, O. Rabi4, S. Saso1, P. Serhal4, J. Ben-Nagi4
B. Abdulrahim1, G. Scotland2, S. Bhattacharya3,
1
A. Maheshwari4 Imperial College London, Department of Surgery and Cancer, London, United
1 Kingdom ;
Aberdeen Fertility Centre, University of Aberdeen, Aberdeen, United Kingdom ; 2
2 University College London, Medicine, London, United Kingdom ;
Institute of Applied Health Science, University of Aberdeen, Aberdeen, United 3
Imperial College NHS Foundation Trust, Clinical psychology, London, United
Kingdom ;
3 Kingdom ;
School of Medicine- Medical Sciences and Nutrition, University of Aberdeen, 4
The Centre for Reproductive & Genetic Health, Fertility, London, United Kingdom
Aberdeen, United Kingdom ;
4
Aberdeen Fertility Centre, NHS Grampian, Aberdeen, United Kingdom Study question: What are women’s motivations and concerns in using pre-im-
plantation genetic testing for aneuploidy (PGT-A), and what are their perceptions
Study question: What are the factors influencing couples’ preferences for fresh after undergoing it as part of in vitro fertilisation (IVF) treatment?
versus elective frozen embryo transfer (ET) in in-vitro fertilisation (IVF)? Summary answer: Most women use PGT-A to optimise the chance of suc-
Summary answer: Accepting that elective frozen ET can delay treatment but cessful embryo transfers, are satisfied after treatment, and would use it in future
reduces the risk of OHSS, couples’ choices are driven by expected pregnancy IVF cycles
and birth outcomes. What is known already: PGT-A facilitates embryo selection by allowing the
What is known already: Elective frozen embryo transfer results in livebirth opportunity to prioritise chromosomally normal embryos for transfer to the
rates which are on par with, or better than those following fresh embryo transfer. uterus. The transfer of embryos adjudged to be euploid using modern PGT-A
In terms of safety, elective embryo transfer reduces the risk of ovarian hyper- methods is associated with higher implantation, ongoing pregnancy and delivery
stimulation (OHSS) and small for gestational age babies but increases the risk rates with reduced pregnancy loss compared to unscreened embryos, thus mit-
of pre-eclampsia and large for gestational age compared to fresh embryo transfer. igating the negative clinical impact maternal age has upon reproductive outcomes.
Given the prevailing uncertainty around a universal policy of elective frozen Despite the upward trend in women utilising PGT-A, there is little data about
embryo transfer for all couples undergoing IVF, user preferences are critical in the women’s motivations and concerns toward using the technology, and per-
informing clinical decision making. ceptions having gone through the process.
Study design, size, duration: Discrete choice experiments (DCEs) are a pref- Study design, size, duration: This was a cross sectional survey which included
erence elicitation technique for assessing the value that individuals derive from 161 women who used PGT-A in a single fertility clinic in the UK between
different aspects of a treatment or service. We asked both partners of 104 IVF 2014-2018.
naïve couples attending a tertiary referral centre to independently complete a Participants/materials, setting, methods: An online questionnaire was
questionnaire with nine hypothetical choices illustrating potential advantages and created to assess motivations and concerns toward PGT-A as well as outcomes
disadvantages of fresh versus frozen embryo transfers in terms of four attributes and perceptions following IVF using PGT-A. Motivating and concerning factors
(live birth rate, miscarriage rate, neonatal complication rate and cost of treatment). were quantified out of 10 depending on perceived significance (0=insignificant;
Participants/materials, setting, methods: Participants were informed that 10=very significant). A mixture of closed and Likert scaled questions were used
frozen ET would delay the transfer of embryos by 6-8 weeks but reduce the risk to assess perceptions to PGT-A after treatment. Descriptive statistical analysis
of OHSS. An opt-out (no IVF treatment) was included for each choice task. was undertaken using mean +/- SD.
Logistic regression was used to analyse the choice response data and estimate Main results and the role of chance: 333 women were invited to partici-
preference weights for fresh and frozen ET and each treatment attribute. pate. The response rate was 48.3% (n=161). The most common reasons to use
Willingness to pay (WTP) was calculated as the marginal rate of substitution PGT-A was advanced maternal age (n=84;35.1%) and repetitive (≥2) unsuccess-
between the cost attribute and each other attribute. ful IVF cycles (n=77;32.2%). The most significant motivating factors to undergo
Main results and the role of chance: A total of 104 couples completed PGT-A were to improve the probability of having a baby per embryo transfer
208 questionnaires. Couples preferred both fresh and elective frozen embryo (9.22+/-1.8) and optimise the chance of implantation (8.85+/-2.52). The most
transfer (Odds ratio 27.65 and 27.7 respectively) compared to no IVF treatment significant concerning factors identified were not having any embryos to transfer
with no strong preference for fresh over frozen. (5.76+/-3.4) and the potential for damage to be caused to embryos (5.45+/-
Couples strongly preferred any IVF treatment options that increased the live- 3.1). More than three quarters (n=122;77.2%) achieved at least one live-birth
birth rate by 5% (OR 1.28; 95% CI; 0.070 - 0.424), reduced the miscarriage rate using IVF with PGT-A. Two thirds (n=81;66.4%) of those who had a livebirth
by 18% (OR 0.54; 95% CI; 0.788 - 0.454) and diminished neonatal complication planned to tell their children regarding the use of PGT-A, whereas a minority
rates by 10% (OR 0.59; 95% CI; 0.697 - 0.357). (n=10;8.2%) did not intend to. The majority of participants were satisfied/very
Respondents were willing to pay an additional £2452 (95% CI; 596 - 4,308) satisfied with their experience with PGT-A (n=117;73.6%), whereas 15.7%
and £7,168 (95% CI 5,053 – 9,283) for a 5% and 15% increase in the live birth (n=25) were dissatisfied/very dissatisfied. Of those that were dissatisfied/very
rate respectively. They required compensation of £5,230 (95% CI; 3,320 -7,141) dissatisfied, almost two thirds (n=16;64%) did not achieve a live-birth. In those
and £13,245 (95% CI; 10,110 – 16,380) for treatments that increased the neo- who were satisfied/very satisfied, just 8.5% (n=10) did not have a successful
natal complication rate by 10% and 25% respectively. Couples valued a 10% live-birth. More than two thirds (n=108;67.9%) would use PGT-A in future IVF
diminution in the risk of neonatal complications and a 10% increase in the live- cycles, and more than three quarters (n=123;77.4%) would recommend it to
birth rate equally. family and friends.

Limitations, reasons for caution: The data from this study was retrospective not achieved due to other reasons (e.g., lack of partner, partner who does not
and from a single centre. want children) and of men.
Wider implications of the findings: PGT-A offers an opportunity to opti- Wider implications of the findings: This study shows that a psychological
mise reproductive outcomes. This study highlights that, in a UK population, intervention based on the 3TM is perceived to be acceptable and useful, sug-
perceptions amongst women who undergo PGT-A as part of IVF are mostly gesting it successfully responds to an unmet need for support from infertile
positive. We clearly identify that women are more satisfied following treatment women with an UPG. Future work will focus on improving the intervention as
if a resultant livebirth is achieved. per participants’ feedback and testing its efficacy.
Trial registration number: not applicable Trial registration number: Not applicable
P-455 Prospective acceptability study of a psychological online P-456 Promoting fertility awareness and preconception health
self-help intervention for individuals with an unmet parenthood using a chatbot: A randomized controlled trial
goal (UPG) E. Maeda1, A. Miyata2, J. Boivin3, K. Nomura1, Y. Kumazawa4,
B. Rowbottom1, S. Gameiro1 H. Shirasawa4, H. Saito5, Y. Terada4
1 1
Cardiff University, School of Psychology, Cardiff, United Kingdom Akita University Graduate School of Medicine, Environmental Health Science and
Public Health, Akita, Japan ;
Study question: How acceptable is a psychological online self-help intervention 2
Dokkyo Medical University, Reproduction Center, Saitama, Japan ;
for individuals with an UPG? 3
Cardiff University, School of Psychology, Cardiff, United Kingdom ;
Summary answer: Overall the self-help intervention was found to be accept- 4
Akita University Graduate School of Medicine, Obstetrics and Gynecology, Akita,
able and useful, providing structured support for individuals with an UPG, with Japan ;
minor amendments suggested. 5
Umegaoka Women’s Clinic, ART Center, Tokyo, Japan
What is known already: Individuals who are unable to achieve the family
they desire, remaining childless or having fewer children than desired, have an Study question: What are the effects of using a fertility education chatbot on
UPG. People with an UPG can experience adjustment problems, resulting in knowledge, intentions to improve preconception behaviour, and anxiety?
poorer mental health and well-being. An in-depth systematic review of how Summary answer: Providing fertility education using a chatbot improved
people adjust to an UPG (Gameiro and Finnigan, 2017) generated the Three fertility knowledge and intention to optimize preconception behaviour without
Task Model (3TM). This proposes three psychological mechanisms that facilitate increasing anxiety.
adjustment to an UPG: acceptance, meaning making and pursuit of new life goals, What is known already: Providing fertility information through educational
leading to better mental health and well-being. To date there is no widely used brochures improves fertility knowledge and changes intentions regarding child-
evidence-based interventions for individuals facing an UPG. bearing, but increases anxiety. Chatbots are automatic conversation programs
Study design, size, duration: A prospective qualitative acceptability study that are increasingly used in health education, but not yet for fertility awareness.
of an online self-help intervention based on the 3TM was carried out. Twelve Study design, size, duration: A three-armed randomized controlled trial
participants were interviewed twice: first to obtain their initial reaction to the was conducted using an online social research panel in Japan in March 2019. All
intervention (T1); and eight weeks later to collect their perspectives of the 927 participants were randomized and exposed to one of three materials: a
intervention once they had engaged with it (T2). The interviews focused on chatbot (intervention group, IG), a document about fertility and preconception
assessing participant’s perceptions of how acceptable (willingness to use inter- health (control group 1, CG1), or a document about an irrelevant topic (control
vention) and important/useful (perceived benefits of using intervention) the group 2, CG2).
intervention is. Participants/materials, setting, methods: Participants were women
Participants/materials, setting, methods: Each participant was inter- aged between 20 and 34 years old who hoped to have (more) children in the
viewed via Skype and audio recorded. At T1, participants were asked to ‘think future. The scores for the Japanese version of the Cardiff Fertility Knowledge
aloud’ to the researcher as they engaged with the intervention for the first time. Scale and intention to optimize eight preconception behaviours among those
A semi-structured interview then took place, followed by a short questionnaire. who did not exhibit that behaviour were assessed immediately before and
The interview and questionnaire were repeated at T2 to gather participants’ after exposure to the material. The scores for the State-Trait Anxiety Inventory
perspective after engaging with the intervention. Interviews were transcribed and the free-text feedback provided by chatbot users were assessed after
and analysed using thematic analysis. exposure.
Main results and the role of chance: The analysis resulted in 8 main themes, Main results and the role of chance: Baseline characteristics were well
organized under 3 higher order themes: acceptability, importance, and other. balanced between groups. A repeated-measures analysis of variance showed
Themes under acceptability reflected that participants considered the inter- significant fertility knowledge gains after the intervention in the IG (+9.1
vention acceptable due to being easy to use, comprehensive and appropriate. points) and CG1 (+14.9 points), but no significant change in the CG2 group
Nearly all participants used the intervention individually, with one using it ver- (+1.1 points). Post-test increases in the intention to take folic acid, to receive
bally with their partner. Just under half engaged with it digitally, the rest printed human papilloma virus vaccination, to obtain a primary obstetrics and gynae-
out the activities. Barriers to acceptability included limited digital access, poor cology doctor, to take oral contraceptives, and to try to get pregnant were
interactivity, and unclear organisation. A minority of participants felt some significantly higher in the IG than in CG2 and were similar to that in CG1.
sections were difficult to understand and some language was not always Post-test state anxiety scores (mean ± SD) were significantly lower (less
appropriate. anxiety) in IG (43.2 ± 9.5) than in CG1 (47.5 ± 9.5) and CG2 (46.2 ± 9.0),
Themes under importance showed that all participants felt the intervention all P <0.001. Three themes emerged from user feedback about the chatbot:
was useful and helpful, given that it provided a structure to organise and clarify technical limitations (e.g., instability of the system such as freezing, low com-
thoughts, whilst providing guidance to move through their loss. Around two prehension of users’ words); pros and cons of using chatbot (e.g., easy and
thirds of participants initially felt concerned that engaging with the intervention convenient versus robotism and coolness); and experiences of learning about
might be challenging. However, a majority experienced no negative effect engag- fertility and preconception health.
ing with the intervention, with three participants reporting they felt upset Limitations, reasons for caution: The possibility of selection bias associated
at times. with the use of a social research panel and volunteer bias toward those more
Themes under other highlighted that participants saw their experience of interested in fertility may limit the generalizability of these findings.
adjusting to UPG as a journey and that the intervention could facilitate this. Finally, Wider implications of the findings: The improvement in fertility knowl-
participants thought that connecting with others is important and that the inter- edge was smaller than that of participants who read a well-written booklet but
vention can support this. applied on a large scale may still warrant implication. Further technical devel-
Limitations, reasons for caution: The study had a small homogeneous opment of the chatbot and exploration of personal affinity for technology is
sample of childless married infertile women and results may not reflect the views required.
of people who have children (but desire more), whose parenthood goals were Trial registration number: UMIN Clinical Trials Registry (UMIN000035736)

P-457 The age of the patient at the time of decision making is the 1
Ghent University Hospital, Department for Reproductive Medicine, Ghent,
only factor associated with the future use of conditionally blocked Belgium ;
donors 2
Universitair Ziekenhuis Brussel, Center for Reproductive Medicine, Brussels,
V. Muyshond1, A. Tolpe1, A. De Baerdemaeker1, K. Tilleman1 Belgium ;
3
1 CHU St Pierre, Center for Reproductive Medicine, Brussels, Belgium ;
Ghent University Hospital, Reproductive Medicine, Gent, Belgium 4
Hôpital Erasme- Université Libre de Bruxelles, Fertility Clinic, Brussels, Belgium ;
5
Study question: Which patient characteristics are predictive for further use Clinique CHC Mont Légia, Centre de Procréation Médicalement Assistée, Liège,
of a sperm donor after the donor has been conditionally blocked? Belgium ;
6
Summary answer: The younger a patient is when confronted with the use CHR Citadelle, Centre de Procréation Médicalement Assistée, Liège, Belgium ;
7
of a quarantined sperm donor, the more likely she will use it for future Chirec-hôpital de Braine l’alleud, Centre de Procréation Médicalement Assistée,
treatments. Braine l’alleud-Waterloo, Belgium ;
8
What is known already: Harm to a fetus via transmission of disease is con- Leuven University Hospital, Leuven university fertility center, Leuven, Belgium ;
9
sidered a severe adverse reaction and needs to be reported through a biovigi- AZ Sint-Jan, Center for Reproductive Medicine, Bruges, Belgium ;
10
lance system. Karyotype screening and carrier screening for cystic fibrosis and Cliniques universitaires Saint-Luc, Gynécologie- andrologie et fertilisation in vitro,
spinal muscular atrophy are mostly performed in sperm donors and although Brussels, Belgium ;
11
this reduces the risk for transmission of the latter diseases, it does not diminish Antwerp University Hospital, Center for Reproductive Medicine, Antwerp, Belgium
the risk for transmission of other conditions (Isley L. et al. 2016). When a donor
child is born with a condition, sperm donors are conditionally blocked, distribu- Study question: What is the dropout rate for couples with remaining cryo-
tion of semen restricted. Patients are notified and decisions on further use of preserved embryos and what are the reasons for postponing or discontinuing
the donor are necessary. the transfer of cryopreserved embryos?
Study design, size, duration: A retrospective descriptive analyses was Summary answer: The dropout rate is 9.0% in patients with remaining
performed of sperm donors imported from EU sperm banks that were blocked cryopreserved embryos. Main reasons to discontinue their treatment are psy-
in the period from 01/01/2010 to 31/12/2019. From this patient population, chological/physical burden and impact on work.
a subgroup was selected based on the fact that these patients had already given What is known already: In fresh ART cycles, the physical and psychological
birth to a child of this donor or were in treatment at the moment of the burden associated with an ART treatment is the major reason for patients to
blocking. This subgroup of patients had to decide to further use the donor delay or even discontinue their treatment. For IVF and ICSI, rates of discontin-
or not. uation vary in Europe from 17 to 70%. In addition, discontinuation of ART
Participants/materials, setting, methods: In total 767 patients had been treatment may contribute to compromised cumulative pregnancy rates in ART.
treated with donors (n=196) that were subsequently blocked. Patients who had In case of the presence of cryopreserved embryos, delaying treatment is prob-
to make a decision on further use of the donor were selected. Variables like ably less frequent given the lower burden of a natural or artificially prepared
country of residence, relation type, children conceived and number of children frozen embryo transfer cycle. Nevertheless, no data are available regarding rates
born, age patient at time of decision and the straws left were included. Mixed and reasons for dropout in this population.
model binary logistic regression and Fisher’s exact test was used for statistical Study design, size, duration: This multicenter study included patients from
analysis (p≤0.05). 11 Belgian fertility centers between 2012 and 2017. Patients were considered
Main results and the role of chance: Of the total population, 292 patients dropouts (n = 1917) in case they underwent an unsuccessful fresh (“fresh group”)
were informed about the condition of the donor and a selected sample size of or cryo (“in between group”) embryo transfer and did not start a subsequent
184 patients had to make a decision on further use of in total 104 blocked cryo cycle within one year.
donors. 87/184(47%) of the patients decided not to use the donor for future Participants/materials, setting, methods: An online anonymous ques-
treatments, compared to 57/184(31%) decided to further use and 40/184(22%) tionnaire, aiming to explore the reasons for dropout, was developed, approved
have not decided yet. by all the ethical committees and sent to the eligible patients. An unsuccessful
At the time of decision making, patients had on average 3.2 straws left of the embryo transfer was defined by no pregnancy, a biochemical pregnancy, an
donor and the average age of the patient was 36.23years. Of the 292 patients, ectopic pregnancy or a miscarriage <12 weeks. Exclusion criteria were cryo-
144 made an informed decision on the further use of the donor. preservation of embryos in case of preimplantation genetic testing, fertility
More than half of the lesbian couples chose to not further use the straws preservation cycles and oocyte donation cycles.
(55% (42/77)), 69%(29/42) of the heterosexual couples and 64%(16/25) of Main results and the role of chance: The overall treatment dropout rate
the single mothers decided not to use the blocked donor in further treatments for couples with remaining cryopreserved embryos is 9.0%. The dropout rates
(p=NS). Binary logistic regression analysis including multiple variables showed in the “fresh” and “in between” group are 9.7% and 8.0%, respectively. The
that only the age of the patient had predictive value for deciding not to further response rate for the online questionnaire was 15.8% (n = 304/1917). The most
use the straws: the average age of a patient deciding not to use the donor (av±SD important reasons for dropout are psychological (50%) and physical (43%) bur-
37.77±5.45) is higher compared to the average age of a patient deciding to den, the impact on work (29%), age of the woman in treatment (25%) and the
further use the donor (av±SD 33.88±4.67) (p=0.02). impact on relationship (25%). The decision of the couple to postpone or stop
Limitations, reasons for caution: Of the selected sample size, 22% of the treatment is influenced by external factors in only 16% of the cases. Seventy-eight
patients did not yet decide on the future use of the donor. There might be more percent of respondents are satisfied with the care delivered by their fertility
intrinsic factors related to the decision making of the patients that are not center, and 92% would recommend the center to peers. Out of suggested
included in the analysis. improvements to the respondents, psychological support before (41%), during
Wider implications of the findings: The future use of a donor is probably (51%) and after (51%) treatment, as well as lifestyle counseling (44%) and receiv-
related to the fact that the genetic link between future siblings is important to ing digital information (43%) are most frequently selected.
the parent(s), regardless of the type of relationship the patient is in. However Limitations, reasons for caution: As questionnaires cannot fully capture
more research into decision making of patients in these cases is very much emotional responses or feelings, open answer boxes were added to the 10-point
needed. Likert scale used for some of the questions. Although the response rate is quite
Trial registration number: B2015/1014, amendment 2020 high compared to similar studies in the field, the results should not be extrapo-
lated to the entire dropout population.
P-458 Discontinuation of treatment in subfertile couples with Wider implications of the findings: Based on our data, patients highly
cryopreserved embryos: a multicenter study on rates and reasons recommend enhanced psychological support throughout the entire ART treat-
of dropout. ment, as well as lifestyle advice and digital communication regarding information
F. Vanden Meerschaut1, C. Blockeel2, S. Blaiberg3, A. Delbaere4, relevant for the treatment. This stresses the need to better integrate these
A. Delvigne5, L. Henry6, R. Imbert7, S. Lie Fong8, A. Van De aspects of patient centered care in our daily ART practice.
Vijver9, C. Wyns10, D. De Neubourg11 Trial registration number: not applicable

P-459 What happens in the counsellor’s room? A qualitative study P-460 Reproductive choices after oocyte vitrification for age-
on the aims of psychological intervention in Italy related reasons.
Z. Donarelli1, G. Lo Coco2, S. Gullo2, V. Oieni1, A. Marino3, J. Nekkebroeck1, L. Leunens1, E. Buyse1, H. Tournaye1, M. De Vos1
A. Allegra3 1
UZ Brussel, Centrum voor Reproductieve Geneeskunde, Jette- Brussels, Belgium
1
Andros Day Surgery Clinic, Psychology Unit, Palermo, Italy ;
2
University of Palermo, Department of Psychology- Educational Science and Study question: What is the relational status of women who returned to the
Human Movement, Palermo, Italy ; fertility clinic after planned oocyte vitrification?
3
Andros Day Surgery Clinic, Reproductive Medicine Unit, Palermo, Italy Summary answer: Of women who returned to the clinic after planned oocyte
cryopreservation, 48% (59/122) had found the right partner to pursue
Study question: What type of intervention, methods and goals guide the childbearing.
Mental Health Professionals’ (MHPs) work with couples in infertility counselling? What is known already: The profile of women seeking planned oocyte
Summary answer: Although Italian MHPs’ interventions follow different vitrification has been quite stable during the past decade. Most women are in
approaches in infertility counselling, there is a consensus on aiming to strengthen their mid-and late thirties, are highly educated and have a career. These women
personal generativity beyond the goal of pregnancy. attribute their childlessness to ‘not having a suitable partner’ rather than to
What is known already: Within the context of infertility, three types of ‘prioritisation of career achievements’. Hence, the main driver for planned
counselling are commonly adopted (implication counselling, support counselling oocyte cryopreservation is to buy more time to find a suitable partner. The
and therapeutic counselling) and ESHRE has recognized the need for providing return rates of these women to the fertility centre and the utilisation rate of the
integrated psycho-social care by the entire staff. Although some evidence-based cryopreserved oocytes are only 7-12%. Little is known about their reproductive
guidelines (which should guide psychological interventions in infertility clinics) choices.
focusing on the themes and aims of psychological counselling have been provided Study design, size, duration: In a single centre, computerized clinical data
in recent years, there is scarce evidence on the MHP’s adherence to these man- were retrieved from women who underwent planned oocyte vitrification for
uals. Moreover, there is a dearth of research on how MHPs really conduct their age related reasons between January 2009 and December 2018 (n=668) and
interventions in countries where clinics are not compelled to include MHPs in from those who returned (n=122) to the centre to discuss their reproductive
their staff. options and/or to proceed with reproductive treatment.
Study design, size, duration: This was a cross-sectional qualitative study of Participants/materials, setting, methods: We collected data from com-
14 experienced MHPs who have been working in public and private infertility puterised clinical charts of women who applied for reproductive treatment after
clinics in different regions of Italy. All professionals were individually interviewed planned oocyte vitrification. We evaluated whether women actually started
during a five-month period from June to October 2019. Semi-structured inter- treatment, assessed their relational status (single or in a relationship) and the
views were held in Italian via Skype or phone and lasted between 60 and 80 utilisation rate of vitrified oocytes and treatment outcome.
minutes. Each interview was transcribed verbatim. Main results and the role of chance: 18.2% (122/668) women returned
Participants/materials, setting, methods: Eighteen invitations to partici- to the fertility centre. Four were beyond the age limit of IVF in Belgium and
pate in an interview-based study were sent to MHPs and 14 agreed to participate. requested transport of their oocytes to a centre abroad. For four women the
The interview guide was devised based on themes identified in the literature cryopreservation period of 10 years had expired. Upon return to the clinic, 55
review. Data were analyzed thematically using NVivo software. A coding frame- women were single and 59 had found a partner. 28.8% (17/59) of women with
work was drafted by the Principal Investigator and reviewed by the team. The a partner gained information but did not proceed with fertility treatment. No
entire data set was then coded by two researchers, using the coding comparison less than 42% (23/55) of single women refrained from fertility treatment after
function in NVivo, reporting fair to good agreement. counselling by a gynaecologist and psychologist. For 60.9% (14/23) of those, a
Main results and the role of chance: An all-encompassing thematic descrip- genetic reference for their child was utmost important whilst only 3/23 requested
tion of the data set was produced. The key themes were classified under the an anonymous sperm donor. 60.7% (74/122) women proceeded with repro-
following thematic headings: providing implication counselling to couples attend- ductive treatment (artificial insemination, IVF or ICSI) of which 42 had a partner
ing infertility treatment or facing repeated failures; the couple as the unit of and 32 were single. The utilisation rate of vitrified oocytes in women who
counselling intervention; strengthening couples’ commitment to the course of returned was 37% (45/122). Of these 45 women who used their vitrified
treatment. oocytes, 17 had an ongoing pregnancy and 4 already delivered a baby.
Almost all MHPs reported facing some challenges, when working with patients Limitations, reasons for caution: Patients who did not return to the centre
who had received donated gametes, such as addressing the need to build an were not included. Hence, the reproductive pathway of only a subset of women
affective parental bond beyond genetic heritage; MHPs reported a different who had their oocytes cryopreserved was analysed. Follow-up of reproductive
stance from physicians when responding to a couple’s requests for advice on outcomes in women who did not return is required for a comprehensive
managing the disclosure issue, often showing a tendency to hold back their appraisal of planned oocyte vitrification.
personal inclinations; moreover, most MHPs experience difficulties in responding Wider implications of the findings: Almost 50% of women who returned
to physicians’ expectations (i.e. to obtain a quick patient response and relief to the fertility centre with a desire for pregnancy had found a partner. After
from anxiety and doubts/fears, or an early evaluation of psychiatric disorders counselling, an important subset (42%) of single women decided not to pursue
to avoid possible legal proceedings in the future). single motherhood.
Overall, the results suggest that MHPs would like to have greater adherence Trial registration number: not applicable
to ESHRE psycho-social routine care guidelines, with a need for further integrated
care by the entire staff.
P-461 Impact of pre-treatment optimism on emotional health
However, MHPs seem to provide counselling with common strategies, pur-
during the waiting period and after treatment results
suing the same aims, albeit with huge differences in treatment types and settings.
Limitations, reasons for caution: The auto-selection of participants may J. Boivin1, H. Ockhuijsen2, R. Morey3, A. Van den Hoogen4,
have resulted in a sample representing those with greater clinical experience A. Domar5, N. Macklon6
1
and having seen a large number of patients. Moreover, the findings may under- Cardiff University, School of Psychology Cardiff Fertility Studies Research Group,
estimate the ways in which infertility counselling is delivered outside infertility Cardiff- Wales, United Kingdom ;
2
Clinics. University Medical Center Utrecht, Women & Baby Unit, Utrecht, The Netherlands ;
3
Wider implications of the findings: This is the first study aiming to explore Cardiff University, School of Psychology, Cardiff- Wales, United Kingdom ;
4
the characteristics of psychological counselling from the MHPs’ perspective. University Medical Centre Utrecht, Birth Center Neonatology, Utrecht, The
Although some heterogeneity across clinical approaches to patients and settings Netherlands ;
5
exists, MHPs do deliver counselling in general attunement with couples’ needs, Harvard Medical School, Obstetrics- Gynecology- and Reproductive Biology,
with efforts to break the barriers of non-integrative care. Boston, U.S.A. ;
6
Trial registration number: Not applicable London’s Women’s Clinic, London’s Women’s Clinic, London, United Kingdom

Study question: What is the association between pre-IVF success expectations Summary answer: As parent’s experiences of ‘fulfilling’ conditions of par-
and wellbeing during and after treatment? enthood vary, so does their willingness to integrate the donor in the parent-off-
Summary answer: Patients were more optimistic than they perceived their spring-donor triad.
doctors to be but this optimism was not harmful to wellbeing during or after the What is known already: In Sweden, offspring conceived with donor gametes
pregnancy test are entitled to obtain identifying information about the donor when reaching
What is known already: Many professionals worry that excessive patient mature age. Since this law was introduced in 1985, very few of the adult offspring
optimism at the start of IVF will negatively impact emotional reactions during who are eligible to obtain donor information have exercised this right. There is
treatment and after a treatment cycle. Staff report managing expectations to be limited knowledge about the experiences of parents with adult offspring following
a major challenge in working with patients. Patients tend to over-estimate the identity-release donation.
chance of success with treatment, even among groups with characteristics that Study design, size, duration: Based on a parallel multicenter study of adult
should reduce expectations of success (e.g., older age). The effect of high opti- offspring who had requested and received donor information at Swedish clinics,
mism on wellbeing during and after treatment results is not known. a purposive sample of their parents were approached during 2018-2019.
Study design, size, duration: The study comprised a secondary analysis of Participants/materials, setting, methods: Participants were 23 parents
data from the 20-month Positive Reappraisal Coping Intervention (PRCI) ran- (15 women and 8 men) with varying educational and socioeconomic background
domised controlled trial (Ockhuijsen 2014). Only data from the two control and from urban and rural areas throughout Sweden. In these families, offspring’s
groups not receiving PRCI were used in analysis. Of 1445 invited, 230 were relation to the donor ranged from no further interest, considering future con-
randomised to the control groups. Participants completed questionnaires prior tact, occasional contact and regularly meeting with the donor. Individual
to treatment (Time 1, T1), 10 days after embryo transfer during the waiting semi-structured interviews with parents in heterosexual couples were con-
period (Time 2, T2), and six weeks after embryo transfer (Time 3 T3). ducted face-to-face or via telephone, transcribed verbatim and analyzed using
Participants/materials, setting, methods: Participants were recruited from thematic analysis.
a fertility clinic if being female gender and undergoing stimulated/cryopreserved Main results and the role of chance: Two overarching themes were con-
IVF/ICSI cycle. Patient optimism was measured as a difference score between structed as (a) Fulfilling conditions of parenthood, and (b) Parents’ handling of
their ratings (0 to 100%) on: “What do you personally think is your chance of the parent-offspring-donor triad. Conditions of parenthood, e.g. the importance
success with your treatment cycle?” and “What do you recall your doctor telling of parent-offspring resemblance or importance of achieving genetic parenthood,
you was the chance of success with your treatment cycle?”. At T1, T2, T3 women are set by parents and society, and are to varying extents fulfilled by the parents.
completed the Hospital Anxiety and Depression Scale. The extent to which parenthood is experienced as ‘fulfilled’ appear to further
Main results and the role of chance: Mean personal chance of success influence how the parent handles the parent-offspring-donor triad. For some,
42.70% (SD=22.76) was significantly higher than recalled doctor chance of preg- the donor is included in the family, as a new father figure, or as an extension of
nancy 30.42% (SD=16.52) (n=157, t(156)=6.57, p < .000). The difference the family. For others, the donor is kept at a distance from the family, perceiving
between self and doctor estimates decreased as doctor estimates were reported his intentions as non-relational, or as a threat to parenthood. I.e., as parents to
to be more optimistic (F(3, 153)=4.58, p <.004). Regression was used to exam- varying extents are confronted with conditions of parenthood, and to varying
ine association between patient optimism and wellbeing (anxiety, depression) extents ‘fulfill’ those conditions, they develop their lines of action to the off-
during and after treatment results (controlling for pre-treatment anxiety and spring’s search for identifying information about his/her sperm donor on a con-
depression). Overall regression showed that after controlling for pre-treatment tinuum from excluding to including the donor in the family. In cases where the
values, patient optimism was not associated with anxiety (F(1, 130)=.497, donor holds more ambiguous roles, or where there is disagreement in regards
p. = 482, B=-.049 [95% CI -.041 - .020]) or depression F(1, 130)=.481, to the importance of the donor between the parent and the child, friction may
p = .489, B=-.044 [95% CI -.031 - .015]) during the waiting period. Similarly, arise within the parent-offspring-donor triad.
after controlling for pre-treatment and waiting period scores, pre-treatment Limitations, reasons for caution: Concerning transferability to other pop-
patient optimism was not significantly associated with post-treatment anxiety ulations, it should be considered that the study was performed, and parents’
(F(1, 116)=.000, p =.993, B=-.001 [95% CI -.008 - .993]) or depression experiences were given, within the context of the Swedish legislation on iden-
(F(1, 116)=.202, p. = .654, B=-.031 [95% CI -.020 - .032]). tity-release donation.
Limitations, reasons for caution: Participants were in the control arms of Wider implications of the findings: The present findings highlight the com-
an RCT testing a psychological intervention. Optimism was measured only in plexity of providing adult offspring access to identifying information about the
reference to what patients believed the doctor said, and not what the doctor donor. Parents following identity-release gamete donation may benefit from
actually said. About 36.5% (n=89) of women reported not being told a chance counselling and support to manage family life with varying genetic linkage within
of pregnancy with treatment. and outside the family unit.
Wider implications of the findings: Pre-treatment optimism may be a Trial registration number: not applicable
cognitive heuristic patients use to initiate treatment that does not markedly affect
wellbeing in treatment. Future research should replicate findings using doctor P-463 Let’s talk about it: exploring attitudes towards engagement
reports and a wider range of wellbeing and psychological measures. Fertility staff and open discussion of (in)fertility and reproductive heath on
should strive for accurate expectation management but not discourage optimism. social media
Trial registration number: not relevant S. Tothill1, B. Grace1, P. Zadeh1, D. Patel1
1
University College London, Institute for Women’s Health, London, United Kingdom
P-462 Parent’s experiences of identity-release sperm donation as
adult offspring obtain donor information Study question: What is the level of engagement of users on (in)fertility and
A. Widbom1, S. Isaksson2, G. Sydsjö3, A. Skoog-Svanberg4, reproductive health topics two popular social media platforms?
C. Lampic1 Summary answer: Level of engagement is high, especially where celebrities
1
Karolinska Institutet, Women’s and Children’s Health- Division of Reproductive are involved.
Health, Stockholm, Sweden ; What is known already: The media remains a powerful influence on
2
Uppsala University, Department of Public Health and Caring Sciences, Uppsala, health-related behaviour, warranting further critical examination. Social media
Sweden ; platforms like Twitter, Facebook, and Instagram have rapidly redefined the pro-
3
Linköping University, Division of Obstetrics and Gynaecology- Department of cess of communication between healthcare workers, patients and public, and
Clinical and Experimental Medicine, Linköping, Sweden ; extremely popular way of disseminating both health information and individuals’
4
Uppsala University, Department of Women’s and Children’s Health, Uppsala, opinions and own health experiences. Yet little is documented about social media
Sweden users’ levels of engagement and discussion of the topic of (in)fertility, traditionally
considered a taboo subject. As of January 2020, there are 1billion Instagram
Study question: How do parents experience and handle their adult offspring’s users and 363million twitter users globally. This study was conducted to explore
search for identifying information about their sperm donor? engagement on fertility topics on these platforms.

Study design, size, duration: Data from both Instagram, a photo and vid- regarding individuals’ reproductive plans. Studies exploring the complex rela-
eo-sharing social networking service and Twitter, a microblogging and social tionship between the perceived threat of infertility and undertaking fertility
networking service were analysed, to understand the engagement of users protective behaviours are needed.
regarding the topic of (in)fertility. Study design, size, duration: For this cross-sectional study, childless women
Participants/materials, setting, methods: In order to find relevant posts, were invited to participate, at private gynaecology clinics and through social
Google searches were initially conducted. The most popular posts were all networks between July 2016 and February 2018. The eligibility criteria were:
celebrities’, which were then searched on the 2 platforms over a 2 month period, being involved in a romantic heterosexual relationship (≥1 year); desiring to have
ending in January 2020 and analysed. Legitimate accounts of public figures are children in the future; being between 20-45 years of age; not having knowledge
confirmed via ‘verified’ badges on Instagram and Twitter. The number of fol- of a fertility problem, and not being actively trying to conceive for ≥12 months
lowers, likes, the topics discussed and comments were all documented, analysed (or 6 months, if women aged ≥ 35).
and categorised into themes. Participants/materials, setting, methods: The sample is composed of
Main results and the role of chance: In the past two years, many celebrities 240 childless women committed in a heterosexual relationship who desired to
such as Amy Schumer, Anne Hathaway, Kim Kardashian-West and Chrissy have children in the future. Women completed a self-reported questionnaire
Teigen, all millennials, have openly discussed their reproductive health; specifically including measures about their reproductive project, barriers, and facilitators to
their fertility issues and trouble conceiving. These women have over 211 million achieving their reproductive goals, and infertility’ susceptibility and severity. A
followers on Instagram and nearly a million on Twitter, showing the incredible mediation model using PROCESS was used to test whether the relationship of
reach their posts have. Their latest Instagram stories and posts regarding their the perceived threat of infertility and intention to have children earlier is medi-
fertility have been liked by over 16 million people, each having thousands of ated by barriers and facilitators.
comments from users showing support, advice, their own personal experiences Main results and the role of chance: Women were on average 28 years
and heartache. Topics such as IVF, Infertility, endometriosis and TTC have been old and were committed in their relationship for an average of 6 years.
hashtagged over 5 million times, with other words such as #IVFJourney, Participants desired to have 2.25 children, the first one at 30.5 years old and the
#InfertilityWarrior and #InfertilitySucks, having hundreds of thousands of posts last at 34.5. Only 25% reported high intention to try to have children earlier than
and comments. Celebrities opening up about such intimate issues has given social planned. Being actively trying to conceive and the number of desired children
media users the confidence to open up about their personal fertility struggles were correlated with higher intention to anticipate childbearing. For this reason,
and realise they are not alone in their journey. Attitudes are changing across these two variables were further introduced in the model as covariates. The
generations, Michelle Obama, only recently publicly discussed her IVF experience mediation model revealed no significant direct effect after including mediators
which happened over 2 decades ago. As Gen-Z-ers start to embark on their (effect=.02; 95%CI [-.013, .044]). Two indirect effects of perceived threat of
fertility journey, social media usage for disseminating (in)fertility information will infertility on intention to have children earlier were found, through perceiving
continue rise. However, so will the risk of spreading inaccurate information. infertility as a strong barrier [estimate for indirect effect: .01 (bias-corrected
Limitations, reasons for caution: A key methodological limitations is that (BC) 95% confidence interval (CI) = .005; .027)], and through being willing to
findings mostly reflect views of western social media users. As use of social use fertility treatment as a facilitator [estimate for indirect effect: .01 (BC 95%
media continues to rise in less developed countries, further studies can provide CI= .001; .016)]. The analysis confirmed a full mediation model and it explained
more geographic and global representations. As not all posts include hashtags, 20% of the variance of intention to have children earlier.
our findings may under-represent levels of engagement. Limitations, reasons for caution: Due to the nature of our sample and
Wider implications of the findings: As attitudes towards discussions of recruitment in several settings, the results need to be interpreted with caution.
fertility and reproductive health continue to evolve, social media and celebrity The cross-sectional design does not allow drawing causal directions; further
influence will continue to be influential tools. This should be considered in the longitudinal studies exploring the role of these variables are needed.
context of improving fertility awareness and effective digital health interventions Wider implications of the findings: Perceived threat of infertility seems to
based on robust scientific evidence. be associated with the intention to anticipate childbearing. Due to lack of fertility
Trial registration number: Not applicable awareness, intervention initiatives might take into account these mediators,
aiming to increase the knowledge on the risk of infertility and to clarify myths
P-464 Perceived threat of infertility and women’s intention to that can hinder people from seeking fertility treatments.
anticipate childbearing: the mediating role of personally perceived Trial registration number: NA
barriers and facilitators
J. Pedro1,2, T. Brandão3, J. Fernandes1, A. Barros4,5, P. Xavier5,6, P-465 Incremental validity of the Psychological Inflexibility
L. Schmidt7, M.E. Costa1,2, M.V. Martins1,2 Scale – Infertility (PIS-I)
1
University of Porto, Faculty of Psychology and Educational Sciences, Porto, A. Galhardo1, M. Cunha1, B. Monteiro2, J. Pinto-Gouveia3
Portugal ; 1
Instituto Superior Miguel Torga- Faculty of Psychology and Educational Sciences of
2
Centre for Psychology at University of Porto-, University of Porto, Porto, Portugal ; the University of Coimbra, Psychology- Center for Research in Neuropsychology
3
Universidade Autónoma de Lisboa Luís de Camões, CIP- Departamento de and Cognitive and Behavioral Intervention CINEICC, Coimbra, Portugal ;
Psicologia-, Lisboa, Portugal ; 2
4 Instituto Superior Miguel Torga, Psychology, Coimbra, Portugal ;
University of Porto, Department of Genetics- Faculty of Medicine, Porto, Portugal ;
3
5
Centre for Reproductive Genetics A. Barros, Porto, Portugal ; Faculty of Psychology and Educational Sciences of the University of Coimbra,
6
University of Porto, São João Hospital, Porto, Portugal ; Center for Research in Neuropsychology and Cognitive and Behavioral Intervention
7
University of Copenhagen, Department of Public Health, Copenhagen, Denmark CINEICC, Coimbra, Portugal
Study question: What are the roles of the perceived threat of infertility (sever- Study question: Does an infertility-related self-report measure of psycholog-
ity and susceptibility), barriers and facilitators on the intention to anticipate ical inflexibility, the Psychological Inflexibility Scale – Infertility (PIS-I) perform
childbearing? better than a generic measure of psychological inflexibility?
Summary answer: Perceived threat of infertility affects intention to anticipate Summary answer: The PIS-I was found to add for the prediction of infertili-
childbearing via the perception of infertility as barrier and willingness to go ty-related mental health outcomes as well as more general mental health
through fertility treatments as facilitator. outcomes.
What is known already: Previous studies showed that people of reproductive What is known already: Psychological inflexibility (PI) has been defined as a
age do not have adequate fertility awareness. Infertility might be partly prevent- core transdiagnostic mechanism involved in the development and maintenance
able (if people try to conceive earlier in their lives) and it has been supported of a broad range of psychological difficulties. This led to the development of
that the postponement of childbearing can also be partly attributable to low several context-related measures of PI (e.g., pain, tinnitus, diabetes), besides its
fertility awareness. Some intervention studies have targeted fertility awareness general measure, the Acceptance and Action Questionnaire (AAQ-II). The PIS-I
to promote it, but very few considered barriers, facilitators or intentions was developed to assess infertility-related PI. Previous studies have suggested

that people facing infertility showed higher scores in PI,when compared to fertile model of the intervention. Data collection took place between April and
couples and couples pursuing adoption. Furthermore, PI showed a mediation October 2019.
role in the relationship of the impact of infertility on several life areas and depres- Participants/materials, setting, methods: The interviewed men were
sive symptoms. single or cohabiting childless men and were all residents in the Capital Region of
Study design, size, duration: The study had a longitudinal design. Participants Copenhagen. The men were between 23 to 32 years old, and almost all were
were recruited through the Associação Portuguesa de Fertilidade (patients’ university students or had a university degree. The interviews were audiotaped,
association). Inclusion criteria were age (18 years or older) and an infertility anonymized and transcribed in full. Data were analyzed using qualitative content
medical diagnosis. Data were collected online through self-report instruments analysis following the method by Graneheim and Lundman.
between June and December 2018. A sub-sample of 53 participants completed Main results and the role of chance: The overall theme regarding men’s
the PIS-I 6 weeks after the first administration. attitudes towards a fertility campaign and fertility awareness in general was:
Participants/materials, setting, methods: A sample encompassing 313 ‘Fertility interventions targeting men’. The subthemes were: ‘Campaign com-
participants (287 women and 26 men) completed online the PIS-I, the AAQ-II, munication’, ‘Campaign exposure and relevance for the target group’, ‘The
the Fertility Problem Inventory (FPI), the Infertility Self-efficacy Scale (ISE) and sender and aim of the campaign’, ‘Considerations of infertility’, ‘Attitudes
the Depression, Anxiety and Stress Scales – 21 (DASS-21). Partial correlations, towards future parenthood’, ‘Fertility knowledge’ and ‘Future initiatives’. Overall,
controlling for the AAQ-II, were calculated to test the PIS-I incremental validity. the campaign had limited impact on the men because they believed the campaign
Moreover, hierarchical regression models were computed to address the pre- was not relevant to their current life situation. Furthermore, the men were
dictive power of the PIS-I regarding infertility-related mental health outcomes confused about the aim and message of the campaign, as they thought the
and more general mental health outcomes. campaign encouraged men to have their sperm quality tested. The men also
Main results and the role of chance: Partial correlations results were all criticized the campaign for making a link between sperm quality and masculinity.
significant, ranging from .16 to .45, suggesting that the PIS-I accounts for import- They recognized the importance of knowledge about reproductive health but
ant variance in several measures after accounting for the related construct of they wanted more accurate information about fertility and risk factors for infer-
general psychological inflexibility. Concerning infertility related mental health tility. According to the men, future initiatives should prioritize accurate dissem-
outcomes, in the first step the AAQ-II predicted infertility-related stress (FPI) ination of fertility information in web-based venues. In addition, the men
and infertility self-efficacy (ISE) with R2values of .40 and .20, respectively. In the suggested that fertility information should be a mandatory part of the (sexual)
second step the PIS-I significantly predicted these dependent variables above education in upper-secondary schools and/or university level.
and beyond the AAQ-II with increases in R2values of .12 and .04. Regarding Limitations, reasons for caution: Most of the interviewed men had a uni-
general mental health outcomes, in the first step the AAQ-II acted as a significant versity degree and wanted more facts instead of humor in the campaign, hence
predictor for each outcome with R2values ranging from .24 to .40. In the second our results may not be directly transferred to a similar age group in the general
step, the PIS-I also significantly predicted the depression, anxiety and stress population.
symptoms with increases in R2values between .01 and .05. Wider implications of the findings: Our study contributes to the under-
Limitations, reasons for caution: The online recruitment tends to recruit standing of how to communicate future fertility awareness campaigns and ini-
more educated participants, with more access to online platforms. The study tiatives. This may be useful in the process of increasing the fertility awareness in
was disseminated through a patients’ association, limiting the inclusion of people the population.
with infertility who do not seek medical treatment. The sample included more Trial registration number: N/A
female participants when compared to male participants.
Wider implications of the findings: Context-specific measures have proved P-467 Taking fertility for granted – A qualitative exploration
to be valuable, allowing to capture more specified features of psychological of fertility awareness among young, childless men in Denmark and
inflexibility. The PIS-I also showed to be a useful context-specific measure of PI Sweden
for people facing infertility, adding to the prediction of other mental health G.M. Malling1, L. Schmidt1, T. Pitsillos2, K. Hammarberg3,
outcomes. T. Tydén2, B. Friberg4, I. Toftelund Jensen1, S. Ziebe5
Trial registration number: N/A. 1
University of Copenhagen, Department of Public Health, Copenhagen K,
Denmark ;
P-466 How do men want to receive information about fertility? 2
University Hospital Uppsala, Department of Women’s and Children’s Health,
An evaluation of a fertility campaign targeting Danish men in Uppsala, Sweden ;
Copenhagen 3
Monash University- Melbourne, Jean Hailes Research Unit- School of Public
A.S.N. Berthelsen1, A.L.N. Gamby1, U. Christensen2, L. Schmidt2 Health and Preventive Medicine, Melbourne, Australia ;
4
1
University of Copenhagen, Department of Public Health, Copenhagen, Denmark ; Lund University, Molecular Reproductive Medicine- Department of Translational
2
University of Copenhagen, Department of Public Health- Section of Social Medicine, Malmö, Sweden ;
5
Medicine, Copenhagen, Denmark Copenhagen University Hospital- Rigshospitalet, The Fertility Clinic, Copenhagen,
Denmark
Study question: What are young men’s attitudes towards the fertility campaign
“How’s your sperm?”, and how do they want to receive fertility information in Study question: How do young childless men in Denmark and Sweden reflect
the future? on their fertility, view fertility treatment, and how aware are they about infertility
Summary answer: The young men found the campaign relevant but it had risk factors?
limited impact on them. They preferred fertility information through web-based Summary answer: Most men had limited knowledge about factors that can
venues and during school. impair fertility, presumed they were fertile, and were positive towards fertility
What is known already: It is estimated that 16-26% of the Danish population treatment if they needed it.
who want children, at some point in their lives, will experience infertility. In What is known already: Almost all men in the Nordic countries want to
Denmark, 40% of young healthy men have decreased sperm quality, and every become fathers and want at least two children. Previous qualitative interview
fifth 50-year old man is childless. The fertility awareness is limited among men. studies with childless men aged 21-46 have shown that most men expect to
There are few fertility awareness initiatives targeting men. In October 2018, the become fathers when they want to. Further, recent reviews have concluded that
Municipality of Copenhagen launched the campaign ’How’s your sperm?’ as a men around the world have limited knowledge about fertility and factors that
tool to increase fertility knowledge among men. In order to identify potential can diminish it. Most previous studies about fertility knowledge and attitudes
barriers for the effect of fertility campaigns targeting men, evaluation of such among men have been based on quantitative methods using questionnaires with
campaigns is needed. fixed-choice response options.
Study design, size, duration: Qualitative focus group interviews with 27 Study design, size, duration: This qualitative study assessed knowledge
men distributed in six focus groups were carried out. Furthermore, three expert about fertility and attitudes towards fertility treatment among childless men in
interviews of the main campaign developers were conducted to identify the logic Denmark and Sweden in their last year of university education or vocational

training. In total, 17 Danish and 12 Swedish male students were interviewed. alongside thematic analysis were used to express and discuss the data collected
The interviews were conducted between February and September 2017 and from the interviews.
ranged between 30 and 90 minutes, with an average length of one hour. Main results and the role of chance: Four main themes were identified:
Participants/materials, setting, methods: Participants were recruited face- my destiny, my Rabbi, my relationships and my identity. My destiny focused on
to-face, through postings on their educational institutions, and by snowball the view women had of God as “controller” and “giver” of their infertility. My
methods. Inclusion criteria were: being male, childless, aged 20-30 years and in Rabbi introduced the deep relationship that exists between some Orthodox
the last year of education. The interview guide included questions probing par- Jewish women and their Rabbis, highlighting the way Rabbis are viewed as rep-
ticipants’ perceptions about their own fertility, fertility treatment, and fertility resentatives and messengers of God. My relationships explored the effect of
risk factors. The interviews were recorded and transcribed and the content infertility on the women’s relationships with their partners, families, friends and
analysed thematically. communities. My identity indicated the way women internalize their infertility
Main results and the role of chance: Study participants were on average due to their feelings of isolation, loneliness and estrangement from their familiar
25 years old (range 20-30). The analysis revealed the following themes: 1) Hope culture, community and beliefs. It became apparent that for some women, who
and Fear; 2) Parenthood and Involuntary Childlessness; 3) Fertility Treatment – did not feel fulfilled as women unless they were able to be a mother, all sense
A great Alternative; 4) Factors Meant to Influence Fertility; and 5) Uncertainty. of self and identity slowly breaks down.
Few participants had considered their own fertility but most were optimistic Limitations, reasons for caution: Due to the small sample size and limited
about their ability to become a parent in the future. Among those who that had geographical spread, the findings cannot claim to be representative of the
thought about their fertility, hope and fear were the most prominent emotions. Orthodox Jewish community of the UK. The lead author’s own Jewish status
Most men were positive towards and would consider having fertility treatment could have influenced the participants’ responses and/or the analysis, although
if they experienced fertility problems. The young men had limited knowledge involvement of other authors mitigated this.
about factors that can potentially impair male and female fertility. On average Wider implications of the findings: This research gives useful insight into
each participant mentioned three different factors they believed influence male an underresearched population. Its findings offer guidance to medical profes-
and female fertility either positively or negatively. However, their responses often sionals, counsellors, policy makers, and religious leaders. Other Orthodox Jewish
entailed uncertainty, illustrated through words like ‘uh’, ‘I think’, ‘maybe’, and women may be helped by knowing they are not alone. The findings suggest that
‘could be’. None of the men mentioned sexual transmitted infections (STIs) as other underresearched populations could experience distinctive difficulties with
a risk factor for infertility but most appeared aware of the effect of increasing infertility and ART.
age on fertility. Trial registration number: n/a
Limitations, reasons for caution: Participants’ responses to the question
about factors that affect fertility may not reflect all aspects of their knowledge P-469 Posttraumatic growth (PTG) in women with a long-standing
on the topic. Further, men who do not want children may have been less likely experience of involuntary childlessness
to participate than men who want children. G. Duraskova1, B. Peterson2
Wider implications of the findings: This study highlights the need for edu- 1
psychologist, Motol University Hospital, Prague, Czech Republic ;
cational strategies to improve young men’s knowledge about fertility and the 2
professor Crean College of Health and Behavioral Sciences, Chapman University,
factors that influence it, particularly about the potential adverse effect of STIs
California, U.S.A.
on fertility. Men also need to be aware that there is no guarantee of success with
fertility treatment. Study question: What aspects of posttraumatic growth (PTG) are found in
Trial registration number: N/A women with a long-standing experience of involuntary childlessness?
Summary answer: Women developed deeper self-recognition and greater
P-468 The infertility and fertility treatment experiences of
humility, they reassessed their existing relationships, strengthened their partner-
Orthodox Jewish women in London
ships, and experienced changes in their life philosophy.
A. Vaz Mouyal1, Z. Gurtin1, J. Leach Scully2, J. Harper1 What is known already: “The term posttraumatic growth refers to positive
1
University College London, Institute For Women’s Health, London, United psychological change experienced as a result of the struggle with highly challeng-
Kingdom ; ing life circumstances” (Calhoun & Tedeschi, 2004). For women experiencing
2
Innovation Institute UNSW, John Goodsell Building, Sidney, Australia infertility, research mainly focuses on the description of negative psychological
Study question: What are the infertility experiences of 26 British Orthodox factors with only few published reports mentioning positive personality devel-
Jewish women? opment in conjunction with the infertility experience (Lee at al., 2009; Daniluk,
Summary answer: Infertility challenges the way Orthodox Jewish women 2001). The lack of studies on PTG in women diagnosed with infertility represents
understand their lives and identities. It further impacts their relationships with a gap in the literature which limits our knowledge about the potential positive
their Rabbis, partners, families, friends and communities. implications that may result from efforts to cope with this stressful life challenge.
What is known already: Procreation has a very important place within Study design, size, duration: The study used a qualitative design interviewing
Orthodox Jewish communities, therefore infertility presents a particularly severe 24 women with a long-standing experience of primary infertility. In-depth
difficulty to Orthodox Jewish women. Jewish law tends to permit the use of semi-structured interviews were conducted in the Czech Republic during 2016
assisted reproductive technologies (ART). Several studies on the use of ART in and lasted an average of 53 minutes. Interviews aimed to further understand
Israel have been carried out. Kahn’s work shows the difficulties single Jewish the possible experiences of PTG in study participants. Participants shared both
women face when seeking solo motherhood in a society that honours mother- positive and negative aspects of the infertility experience. Data were analyzed
hood and family life (Kahn, 2000), while Ivry (2010) describes the koshering of using thematic analysis.
medical care in rabbinically mediated fertility treatment. The research presented Participants/materials, setting, methods: Invitations to participate in the
here is the first in the field carried out in the UK. study were sent to 28 women who experienced involuntary childlessness (pur-
Study design, size, duration: A total of 26 Orthodox Jewish women who posive selection). The final sample consisted of 24 women (4 declined to par-
had experienced infertility were interviewed. All interviews were conducted ticipate). The average age of participants was 37,3 years, and their average length
between 2017 and 2018. Women were recruited via adverts in local Jewish of experience with infertility was 6,1 years. Women were asked how infertility
magazines, Facebook groups and via snowballing. The study design resulted affected/changed their partner relationships, sexual life, job, future plans, attitude
from previous research carried out in Israel and the UK as part of PhD to children/values/faith, and leisure time.
research. Main results and the role of chance: Five main themes of posttraumatic
Participants/materials, setting, methods: Semi-structured qualitative growth were identified: greater humility, deeper self-recognition, overall reas-
interviews were undertaken. The women were mostly interviewed in their sessment of relationships, changes in philosophy of life, and strengthening part-
houses. Interviews were conducted in English and lasted between an hour and nership. The experience of infertility taught these women to have more respect
an hour and half. All were transcribed verbatim and any identifiable information to life, reassess and find their personal value, recognize their inner strength, be
was changed to protect the women’s identity. Qualitative descriptive methods more emphatic, develop their spiritual lives, and be responsible for their own

health. Women connected these changes with their experience of infertility. Women described feeling empowered to make informed decisions about
Without a control group of participants, we cannot exclude the possibility that their fertility after attending the FAC clinic. However, when considering a second
women may have experienced these changes without experiencing infertility. pregnancy several years later, women experienced the pressure of time and
Limitations, reasons for caution: The study findings are limited by the their age and uncertainty about their fertility. They wished for repeated coun-
qualitative study design, sampling procedure, and number of participants. selling and advice to aid decision-making after a first pregnancy, or if several
Because of this, we cannot generalize our results for all women with long-term years had passed since attending the FAC clinic.
infertility experience. In addition, PTG is a subjective phenomenon and is limited Limitations, reasons for caution: There may be a selection bias given that
by retrospective self-report and reflection. participants were self-selected from women who attended the FAC clinic with
Wider implications of the findings: The findings of our qualitative study 25% expressing an interest in the study after receiving an invitation. That said,
show what aspects of PTG are experienced by women experiencing infertility. the data from 24 women presented a broad spectrum of fertility decision-making
Medical and mental health providers can use our findings to facilitate the PTG experiences and trajectories.
process in patients. Because the study only sampled women, it is unknown if/ Wider implications of the findings: When providing education and guidance
how PTG is experienced by men. to women regarding future fertility plans, consideration of their “family clock”
Trial registration number: not applicable should be included so women can make informed fertility decisions about timing
of pregnancy and achieve all their family goals. Additional counselling and advice
P-470 The importance of the “family clock”: Women’s experience after pregnancy or several years have passed may be warranted.
of fertility decision-making 6 years after attending the Fertility Trial registration number: Not applicable
Assessment and Counselling clinic
E. Koert1, R. Sylvest1, I. Vittrup2, H.W. Hvidman2, K. Birch P-471 What guidelines should be followed for subfertile patients
Petersen3, J. Boivin4, A. Nyboe Andersen2, L. Schmidt1 to achieve spontaneuos pregnanycy? A randomized controlled trial
1 of couples trying to conceive
University of Copenhagen, Section of Social Medicine- Department of Public
Health, Copenhagen, Denmark ; M. Martins, M.Sc.- Ph.D.1,2, J. Fernandes2, J. Pedro1,2, P. Xavier3,4,
2
Copenhagen University Hospital - Rigshospitalet, Fertility Clinic, Copenhagen, A. Barros4,5, E. Costa1,2, L. Schmidt6
1
Denmark ; University of Porto, Faculty of Psychology, Porto, Portugal ;
3 2
The Fertility Partnership, Fertility Clinic, Copenhagen, Denmark ; University of Porto, Centre for Psychology at University of Porto, Porto, Portugal ;
4 3
University of Cardiff, School of Psychology, Cardiff, United Kingdom Porto Clínica, Reproductive Endocrinology, Porto, Portugal ;
4
University of Porto, Faculty of Medicine, Porto, Portugal ;
Study question: What is women’s lived experience of making fertility decisions 5
Centre for Reproductive Genetics A. Barros, Reproductive Endocrinology, Porto,
in the six years after receiving personalized fertility education, assessment and Portugal ;
counselling? 6
University of Copenhagen, Public Health, Copenhagen, Denmark
Summary answer: Women’s “family clock” (preferred start time, number
and spacing of children) was an important aspect of their lived experience of Study question: Do educational interventions on timing intercourse improve
fertility decision-making. psychosocial adjustment and pregnancy rates in unselected subfertile individuals
What is known already: Research demonstrates that fertility education is over the course of 12 months?
needed to support informed and satisfying fertility decision-making. The Fertility Summary answer: No significant psychosocial adjustment differences were
Assessment and Counselling (FAC) clinic in Copenhagen, Denmark (opened in found between groups over one year. However, the ‘fertile window’ monitoring
2011) is a personalized fertility education, assessment and counselling interven- group achieved a higher pregnancy rate than CG.
tion aimed at increasing knowledge through provision of general fertility infor- What is known already: It is well-know that general knowledge on fertility
mation and a clinical examination and evaluation of personal risk factors and fecundity is low. Recent evidence also suggests that both patients and pro-
(Hvidman et al., 2015). A one-year follow up study of women attending the FAC fessionals lack confidence in natural conception or expectant management and
clinic found that the intervention impacted women’s fertility decisions and was that education is needed. There is solid evidence that that time to pregnancy
a catalyst for change in their lives. Additional long-term follow-up is needed to can be significantly shortened by targeting the fertile period with cervical mucus
explore women’s experiences in detail. monitoring or the use of ovulation predictor kits. However, reproductive care
Study design, size, duration: A qualitative 6-year follow-up study with 24 guidelines recommend instead the practice of intercourse every other day for
women who attended the FAC clinic between January and June 2012. Women being less stressful to the couple. At present, there are no studies comparing
were interviewed in person between February and March 2018 at Rigshospitalet, the recommendation of these two strategies against a control group.
Copenhagen, Denmark. Interviews ranged between 60 to 94 minutes (average Study design, size, duration: We conducted a prospective, double-blind,
73 minutes) and were conducted in English. three-arm parallel RCT, recruiting between July 2016 and June 2018. Subjects
Participants/materials, setting, methods: Invitations were sent to 141 (n=414, 263 women) were randomly allocated into ‘fertile window monitoring’
women who attended the FAC clinic in 2012. Ninety-five women opened the (FWM, n = 137), ‘every other day’ (EOD, n = 124), and control group (CG, n
invitation, 35 were interested in participating, 25 interviews were booked and = 153), and were assessed before (T0), 4-weeks (T1), 6-months (T2), and
24 interviews held. Interviews were semi-structured and examined the women’s 12-months (T3) after the intervention. Interventions consisted of short animated
perceptions and experience of the intervention six years after attending. This videos addressing fecundity knowledge and the correspondent trying-to-conceive
abstract focuses on data specific to the women’s lived experience of making (TTC) strategy.
fertility decisions after attending the FAC clinic. Data was analyzed using thematic Participants/materials, setting, methods: Subjects were recruited via
analysis. social media advertising and brochures with an accompanying link to the survey,
Main results and the role of chance: At the time of the follow-up interview, or via gynecology and fertility clinics, where patients received tablet PCs with
women were on average 39.5 years old (range 31-45). Ten were single and 14 headsets. Individuals trying to conceive and in a marital relationship for > 1 year
were partnered. The majority (21 women, 88%) became parents in the six years were enrolled in the RCT, and completed self-report questionnaires measuring
after attending the FAC clinic, with almost half (9 women) starting to try to psychosocial adjustment besides a specific questionnaire. Mixed models ANOVA
become pregnant in the first year. tested effects on psychosocial variables, and OR and RR were estimated for
The women’s “family clock” played an important role in their fertility decisions pregnancy rates.
– beliefs regarding when to start trying to become pregnant, their preferred Main results and the role of chance: Participants were in their thirties
number of children and spacing between them. All who tried to become pregnant (33.12±4.46), in a relationship for around eight years (8.81±5.19) and were
had given birth to at least one child. However, all of the women with one child TTC for around two years (2.23±2.53). One participant gave-up TTC at T1,
regretted that they had “run out of time” and not achieved their desired family and 331 were lost to follow-up. Eighty-three subjects completed all four moments
size (two or three children), commonly attributed to their age, age-related fer- and were included in the longitudinal analysis having psychosocial adjustment as
tility decline, and/or single parenthood. DVs. Baseline characteristics were well balanced between groups. No significant

interaction or main effects of the interventions were found for stress (PSS), rate (FR) was percentage of transformation of micro injected oocytes into two
depression, anxiety (HADS), and sexual functioning (FSFI/IIEF). Results remained pronuclei.
non-significant after controlling for pregnancy (P > .05). Amongst the 263 The number of patients in each group were A(127/196) B(39/196) and
women, 55 reported not being pregnant at follow-ups and 46 reported a preg- C(30/196)
nancy. Of these, fourteen had a positive pregnancy test after IVF/ICSI and were The average hours of sleep in the 3 groups were 7.9 hours,7.3 hours and
excluded from analysis. The final sample for pregnancy rate analysis included 101 6.3 hours
women. Individuals in the FWM group had significantly higher pregnancy rates Fertilization rates in the Groups A,B and C were 68%,62% and 46% , and
than those in the CG (60.7 vs 35.9%, P = .038); OR 2.8 (95% CI 1.058-7.415). clinical pregnancy rates 44%,39% and 23% respectively with a significant p value
There were no differences in pregnancy rates between the EOD and CG sub- of <0.001.
jects, nor between EOD and FWM. Also it was found that the prevalence of co-existent psychological issues like
Limitations, reasons for caution: This study is prone to bias as both undi- anxiety, depression and OCD were higher among the C group with severe sleep
agnosed and individuals in fertility treatment were included. The analyses did difficulties. 6 of them required psychotherapy . All members in group C were
not include the monitoring of the use of the TTC strategy and did not use clinical advised to attend our classes on YOGA and meditation.
pregnancy as an outcome. Limitations, reasons for caution: Though the prevalence of Insomnia and
Wider implications of the findings: Results show that the EOD strategy sleep disturbances are common, no major studies have been done on their
does not lead to better adjustment nor less stress, and hence guidelines need impact on reproductive outcome in ART. Studies using larger population and
to be reviewed. Further studies should use large samples of untreated couples multicentric trials is necessary .
TTC<12 months to confirm the potential of FWM education as a tool to Although the outcomes were adjusted for confounding factors, some unknown
empower the self-management of reproductive health. confounders may affect the outcomes.
Trial registration number: Study partially funded by the Portuguese Wider implications of the findings: The importance of sleep on reproduc-
Foundation for Science and Technology [FCT PhD grant SFRH/BD/103234/2014] tive outcome must be reiterated and insomnia ,if detected must be treated
aggressively. Simple methods such as yoga,meditation and psychological support
P-472 Sleep - an underrated fertility booster? A questionnaire from family and peers would go a long way in helping these women cope with
survey on the pattern of sleep among IVF patients and their the stress of anxiety and insomnia during IVF treatment.
reproductive outcome Trial registration number: not applicable
R. CHELLADURAI1, G. Pm1, G.D. Ss1, H. Vaidyanathan1
1 P-473 Responses to and communication about infertility among
SIMS Hospital, OBG and IVF, Chennai, India
couples treated at fertility clinic: Implications for depressive
Study question: Evaluating the impact of quality and quantity of sleep on symptoms
fertilisation rate and clinical pregnancy rate in patients undergoing IVF in SIMS T. Matitashvili1, K. Slauson-Blevins2, A. Greil3, S. Bocca1
hospital,Chennai,India 1
Jones Institute for Reproductive Medicine- Eastern Virginia Medical School,
Summary answer: Fertilization rates and clinical pregnancy rates were signifi- Reproductive Endocrinology and Infertility, Norfolk, U.S.A. ;
cantly higher among patients with PQLI (Pittsburg sleep quality index) score of 2
Old Dominion University, Department of Sociology and Criminal Justice, Norfolk,
less than 9 (no/mild sleep difficulties) U.S.A. ;
What is known already: High levels of psychological stress, anxiety, insomnia 3
Alfred University, Division of Social Sciences, Norfolk, U.S.A.
and depression have been reported in IVF patients.Sleep disturbance is a frequent
but underrated symptom in IVF patients and aggravate the existing psychological Study question: Do couple’s responses to infertility depend on the source of
problems .Secretion of Melatonin ,the hormone responsible for improving quality the infertility and how do responses to infertility affect one’s own and partner’s
of oocytes by acting as an antioxidant, is increased during sleep. Also,sleep depressive symptoms?
promotes pulsatility of gonadotrophins and the sleeP-wake cycle maintains the Summary answer: Generally couples responses to infertility are not influenced
normal hormonal balance. However in most places, sleep disturbance is disre- by the source of the problem. One’s own responses are typically associated with
garded and ignored even during routine psychological counselling sessions. It one’s own depressive symptoms.
remains an under-investigated modifiable target that may provide a non-phar- What is known already: Most studies about psychological responses to
macological ,cost-effective method to improve IVF outcome. infertility have used the individual rather than the couple as the unit of analysis.
Study design, size, duration: A questionnaire survey conducted at SIMS Several studies have reported on the coping strategies employed by each partner
hospital Chennai between Jan 2017-Dec 2019. A total of 196 women undergoing in an infertile couple. Less, however, is known about the associations between
IVF were given the Pittsburg sleep quality index questionnaire on the day of difference in perceptions of the experience of infertility and communication and
stimulation to evaluate their sleeping patterns in the previous month by mea- each partner’s well-being. In addition, a number of studies have examined
suring hours of sleep , latency, having bad dreams disturbing sleep, snoring, sleep whether there is difference in the experience of infertility depending on which
disturbances like somnambulism ,whether requiring medications to induce sleep partner is being diagnosed with infertility, but, so far, no clear conclusion can
and job related sleep disturbances like shift-work or part-time. be drawn.
Participants/materials, setting, methods: Patients were divided into three Study design, size, duration: This was a cross-sectional study administered
categories on the basis of PQLI: (A)a total score of ‘‘5’’ or less indicated no by paper questionnaire. Both partners of infertile couples attending a fertility
sleep difficulties; ‘(B)‘6-8’’ indicated mild difficulties; ‘(C) ‘9’’ or more indicated clinic filled out identical questionnaires. The questionnaires included background
severe difficulties. Fertilisation rates and clinical pregnancy rates were compared information, reproductive history, measures of life satisfaction and depressive
between the 3 groups. symptoms, and scales measuring perceptions of various aspects of the infertility
Normal responders undergoing stimulation using Antagonist protocol were experience. Information about medical diagnosis was obtained from medical
included. records. The study involved 156 men and women, including 58 couples, who
Exclusion criteria included poor and hyper responders, male factor, female are the main focus of this paper.
partner age >35 years, endometriosis, PID, using Donor gametes. Participants/materials, setting, methods: Study received IRB approval
Main results and the role of chance: Based on the collected responses to from Eastern Virginia Medical School and Old Dominion University. Patients who
the above questions, a statistical comparison of ART outcomes, in three PSQI attended the fertility clinic from December 2013 to April 2015 were asked to
categories, was performed by Mann-Whitney U test and chi-square test. Logistic fill out a questionnaire containing 161 questions. Validated scales were con-
regression analysis was subsequently conducted to assess the association of the structed measuring importance of children, communication about fertility, per-
conditions in the questions with ART outcomes, as represented by fertilization ceived fertility-specific relationship power and fertility-specific sexual satisfaction.
rates and clinical pregnancy rates. Results were analyzed via paired-samples t-tests, one-way ANOVAs, mixed
Clinical pregnancy was determined by observation of a gestational sac with ANOVAs, and OLS regression. P < 0.05 was regarded as statistically
fetal heart beat by transvaginal ultrasound at 6 weeks of pregnancy. Fertilization significant.

Main results and the role of chance: Women reported higher levels of The researchers provided stress management consisting of educational sessions
importance of children and fertility-related sexual problems than their part- about the treatment and relaxation techniques.
ners. Men reported higher general relationship satisfaction, fertility-specific Participants/materials, setting, methods: Literate couples with a
relationship satisfaction, and perceived fertility-specific relationship power female partner < 38 years old, using their fresh gametes, and without psychiatric
than their partners. With one exception, scale scores were not associated disorders were included. Stress levels were assessed on the first day of
with which partner had a fertility problem. Women scored higher than their ovarian stimulation and on the day of embryo transfer using the Perceived
partners on the importance of children scale except among couples with Stress Scale (PSS 10). Luteal granulosa cells were collected on the day of
infertility of unknown cause. For women, higher scores on importance of oocytes pick-up. Granulosa vitality and morphology were evaluated and
children, satisfaction with fertility–related communication and fertility-related mtDNA levels were quantified using Ion Torrent next generation sequencing
sexual problems were associated with higher levels of one’s own depressive technology.
symptoms. For men, higher scores on satisfaction with fertility–related com- Main results and the role of chance: During the first day of ovarian
munication, perceived fertility-related power, fertility-related relationship stimulation, statistically significant higher scores of PSS (PSS initial) were
satisfaction, general relationship satisfaction and fertility-related sexual prob- detected in women of the study group (23.1 ± 5.1) compared to the control
lems were associated with higher levels of one’s own depressive symptoms. group (19.3 ± 7.01) (p < 0.05). However, ΔPSS (PSS initial – PSS on the day of
For women, fertility-related sexual problems was associated with higher transfer) showed significant reduction in perceived stress in women of the study
levels of partner’s depressive symptoms. For men, higher scores on impor- group compared to the control group (p < 0.01). At the ovarian level, trypan
tance of children was associated with higher levels of partner’s depressive blue staining indicated that the percentage of viable granulosa was significantly
symptoms. The higher women score on the importance of children compared higher in the study group compared to the control group (p < 0.05). In addition,
to their partners, the higher their level of depressive symptoms. The higher a statistically significant higher percentage of granulosa cells with normal mor-
men score on fertility-related sexual problems and satisfaction with fertili- phology were detected in women of the stress management group compared
ty-related communication, the higher their level of depressive symptoms (all to the control group (p < 0.05). Furthermore, mtDNA levels were significantly
P < 0.05). higher in luteal granulosa cells of control group compared to the study group
Limitations, reasons for caution: Because study was cross-sectional, it is (p < 0.05).
impossible to make causal inferences. The findings of this study are limited in Limitations, reasons for caution: The study excluded subjects with severe
generalizability as all patients were recruited from a single site and small sample infertility factors. Consequently, the efficiency of this stress management pro-
size prevented us from including more variables in each analysis. gram should be also tested in these subjects before automatically generalizing it
Wider implications of the findings: These findings have important impli- to all infertility cases.
cations for counseling infertile couples. It is especially important to note that Wider implications of the findings: This stress management program seems
relationship satisfaction and satisfaction with communication does not always to be able to reduce psychological stress among IVF patients thus improving their
have a positive effect on well-being. Fertility-related sexual problems appear to mental health. It also influences the ovarian physiology through the granulosa
have implications for well-being among both men and women. cells. Hence, further investigations are needed to evaluate whether applying this
Trial registration number: n/a program in clinical settings could affect other ICSI related outcomes.
Trial registration number: LBCTR2019101289
P-474 Luteal granulosa cells from women undergoing a
stress management program during ICSI cycles exhibit lower
mitochondrial DNA levels: a randomized clinical trial P-475 Seeking pregnancy through assisted reproduction
technology treatments (ART): A comparison between couples
G. Raad1, J. Tanios2, N. El Salibi3, S. Kerbaj4, F. Shamas5, J. Azouri6,
receiving donated gametes and couples using their own gametes
Y. Mourad7, C. Fakih8
1
F. Andrei1, P. Salvatori1, L. Cipriani2, G. Damiano2, N.C. Rossi1,
Al Hadi Laboratory and Medical Center, ivf, Beirut, Lebanon ;
2
E. Porcu2
Lebanese University, Master In Cognition- Behavior- and neuroscience, Fanar, 1
University of Bologna, Department of Psychology, Bologna, Italy ;
Lebanon ; 2
3 Infertility and IVF Unit, University of Bologna-Sant’Orsola Hospital, Bologna, Italy
King Abdullah University of Science and Technology KAUST, Counseling Center,
Saudi Arabia, Saudi Arabia ;
4
Lebanese University, Master in psychosomatic therapy, Fanar, Lebanon ; Study question: What are the psychological differences between couples
5
Al Hadi Laboratory and Medical Center, Molecular Biology, Beirut, Lebanon ; waiting for gametes donation and those using their own gametes, considering
6
Mount-Lebanon Hospital Lebanon, Azoury IVF clinic, Hazmieh, Lebanon ; previous ART attempts and cause of infertility?
7
Al Hadi Laboratory and Medical Center, Medical Laboratory, Beirut, Lebanon ; Summary answer: Differences in infertility self-efficacy were found between
8
Al Hadi Labratory and Medical Center, Ivf, Beirut, Lebanon the two groups, while no significant differences with regards to psychological
symptoms and quality of life emerged.
Study question: What impact does stress management have during intracy- What is known already: Involuntary childlessness often produces overwhelm-
toplasmic sperm injection (ICSI) cycles on mitochondrial DNA (mtDNA) levels ing feelings of anger, shame, grief, and guilt. For those couples who decide to
in luteal granulosa cells? undergo ART, such emotional reactions may persist, if not worsen, throughout
Summary answer: Women undergoing stress management during ICSI cycles the different phases of the treatment. Higher levels of psychological distress and
had lower mitochondrial DNA levels in their luteal granulosa cells compared to lower levels of quality of life seem to affect especially couples who experienced
the control group. repeated failures. Additionally, the only published study comparing couples by
What is known already: ICSI cycles were demonstrated to be psychologically the severity of infertility shows that couples who use their own gametes seem
stressful for most couples. In parallel, growing evidence supports the implication to have higher depression, anxiety, infertility stress, and infertility-related sexual
of mitochondria in the stress response. Furthermore, mitochondria are consid- concerns than couples awaiting for gametes donation.
ered as potential regulators of oocyte- granulosa cells cross-talk. However, the Study design, size, duration: Data were collected cross-sectionally from
effect of psychological stress during ICSI cycles on mtDNA levels in luteal gran- October 2018 to October 2019. One hundred seven couples seeking treatments
ulosa cells and subsequent oocyte competence was never fully investigated nor to conceive took part to the study.
well understood. Participants/materials, setting, methods: Data were collected from 107
Study design, size, duration: A randomized controlled trial, with pre- couples at their first access to the Infertility and IVF Unit of the Sant’Orsola
test-posttest experimental design was conducted on 60 couples undergoing Hospital, University of Bologna (Italy). During a first consult, clinical information
ICSI cycles, between May 2018 and May 2019, at Mount-Lebanon hospital. were recorded and the Infertility Self-Efficacy Scale, Fertility Quality of Life, and
On the first day of ovarian stimulation, the couples were randomized using Symptom Check-List were given to participants. During a second consult, couples
sealed opaque envelopes into control group (n=30) and stress management were placed on a waiting list for either gametes donation (n=42) or IVF with
group (n=30). The physicians and embryologists were blinded to the allocation. their own gametes (n=65), and questionnaires were returned.

Main results and the role of chance: Univariate ANOVAs showed a sig- women reported mild regret and 26 (16%) moderate to severe regret. Increased
nificant interaction between previous ART attempts (yes/no) and awaited decision regret was associated with perception of inadequate emotional support
treatment (IVF with gametes donation/IVF with the couple’s own gametes) with during OC (p=0.045) and low expectations about the efficacy of the proce-
regards to infertility self-efficacy. Couples who used their own gametes showed dure(p=<0.001). Decision regret was not associated with age, number of frozen
higher levels of self-efficacy if they were at their first ART attempt compared eggs, perceived adequacy of information prior to OC, patient-estimated prob-
to those who already tried to conceive through ART without succeeding. ability of achieving a live birth and use of banked eggs, attitude towards alter-
Gametes receiving couples showed an opposite pattern (F = 6.54, P < .01). native child bearing methods, current relationship status or time elapsed since
With regards to quality of life, we found a main effect of the variable previous OC. One hundred sixty-seven women (88%) reported increased sense of con-
ART attempts, showing that couples who already underwent ART treatments trol over future reproductive planning following OC. One hundred eighty-three
had overall lower levels of quality of life (F = 2.68, P < .01). No differences women (89%) indicated they would be satisfied with their decision, even if they
were found with regards to levels of anxiety, depression and anger/hostility (all never used their frozen eggs.
Ps > .05). Limitations, reasons for caution: One of the main limitations of our study
Limitations, reasons for caution: The sample size and the cross-sectional is that these results reflect the attitudes of highly educated Turkish women with
design allowed us to draw conclusions only about the associations between study DOR towards OC, which should be extrapolated to other populations with
variables. We could not include additional relevant variables, such as time from caution. The other limitation is the relatively short and variable follow-up dura-
infertility diagnosis and parity, which may have affected our results. tion of the study.
Wider implications of the findings: The differences between gametes Wider implications of the findings: Findings of this study indicate that the
receiving couples and couples who used their own gametes in self-efficacy may prevalence of regret among patients with DOR undergoing OC is low. Hence,
be determined by both the difficulties each couple face during ART treatment the number of patients who return to use their oocytes is low, this restricts our
and the infertility history. Shifting to gametes donation may give new hope to ability to assess the impact of reproductive failure. Long-term studies including
those couples who experienced repeated failures with ART. patients with failure are necessary.
Trial registration number: CE: 273/2018/Sper/AOUBO Trial registration number: None
P-476 Factors associated with decision regret following oocyte P-477 Assessment of the psycho-affective repercussions of male
cryopreservation for diminished ovarian reserve and/or age- infertility
related fertility decline F. Hammami1, A. Zouari2, S. Ellouze2, G. Rim2, M. Chibani3,
T.A. Boza1, A.P. Çil1, L.S. Karakis2, R. Abalı2, M. Ceyhan1, R. Rachdi3, S. Jellad1
E.G. Aksakal1, M. Bahçeci2, B. Urman1 1
Military hospital of Tunis, Laboratory of Reproductive Biology- Unit of Assisted
1
American Hospital, Womens’ Health Unit, Istanbul, Turkey ; Medical Procreation, Tunis, Tunisia ;
2 2
Bahceci Health Group, IVF Centre, Istanbul, Turkey Razi hospital, Department of Psychiatry B, Mannouba, Tunisia ;
3
Military hospital of Tunis, Unit of Assisted Medical Procreation Military Hospital
Study question: What are the factors associated with decision regret following of Tunis, Tunis, Tunisia
oocyte cryopreservation (OC) in women with diminished ovarian reserve (DOR)
and/or age-related fertility decline? Study question: This study aims to evaluate the prevalence of anxiety and
Summary answer: The perception of inadequate emotional support during depressive disorders in men after diagnosis of their infertility.
the procedure and low expectations regarding its efficacy were associated with Summary answer: Diagnosis of infertility is associated with important prev-
increased decision regret. alence of anxiety and depressive disorders in men. Severity of these disorders
What is known already: Recently we published our data on cycle character- increases with age and infertility’s duration.
istics and reproductive outcomes of women with DOR undergoing OC. Since What is known already: Studies comparing women and men found that
cycle cancellation rates are high and reproductive outcomes are poor in such women react more strongly overall to infertility. The impact of psychological
patients, we aimed to counsel these patients about their expectations of OC wellness in infertile women has been widely studied unlike the consequences of
and determine factors which may be associated with future decision regret. male psychological disorders that are not well explored. Similar to their female
Decision regret is considered as an overall indicator of quality of health decisions. counter parts, men seeking fertility treatments have also an increased prevalence
Although there are a few studies on decision regret following elective OC in of anxiety and depression.
women with normal ovarian reserve, no studies to date have addressed factors Study design, size, duration: A cross sectional study was conducted between
associated with decision regret in women with DOR. June and September 2019 in the laboratory of reproductive Biology and Unit of
Study design, size, duration: A cross sectional survey study was conducted assisted medical procreation of Military Hospital of Tunis.
involving 552 women with DOR and/or age-related fertility decline who under- Participants/materials, setting, methods: Data of 108 men
went OC in two private ART centers (American Hospital and Bahceci IVF consulting for couple’s infertility exploration was included. All patients were
Centre) from 2015 to 2019. Following IRB approval, women were contacted by interviewed on sociodemographic characteristics, medical and surgical history.
phone and asked whether they would be willing to participate in a standardized We specified also the type and the duration of the infertility and we analyzed
online questionnaire to evaluate decision regret, their current relationship status, different semen parameters for each patient. Sperm abnormalities were clas-
satisfaction level with the procedure, and their attitude towards future sified as normal, moderate, severe, extreme and azoospermia. We used
childbearing. Hospital Anxiety and Depression Scale (HADS) to assess anxiety and
Participants/materials, setting, methods: Of the 552 women who were depression.
contacted, 468 accepted the invitation, and 162 (34.6%) responded to the survey. Main results and the role of chance: The mean age of participants was
The primary outcome of the study was decision regret measured using the 36,8 years. Eleven patients had a history of varicocele and six of them suffered
validated Decision Regret Scale (DRS). The associations between decision regret from associated erectile dysfunction. Infertility was primary in most of patients
and level of patient satisfaction, reproductive expectations and desire for par- (77,8%) with an average duration of 3,32 years. About 25% of the patients
enthood were assessed. DRS was interpreted as follows: 0: no regret; 1-25: mild had at least one previous failed assisted reproductive attempt. Spermogram
regret, and 25-100: moderate to severe regret. abnormalities were found in 78,7% of patients. 8,33% of them suffered from
Main results and the role of chance: Mean age at the time of OC and azoospermia. Depression was diagnosed in 29,7% of patients and anxious
survey submission was 37.8±4.6 and 40 ±1.0, respectively. The total number state in 34,2% of them. The severity of depression was correlated to anxiety
of oocytes cryopreserved was <10 in 70% of women. The average follow-up (P < 0,001), older age of patients (P=0,034), and high infertility duration
time interval from OC to survey submission was 2.5 years. Five patients reported (P=0,037). However, the severity of sperm abnormalities was not correlated
pregnancy after OC; 4 spontaneous and one using cryopreserved oocytes. The to anxiety or depression. No associations were found between psychological
median and mean DRS scores were 10 (interquartile range: 25) and 13.4 (range distress and erectile dysfunction or previous failed assisted reproductive tech-
0-70), respectively, indicating a low prevalence of regret. Eighty-five (52.5%) nique attempt.

Limitations, reasons for caution: Diagnosis was established according to a P-479 Answer patterns by recurrent pregnancy loss patients on
self-reported scale without psychiatric consultation. Most of the couples had a depression and stress scales
feminine infertility factor, which might have generated a sample bias. A. Slot1, M.C. Krog2, S. Bliddal3, L.R. Olsen4, H.S. Nielsen5,
Wider implications of the findings: Anxiety and depression should be A.M. Kolte6
diagnosed and treated in men seeking for fertility treatment, especially that it 1
Copenhagen University Hospital- Rigshospitalet, The Recurrent Pregnancy Loss
can lead to accelerated cellular aging, poor quality gametes, and other compli- Unit- The Fertility Clinic, Copenhagen, Denmark ;
cations. Systematic screening for these psychiatric disorders in infertile men 2
Copenhagen University Hospital- Rigshospitalet, The Recurrent Pregnancy Loss
population is therefore essential, in order to ensure multidisciplinary and ade- Unit- The fertility Clinic and Department of Clinical Immunology, Copenhagen,
quate care. Denmark ;
Copenhagen University Hospital, Departments of Medical Endocrinology,
Copenhagen, Denmark ;
4
P-478 Transgender and Fertility preservation: how to assist young Mental Health Services in the Capital Region of Denmark, Child and Adolescent
adults and adolescents transgender pathway Mental Health Center, Copenhagen, Denmark ;
5
University of Copenhagen- Rigshospitalet and Amager-Hvidovre Hospital, The
N. Mendes1,2, A. Condat2, J. Chambry3, B. Bennanni Smires1,
Recurrent Pregnancy Loss Unit and Department of Gynaecology and obstetrics,
S. Sarandi1, L. Chobe Christian1, L. Martinerie4, F. Eustache1
1
Copenhagen, Denmark ;
Service de Biologie de la Reproduction- CECOS- Hôpital Jean-Verdier- Avenue du 6
Copenhagen University Hospital, Recurrent Pregnancy Loss Unit- The Fertility
14 Juillet- 93140 Bondy- France, Fertility center, Bondy, France ;
2
Clinic, Copenhagen, Denmark
Service de Psychiatrie de l’Enfant et de l’Adolescent- Hôpital Pitié-Salpêtrière- 47-
83 boulevard de l’Hôpital- 75013 Paris- France, Psychiatry, Paris, France ; Study question: Are there specific patterns in how women with recurrent
3
Centre interhospitalier d’accueil permanent pour adolescents Paris rive droit- pregnancy loss (RPL) answered the Major Depression Inventory (MDI) and the
Hôpital Maison Blanche- Paris- 75018- France, Psychiatry, Paris, France ; Perceived stress scale (PSS)?
4
Service d’Endocrinologie et de Diabétologie Pédiatrique- Hôpital Robert Debré & Summary answer: Women with RPL often feel guilt, angered and lack
Université de Paris- Paris- 75019- France., Endocrinoly, Paris, France self-confidence. Women with primary RPL are a particularly sensitive group.
What is known already: RPL affects between 1-3% of all couples trying to
Study question: Do our national counseling protocol and social insurance conceive a child and the grief of experiencing pregnancy losses are to some
system improve access of fertility preservation (FP) prior hormonal therapy or women comparable to the grief of experiencing a neonatal death. Women with
sex reassignment surgery for transgender adolescents in comparison to inter- RPL more often suffer from psychological comorbidities such as anxiety, stress,
national experience? and depression compared with other women trying to have a child. Recently, a
Summary answer: Two-thirds of the cases from our fertility center desire to qualitative study underlined the need for all involved health care professionals
FP. The obstacle in France seems to be the impossibility of using preserved to be sensitive to the couples’ history of pregnancy losses and to acknowledge
gametes. their grief.
What is known already: International guidelines recommend that healthcare Study design, size, duration: This is a secondary analysis of a cohort study
personnel should discuss FP with transgender individuals before starting gender including women referred to the RPL Unit at the Fertility Clinic at Rigshospitalet
affirmation therapy in adults and adolescents. Unfortunately, low access rates in Denmark June 2010 -June 2013. In total, 302 women completed the MDI and
to FP center are reported. Many factors can affect the decision to undergo a FP, the PSS. The prevalence of psychological stress and depression as summarized
as laws, financial status, social climate and attitude of medical professionals ... scores was published by our research group in 2015. Here we investigated if
Several studies indicate the risk of regret after a potential irreversible treatment. there were patterns in responses to the individual items on the MDI and the PSS.
The nation social insurance system improve access to fertility preservation in Participants/materials, setting, methods: RPL was defined as ≥3 consec-
France. It is important that transgender adolescents and adults be advised in the utive pregnancy losses, and 162 women had primary RPL (experienced RPL
light of their vulnerability. without having a child), 136 had secondary RPL (having a child prior to experi-
Study design, size, duration: From january 2018 to january 2020, 67 trans encing RPL), and four had tertiary RPL. The women’s answers were reviewed
people (adults and adolescents), AMAB (assigned male at birth) (n=38) and and the cut-off for a “confirmative” answer was “slightly less than half the time”
AFAB (assigned female at birth) (n=29), were seen at our ART center. or more often on the MDI and “sometimes” or more often on the PSS.
Participants/materials, setting, methods: All trans people were seen first Main results and the role of chance: The most frequent answers to the
by a medical biologist and a psychologist to discuss and explain FP processes and MDI were the three core symptoms of depression (42% lack of energy, 35%
the French law. In line with the psychiatrists and endocrinologists, a reflection feeling sad, 35% loss of interest,) along with feeling less self-confident (28%) and
period was proposed, then FP was performed whenever the person was willing guilty (26%). The cohort included 26 women who fulfilled the criteria for mod-
to and organized in accordance with their choice. Comparison between adults erate to severe depression, 19 had primary RPL and 7 had secondary RPL. Of
and adolescents wish for fertility preservation and the method chosen were the 276 women who did not fulfill the criteria for moderate/severe depression,
analyzed using student t test. 9 women (3%) “felt that life wasn’t worth living”. The most frequent answers in
Main results and the role of chance: Mean age was 20.7 +/-6.7 years. Of PSS evaluation were being “angered because of things outside their control”
all trans people, 75% were willing to undergo FP but only 42% actually went (76%) and not feeling “things were going your way” (71%). Women with primary
through the process. Almost 20% remained undecided. Among AMAB, 82% RPL answered that they felt less self-confident (32% vs 18%, p=0.007), restless
wished to undergo FP (mean age 23.4 +/-7.7), in comparison to 66% of AFAB (17% vs. 9% p=0.033), had concentration problems (25% vs. 10%, p=0,001) and
(mean age 17 +/-1.8). No difference was found in the desire for FP between that life wasn’t worth living (13% vs. 3%, p=0.002) significantly more often com-
adults and adolescents (p=0.503). However, adults were more likely to actually pared with women with secondary RPL. In the PSS evaluation women with
go through the process of FP than adolescents (p<0.01). Qualitative data col- primary RPL more often reported “being unable to control important things”
lected from psychological and biological evaluations suggested this result what (69% vs 55%, p=0.018) and “unable to handle personal problems” (38% vs. 25%,
related to the difficulty, consistent with the French Law, to reuse the preserved p=0.020) compared with the women experiencing secondary RPL. There were
gametes after change of civil gender status. no significant differences between the groups regarding the remaining items.
Limitations, reasons for caution: These first results will need to be con- Limitations, reasons for caution: The questionnaires were constructed to
firmed on a larger cohort. result in a combined score or as a clinical tool for diagnosing, and not to analyse
Wider implications of the findings: Psychological counseling is in need of each item individually. Confirmatory answers on the MDI in the study presented
more data to assist trans people in the pathway decision. If financial aspect is here does not equal clinical depression but reflect prevalent feelings among
not an argument to understand the difference between adults and adolescents women with RPL.
fertility decision, is important to investigate further tracks. Wider implications of the findings: Women with RPL commonly experi-
Trial registration number: not applicable ence guilt, lack of self-confidence, and being angered. Women with primary RPL

may be a particularly sensitive group. Treating clinicians need to understand the P-481 Social aspects of infertility in Europe: a patient survey
psychological impact of RPL and could try to encourage behavior to empower M. Lopez-Teijon1, M. Benigna2, A. Garcia-Faura1, B. Marques1,
the women. S. Novo1, F. Garcia1, C. Carolina1
Trial registration number: Not Applicable 1
Institut Marques, Reproductive Medicine Service, Barcelona, Spain ;
2
Institut Marques, Reproductive Medicine Service, Roma, Italy
P-480 Understanding parents’ intention to share information
about the donor-conception with their offspring by application of Study question: Do european patients have a feeling of social rejection to
the Theory of Planned Behavior infertility? Has it changed during the last years?
C. Lampic1, A. Skoog Svanberg2, K. Sorjonen3, G. Sydsjö4 Summary answer: 63,34% of patients believe that fertility treatments aren’t
1
Karolinska Institutet, Women’s and Children’s Health, Stockholm, Sweden ; accepted by the society in which they live, and their feeling of rejection by society
2
Uppsala University, Women’s and Children’s Health, Uppsala, Sweden ; is increasing.
3
Karolinska Institutet, Clinical Neurosciences, Stockholm, Sweden ; What is known already: Each country offers a different cultural context in
4
Linköping University, Clinical and Experimental Medicine, Linköping, Sweden which to view infertility, and legislation controlling fertility treatments also forms
opinion. In Europe the law differs very widely between countries, with some
Study question: Does the Theory of Planned Behavior contribute to under- countries being extremely restrictive (such as Germany) and others, such as
standing parents’ intentions to share information about genetic origin with Great Britain or Spain, where infertility treatments are far more visible in society.
their child? Study design, size, duration: Between June-December 2019 questionnaires
Summary answer: Parents’ intention to start disclosure was influenced by were filled in by 1733 patients undergoing fertility treatment, from 10 different
beliefs that disclosure would have desired consequences and a desire to act in countries. We asked 3 questions:
accordance to societal norms. Do you think that in your country fertility treatments are taboo?
What is known already: There is growing consensus on donor-conceived Have you told the people in your immediate environment that you are under-
persons’ right to information about their genetic origin. As a result, there an going fertility treatment?
increased use of identity-release donation and parents are encouraged to share Would you agree to be interviewed in the media in order to help other people?
information about the donor conception with their child from an early age. We compared the results to those obtained from the same questionnaire
Nevertheless, disclosure to the child remains a challenge for many parents, par- in 2013.
ticularly heterosexual couples, who are concerned about disrupted relationships Participants/materials, setting, methods: The questionnaire was com-
and uncertain about when and how to talk with the child. The Theory of Planned pleted by patients currently in treatment at our clinics in Spain, Italy and Ireland.
Behavior (TPB) has been shown to explain various health-related behaviors and They were completed by female patients (either single, with a male partner or
may be applied to increase the understanding of parents’ decision-making regard- with a female partner). We performed a descriptive analysis of the results.
ing disclosure. Main results and the role of chance: For 63,3% of patients fertility treat-
Study design, size, duration: The present study is part of the prospective ment is a taboo subject. In 2013, this percentage was 52,3%, indicating that
longitudinal ‘Swedish Study on Gamete Donation’ (SSGD) including all fertility infertility as a taboo subject increased in nearly all countries.
clinics performing gamete donation in Sweden. Consecutive groups of recipient However, the social perception of infertility within Europe was very variable.
couples were recruited 2005-2008 and followed with individual questionnaires The greatest feeling of rejection was found in Italy (74%) and Germany (74%).
before and after treatment with oocyte and sperm from identity-release donors. Ireland (56%), Switzerland (55%), France (54%), Spain (52%), Sweden(50%) and
The present study includes data from the follow-up assessment of the SSGD UK(47%) were in the middle. Belgium was the most tolerant (36%).
when participants’ offspring had reached age 7. It is interesting to note that the most tolerant countries in 2013 no longer are
Participants/materials, setting, methods: Eligible were heterosexual recip- so; Spain increased by 17% and the UK by 11%.
ients with 7-year old children following identity-release oocyte donation (OD, 87% of patients did share their experience of infertility with their closest family
n=147) and sperm donation (SD, n=174). A total of 83 OD-parents (56% and friends but without giving any details of the treatment. This was 12% more
response) and 113 SD-parents (65% response) responded to a postal survey. than in 2013. The family continues to provide a point of refuge but now 37%
They individually completed the study-specific TPB-Disclosure Questionnaire more patients inform everyone in their immediate environment of their
assessing behavioral, normative and control beliefs and the intention to talk with treatment.
child about the donor conception during the upcoming year. Data were analyzed With regard to being willing to discuss their infertility with the media, 68%
with Path analysis. would now agree to do so (in 2013 55% agreed). Maintaining their anonymity
Main results and the role of chance: More than half of participants had is still an indispensable factor for the majority of patients.
already started talking with their 7-year old child about his/her donor conception There is very little variability in reponses between patients from different
(OD 61%, SD 58%). All components of the Theory of Planned Behavior signifi- countries for the last two questions.
cantly influenced parents’ intention to talk with their child about his/her donor Limitations, reasons for caution: The nature of the questions and the
conception during the upcoming year. Among parents who had not yet started inclusion of personal values regarding the concept of a ‘taboo’ issue could give
the disclosure process, their intention to talk with their child was influenced by rise to ambiguity and therefore limit the validity of some replies to the
beliefs that disclosure would have desired consequences and a desire to act in questionnaire.
accordance to perceived positive attitudes towards disclosure in society. Wider implications of the findings: The results allow us to better define
Interestingly, a higher level of control beliefs, i.e. feeling confident and in control the psychological needs of infertile patients, and to see how the context of their
over the disclosure process, was not significantly related to a stronger intention country of origin may affect them.
to talk with the child during the upcoming year. Type of treatment (OD/SD) It’s surprising that the increasing success of techniques in reproductive medicine
and the existence/absence of a genetic link to the child did not directly influence isn’t accompanied by a similar increase in social acceptance of the treatments.
parents’ intention to talk with the child about the donor conception. Trial registration number: Not applicable
Limitations, reasons for caution: The study was performed with hetero-
sexual couples within the context of the Swedish legislation that mandates iden-
P-482 Characteristics and intentions of heterosexual couples
tity-release donation, which limits the generalizability to other populations. Also,
comprised of a transgender man and his cisgender woman
attrition may have introduced selection bias to the study findings.
awaiting sperm donation to conceive a child.
Wider implications of the findings: The present results indicate that the
Theory of Planned Behavior may contribute to understanding disclosure deci- S. Bonan Benchetrit1, N. Mendes2, E. Chapel-Lardic1,
sions among heterosexual couples building a family with donor gametes. Parents O. Rosenblum2, C. Dudckiewicz-Sibony1, J.P. Wolf1, A. Condat2,
may benefit from opportunities to discuss perceived consequences of disclosure C. Chalas1, C. Patrat1, V. Drouineaud1
1
in order to support family life following donor conception. Assistance Publique – Hôpitaux de Paris AP- HP Centre- Université de Paris-,
Trial registration number: not applicable Service de Biologie de la Reproduction - CECOS-, Paris, France ;

2
Assistance Publique-Hôpitaux de Paris AP-HP- Groupe Hospitalier de La Study question: Should a good morphology day 6 embryo be chosen over
Pitié-Salpétrière-, Service de Psychiatrie de l’enfant et de l’adolescent-, Paris, France day 5 poor morphology in a limited options scenario of PGT-A embryo transfer?
Summary answer: Day 6 PGT-A embryos produced less pregnancies in our
Study question: What are the main characteristics and intentions of couples settings even when compared to the day 5 “BB grade only” embryo transfers.
comprised of a transgender man and his cisgender woman awaiting sperm What is known already: To date, many studies have shown that day 6
donation? embryos seem to have a reduced potential for achieving pregnancy when com-
Summary answer: The couples often met before the man’s transition, shared pared to day 5 embryos. However, most of those included data of non-PGT-A
a common life of several years, intend to inform their child of sperm donation embryos, while day 5 embryos are well known for having higher euploidy rates.
and transidentity. If PGT-A embryos were compared, authors usually were unable to conclude
What is known already: It has been reported that more than half of trans- any differences in pregnancy rates between euploid day 5 and day 6 embryos.
gender men (female-to-male transgender persons) desired to have children. So, during the laboratory routine there is no obvious choice if only good mor-
Since 1994, Assisted Reproductive Technologies (ART) are strictly offered to phology day 6 and impaired morphology day 5 embryo are available for PGT-A
heterosexual couples in France. Indeed, in order for a transgender man to have transfer.
access to ART to conceive a child, he must be in a relationship with a woman Study design, size, duration: This was the retrospective analysis of 1331
and have changed his legal sexual identity from female to male. Very little data PGT-A single embryo transfers in regard to embyo culture day, morhphological
have been previously reported about the characteristics and intentions of het- score and pregnancy outcome. When available the time-lapse analysis data were
erosexual couples in which the man is transgender and awaiting sperm donation included in the comparison.
to conceive. Participants/materials, setting, methods: PGT-A analysis was performed
Study design, size, duration: A retrospective analysis of unit records of 43 by aCGH method in
heterosexual couples comprised of a transgender man with his cisgender female Main results and the role of chance: From 1331 PGT-A single embryo
partner, who apply for ART with donor sperm to conceive a child, was performed. transfers the day 6 embryos were used in 342 cases (25.7%), day 5 “BB grade
Couples were recruited at the Centre de Conservation des Oeufs et du Sperme only” in 101 cases (7.6%). Surprisingly, corresponding clinical pregnancy rates
(CECOS) (APHP, Cochin, Paris) between October 2010 and December 2019. were 45.0% and 54.5%.
Couples were asked questions of qualitative and quantitative nature. When morphology of embryos transferred after biopsy at day 6 was analysed,
Participants/materials, setting, methods: The study was set in the CECOS we observed that 41.6% of those had AA grade, 31.1% were AB or BA and only
of (APHP, Cochin, Paris). Our study reported here sought to analyze: socio-de- 28.3% were BB scored. Thus, this group had obviously better scores than day
mographic characteristics of the couples, personal history of the transgender 5 “BB grade only” cohort and difference was not caused by poor morphology.
man, sex transition, meeting and life of the couple, intentions regarding disclosure After we compared the distribution of embryo transfer orders for day 6 AA
of sperm donation and father’s transidentity to the child. and day 5 “BB grade only” transfer, we can conclude that in most cases the
Main results and the role of chance: Men and women were 32±6.6 and reason for day 6 embryo selection was not the fact that all day 5 euploid embryos
29.7±4.6 years old, respectively. 37% of transgender men had a higher level than were already transferred in the previous attempts. In fact, the day 5 “BB grade
the bachelor degree. Most men (65%) and women (67%) were employees. only” transfers occurred after several unsuccessful transfers more often.Time-
Transgender men often reported first manifestations during early childhood lapse data indicates that day 6 embryos had slower development starting at
(54%), difficult experience of breast development (70%) and menstruations 6 cell stage with increasing lag onward, otherwise performing similar.
(79%). 40% of men reported difficulties during the school and an unstable pro- This data leads us to conclusion that day 6 embryos produce less pregnancies
fessional career. Sex transition process was often triggered by a television pro- due to their intrinsic values other than morphology and chromosomal
gram (28%) or following web/social networks use (28%). Importantly, in 77% constitution.
of the cases, the couple met before the man’s transition. Most women were Limitations, reasons for caution: This is a retrospective analysis and thus
heterosexual (60.5%), some identified as lesbian (2.3%) [CP1] or bisexual (14%). it may be subject of bias.
81% of women were comprehensive at the announcement of transidentity. The Wider implications of the findings: Our data seem to suggest that it may
couples were often stable with a good socio-professional integration and a com- be better to avoid culture of embryos until day 6 if biopsy is possible at day 5 .
mon life of at least 3 years in 80% of the cases. 4.6% of men and 11.6% of women Trial registration number: Not applicable
previously have had a child with another partner. 7% of couples had already a
child conceived by sperm donation and 54% had a child project for less than 2 P-484 FMR1 and AKT/mTOR signaling in human granulosa cells:
years and 41% for 3 to 5 years. Almost all couples intended to inform their child functional interaction and impact on ovarian response
of sperm donation (95%) and father’s transidentity (90%). J. Rehnitz1, E. Capp2, B. Messmer1, A. Germeyer3, J.E. Dietrich3,
Limitations, reasons for caution: The profile of our couples probably does T. Strowitzki3, P.H. Vogt1
not reflect that of all couples consisting in a transgender man and a cisgender 1
University Women´s Hospital, Gynecological Endocrinology and Fertily Disorders
woman. The study spends a long period and couples’ characteristics could prob-
- Division of Reproductive Genetics, Heidelberg, Germany ;
ably change over the time. The French legislation is still restrictive and influences 2
Universidade Federal do Rio Grande do Sul, Department of Obstetrics and
the profile of those couples
Gynecology- Medicine School, Porto Alegre, Brazil ;
Wider implications of the findings: The future Bioethics law should allow 3
University Women´s Hospital, Gynecological Endocrinology and Fertily Disorders,
the access to ART for single women and lesbian couples and will probably allow
Heidelberg, Germany
the profile of couples composed of a transgender person to be diversified.
Study question: Does FMR1 impact AKT/mTOR-signaling-pathway in human
granulosa cells (GC) and is this relevant for ovarian response?
Summary answer: FMR1-expression significantly correlates with AKT-, TSC2-,
POSTER VIEWING SESSION mTOR- and S6K-expression in GC of women with different ovarian-response
indicating a functional effect of FMR1/FMRP on AKT/mTOR-regulated
follicular-maturation.
REPRODUCTIVE (EPI)GENETICS What is known already: FMR1 controls folliculogenesis and human
oocyte-maturation. Its transcript and protein (FMRP) is highly expressed in wom-
en´s GC. Increased FMR1 expression levels lead to reduced FMRP production
P-483 Embryo selection after PGT-A. Day 5 embryos should be
(i.e. negative feed-back loop) causing premature ovarian insufficiency/failure
preferred to day 6, even if worst by morphology and time-lapse.
(POI/POF) in 20% of cases.
V. Nagornyy1 FMRP is supposed to control translation of Tsc2 and mTor, both members
1
Clinic of Reproductive Medicine ‘NADIYA’, Embryology department, Kiev, of the AKT/mTOR signaling-pathway, via RISC-(RNA-induced-silencing-
Ukraine complex)-formation.

AKT/mTOR-signaling controls primordial follicle activation and therewith ovar- Study question: Does NIPGT-A have lower false positive rates (FPR) than
ian-reserve. FMR1/FMRP expression and mTOR/AKT-signaling are putatively invasive PGT-A?
linked in human GCs as their expression was altered in cell culture after FSH- Summary answer: When DNA sequencing from whole embryo cells was
stimulation or inhibition of AKT with MK-2206-2HCl or mTOR with rapamycin, used as the gold-standard, the FPR of NIPGT-A was 3.32-times smaller than
respectively. that obtained with invasive PGT-A.
Study design, size, duration: 297 fertile women undergoing controlled ovarian What is known already: After many years of using PGT-A, there are still many
stimulation for IVF/ICSI-treatment were recruited from 2013 to 2019 in our out- concerns, such as risks of invasive action and difficulties in the correct interpretation
patient clinic in Heidelberg. They were divided according to their ovarian response of mosaicism, which could lead to errors in the interpretation of false positive and
into either NOR: normal (n: 217) or POR: poor responders (n: 80) (according to negative results. Recently, a new technology (NIPGT-A) has arisen using cell-free
ESHRE guidelines).This study was approved by the local ethical committee and DNA present in the spent culture media of human blastocysts. Unlike invasive
conducted according to the principles of the Declaration of Helsinki. PGT-A that uses only trophoblastic cells, NIPGT-A reflects the ploidy status of
Participants/materials, setting, methods: All patients provided written trophoblastic cells and inner cell mass, suggesting that this new technology could
informed consent and completed a clinical questionnaire. be less prone to errors, being more reliable than invasive test.
mRNA was extracted from GCs after follicular aspiration and oocyte sepa- Study design, size, duration: This cohort study included a total of 37 blas-
ration; quantitative expression analyses of FMR1, AKT, TSC2, mTOR, S6K, FOXO3, tocysts vitrified on day 5 that were previously biopsied for invasive PGT-A and
FOXO1 genes were performed with specific TaqMan-Assays. presented a diagnosis of aneuploidy. The embryos were donated under informed
Statistical analyses with SPSS; significance set p<0.05. consent by patients following the Human Medical Authority regulations.
Main results and the role of chance: Rate of FMR1 expression significantly Blastocysts were thawed and cultured in 15µl drops of culture medium under
correlates with AKT-, TSC2-, mTOR-, S6K-expression levels (p< 0.001 for all; oil. After their expansion (4-8hours), the blastocysts were transferred to NGS
correlation coefficients (cc): 0.515 for AKT, 0.458 for TSC2, 0.542 for mTOR and tubes and their corresponding spent media were collected for analysis.
0.465 for S6K) suggesting a functional linkage. In patients with POR those effects Participants/materials, setting, methods: The DNA of all samples (spent
were even more prominent with the biggest increase for mTOR and S6K (cc: culture medium and whole embryo) was amplified by the MALBAC® technology
0.763 for mTOR; 0.656 for S6K; all p<0.001). (Yikon Genomics). The samples were subjected to next-generation sequencing
Functional interaction between FMRP and transcripts of these members of (NGS) using Illumina MiSeq® System. The ploidy status results obtained from
the AKT/mTOR signal pathway seem therefore to be required for follicular ChromGo™ software (Yikon Genomics) for culture medium and whole embryo
oocyte maturation. were compared to determine the accuracy of NIPGT-A for screening chromo-
Interestingly, corresponding expression analyses of some downstream mem- somal abnormalities in each embryo.
bers of this signal pathway, FOXO3 and FOXO1, in NOR and POR samples did Main results and the role of chance: DNA from all 37 spent media samples
not display association with FMR1 expression levels. But they demonstrated and whole embryos were successfully amplified. Comparing the results of
higher expression of FOXO1 in NOR and lower in POR, associated each with NIPGT-A and whole embryos sequencing, the positive predictive value (PPV)
inverse AKT-levels. This suggests a negative feedback mechanism for these genes was 93.5% and the FPR was 6.5%. On the other hand, comparing the whole
also with a putative impact on women´s ovarian reserve. embryo and invasive PGT-A results, the PPV was 78.4%, and the FPR was 21.6%
Limitations, reasons for caution: Control of human folliculogenesis and (Table 1). Both NIPGT-A and invasive PGT-A had a negative predictive value
oocyte maturation by AKT/mTOR-signal-pathway is well known. To prove pro- (NPV) of 100% and a false negative rate (FNR) of 0%. In the eight cases of
posed functional interactions to the molecular control of FMR1/FMRP- disagreement the results are presented in the Table 2.
expression in GC significant numbers of NOR- and POR-samples in different
age groups have to be collected to exclude age dependent expression variabilities
of these genes. Table 1. NIPGT-A and Invasive PGT-A results
Wider implications of the findings: Results confirm our earlier
expression analyses performed in granulosa cell cultures (COV434) pointing Whole Embryo Whole Embryo
Invasive
already to a linkage of FMR1/FMRP expression with the mTOR/AKT-signaling NIPGT-A
PGT-A
pathway. Aneuploidy Normal Aneuploidy Normal
Results will also help to improve POR-diagnostic in women before controlled
Aneuploidy 29 2 Aneuploidy 29 8
stimulation for subsequent IVF/ICSI-procedures to raise their chances for suf-
ficient mature follicles. Normal 0 6 Normal – –
PPV: 93.5% FPR: 6.5%
P-485 Noninvasive preimplantation genetic test for aneuploidy PPV: 78.4% FPR: 21.6%
(NIPGT-A) has a lower false positive rate than that of the invasive
PGT-A
L. Vagnini1, C.G. Petersen1,2, A. Renzi1, J.B.A. Oliveira1,2,
Table 2. Disagreement results of whole embryo,
A.H. Oliani3, R. Nakano4, C.G. Almodin5, C. Marcondes6,
NIPGT-A and invasive PGT-A
A. Ceschin7, A. Amaral8, E. Borges Jr9, A. Castelo Branco10,
J.B. Soares11, J. Lopes12, J.G. Franco Jr.1,2
Whole
1
Paulista Center for Diagnosis- Research and Training, Research, Ribeirao Preto, Brazil ; NIPGT-A Invasive PGT-A
embryo
2
Centre for Human Reproduction Prof. Franco Jr, Research, Ribeirao Preto, Brazil ;
3
Sao Jose do Rio Preto School of Medicine FAMERP, Research, Sao Jose do Rio 46,XY 46,XY XY,+1q(x3);+3q(x3)
Preto, Brazil ;
4
46,XY 46,XY XY,-2(x1)
Ferticlin Human Fertility Clinic, Research, Sao Paulo, Brazil ;
5
Materbaby, Research, Maringa, Brazil ; 46,XY XY,-1(x1);-9q(x1) XY,+9q(x3)
6
Santista Nucleus of Human Reproduction, Research, Santos, Brazil ; 46,XX 46,XX XX,+9q(x3)
7
Feliccita Fertility Institute, Research, Curitiba, Brazil ;
8
46,XX 46,XX XX,-4(x1)
Genesis Human Reproduction Assistance Center, Research, Brasilia, Brazil ;
9
Fertility Medical Group, Research, Sao Paulo, Brazil ; 46,XY 46,XY X0, multiple abnormalities
10
Art Fertil Human Reproduction Clinic, Research, Recife, Brazil ; 46,XX 46,XX XX,+13(x3)
11
Alpha Project - Alliance of Assisted Fertilization Laboratories, Research, Sao
46,XY XY,-1(x1);-9(x1),-19(x1);-21(x1) XY,-9(x1)
Paulo, Brazil ;
12
CENAFERT, Research, Salvador, Brazil

Limitations, reasons for caution: The sample size was relatively small, P-487 The polymorphism Ala307Thr of the Follicle Stimulating
however comparative analysis between the results of invasive and noninvasive Hormone Receptor (FSHR) gene is associated with different doses
PGT-A with whole embryo are rare. All donated embryos were classified as of recombinant FSH received during IVF/ICSI treatment.
aneuploidy by invasive PGT-A. Additionally, the cut-off for aneuploidy in cases F. Dieamant1,2, C.G. Petersen1,2, L.D. Vagnini2, A. Renzi2,
of invasive PGT-A could be variable (multicenter study). M. Canas2, B. Petersen1,2, A.L. Mauri1,2, M. Mattila1, J. Ricci1,
Wider implications of the findings: NIPGT-A has a lower FPR than A. Nicoletti1, C. Zamara1, A.H. Oliani3, J.B.A. Oliveira1,2,
invasive PGT-A and does not require micromanipulation skills, avoiding tro- J.G. Franco Jr.1,2
phectoderm biopsies trauma and seems to provide more accurate results 1
Centre for Human Reproduction Prof Franco Jr, Research, Ribeirao Preto, Brazil ;
corresponding to the ploidy status of the whole embryo. Thereby NIPGT-A 2
Paulista Center for Diagnosis- Research and Training, Research, Ribeirao Preto,
should be considered as the test of choice for genetic evaluation of Brazil ;
the embryo. 3
Sao Jose do Rio Preto School of Medicine FAMERP, Research, Sao Jose do Rio
Trial registration number: Not Applicable Preto, Brazil
P-486 Does day of PGT-A biopsy predict mosaicism rates? Study question: Is there an association between FSHR gene Ala307Thr poly-
morphism (rs6165) and ovarian reserve, ovarian response or clinical results in
B. McGuinness1, B. Abittan2, A. Pratt3, W. Shan4, S. Brownridge2,
IVF/ICSI treatment?
R. Goldman2, M. Christine2
Summary answer: Ala/Ala genotype was associated with the use of higher
1
NYU Winthrop Hospital, Obstetrics and Gynecology, Mineola, U.S.A. ; doses of recombinant FSH(r-FSH), suggesting that homozygosis of this allelic
2
Northwell Health Fertility, Reproductive Endocrinology and Infertility, Manhasset, variant(Ala) provides lower sensitivity to r-FSH.
U.S.A. ; What is known already: Follicle-stimulating hormone (FSH) is essential
3
Sanford H. Calhoun High School, General, Merrick, U.S.A. ; for folliculogenesis and acts through the FSHR that is present on the mem-
4
The Feinstein Institute for Medical Research, Biostats Unit, Manhasset, U.S.A. brane of granulosa cells. Polymorphisms in the FSHR gene may lead to an
altered pattern of receptor expression on the cell surface or to changes in
Study question: Does the rate of mosaicism differ between blastocysts affinity for FSH. The Ala307Thr polymorphism is located in the extracellular
biopsied on day 5 (D5) versus day 6 (D6) using NGS (Next-Generation domain within the hormone binding region, which can influence the response
Sequencing) technology for PGT-A (Preimplantation Genetic Testing for to endogenous and exogenous FSH stimulation. However, the influence of
Aneuploidies)? the FSHR Ala307Thr polymorphism on ovarian function is still
Summary answer: Although rate of aneuploidy is higher with embryos biop- controversial.
sied on D6 versus D5, rate of detected mosaicism does not vary based on Study design, size, duration: This prospective cohort study included
blastocyst biopsy day. 450 women who underwent IVF/ICSI cycles. The enrolled individuals met
What is known already: Advancing technologies in the field of assisted repro- the following inclusion criteria: age of ≤37 years; normal karyotype; presence
ductive technology has sensitized our methodologies with the utilization of NGS of two ovaries as observed by ultrasound examination; and no history of
in identifying trophectoderm (TE) mosaicism (≥ 2 distinct cell lines within a TE ovarian surgery, endometriosis, hydrosalpinx, infection, or endocrine
biopsy). The timing of post-fertilization mitotic errors affects the percentage of disorders.
abnormal cells present within a mosaic embryo, but it is not known if the rate Participants/materials, setting, methods: DNA was extracted from
of mosaicism is related to the day of TE biopsy. peripheral blood, and the Ala307Thr FSHR polymorphism (rs6165) was geno-
Study design, size, duration: This is a retrospective cohort study of patients typed using TaqMan SNP genotyping assay. The results were associated with
who underwent IVF/PGT-A using NGS technique from January 2018 to July age, anti-Müllerian hormone (AMH) levels, antral follicle counts (AFC), total
2019. A total of 663 patients underwent 797 IVF/PGT-A cycles. 50 cycles were dose of r-FSH, follicle size, number of retrieved oocytes, and clinical outcome
excluded due to incomplete data for analysis. of IVF/ICSI cycles. The statistical analyses were performed using Fisher’s exact
Participants/materials, setting, methods: The numbers of euploid, test and Kruskal-Wallis test.
aneuploid, low and high-level mosaic embryos were collected. Of the Main results and the role of chance: An association between the genotypes
included cycles, 257 cycles (34.4%) had embryos biopsied on both day 5 of the FSHR (Ala307Thr) polymorphism and different doses of r-FSH was
(D5) and day 6 (D6). These cycles were analyzed using paired non-parametric observed. Patients with the Ala/Ala genotype received a higher r-FSH dose than
testing to compare the D5 and D6 rates of mosaicism. The Wilcoxon rank- patients with the Ala/Thr (P=0.0002) and Thr/Thr (P=0.02) genotypes. No
sum was used to analyze rates of euploidy, aneuploidy, total mosaicism, other correlation was observed. Table 1 summarizes the results.
low-level mosaicism, and high-level mosaicism. Significance was defined by Limitations, reasons for caution: Possible limitation is the cross-sectional
p < 0.05. nature of the data. Differences in the genetic backgrounds of various ethnic
Main results and the role of chance: The rates of total mosaicism (15.7% populations might also be considered.
vs. 14.2%; p=0.19), low-level mosaicism (61.5% vs. 58.9%; p=0.14), and high- Wider implications of the findings: The results suggest that FSHR
level mosaicism (38.5% vs. 41.1%; p=0.09) did not differ significantly between Ala307Thr (rs6165) gene polymorphism is related to ovarian response but not
D5 and D6 biopsies respectively. Embryos biopsied on D5 were less likely to to ovarian reserve. This SNP can be used as an additional tool in the individual-
be aneuploid and more likely to be euploid than those biopsied on D6 (Aneuploid: ization of ovarian stimulation protocols.
33.3% vs. 44.2%; p=<0.001. Euploid: 48.8% vs. 38.8%; p=<0.001). Comparison Trial registration number: Not Aplicable
of matched samples determined that D5 and D6 blastocytes have the same
distribution of mosaicism and do not have the same distribution of aneuploidy P-488 The live birth rate and the position of breakpoints
or euploidy. in patients with recurrent miscarriage caused by reciprocal
Limitations, reasons for caution: This study is limited by small sample size. translocation
797 IVF/PGT-A cycles occurred but only 257 cycles had embryos biopsied on T. Sato1, F. Ozawa1, C. Saito1, H. Yosihara1, C. Banno1, Y. Sawada1,
both D5 and D6 and complete data for analysis. Further research is needed to Y. Matsumoto1, M. Sugiura-Ogasawara1
better understand the in vitro development of mosaic embryos and the under- 1
Nagoya City University Graduate School of Medical Sciences, Department of
lying mechanisms of blastocyst formation. Obstetrics and Gynecology, Nagoya, Japan
Wider implications of the findings: Although studies have examined the
detection of blastocyst stage mosaicism with use of PGT, no study has used Study question: Can we estimate the frequency of alternate segregation from
paired non-parametric testing to compare D5 and D6 rates of mosaicism as was the position of breakpoints in patients with recurrent miscarriage (RM) caused
done here. This study supports the argument that timing of PGT biopsy does by reciprocal translocation?
not affect rate of detected mosaicism. Summary answer: We could find no association between the frequency of
Trial registration number: not applicable alternate segregation and the position of breakpoints.

Table 1. Results
FSHR (rs6165) genotypes
Thr/Thr Thr/Ala Ala/Ala P
n (%) 141 (31.3%) 213 (47.4%) 96 (21.3%)

Age 35.0±3.8 34.5±4.1 34.5±4.7 0.57
BMI (kg/m2) 24.3±4.0 24.3±4.0 24.4±4.7 0.81
AMH (ng/ml) 1.7±2.3 1.9±2.1 1.6±2.1 0.26
AFC (n) 12.6±8.7 13.4±7.4 12.3±9.0 0.11
Total dose FSH (UI) 2085±922a 1946±955b 2364±1060a,b a
0.02; b0.0002
Time of stimulation (days) 10.3±2.5 10.1±2.4 10.5±2.2 0.17
Follicles (n): Total 11.4±6.6 12.0±7.4 11.6±7.4 0.86
Follicles (n): ≥18 mm 3.8±2.2 5.7±2.0 3.6±2.5 0.64
Retrieved oocytes: Total (n) 8.2±5.4 8.1±5.1 8.2±5.4 0.98
Retrieved oocytes Metaphase II (n) 6.1±4.3 5.9±4.1 6.1±4.8 0.92
Fertilization rate 64.5±26.4% 64.6±26.5% 64.4±29.9% 0.96
Implantation rate 23.9% 26.1% 25.7% 0.82
Pregnancy rate/transfer 34.6% 39.9% 39.8% 0.65
Pregnancy rate/patient 31.2% 35.2% 34.4% 0.79
What is known already: Preimplantation genetic testing for structural rear- Limitations, reasons for caution: The findings of the present study were
rangements (PGT-SR) is performed to prevent miscarriage worldwide, but it limited by the relatively small sample size. In particular, more samples will be
cannot improve the live birth rate in patients with RM. In carriers of reciprocal needed to evaluate the relationship between the form of the tetravalent chro-
translocations, it has been reported that the proportion of each meiotic segre- mosome structure in meiosis and the frequency of alternate segregation.
gation mode depends on certain characteristics, including the gender of the Wider implications of the findings: It might be possible to estimate the
carrier, the position of the breakpoints and the involvement of acrocentric chro- miscarriage rate because the frequency of adjacent-1 segregation which tends
mosomes. However, there is no method for using the karyotype to estimate the to end in miscarriage could be speculated from the position of the breakpoints.
frequency of alternate segregation ending in a live birth. This information may be useful in the genetic counseling of couples who are
Study design, size, duration: This is a retrospective study to evaluate the reciprocal translocation carriers before performing PGT-SR.
proportion of meiotic segregation modes in PGT-SR performed at our hospital Trial registration number: Not applicable
between 2007 and 2019. A total of 469 day 3 or day 5 embryos from 67 cycles
of PGT-SR for 20 couples with RM caused by reciprocal translocation were
examined by fluorescence in situ hybridization (FISH) or array comparative P-489 Is there a role for mitochondrial DNA quantification as a
genomic hybridization (aCGH). biomarker to select euploid blastocysts with high implantation
Participants/materials, setting, methods: For PGT-SR, FISH or aCGH potential?
was performed on biopsied samples of day 3 or day 5 embryos. According to V.C. Moreno1, R.C.S. Figueira1, S. Verza Jr1, S.C. Esteves2
the results, the meiotic segregation mode was classified, and the effects of the 1
Androfert – Andrology and Human Reproduction Clinic, Laboratory, Campinas/SP,
gender and age of the carriers, the size of the translocated segments, the ratio Brazil ;
of the translocated segment to the size of the arm with the breakpoint and the 2
Androfert – Andrology and Human Reproduction Clinic, Clinical Board, Campinas/
form of the tetravalent chromosome structure in meiosis were evaluated. SP, Brazil
Main results and the role of chance: Overall, 26% of 391 embryos with
results were diagnosed with alternate segregation, 18% with adjacent-1 segre- Study question: Is mitochondrial DNA (mtDNA) quantification in trophec-
gation, 8% with adjacent-2 segregation, 37% with 3:1 segregation, 2% with 4:0 toderm cells a useful biomarker to select euploid blastocysts with a high implan-
segregation and 9% remained undiagnosed. There was no significant difference tation potential?
in the alternate segregation frequencies between 10 patients with more than 10 Summary answer: Although low mtDNA levels are associated with higher
embryos with the meiotic segregation mode results. The frequency of alternate implantation rates, blastocyst morphology is more informative than mtDNA
segregation was identical between male and female translocation carriers (26.1 quantification to identify euploid embryos that implant.
vs 26.1%), while the incidence of the 3:1 segregation was significantly higher in What is known already: Transfer of euploid blastocysts does not invariably
female carriers than in male carriers (33.7 vs 19.1%, p<0.05). The age of the result in ongoing pregnancies, thus indicating that other factors impact embryonic
carrier did not affect the proportion of the meiotic segregation mode.No cor- implantation potential. Factors such as metabolic rates and adenosine triphos-
relation was observed between the size of the translocated segments or its ratio phate content have been shown to vary significantly in oocytes and embryos
to the size of the arm with the breakpoint and the frequency of alternate seg- and could affect embryo viability. On this basis, embryo function, indirectly mea-
regation.The frequency of adjacent-1 segregation was significantly higher in sured by mtDNA copy number, emerged as a potential quantitative biomarker
patients in which the sizes of both translocated segments were less than 50% of for embryonic selection prior to transfer. Initial data have shown that euploid
the size of each arm with the breakpoint compared with others (27.1 vs 12.1%, blastocysts with elevated mtDNA copy numbers rarely result in successful preg-
p<0.01).The form of the tetravalent chromosome structure in meiosis did not nancies. However, subsequent research could not confirm these observations
correlate with the frequency of alternate segregation. and the literature remains equivocal.

Study design, size, duration: A prospective observational study was per- Study design, size, duration: Observational study including all trophecto-
formed between September 2017 and September 2019 to analyze the contri- derm biopsy-based PGT-A cycles (n=2236 couples;2676 treatments;8151
bution of mtDNA levels for embryo implantation in 160 euploid blastocysts embryos) conducted via comprehensive-chromosome-testing techniques (CCT:
obtained from 123 infertile couples undergoing preimplantation genetic testing qPCR or NGS;Igenomix) between 2013 and 2019 at a private Italian clinic
for aneuploidy (PGT-A). Implantation outcomes were assessed based on (Genera Rome). The m-ER was investigated according to couples’ previous
mtDNA levels and blastocyst morphology criteria to determine the usefulness reproductive history (previous live birth(s):no/yes; previous failed IVF cycle(s):-
of these biomarkers to predict successful implantation in a single euploid blas- none/1/2/>2; previous miscarriage(s):none/1/>1). For PGT-A cycles following
tocyst transfer program. previous attempts (n=440/2676,16.4%), the m-ER was also investigated accord-
Participants/materials, setting, methods: Trophectoderm biopsies were ing to the former results (absence/presence of euploid blastocyst(s) in the
subjected to whole genome amplification followed by comprehensive chromo- previous cohort(s)).
some analysis via next-generation sequencing (NGS). mtDNA quantification was Participants/materials, setting, methods: The m-ER followed a distribu-
also performed using NGS. A mitochondrial score (Low - High) using the mito- tion different from a Gaussian (Kolgomorov-Smirnov and Shapiro-Wilk
chondrial-to-nuclear DNA ratio (mt/gDNA) was applied. Statistical analyses tests<0.01). Therefore non-parametric tests were adopted for the investigation
included comparative analyses, correlation coefficients, and generalized linear (i.e. Mann-Whitney U or Kruskal-Wallis tests). Generalized-linear-models were
models. Areas under the ROC curve (AUC) were estimated to assess the ability adopted to adjust for confounders. Maternal age (mean:39.3±3.3yr) was the
of the mt/gDNA and blastocyst morphology to predict implantation outcomes. only variable significantly associated with the m-ER (partial-eta-
Main results and the role of chance: Implantation rates were significantly squared:0.19,p<0.01 and power>0.99) and all the analyses were thus conducted
higher in Low-score (n=140) than High-score (n=20) euploid blastocysts among ranges of maternal age (<35yr,n=208 m-ER:66%±31%; 35-37yr,n=533
selected for transfer (48.6% vs. 25.0%, p=0.048). Correlational analysis showed m-ER:58%±33%; 38-40yr,n=909 m-ER:43%±35%; 41-42yr,n=574
a significant but weakly positive correlation between mt/gDNA and successful m-ER:28%±34%; >42yr,n=452 m-ER:17%±31%).
implantation (Spearman’s correlation, r=0.157, p=0.04). By contrast, a strong Main results and the role of chance: The m-ER was independent from
negative correlation was observed between blastocyst morphology and implan- previous live births (<35yr:no, n=195 67%±31%/yes, n=13 54%±36% p=0.2;
tation occurrence (Spearman’s correlation, r=-0.283, p < 0.0001). The logistic 35-37yr:no, n=500 58%±32%/yes, n=33 58%±38% p=0.8; 38-40yr:no, n=795
regression analyses showed no association between mt/gDNA and implantation, 43%±35%/yes, n=114 41%±33% p=0.6; 41-42yr:no, n=510 28%±34%/yes,
whereas blastocyst morphology had a positive influence on the likelihood of n=64 29%±35% p=0.5; >42yr:no, n=387 17%±30%/yes=65 21%±36% p=0.6),
euploid blastocyst implantation (OR=5.103, 95% IC: 1.027-25.357, p=0.04). as well as from the number of previous failed IVF cycles (<35yr:none, n=119
ROC analyses suggested that blastocyst morphology alone was able to better 65%±31%/1, n=53 67%±30%/2, n=21 78%±23%/>2, n=15 54%±39% p=0.3;
rank the embryos with positive implantation outcomes than mt/gDNA alone 35-37yr:none, n=301 59%±32%/1, n=102 59%±32%/2, n=62 57%±33%/>2,
(AUC = 0.72635, p=0.05). The combination of both parameters conferred a n=68 51%±35% p=0.4; 38-40yr:none, n=466 44%±35%/1, n=187 42%±34%/2,
small increase in the implantation predictive ability (AUC= 0.74725, p=0.04). n=122 41%±34%/>2, n=134 40%±35% p=0.5; 41-42yr:none, n=277
Limitations, reasons for caution: Our findings are based on a relatively 27%±34%/1, n=138 27%±34%/2, n=76 35%±38%/>2, n=83 27%±29% p=0.4;
small dataset. Another limitation is the lack of randomization as embryo mor- >42yr:none, n=235 15%±28%/1, n=107 15%±28%/2, n=54 28%±39%/>2,
phology was the primary criterion for selecting euploid blastocysts for single n=56 23%±34% p=0.1) and/or miscarriages (<35yr:none, n=171 66%±31%/1,
embryo transfer. Thus, there was a difference in the number of embryos among n=25 69%±27%/>1, n=12 63%±40% p=0.9; 35-37yr:none, n=416 58%±33%/1,
the mt/gDNA score groups. n=75 56%±30%/>1, n=42 60%±36% p=0.6; 38-40yr:none, n=674 42%±35%/1,
Wider implications of the findings: Although an association exists between n=158 44%±34%/>1, n=77 44%±35% p=0.9; 41-42yr:none, n=404
mtDNA levels and implantation rates, mtDNA cannot be used as an independent 29%±35%/1, n=99 29%±33%/>1, n=71 20%±27% p=0.3; >42yr:none n=324
biomarker to identify euploid blastocysts that result in successful implantation. 18%±31%/1, n=79 17%±33%/>1, n=49 14%±25% p=0.8). Lastly, for the 440
Our findings suggest that at present blastocyst morphology should be prioritized cycles performed after former completed PGT-A cycles, the absence/presence
in the clinical settings to rank euploid blastocysts with high implantation potential. of euploid blastocysts in the previous cohort(s) did not associate with the m-ER
Trial registration number: None (<35yr:absence, n=5 69%±19%/presence, n=203 66%±31% p=0.3; 35-37yr:ab-
sence, n=23 40%±33%/presence n=510 59%±32% p=0.1 ; 38-40yr:absence,
P-490 The mean euploidy rate per cohort of biopsied blastocysts n=84 38%±33%/presence, n=825 43%±35% p=0.25; 41-42yr:absence, n=75
is independent of couples’ previous reproductive history: an 29%±35%/presence, n=499 28%±34% p=0.4; >42yr:absence, n=78 20%±31%/
analysis of 2676 cycles with preimplantation-genetic-testing-for- presence, n=374 17%±30% p=0.7).
aneuploidies. Limitations, reasons for caution: Segmental and allegedly-mosaic aneuploi-
D. Cimadomo1, A. Capalbo2, A. Vaiarelli1, S. Colamaria1, dies were not reported. The sample size among the treatments following a
R. Maggiulli1, L. Albricci1, L. Dovere1, M. Stoppa1, L. Tacconi1, former completed PGT-A cycle should be increased. The data should be con-
F. Sanges1, D. Soscia1, A. Giancani1, E. Scepi1, F.M. Ubaldi1, firmed from a multicenter perspective. For couples who experienced former
L. Rienzi1 IVF failures and/or miscarriages, the implantation rate after euploid blastocyst
1 transfer should be investigated.
Clinica Valle Giulia, G.EN.E.R.A. centers for reproductive medicine, Roma, Italy ;
2 Wider implications of the findings: These data are critical to counsel infer-
Igenomix, Igenomix Italy, Marostica, Italy
tile couples before and after PGT-A cycles. Apparently, their previous repro-
Study question: Is the mean euploidy rate per cohort of biopsied blastocysts ductive history does not affect the m-ER beyond the maternal-age-effect. For
(m-ER) associated with the previous reproductive history of the couples under- idiopathic repeated-IVF-failures and recurrent-pregnancy-losses, the causes might
going preimplantation-genetic-testing-for-aneuploidies (PGT-A)? be related to endometrial receptivity issues. Yet, transferring only euploid blas-
Summary answer: If adjusted for maternal age, couples’ previous reproduc- tocysts minimizes embryo-derived adverse reproductive outcomes.
tive history has no effect on the m-ER after PGT-A. Trial registration number: None
What is known already: Several studies investigated the association between
the euploidy rates after PGT-A and i) patients’ characteristics (e.g. maternal/ P-491 Cleavage biopsy, same results as trophectoderm biopsy but
paternal age, sperm factor), ii) ovarian stimulation strategy (e.g. protocol, dose without mosaicism
of gonadotrophins, trigger of ovulation), iii) IVF cycle characteristics (e.g. incu- C. Encinas1, S. Novo1, A. Brualla2, Á. García-Faura1, B. Marquès1,
bator, culture strategy), iv) blastocysts’ features (e.g. morphological quality, F. García1, C. Castelló1, M. López-Teijón1
developmental rate). Conversely, the expected m-ER after PGT-A due to cou- 1
Institut Marquès, Reproductive Medicine Service, Barcelona, Spain ;
ples’ previous reproductive history (previous cycles, miscarriages, live births and 2
Institut Marquès Ireland, Reproductive Medicine Service, Clane, Ireland
lack of euploid blastocysts identified during prior cycles) still needs to be outlined.
Yet, this information is critical to counsel the patients towards a first PGT-A Study question: Analysing the evolution of PGT-A, does the biopsy at blas-
cycle, or after former adverse outcomes. tocyst stage bring great advantages?

Summary answer: Both cleavage and blastocyst stage biopsy provide optimal more research is needed to further our understanding of the clinical usefulness
and equivalent clinical results. of mitoscore as a biomarker for embryo viability
What is known already: Recently, trophectoderm biopsy has been consid- Study design, size, duration: This is a retrospective study involving analysis
ered the gold standard for Preimplantational Genetic Testing (PGT). The main of 132 term babies born from transfer of a single euploid blastocyst from
reason is that the higher number of analysed cells, the better representation of February 2018 to march 2019 at our center
embryo’s chromosomal constitution should be achieved. However, this technique Participants/materials, setting, methods: We performed preimplantation
has also made the mosaic embryo diagnostic group appear, a result that can be genetic testing for aneuploidy (PGT A) for patients with advanced maternal
considered as ambiguous. The possibility of getting a confusing result is being age, recurrent pregnancy losses and recurrent implantation failure.Trophectoderm
used by some groups as an argument to challenge the usefulness of blastocyst biopsy was done and the samples were analysed by comprehensive chromosomal
biopsy and PGT-A. This study aims to assess the differences in clinical outcomes screening. Based on mitoscore value two groups were formed, group A
between treatments were cleavage or trophectoderm biopsies have been (< 25 ) group B (> 25). Clinical pregnancy rates(CPR), live birth rates (LBR),
performed. hypertensive disorders in the antenatal period, birth weights and anomaly rates
Study design, size, duration: This is a retrospective study containing data in the two groups were compared.
from 681 single blastocysts transfer PGT-A cycles between 2017 and 2019 using Main results and the role of chance: There was no difference in baseline
Next Generation Sequencing (NGS). On one hand, D3-group: 229 cycles with characters between the two groups. Out of 132 euploid blastocysts 64 (48.4%)
cleavage stage embryos biopsied at day 3 of culture, cultured up to blastocyst of the blasts had a mitoscore< 25 and 68 (51.6%) had a mitoscore of > 25.
stage and fresh transferred. On the other hand, D5/6-group: 452 cycles, with There was no statistically significant difference in CPR (68.7 % vs 66.1%) and
embryos cultured to blastocyst stage, biopsied and vitrified. LBR (57.8% vs 55.8%).Hypertensive disorders in the antenatal period were sig-
Participants/materials, setting, methods: In both groups, embryos were nificantly higher in the group with mitoscore > 25 ( 37.6% in group B vs 12.2%
cultured using single step media in time-lapse incubators up to blastocyst stage. in group A). %). Birth weights of babies in the group with mitoscore more than
Patient characteristics and demographic data were homogenous and similar 25 (2.54 +/- 0.46) was significantly lower when compared to the group with
between both groups. The number of embryos biopsied, blastocyst rate, results mitoscore less than 25 (2.19 +/- 0.36) (p=0.025). There was no difference in
of NGS analysis and pregnancy and miscarriage rates were compared between anomaly rates in both the groups
both groups. The Chi-square test was used for statistics. Limitations, reasons for caution: This is a retrospective data and sample
Main results and the role of chance: D3-group had significantly more size was small and indications of PGT A are still debatable. Further research is
biopsied embryos (n=1097/1287; 85.2%) than D5/6-group (n=1715/3233; necessary.
53%; p<0.05). However, the good quality blastocyst rate was equivalent between Wider implications of the findings: This study provides evidence that mito-
groups (D3=50.2%; D5/6=53%; p=0.086), demonstrating that cleavage stage score has no role in predicting the implantation potential of an embryo. Higher
embryo biopsy did not compromise embryonic development. mitoscore embryos were associated with hypertensive disorders in pregnancy
To compare NGS results between groups, all biopsied embryos were included and lower birth weights. Therefore further research is required to understand
in D5/6 group, while in D3-group only the biopsied embryos with good quality the implications of mitoscore and its effect on reproductive outcomes
in day 5/6 were considered (≥3BB Gardner score). Aneuploidy rates proved to Trial registration number: not applicable
be similar (D3:48%; D5/6:46.8%; p=0.262) as well as the non diagnostic result
(D3=5.1%; D5/6=1.2%; p=0.091). Nevertheless, there were statistically signif-
P-493 Prospective analysis of multiplatform preimplantation
icant differences between euploidy rates, being higher in D3-group (46.9%) than
genetic testing for aneuploidy (PGT-A) confirmation of
in grouP-D5/6 (34.5%; p<0.05). After genetic counselling, and considering some
trophectoderm re-biopsy from aneuploid embryos and
mosaic embryos as transferable, a total of 45.3% of embryos could be considered
corresponding whole embryos
for transfer matching the values obtained in D3-group.
In both groups, a single blastocyst was transferred. There were 144 embryo M. Abae1, N. Arrach2, Y. Ying3, X. Zhang4, A. Goodman5, K. Abae6
1
transfers in D3-group and 262 in D5/6 group. Pregnancy rates (D3=68.8%; Fertility & Genetics, Reproductive Endocrinology, Plantation- FL, U.S.A. ;
2
D5/6=72.5%; p=0.152), ongoing pregnancy rates (D3:=52.8%; D5/6=52.3%; Progenesis, Progenesis, San Diego, U.S.A. ;
3
p=0.411), and miscarriage rates (D3=8.1%; D5/6=8.4%; p=0.356), were equiv- Univeristy of Soujht Florida, Obstetrics & Gynecology, Tampa, U.S.A. ;
4
alent between groups. Fertility & Genetics, Embryology, Plantation, U.S.A. ;
Limitations, reasons for caution: Although the considerable data set pro- 5
NXGENMDX, Pgt, Grand Rapids, U.S.A. ;
6
vided, live birth rates should be analysed. Every D3-group embryo was trans- Fertility & Genetics, Fertiility & Genetics, Plantation, U.S.A.
ferred in the fresh cycle, but all D5/6-group embryos were transferred in a cycle
were no ovarian stimulation was performed. This could be a source of bias. Study question: How do results of PGT-A compare between different con-
Wider implications of the findings: Considering that pregnancy and mis- temporary testing platforms, next generation sequencing (NGS) and single
carriage rates were similar, both strategies can coexist and it is not mandatory nucleotide polymorphism (SNP) from trophectoderm (TE) re-biopsy samples
to biopsy at blastocyst stage. As it is done in our clinic, each case should be and corresponding whole embryos (EM)
evaluated to decide which is the best strategy for the patient. Summary answer: SNP had moderate correlation with NGS. Discrepancy
Trial registration number: doesn’t apply between TE and whole EM results was not associated with platforms, but to
sample variability and potentially mosaicism.
What is known already: PGT-A improves successful pregnancy outcomes
P-492 Mitoscore in a euploid embryo- can it be a game changer ?
following elective single embryo transfer. However, significant clinical uncertainty
S.S. Garimella1, K. Mantravadi1, D.R. Gedela1 remains about the accuracy of mosaicism in embryos undergoing PGT-A testing.
1
oasis fertility, reproductive medicine, hyderabad, India Furthermore, re-biopsy of abnormal embryos have occasionally led to euploid
results adding new challenges to the PGT-A management and patient expectation.
Study question: Does mitoscore value predict implantation potential of an Study design, size, duration: Thirty pre-tested aneuploid embryos that were
embryo and have an impact on antenatal outcomes and neonatal birth weight? donated by couples who had a previously confirmed viable pregnancy after
Summary answer: Mitoscore has no role in predicting implantation potential transfer of single euploid embryo following PGT-A. Discarded genetically abnor-
of an embryo, but high mitoscore was associated with hypertensive disorders mal embryos that were consented for donation for research were analyzed in
in pregnancy and lower birth weight. this exempt IRB approved blinded study. The inclusion criteria did not consider
What is known already: Mitoscore is a mitochondrial biomarker which is an factors such as cause of infertility, age, race, or ethnicity. This study excluded
indicator of energy status of embryo. Some studies indicate that increase in the any untested or genetically normal embryos.
mitochondrial DNA (mtDNA) in the embryo is indicative of insufficient level of Participants/materials, setting, methods: Thirty pre-tested aneuploidy
energy and a low implantation potential. But recently there is a debate about embryos with SNP that were donated for this study were re-biopsied. TE and
the usefulness of mitoscore as a predictor of successful implantation .Therefore, the corresponding EM were loaded separately. Sixty blinded samples were

subject to whole genome amplification followed by NGS. Each DNA sample non-synonymous variants with consequence at protein level were considered
was tested with two sequencing platforms: VeriSeq-PGS and Reproseq. for case-control comparison.
Main results and the role of chance: Of 30 embryos, 23 were successfully Main results and the role of chance: Genes previously associated with
amplified/sequenced in both TE and EM. The two NGS platforms had 100% ovarian failure accumulated less gene variants than any selection from the total
correlation when same source DNA is used. Each sequencing platform showed of the exome (t test, p value =2.2e-16). Analysis of the clinical exome (18311
96% correlation between TE and EM. SNP and NGS had 79% correlation at TE genes) revealed more than 47,000 variants were patient specific, therefore asso-
level and 84% at EM. Discrepancy between TE and whole EM results was not ciated with individual genetic variation. We highlighted 66 novel variants that
associated with the platforms, but to sample variability, potentially due to mosa- were at least shared by 10 % of cases and absent in controls, as a criterion to
icism. There was 17.4% (4 out of 23) of re-biopsied TE and whole EM found to be related to ovarian failure. This profile of 66 variants altogether identified the
be euploid by the two NGS platforms compared to SNP. This discrepancy could 100% of cases stratifying into two main types of ovarian failure, finding inside
be related to the technology used, biopsy quality or to embryonic mosaicism. one of them two clear subtypes. With two variants, DNAH6, a gene related to
Limitations, reasons for caution: The two NGS platforms were performed microtubule activity, was the major contributor to the stratification. Furthermore,
on the same DNA, while SNP array was performed with separate biopsy. 6 of these 66 variants were presented in at least 20 cases, one particularly
Difference in biopsy quality and/or potential mosaicism can lead to a discrepancy affecting 31 cases of ovarian failure including both primary ovarian insufficiency
between NGS and SNP. and occult ovarian failure. Genes and biological processes affected by these new
Wider implications of the findings: PGT-A has shown to improve preg- variants included: MUC6 for cytoprotection of epithelial surfaces; BRPF3,
nancy outcomes with a single embryo transfer. However, the choice of PGT-A involved in chromatin organization; AP5M1,which could play a role in apoptosis;
technology platform presents a challenge from genetic counselling perspective. CRISPLD2, that promotes extracellular matrix assembly; GALNS, participates
The impact of potential mosaicism, PGT-A technological limitation, and variability in degradation of glycosaminoglycans; and MCM5, a component of the complex
across biopsies should be taken into account when counseling patients needed for DNA replication initiation.
with PGT-A. Limitations, reasons for caution: To prove the preventive value of this
Trial registration number: Not Applicable variant profile, an independent clinical validation should be performed. A higher
sample size would help in the patient stratification of subtypes of ovarian failure
P-494 Clinical exome sequencing from peripheral blood reveals due to the genetic heterogeneity of this disorder; however, we have detected
novel gene variants to prevent ovarian failure a common variant in 31 cases.
I. Henarejos Castillo1,2, A. Aleman1, B. Martinez-Montoro3, Wider implications of the findings: This study elucidates the underlying
F. Javier Gracia4, P. Sebastian-Leon1,5, J.A. RemohI2,6, A. Patiño4,7, genetic cause of idiopathic ovarian failure, identifying new variants by blood
P. Royo3, G. Alkorta-Aranburu4, P. Diaz-Gimeno1,5 sequencing that could become a preventive biomarker and create a new
1 taxonomy for this disorder. Additionally, our results highlighted that studies
IVI-RMA IVI Foundation, Department of Genomic & Systems Reproductive
based only on disease associated genes could overshadow discovery of new
Medicine, Valencia, Spain ;
2 variants.
Universidad de Valencia, Department of Pediatrics- Obstetrics- and Gynaecology,
Trial registration number: Research supported by IVI Foundation, IVI-RMA
Valencia, Spain ;
3 Global. I. Henarejos-Castillo is financially supported by the Conselleria de
IVI-RMA Pamplona, Reproductive Medicine, Pamplona, Spain ;
4 Innovación, Universidades, Ciencia y Sociedad Digital (ACIF/19/148). Research
CIMA Lab diagnostics, CIMA lab, Pamplona, Spain ;
5 co-financed by Navarra Goverment and European Regional Development’s funds
Instituto de Investigación Sanitaria Hospital Universitario y Politécnico La Fe,
(FEDER): 0011-1365-2017-000265. Begoña Montoro is financially supported by
Instituto de Investigación Sanitaria Hospital Universitario y Politécnico La Fe,
Navarra Goverment: 0011-1408-2018-000011
Valencia, Spain ;
6
IVI-RMA Valencia, Reproductive Medicine, Valencia, Spain ;
7 P-495 Correlation between blastocyst morphology, euploidy and
Universidad de Navarra, Laboratorio de Pediatría- Unidad de Genética Clínica,
implantation in a PGT-A program of 1312 screened blastocysts. Is
Pamplona, Spain
it worth biopsing a grade C embryo?
Study question: Could the identification of specific sequence variants in DNA C. Escriba1, M. Alaves1, M. Benavent1, C. Miret1, A. Garcia1,
stratify individuals with ovarian failure? N. Costa-Borges2, G. Calderon2, J. Crespo1, J. Teruel1
Summary answer: This prospective study has discovered 66 DNA variants 1
Juana Crespo, IVF Laboratory, Valencia, Spain ;
associated with ovarian failure. 2
Embryotools, Research and quality, Barcelona, Spain
What is known already: Around 1% of women under the age of 40 develop
premature ovarian insufficiency (POI), which includes accelerated attrition of Study question: What is the reproductive potential of grade C blastocysts?
the ovarian follicle reserve and amenorrhea. An unknown percentage of women Are we really helping the patients by biopsing them?
suffer occult ovarian failure, a less severe form of ovarian insufficiency that Summary answer: Grade C blastocysts have significantly poorer reproductive
increases infertility due to a diminished ovarian reserve. In most cases, the genetic potential than grade A and B blastocysts regarding aneuploidy and implantation
cause or origin of this heterogeneous and complex disorder is unknown. rates but still result in ongoing pregnancies.
Interestingly, whole exome sequencing based studies have started identifying What is known already: Several studies show the benefit of trophectoderm
new POI-associated sequence variants even though most of those studies suffer biopsy combined with chromosomal screening, a good vitrification program and
from lack of control or small sample size. subsequent transfer of euploid embryos. However, the indications as to when
Study design, size, duration: A 150 case-control study was conducted to biopsy poor quality embryos and their clinical contribution to patients is still
between 2017 and 2019: 118 cases diagnosed with primary ovarian insufficiency unclear or controversial.
(n=35) or occult ovarian failure (n=83) and 32 controls based on AMH levels Some studies suggest that grade C embryos have a higher aneuploidy rate,
and/or antral follicle counts. Next, whole-exome sequencing of DNA from 150 but euploid ones have a similar implantation rate to better quality embryos.
peripheral blood samples was performed using SureSelect Clinical Research However, this contradicts the establish fact that embryo quality is predictive of
Exome V2 (Agilent Technologies) and Illumina sequencing. The study was implantation.
approved by the institutional review board of Instituto Valenciano de Infertilidad Study design, size, duration: This is a retrospective observational study
(1709-PAM-090-PR). performed in a private centre between May 2017 and September 2019. The
Participants/materials, setting, methods: In order to identify variants study includes the data analysis of 1312 blastocysts undergoing PGT-A (mean
more likely associated with ovarian failure, we excluded variants with a population maternal age, 39.4 ± 3.76 years) obtained from 270 patients following 339 PGS
frequency higher than 5% - considering the negative impact of variants associated cycles. 232 euploid blastocysts were transferred in 178 frozen embryo transfers.
with ovarian failure during natural selection. In addition, only variants in genomic Of this, it was possible to track the implantation outcome of 226 of them.
positions with >100x were considered. Furthermore, variants were filtered Participants/materials, setting, methods: PGT-A was offered to patients
according to their absence/presence in IGSR, to identify new variants and only of advanced maternal age and/or with repeated IVF failures. Patients underwent

ICSI cycles with day 3 assisted hatching. Trophectoderm biopsies were per- biopsy could be the main drawbacks of most of the published studies evalu-
formed on day 5 and/or day 6 embryos, with laser assistance. ating ICE.
Blastocyst morphology was scored in 3 groups: A: excellent (AA, AB, BA), B: Study design, size, duration: This retrospective study includes 95 RecT (124
good (BB), C: average and poor-quality embryos (BC, CB, CC). (Gardner- cycles) and 36 RobT (49 cycles) carriers undergoing PGT-SR between March
Schoolcraft classification) 2016-July 2019. Array Comparative Genomic Hybridization (aCGH) or Next
Relationships between these groups regarding euploidy, implantation and mis- Generation Sequencing (NGS) were the techniques used coupled with trophec-
carriage rates were assessed. toderm biopsy (TB). Only cases with female age under 37 were included. A total
Main results and the role of chance: The euploidy rate was 38.2 of 532 blastocyst embryos (female RecT;200, male RecT;176, female RobT;78
(n=157), 29.7(n=853) and 26.8% (n=302) in the A, B and C blastocyst mor- and male RobT;78) were evaluated.
phology groups, respectively, showing that in group A there were significantly Participants/materials, setting, methods: One hundred and seventy-three
more euploid embryos than in groups B and C. There are no differences PGT-SR cycles with 532 blastocyst-stage embryos were evaluated according to
between the mean age of the 3 groups. (38.16, 38.26 and 38.46 years, the gender of the carrier partner, the type of rearrangement, chromosomes
respectively). involved in the translocations as well as the position of breakpoints on the
The mosaicism rate does not differ between groups. (14, 15 and 13.6% in A, incidence of balanced/unbalanced embryos and ICE. Multi-level generalized
B and C groups, respectively). linear mixed model was used to for statistical comparisons of the probability of
Chi-square tests were used to assess the relationship between blastocyst achieving a euploid embryo.
morphology and euploidy rates and between blastocyst morphology and implan- Main results and the role of chance: The median female and male ages,
tation/miscarriage rates of euploid blastocysts. retrieved oocytes, matured oocytes, number of fertilized oocytes and num-
The implantation rate of euploid blastocysts was 80 (n=45),61.15 (n=139) ber of biopsied embryos were similar among the female RecT and RobT,
and 35% (n=42) in the A, B and C blastocyst morphology groups, respectively, and male RecT and RobT carriers. The euploidy rate was also similar in RobT
showing high significant differences among the three groups. compared with RecT carriers [57/156(36.5%) vs 112/376(29.8%), p=0.127].
Focusing only on poor-quality (CC) blastocysts within the grade C embryos Pure ICE (aneuploid balanced) rate was significantly higher in RobT
population we observe that the implantation rate drops to 25% (n=24) from [48/156(30.8%) vs 53/376(14.1%), p<0.001]. In contrast, combined ICE
the 35% of overall implantation rate of the group (BC, CB, CC), highlighting the (aneuploid unbalanced) was significantly higher in RecT [72/376(19.1%) vs
importance of embryo quality as a predictor of the implantation potential even 16/156(10.3%), p=0.011]. No significant differences were observed in the
if euploid. total percentage of pure ICE plus combined ICE between RobT and RecT
Miscarriage rates were inversely correlated to quality (8 (n=36), 13 (n=85) [64/156(41%) vs 125/376(33.2%), p=0.088]. A gender-based subgroup
and 26% (n=15)) but did not achieve statistical significance. analysis also shown no differences in euploidy rate between male RecT and
Limitations, reasons for caution: The study is limited by its retrospective female RecT carriers [58/176(33%) vs 54/200(27%), p=0.207] and male
nature and the low number of grade C blastocysts transferred as they are the RobT and female RobT carriers [28/78(35.9%) vs 29/78(37.2%), p=0.86].
last option for transfer. Additionally, it is common to transfer more than one No significant differences were observed in euploidy rates, pure ICE and
grade C embryo to increase the chances of pregnancy, losing implantation track combined ICE according to the length of the translocated fragment, the
of some of them. chromosome group, and the position of the breakpoints in RecT cases.
Wider implications of the findings: It is uncontested that grade C embryos Multi-level generalized linear mixed model using binomial distribution
have a lower reproductive potential, but we managed to quantify this potential, revealed that and female age was the factor related to the incidence of
providing clinicians with useful information to manage the cycle and inform euploid embryo.
patients accordingly. Limitations, reasons for caution: The size number, the retrospective nature
Trial registration number: Not applicable of the study and the absence of age-matched controls are the main
limitations.
Wider implications of the findings: Limited studies evaluated PGT-SR
P-496 The Structural Rearrangement-specific Characteristics based on TB and 24 chromosomes analysis. Reported studies included limited
Have Limited Effect on Ploidy Status in Young Couples embryo yield; female age, which caused bias, is not restricted. Our data provide
Undergoing Preimplantation Genetic Testing for Structural a deeper insight about the effect of the translocation type, carrier-gender,
Rearrangements (PGT-SR) by aCGH/NGS chromosomes involved in rearrangement and position of breakpoints on
M. Gultomruk1, F.K. Boynukalin2, C. Rubio3, L. Rodrigo3, PGT-SR results.
E. Turgut2, Z. Yarkiner4, S. Ecemis5, G. Karlikaya2, N. Findikli1, Trial registration number: None
M. Bahceci2
1
Bahceci Fulya IVF Centre, IVF Laboratory, Istanbul, Turkey ;
2 P-497 Genetic and Clinical Outcomes of Preimplantation Genetic
Bahceci Fulya IVF Centre, Infertility Clinic, Istanbul, Turkey ;
3 Testing for Aneuploidy (PGT-A) by Using Next Generation
Ignomix S.L, Reproductive Genetics, Valencia, Spain ;
4 Sequencing (NGS) in Young Couples With Idiopathic Recurrent
Cyprus Science University, Biostatistics, Kyrenia, Cyprus ;
5 Miscarriage
Bahceci Ankara IVF Centre, Infertility Clinic, Ankara, Turkey
F.K. Boynukalin1, M. Gultomruk2, E. Turgut3, S. Ecemis4,
Z. Yarkiner5, R. Abali3, G. Karlikaya3, N. Findikli6, M. Bahceci7
Study question: What are the possible effects of carrier and rearrange-
1
ment-specific characteristics on the ploidy status of embryos and presence of Bahceci Health Group, Reproductive Endocrinology, Istanbul, Turkey ;
2
interchromosomal effect(ICE) in cases undergoing PGT-SR? Bahceci Fulya IVF Centre, IVF Laboratory, Istanbul, Turkey ;
3
Summary answer: Euploidy and ICE rates are similar in Robertsonian and Bahceci Fulya IVF center, Infertility Clinic, Istanbul, Turkey ;
4
reciprocal translocations. Euploidy rates are independent from carrier and struc- Bahceci Ankara IVF Center, Infertility Clinic, Ankara, Turkey ;
5
tural rearrangement-specific characteristics, except female age. Cyprus Science University, Biostatistics, Kyrenia, Cyprus ;
6
What is known already: Structural chromosome rearrangements such as Bahceci Fulya IVF Center, IVF Laboratory, Istanbul, Turkey ;
7
reciprocal translocations (RecT), Robertsonian translocations (RobT) and inver- Bahceci Fulya IVF Center, Infertility, Istanbul, Turkey
sions are associated with increased risk of fertility problems, recurrent miscar-
riages and progeny congenital abnormalities due to the production of unbalanced Study question: Are clinical outcomes of preimplantation genetic test for
gametes during meiosis. It has long been suggested that chromosomes with aneuploidy(PGT-A) satisfactory in idiopathic recurrent miscarriage(iRM) and
structural rearrangements can also impair the segregation of other chromo- is there a characteristic distribution for aneuploidy screening data?
somes which is known as ICE. ICE has previously been evaluated in several Summary answer: Although ~50% live-birth per embryo transfer(ET) is
studies, but the results are conflicting. The limited number of chromosomes acquired, high miscarriage rate(MR) is observed in iRM with PGT-A. Aneuploidy
analyzed in fluorescence in situ hybridization(FISH) technique and blastomere type differs compared to product of conception.

What is known already: Aneuploidy account for 42% of miscarriages in Summary answer: Good embryo quality is related to euploidy while day-6
RM. The theoretical goal of PGT-A was to identify and transfer euploid biopsied embryos and hatched blastocysts have fewer chances to survive after
embryos and decrease the rate of miscarriage. There is paucity of data regard- freezing.
ing the role of PGT-A in RM. Although, several retrospective analysis per- What is known already: In Preimplantation Genetic Testing (PGT) cycles,
formed on RM patients demonstrated a trend toward decreased MR and blastocyst formation has long been related to euploidy and some authors have
increased live birth rate(LBR), such results are usually blurred and biased by detected differences in the ploidy depending on the embryo quality or whether
the presence of an additive aneuploidy risk as advanced maternal age. In the biopsy has been performed on day-5, day-6, or day-7 of development.
addition, these studies used chromosomes analyzed in fluorescence in situ However, very little has been published about differences in the survival rate
hybridization(FISH) technique and blastomer biopsy which are the main draw- related to the quality of the Inner Cell Mass (ICM) and the trophectoderm (TE),
back of the studies. the day of biopsy or the hatching status of the embryo.
Study design, size, duration: This is a retrospective single center study to Study design, size, duration: Retrospective study including 2538 biop-
analyze the PGT-A results, using trophectoderm biopsy and NGS, of iRM cases sied blastocysts from 856 PGT cycles performed in the same center between
in which female age is <35. Data of 133 patients undergoing PGT-A with their February 2018 and October 2019. Biopsied embryos were divided according
423 blastocyst embryos between February 2016-January 2019 were evaluated. to ICM and TE quality Gardner’s evaluation (Best AA; Good AB, BA, BB;
Patients were sub-grouped according to the number of previous miscarriages Poor AC, CA, BC, CB, CC) and the day of biopsy (day-5 or day-6).
as; 2, 3, and >3. The effect of patient and embryo characteristics on ploidy Completely hatched blastocysts (HB) were analyzed separately. The inde-
status, LBR and MR were assesed. pendent relationship between each group and both euploidy and survival
Participants/materials, setting, methods: Multi-level generalized linear rate was assessed.
mixed model was used to analyze the effect of previous live birth, previous Participants/materials, setting, methods: For each cycle, serial biopsies
number of miscarriages, body mass index, female age, male age and cycle char- were performed depending on the herniation status of each embryo on day-5
acteristics on probability of achieving a euploid embryo. Binary Logistic or on day-6 of development. TE biopsy was immediately followed by vitrification
Regression was used to analyze affect of factors on abortion compared to live of all biopsied embryos. Comprehensive chromosome screening PGT was per-
birth. The distribution of monosomies, trisomies and partial aneuploidies affect- formed using high resolution next generation sequencing (NGS) methodology.
ing each chromosome were detected in embryo yield. Embryos were thawed and replaced in case of both normalcy and survival evi-
Main results and the role of chance: The overall incidence of euploid dence following an endometrial preparation cycle. Statistical report was per-
embryo was found to be 62.4%(264/423). The distribution of aneuploidy type formed under R version 3.6.1. (2019-07-05).
was as; monosomy 27%(43/159), trisomy 22%(35/159), segmental Main results and the role of chance: A total of 2534 embryos resulted
17%(27/159), double aneuploidy 15.7%(25/159), complex aneuploidy informative after genetic analysis (99.84%). Embryo ploidy showed a rela-
11.9%(19/159) and chaotic 6.3%(10/159). The most frequently involved chro- tionship with embryo quality as significant differences were found among
mosomes in aneuploidies were 16(14%), 21(10.1%) and 22(9.4%). The euploidy the percentage of euploid embryos of the three quality groups (Best 61.56%
rate of embryos was not statistically different between 2, 3 and >3 number of n=294; Good 44.52% n= 1642; Poor 31.09% n= 595) (p<0.001).
miscarriages (64.7%(124/207), 59.4%(82/138), 61.5%(48/78); respectively, Euploidy did not correlate with the biopsy day (hence, the day of herniation)
p=0.598). Multi-level generalized linear mixed model revealed that female age independently of the embryo quality or with the hatching status of the
was the only significant factor that effects the probability of euploidy (OR:0.87, embryo. Thus, no differences were found between the euploidy rate of
95%CI:0.78-0.98, p=0.009). Clinical pregnancy rate (CPR), LBR and MR for day-5 (44.15%; n=1838) and day-6 (40.72%; n=679) (odds ratio [OR]=0.87;
patients per ET was 64.3%(83/129), 48.8%(63/129) and 24.1%(20/83) respec- p=0.1191) or between the chromosomal status of the hatched (47.19%;
tively. Although there is a trend toward increased MR with increasing number n=310) versus non-hatched embryos (42.79%; n=2228) (OR 0.84;
of previous miscarriage, CPR, LBR and MR were not significantly different in p=0.1375). Regarding survival rate, results showed no relationship with
subgroups regarding 2, 3, >3 (CPR: 62.3%(43/69), 75.7%(28/37), 52.2%(12/23), embryo quality even if a trend was detected against Poor embryo quality
p=0.16; LBR: 52.2%(36/69), 51.4%(19/37), 34.8%(8/23), p=0.33; MR: group compared to Best (OR=0.38; p=0.1670) and Good (OR=0.36;
16.3%(7/43), 32.1%(9/28), 33.3%(4/12), p=0.23 respectively). MR is higher p=0.0779). On the other hand, non-hatched day-6 biopsied embryos had
than reported studies for infertile patients in the PGT-A and lower than expected lower survival rates (82.19%) than non-hatched day-5 embryos (91.46%)
MR for RM patients with natural conception. Binary logistic regression analysis (OR=0.12; p=0.0111) independently of the embryo quality. Concerning
showed that BMI (p=0.004, OR:1.22, 95%CI[1.06-1.4]), embryo quality (poor the hatching status of the embryo, HB had fewer chances to survive (69.01%)
quality, p=0.082, OR:4.6, 95%CI[1.2-17.1]) were significantly correlated with than non-hatched blastocysts (91.27%) (OR=4.69; p<0.001).
miscarriage. Limitations, reasons for caution: This study is limited by its retrospec-
Limitations, reasons for caution: The retrospective nature of the tive nature. Even if embryologists do pass routine evaluation controls, qual-
study is the main limitation. In addition, the study did not contain a con- itative blastocyst morphology grading is always confined by its
trol group. subjectivity.
Wider implications of the findings: Until now, there is paucity of data Wider implications of the findings: Knowing that embryo quality correlates
to recommend PGT-A as part of routine use for RM. However the data are to euploidy can help improving the counselling to PGT and non-PGT patients.
compelling that this technology can enhance outcome by reducing the risk of Furthermore, finding that day-6 non-hatched blastocysts and HB have fewer
clinical miscarriages and ongoing aneuploidy pregnancies. Well-designed studies chances to survive can lead to a more continuous evaluation of the embryos on
is needed to analyze efficacy and safety of treatment. day-5 in case of blastocyst biopsy.
P-498 Relationship between both euploidy and thawing survival

rates and embryo quality, day of biopsy or hatching status in P-499 Polar body analysis in preimplantation genetic testing
Preimplantation Genetic Testing cycles for monogenic diseases (PGT-M) – a genome wide approach for
haplotype phasing and assessment of ploidy
M. Florensa Bargalló1, A. Cladellas Artola1, M. Bellés Fernandez1,
M. Jimenez Blasco1, J. Herreros Cuesta1, H. Huete Ferriz1, P. Forsblom1, J. Pricelius1, M. Stecher1, C. Bayerl1, D. Gutknecht2,
N. Garrido Puchalt2, M. Esbert Algam1 B. Poost3, Y. Charron4, C. Meyer5, A. Jantke4, D. Noss3,
1 M. Bals-Pratsch2, C. Nevinny-Stickel-Hinzpeter1
IVI Barcelona, IVF laboratory, Barcelona, Spain ;
1
2
IVI Foundation, Research administration, Valencia, Spain SYNLAB Human Genetics Munich, Preimplantation diagnostics and chromosomal
microarrays, Munich, Germany ;
2
Study question: In blastocyst biopsy cycles, is there any association between Profertilita Centre for Reproductive Medicine, IVF clinic, Regensburg,
embryo quality, biopsy day and embryo hatching status and both embryo ploidy Germany ;
3
or post-thawing survival rates? Centre for Reproductive Medicine, IVF clinic, Munich, Germany ;

4 2
Centre for Assisted Fertility Treatment and Fertility Preservation, IVF clinic, Berlin, Tianjin Medical University General Hospital, Department of Gynecology and
Germany ; Obstetrics, Tianjin, China
5
Cryos International, Denmark ApS, Aarhus, Denmark
Study question: To identify the genetic cause of repeated fertilization failure
Study question: Can single nucleotide polymorphism (SNP) arrays be suc- even after ICSI combined with artificial oocyte activation (ICSI-AOA) in a
cessfully utilized on polar bodies for simultaneous PGT-M and assessment of non-consanguineous family.
the chromosomal status of the oocyte? Summary answer: With whole-exome sequencing, a novel compound het-
Summary answer: Genome-wide SNP array provide a nearly universal tool erozygous mutation in WEE2 gene was identified in an infertile female who
for PGT-M on polar bodies. The oocyte ploidy can be assessed in the same experienced with fertilization failure after ICSI-AOA
analytical procedure. What is known already: The genetic factors play an important role in oocyte
What is known already: Polar bodies can be sourced as an alternative sample activation defect and likely result in repeated human fertilization failure even
material for PGT-M when the analysis of blastomeres or trophectoderm cells is after ICSI-AOA. In mouse oocyte at metaphase II stage, Wee2 inhibited phos-
not possible due to ethical, medical, religious or legal reasons. Traditionally, the phorylation of Cdc2, which was required for metaphase II exit. Down regulation
analysis has been based on short tandem repeats (STRs) and linkage analysis of Wee2 during egg activation leads to failure of pronucleus formation. Recently,
which is time consuming and costly as the assay has to be developed for each mutations in WEE2 were identified to result in failure of pronuclei formation
family separately. As only the disease locus is assessed in the STR-based linkage and human fertilization in infertile women.
analysis the chromosomal status of the oocyte remains unclear. Thus, there is Study design, size, duration: We have characterized a patient with 2-year
room for improvement in the current methodology. history of primary unexplained infertility belonging to a non-consanguineous
Study design, size, duration: Retrospective study in a setting of a commercial family from China. During 3 assisted reproduction attempts (IVF+ rescue ICSI,
diagnostic laboratory. The phenotypes segregating in the families were inherited ICSI and ICSI-AOA), the woman presented a repeated fertilization failure for
as autosomal dominant (inherited breast and ovarian cancer, spinocerebellar MII oocyte retrieval after controlled ovarian hyperstimulation. We performed
ataxia type 3), autosomal recessive (puryvatkinase deficiency) and X chromo- whole-exome sequencing and sanger sequencing in the patient and her family
somal (ornithine transcarbamylase deficiency, Renpenning syndrome) traits. members.
Altogether five patients were included in the study. The work was conducted Participants/materials, setting, methods: Genomic DNA samples of
between February 2018 and December 2019. the patient and her family’s members were extracted from peripheral blood.
Participants/materials, setting, methods: Sequentially biopsied polar bodies Whole-exome capture and sequencing were performed following the stan-
from oocytes fertilized with ICSI were subjected to whole genome amplification dard protocols provided by BGI Genomics (BGI-Shenzhen). The sequencing
(Repli-g Single Cell Kit; Qiagen). Genome-wide genotyping of the amplified samples depth of WES was 100× . The pathogenicity of variants was assessed by
as well as DNA samples of the mother and at least one reference individual was four software: SIFT, PolyPhen2, Mutation Taster and Human Splicing Finder.
carried out (Human KaryomaP-12 SNP array; Vitrolife). A direct analysis of the The structure prediction of wide type and mutant type WEE2 were per-
disease-causing variant was included in the assay when applicable. BlueFuseMulti formed by PyMol software.
software (Illumina) with modified settings was utilized for the data analysis. Main results and the role of chance: Three ART cycles were conducted
Main results and the role of chance: Here we report the successful use of for this couple and even ICSI-AOA did not rescue fertilization failure phe-
genome-wide genotyping on polar bodies allowing the simultaneous assessment notype. Aneuploidy or pathogenic microdeletion/microduplication
of the maternally inherited disease and the oocyte ploidy. Polar body sets of 19 (>100Kb) were not found in the couple. We identified a novel compound
zygotes were analysed: eleven zygotes were non-affected, seven affected and heterozygous mutation c.1535+3A>G and c.946C>T (p. Leu316Phe) in the
one couldn’t be interpreted because the second polar body delivered no data. WEE2 gene in female proband who experienced fertilization failure. The
The genome-wide analysis showed aneuploidy in four of the non-affected allele frequencies of variation c.1535+3A>G and c.946C>T were not found
zygotes. Three patients had one zygote suitable for culture and transfer whereas in the 1000genome database or ExAC database. Trios analysis revealed that
two patients had two suitable zygotes. One double-embryo transfer and two the detected variations follow an autosomal recessive pattern. Variation
single embryo transfers ensued resulting in one clinical pregnancy and birth of c.1535+3A>G in WEE2 was predicted to break WT donor site and probably
a healthy baby. Two non-affected zygotes didn’t survive warming. One patient affects splicing. With SIFT, PolyPhen-2 and Mutation Taster software, it is
with one suitable zygote is awaiting for warming and transfer. predicted that the missense mutation c.946C>T (p. Leu316Phe) of WEE2
The set-backs in polar body analysis are considerable. When the disease is is pathogenic. Prediction with PyMOL software indicated that the mutation
inherited in autosomal recessive or X-chromosomal manner the sole use of could change the structure of WEE2 protein.
polar body analysis results in wasting of a significant number of zygotes as pater- Limitations, reasons for caution: The functional analysis of oocytes is
nally inherited alleles and the sex of the embryo cannot be determined. In not performed yet to detect the local expression of this variant. A causal
comparison to trophectoderm cells the analysis of polar bodies inherently con- relationship between the mutation and the fertilization failure need to be
tains a higher fall out rate, although in this patient series only two polar bodies established.
failed the amplification or analysis. Wider implications of the findings: This case report expanded the
Limitations, reasons for caution: This is a retrospective study with a small spectrum of WEE2 gene mutations responsible for human fertilization failure
sample size. Thus, further studies are needed to establish the clinical and diag- and provided genetic evidence for fertilization failure.
nostic utility of the procedure. Only maternally inherited diseases can be analysed Trial registration number: None
and both polar bodies are needed for reliable diagnostics.
Wider implications of the findings: In situations where PGT-M on embry-
onal cells isn’t allowed or desired polar body analysis is a worthy alternative. In P-501 Model based on maternal age to determine the optimal
these cases SNP arrays allow the testing of a wide range of genetic diseases with number of blastocysts required to retrieve at least one euploid
the simultaneous detection of oocyte ploidy, particularly of interest for patients embryo
of advanced maternal age. M. Ferrández Rives1, M. Enciso1, L. Ortega2, Y. Galiana2,
Trial registration number: Not applicable B. Rodriguez-Estrada1, I. Jurado1, M. Berruezo1, A.B. Climent3,
J. Sarasa1, J. Aizpurua4
1
iGLS, Genetic Laboratory, Alicante, Spain ;
P-500 Novel compound heterozygous mutations in WEE2 gene 2
IVF Spain, Embryology department, Alicante, Spain ;
is associated with fertilization failure: a case report about an 3
IVF Spain, Statistics department, Alicante, Spain ;
infertile woman 4
IVF Spain, Gynecology, Alicante, Spain
Y. Tian1, B. Xiaohong2
1
Tianjin Medical University General Hospital, Department of Gynecology and Study question: Is it possible to anticipate the number of blastocysts and/or
Obstetrics-, Tianjin, China ; cycles needed to retrieve at least one euploid embryo based on maternal age?

Summary answer: The probability of finding at least one euploid embryo can 6
Valencia University and INCLIVA, Department of Obstetrics and Gynecology,
be estimated based on age and the number of blastocysts available. Valencia, Spain ;
What is known already: Aneuploidy in human preimplantation embryos plays 7
Harvard School of Medicine- Harvard University- MA- USA, Department of
an important role in assisted reproduction treatments, being one of the main Obstetrics and Gynecology, Boston, U.S.A.
reasons for implantation failures and miscarriages. The impact of female age on
euploidy rate is well-known, as with advancing maternal age, the risk of chro- Study question: What is the impact on clinical outcome of putative low-grade
mosomal abnormalities in embryos increases. Although patients undergo pre- mosaicism diagnosis?
implantation genetic testing because of the clinical significance of aneuploidy, Summary answer: Blastocysts diagnosed with putative low-grade mosaicism
unfortunately the retrieval of euploid embryos is not guaranteed and limited produce comparable clinical outcomes to uniform euploid blastocysts in terms
information about the probability of obtaining euploid embryos is available. The of sustained implantation and miscarriage rates.
estimation of the probability of euploid embryo retrieval would enable effective What is known already: Highly sensitive NGS-based technologies allow precise
treatment plans and accurate counselling of patients. discrimination of subtle chromosomal copy number variations (CNVs) in multicel-
Study design, size, duration: This is a retrospective study including data lular biopsies.These alterations are often interpreted as evidence of an embryo’s
from a total of 1048 cycles (476 own-oocyte cycles and 572 donor-oocyte chromosomal mosaicism. Because of the unknown clinical impact of mosaicism,
cycles) undergoing PGT-A (Preimplantation Genetic Testing for Aneuploidy) at uniformly euploid embryos are prioritized for transfer, whilst putative mosaic ones
blastocyst stage between January 2017 and January 2020. A total of 3940 are given low priority or even discarded. Presently, transfer outcomes of putative
embryos were studied. Relationship between maternal age, total number of mosaic embryos have only been compared retrospectively in selected subpopula-
blastocysts biopsied and embryo euploidy was assessed. tions of patients that failed to get pregnant with previous euploid embryos. This
Participants/materials, setting, methods: Patients referred for PGT-A prospective non-selection study was designed to avoid/minimize confounding
analysis for a variety of reasons including advanced maternal age, recurrent factors and provide robust evidence on putative mosaicism clinical predictive values.
implantation failure, recurrent miscarriage and male factor infertility (aged 24-54 Study design, size, duration: This is an interim analysis of a multicenter
years) were included in the study. All embryos were cultured to blastocyst stage, prospective non-selection study of consecutive cases performed at five IVF
biopsied and PGT-A-tested for all 24 chromosomes using Next Generation clinics between Sept2018 and July2019. Trophectoderm biopsies showing inter-
Sequencing (Illumina, VeriSeq protocol). Outcome measures were: number of mediate chromosome CNVs consistent with low mosaicism (20-50%) were
blastocysts (BT), euploidy rate (ER) and the probability of finding at least one blindly reported as euploid. The presence of low-grade alterations did not influ-
euploid embryo for transfer (POE). ence the embryo selection process, thus allowing an unbiased comparison of
Main results and the role of chance: Our results show a negative influence clinical outcomes between fully euploid and putative mosaic embryos. Ethical
of female age on euploidy rate and the number of blastocysts per cycle committee approvals were obtained at each site.
(p<0.001). Moreover, the probability of finding at least one transferable euploid Participants/materials, setting, methods: Consecutive patients (female
embryo (POE) is strongly associated to age and the number of blastocysts age 35-44) undergoing homologous IVF cycles with blastocyst-stage PGT-A and
available. POE is highest in egg-donation cycles (96%) and it reached 100% in single frozen euploid embryo transfer (SEET) were enrolled. Main exclusion
cohorts of >3 blastocysts. Nevertheless, POE decreases with age (approximated condition was blastocyst of the worst morphological class. Sustained implantation
by the following quadratic function: y = -0.003x2 + 0.1785x – 1.6825, R2=0.91) rate (>20weeks) was the primary outcome measure. A sample size of 878 SEET
and this decline is very sharp after 40 years old. was planned (beta=0.80; alfa=0.05) assuming 47% sustained implantation rate
In women <35 years old, the mean number of blastocysts per cycle (4.3) in the control group and 12% variation. This sample-size is also powered to
guarantees one euploid embryo in 96% of cases. Meanwhile, women aged detect meaningful differences in the miscarriage rate.
35-40 and over 40 produce 3 and 2.2 blastocysts per cycle, respectively. In Main results and the role of chance: This study evaluates transfer outcomes
these groups, the availability of at least one euploid embryo is guaranteed in of 368 SEET: 197 from uniform euploid embryo group (group A), 94 from
65% and 28% of cycles, respectively. POE decline with age can be mitigated putative very-low mosaic (20-30%, group B) and 77 from the putative low mosaic
by the banking of embryos from various cycles. This embryo banking strategy group (30-50%; group C). Positive pregnancy test rate was 58.4% (95%CI=51.2%-
significantly increases the likelihood of euploid embryo transfer in advanced 65.3%), 60.6% (95%CI=50.0%-70.6%) and 58.4% (95%CI=46.6%-69.6%) for
maternal age groups. The presence of one euploid embryo in the group of group A, B and C, respectively (P=NS). Biochemical pregnancy loss rate was
patients over 40 studied can be guaranteed in cohorts of >7 blastocysts (~3-4 9.6% (95%CI=4.9%-16.5%),10.5% (95%CI=4.0%-21.5%) and 13.3%
cycles). (95%CI=5.0%-26.8%) for A, B and C respectively (P=NS). Miscarriage rate was
Limitations, reasons for caution: This model can serve as a valuable tool to also not significantly different across groups: 12.5% (95%CI=6.8%-20.4%), 13.7%
improve assisted reproduction treatment results, however, other variables such (95%CI=5.7%-26.3%) and 12.8% (95%CI=4.3%-27.4%) for A, B and C, respec-
as the number of oocytes retrieved, sperm quality parameters or AMH levels may tively (P=NS). Sustained implantation rate was also similar: 46.2% (95%CI=39.1%-
also need to be considered as potential factors with influence in the model. 53.4%), 48,8% (95%CI=36.4%-57.4%) and 44.2% (95%CI=32.8%-55.9%), for
Wider implications of the findings: Our results confirm the well-known A, B and C respectively (P=NS). Multivariate logistic regression analysis including
negative effect of female age on embryo euploidy. We have identified a model main and potentially relevant patient and cycle factors showed slower embryo
based on maternal age able to predict the likelihood of euploid embryo transfer. development (embryo developed to blastocysts on day 7) as the only covariate
This model can help doctors to plan their treatments and counsel patients about associated with sustained implantation outcome (R=0.19; 95%CI=0.05-0.7). In
their options of success. the multivariate analysis, after adjusting for confounding factors, the presence
Trial registration number: not applicable of a PGT-A analysis consistent with mosaicism was not significantly associated
with either primary or secondary outcome measures (OR=1.001; 95%CI=0.61-
1.06). When groups B and C were combined, the lack of association persisted.
P-502 Low-degree mosaicism profiles do not provide clinically Limitations, reasons for caution: Although current interim data are suffi-
useful predictive values: interim results from the first multicenter ciently powered to exclude clinically relevant differences between study groups,
prospective non-selection study on the transfer of mosaic embryos the planned sample size isn’t yet fully met. Study’s main limitation consists in the
A. Capalbo1, M. Poli1, D. Cimadomo2, F. Benini3, C. Patassini1, lack of cytogenetic analysis follow-up of putative mosaic embryos pregnancies.
C. Rubio4, E. Albani5, L. Sacchi5, L. Rienzi2, C. Livi3, P.E. Levi-Setti5, However, indication for invasive prenatal testing couldn’t be justified by evidence.
M.U. Filippo2, C. Simón1,4,6,7 Wider implications of the findings: This is the largest prospective non-se-
1
Igenomix, Reproductive Genetics, Marostica, Italy ; lection study evaluating the outcome of putative mosaic embryo transfers. If
2
Genera, Reproductive Medicine, Rome, Italy ; confirmed, these results demonstrate that intermediate CNVs consistent with
3
ART Center DEMETRA, Reproductive Medicine, Florence, Italy ; low-grade mosaicism don’t provide any clinically useful diagnostic criteria for
4
Igenomix Foundation, Reproductive Genentics, Valencia, Spain ; defining embryonic reproductive competence. Re-assessment of aneuploidy
5
Humanitas Clinical and Research Center- IRCCS, Division of Gynecology and categorization and embryo selection schemes will be required.
Reproductive Medicine- Humanitas Fertility Center, Milan, Italy ; Trial registration number: NCT03673592

P-503 Assessing egg utilization efficiency from the outcome of Study question: Does increased Body Mass Index (BMI) affect blastocyst
Preimplantation Genetic Testing cycles ploidy and cycle outcomes in IVF/ICSI patients with Preimplantation Genetic
W. Wu1, X. Yang2 Testing for Aneuploidy (PGT-A)?
1 Summary answer: Increased BMI has no correlation with blastocyst ploidy;
First Affiliated Hospital of Nanjing Medical University, Reproductive Medical
but decreases the number of mature oocytes per large follicles and per total
Center, Nanjing, China ;
2 doses of stimulation drugs.
Nanjing Medical Univesity, First Clinical Medical College, Nanjing, China
What is known already: World Health Organization (WHO) identified obe-
Study question: In patients using preimplantation genetic testing (PGT), the sity as a global epidemic as its prevalence has tripled since 1975. It is a major risk
average number of eggs needed to obtain 1, 2, and ≥3 euploid/balanced factor for many diseases including infertility, with a prevalence of 71% of the
embryos was calculated for populations with different characteristics. female Middle Eastern population being affected. Obesity has a detrimental effect
Summary answer: Couples with one or both reciprocal translocation carriers on female fertility by causing hypothalamic-pituitary dysfunction which may lead
need more eggs to obtain the same number of euploid/balanced embryos,espe- to anovulation and impaired follicular development. Increased BMI has a negative
cially those with general balanced translocation carriers. prognosis factor for cycle cancellation, the number of oocytes collected, the
What is known already: Chromosome rearrangement can significantly affect number of embryos available, clinical pregnancy rate and abortion rate which
fertility and increase the risk of miscarriage. Although there are theoretical mod- may be due to both impaired oocyte quality and uterine function.
els for the probability of normal gamete formation among balanced translocation Study design, size, duration: A retrospective cohort study including 1115
carriers, more clinical data are needed to support the average number of eggs fresh IVF/ICSI cycles from 807 patient was conducted in a single private fertility
required to obtain the expected number of euploid/balanced embryos during clinic in Abu Dhabi, UAE between March 2017 and December 2018. According
ovarian stimulation process. to WHO, patients were stratified as: underweight, normal, pre-obese, obese I,
Study design, size, duration: From June 2011 to July 2019, 1,659 PGT obese II and obese III when they have a BMI of: <18.50, 18.50–24.99, 25.00–
retrieval cycles were collected.There were 368 cycles in PGT for aneuploid- 29.99, 30.00–34.99, 35.00-39.9 and ≥40 kg/m2, respectively. ANOVA test was
ies(PGT-A group), 105 cycles in PGT for monogenetic(PGT-M group) and used to compare means of different parameters.
1186 cycles in PGT for structural rearrangements which including 829 cycles Participants/materials, setting, methods: Females were ≥18 years old
of general translocation group and 357 cycles of Robertson transloca- who underwent fresh IVF/ICSI treatment with PGT-A. Routine work included
tions group. ovarian stimulation using different types of protocols. Fresh oocytes were insem-
Participants/materials, setting, methods: All patients were treated with inated either by IVF or ICSI and the resulted fertilized oocytes were cultured till
appropriate ovarian stimulation protocol.When the diameter of the dominant blastocysts were biopsied on day 5, 6 and 7 of development and tested for
follicles reached about 18mm, human chorionic gonadotropin (HCG) was PGT-A using NGS. blastocysts were vitrified and euploid ones were warmed
injected and ovum extraction was conducted under the guidance of transvaginal and transferred in a subsequent natural or hormonal replaced cycle.
ultrasound 36 hours later. Main results and the role of chance: We stratified our population regarding
Embryo testing: blastocyst or blastomere biopsies are performed using array- BMI into the following groups: Normal, underweight, preobese, obese I, obese
based comparative genomic hybridization (aCGH) or next-generation sequenc- II and obese III with the following percentages: 35.8%, 1.8%, 37.9%, 18.8%, 5%
ing (NGS). and 0.7% (n= 400, 20, 422, 209, 56 and 8) respectively. Our study demonstrated
Main results and the role of chance: The average number of eggs needed no significant difference between the groups regarding number of stimulation
to obtain1, 2 and ≥3 euploid/balanced embryos in the PGT-A group was 7.65, days (p=0.074), oocytes retrieved (p=0.66), mature oocytes (p=0.93), fertilized
10.38, and 12.95, respectively; the average age of the woman was 33.83 ± 4.62, oocytes (p=0.72), biopsied (p=0.21) and euploid blastocysts (p=0.22). The
and the average age of the man was 35.40 ± 5.67. The average number of eggs same non-significant difference was also reported regarding fertilization and
needed to obtain1, 2 and ≥3 euploid/balanced embryos in the PGT-M group euploidy rates (p=0.26 and p=0.19). Interestingly, negative correlations were
was 7.4, 10.77, and 14.33, respectively; the average age of the woman was 31.45 found between BMI and (i) the number of mature oocytes per follicles >11mm
± 4.04, and the average age of the man was 32.53 ± 4.87. The average number (p= 0.044), (ii) aspirated oocytes per total stimulation doses (p= 0.020) and (iii)
of eggs needed to obtain 1, 2 and ≥3 euploid/balanced embryos in the PGT-SR mature oocytes per total stimulation doses (p= 0.029). Correcting for BMI as
general translocation group was 11.1, 13.32, and 15.59, respectively; the average a confounder and applying multivariate regression and Poisson models resulted
age of the woman was 28.97 ± 3.86, and the average age of the man was 30.32 in a negative correlation between euploidy rate and age and a positive correlation
± 4.95. The average number of eggs needed to obtain 1, 2 and ≥3 euploid/ between euploidy rate and the number of mature oocytes (both p<0.001).
balanced embryos in the Robertson translocations group was 10.01, 12.49, and Applying a multivariate regression model and correcting for age and number
15.32, respectively; the average age of the woman was 29.30 ± 4.04, and the of mature oocytes collected revealed that increased BMI did not have any impact
average age of the man was 30.84 ± 4.81. on implantation (51.2% vs 50.8%p=0.079) or clinical pregnancy rates(47.4% vs
Limitations, reasons for caution: Patients using PGT-M are first tested for 46.7%, p= 0.092) respectively.
a single gene of the relevant disease.Aneuploidy screening will not be continued Limitations, reasons for caution: Our results were based on retrospective
if they carried the pathogenic gene,which causing the average number of eggs data from a single fertility clinic, which does not permit to control for various
needed to obtain euploid/balanced embryos increased.The number of patients factors leading to the selection of a stimulation protocol by a physician for each
included in the PGT-M group was relatively small. patient.
Wider implications of the findings: The study provides clinical data on the Wider implications of the findings: Although increased BMI doesn’t affect
expected number of eggs required to obtain euploid/balanced embryos in dif- euploidy rate or cycle outcomes of patients undergoing IVF/ICSI with PGT-A,
ferent patient populations with different characteristics, and provides a reference obese patients need to be administered more stimulation doses for them to
for clinicians to determine the optimal ovarian stimulation protocol for patients. have the same number of mature oocytes as the normal weight patients, which
Trial registration number: without decreases the cost-effectiveness of their treatment cycles.
P-504 Increased Body Mass Index and Its Impact on Blastocyst P-505 BCORL1, USP9x and POF1B copy number variation in
Ploidy Status premature ovarian failure: A Preliminary study
A. El-Damen1, I. Elkhatib1, A. Bayram1, A. Arnanz1, A. Abdala1, C. Furtado1, F. Gennaro2, G. Kogure3, R. Ferriani4, R. Reis3
N. Nobrega1, S. Samir2, L. Melado3, B. Lawrenz3, N. De Munck1, 1
Federal University of Ceara, Drug Research and Development Center-
H. Fatemi3 Postgraduate Program in Medical and Surgical Sciences, Fortaleza, Brazil ;
1
IVI Middle East Fertility Clinic, IVF Laboratory, Abu Dhabi, United Arab Emirates ; 2
Ribeirao Preto Medical School- University of Sao Paulo, Obstetrics and
2
IVI Middle East Fertility Clinic, Administration, Abu Dhabi, United Arab Emirates ; Gynaecology, Ribeirão Preto, Brazil ;
3
IVI Middle East Fertility Clinic, Clinical Department, Abu Dhabi, United Arab 3
Ribeirao Preto Medical School- University of Sao Paulo, Obstetrics and
Emirates Gynaecology, Ribeirao Preto, Brazil ;

4
Ribeirao Preto Medical School- University of Sao Paulo-, Obstetrics and Main results and the role of chance: We used an in silico baseline derived
Gynaecology, Ribeirao Preto, Brazil from amplicon representation from normal samples and a custom CNV calling
pipeline to determine both i) the mean copy numbers from amplicons with 100%
Study question: Copy number variation (CNV) in X-chromosome linked genes identity between homologous portions of the genes of interest and ii) the homo-
are related to idiopathic premature ovarian insufficiency (POI)? log-specific copy numbers for amplicons with distinguishing sequence variants
Summary answer: POI presented alteration in the number of copies for the between the primers or sequence variants under the primers which enriched
X-linked genes USP9X and POF1B. amplification in one member of a gene pair. Using this information and orthog-
What is known already: X-linked genes are strongly associated with ovarian onal microarray results taken as “truth,” we were able to obtain SMN1 carrier
function and have several POI candidate genes. Both amplifications and deletions status with 100% sensitivity (n=4) and 98.9% specificity (n=87). Follow up will
can alter the level of expression of clusters of genes associated with POI, includ- be needed from individual labs employing our test to determine reproducibility
ing genes involved in pairing and segregation such as POF1B and USP9X and in with their own samples of interest.
response to apoptosis BCORL1. Consequently, those alterations may promote For Research Use Only. Not for use in diagnostic procedures.Limitations,
changes in the XCI pattern, such as nonrandom (skewed) X inactivation in female reasons for caution: We tested our method on a limited set of gDNA samples
genome and maybe an important feature of POI. isolated from cell lines. Factors that could compromise performance for other
Study design, size, duration: Cross-sectional study, in which 37 non-syn- samples include amplification and sequencing variation not accommodated by
dromic POI women were included. our CNV baseline and SNPs falling under primers resulting in decreased ampli-
Participants/materials, setting, methods: POI (FSH>40 IU/l) with 46,XX fication efficiency that mimics a copy number loss.
karyotype participated in this study. Age, body mass index (BMI) were analyzed. Wider implications of the findings: Our panel design and analysis methods
The CNVs in the BCORL1, USP9X and POF1B was measured by quantitative real enable the determination of SMN1/2 copy number as part of a much broader
time PCR and the reference genes were the HPRT1 (x-linked) and the MFN2 NGS assay of hundreds of genes responsible for inherited diseases, enabling the
(autossomal). A control DNA (male and female) was used in the comparisons. development of more convenient future tools.
XCI was based in the Human Androgen Receptor (HUMARA) and X-linked Trial registration number: not applicable
retinitis pigmentosa 2 (RP2) assays.
Main results and the role of chance: The mean age was 32.10 ± 6.81 and P-507 DNA detection in the blastocoelic fluid (BF) from expanded
BMI was 25.41 ± 4.64. Most POI women presented number of copies (> 0.75) blastocysts generated by conventional IVF cycles: clinical
close to the female control (46, XX) for the BCORL1 (97.2%), USP9X (55.6%) implications
and POF1B (58.3%). Reduced number of copies similar to the male control C. Albanese1, M.C. Magli1, S. Azzena1, M. Piiffanelli1, B. Balicchia1,
(<0.75) was observed for the USP9X (44.4%) and POF1B (41.17%). Skewed XCI C. Tabanelli1, L. Gianaroli1
(≥75%) was observed in 24,32% (n=9) POI. From those POI with copy number 1
S.I.S.Me.R., Reproductive Medicine Unit, Bologna, Italy
alterations only 10% presented skewed XCI.
Limitations, reasons for caution: Limited number of cases of the study Study question: Does the presence or absence of DNA in the BF correlate
group. An increase of sample size is recommended to make the data more robust with blastocyst viability?
and confirm the results. Summary answer: After whole genome amplification (WGA) of BFs, failure
Wider implications of the findings: Alterations in X-linked genes and to detect DNA correlates with significantly higher implantation rates when com-
skewed XCI was observed in POI women, a genetic condition that is frequently pared to blastocysts with positive DNA amplification.
observed in ovarian dysfunctions, suggesting a possible role in POI etiology. What is known already: The presence of DNA in the BF could represent a
Trial registration number: 13305/2012 strategy towards a euploid condition through the extrusion of aneuploid cells
into the blastocoelic cavity. This event could not occur at a measurable level
P-506 Determining copy number variation in SMN1 using a when the blastocyst is fully or prominently euploid, whereas positive BF DNA
highly multiplexed Next Generation Sequencing panel amplification could indicate a blastocyst with aneuploid cells trying to correct a
A. Harris1, G. Del Mistro1, T. Fahland1, H. Xie1, J. Bishop1, defective chromosome condition. Especially in the case of low to moderate level
J. Veitch2, V. Mozhayskiy1 of mosaicism, the capacity of extruding the majority of abnormal cells into the
1
Thermo Fisher Scientific, Clinical Next Generation Sequencing, Carlsbad- CA, U.S.A. ; BF would preserve the blastocyst potential of further development after transfer.
2
Thermo Fisher Scientific, Clinical Next Generation Sequencing, South San Study design, size, duration: This prospective cohort study included 184
Francisco- CA, U.S.A. conventional IVF cycles performed from January 2016 to October 2019. In 91
cycles (Study-group), the BF was collected from expanded blastocysts before
Study question: Can a Next Generation Sequencing (NGS) panel be used to vitrification and stored at -80°C. The control-group included 93 cycles, whose
accurately assess samples for copy number variation (CNV) in genes that have blastocysts were vitrified without blastocentesis. Single blastocysts were trans-
high homology such as SMN1 and SMN2? ferred by selection according to morphology. BFs were later submitted to WGA
Summary answer: We were able to characterize test samples for functional by operators blinded to the clinical outcome. WGA results were then related
SMN1 copy number with 97.8% accuracy and SMN2 copy number with 95.6% to implantation.
accuracy (n=91). Participants/materials, setting, methods: Maternal age and indications to
What is known already: Loss of SMN1 activity is a cause of spinal muscular IVF were comparable between the two groups. The same protocol of vitrification
atrophy (SMA). SMN1 and SMN2 are highly homologous genes that are difficult was applied and only cycles with expanded blastocysts of high grade were
to distinguish on NGS platforms due to relatively short read lengths. Typically included in the study. Amplification after WGA was evaluated by loading an
the copy number determination of these genes is assessed with other technol- aliquot of the amplified product onto a 1.5% agarose gel. A clinical pregnancy
ogies such as Multiplexed Ligation Probe Assays (MLPA) and microarray. rate was defined by the presence of fetal heart-beat. The ongoing pregnancy
Study design, size, duration: We tested 91 gDNA samples from Coriell cell rate was a pregnancy ongoing after 12 week gestation.
lines with functional SMN1/2 copy number status known from microarray. The Main results and the role of chance: The clinical outcome was comparable
samples were tested as part of a characterization of the NGS panel in develop- in the study-group and control-group with similar clinical pregnancy rates (51
ment. Results needed to meet pre-defined quality control criteria to be included and 55% respectively) and ongoing pregnancy rates (47 and 47% respectively)
in the assessment. suggesting that the procedure of blastocentesis per se had no impact on blas-
Participants/materials, setting, methods: Samples were amplified using tocyst viability. When the study-group was subdivided according to WGA results,
a two-pool primer panel comprised of approximately 14,000 PCR primer pairs we had 55 cases where BF failed to amplify (sub-group with failed BF-WGA)
targeting coding sequences and known intronic variants in 420 genes implicated and 36 with positive amplification (sub-group with positive BF-WGA). Maternal
in recessive autosomal and X-linked diseases. The resulting DNA barcoded age was 36.4±3.2 and 35.5±3.5 years respectively. When looking at the transfer
libraries were sequenced in a multiplex of 16 per Ion 540™ Chip using the Ion outcome in the two sub-groups, the clinical pregnancy rate was significantly
S5™ Sequencing System. higher in the sub-group with failed BF-WGA (75%) when compared to the

sub-group with positive BF-WGA (25%, P<0.001). The ongoing pregnancy rate for AA vs. GA/GG genotype of rs939443 (SENP7). 2) FR: lower for MAs of
showed the same trend and was 69% and 11% respectively (P<0.001). rs1059476 and rs2241909 of AURKB and higher for MAs of rs61736572 and
Limitations, reasons for caution: The study is a prospective cohort study, rs61736575 of AMH, higher for MA of rs56043017 of PLK4. 3) BL/MII: higher
but not prospectively randomized. for MA of rs1801133 of MTHFR, lower for MAs of rs7616677 and rs939443 of
Wider implications of the findings: Failure to detect DNA after BF ampli- SENP7 and higher for MA of rs2433031 of SENP7. 4) BL/2PN: lower for MA
fication could represent an additional criterion to select viable embryos for of rs35518193 of HSPA4L, higher for MA of rs2433031 of SENP7. No significant
transfer in conventional IVF cycles. If confirmed in a prospective randomized associations were revealed for rs1801131 and rs2305957. Where comparisons
study, this approach would improve the clinical outcome of vitrified blastocysts were possible, these findings remained significant independent of age and stim-
by using a simple, non-invasive and moderately expensive approach. ulation protocol. Additional associations were revealed for rs3746964, rs878081,
Trial registration number: Not applicable rs1800521 (AIRE), rs10407022 (AMH) and rs175080 (MLH3) when stratifying
for age and stimulation protocol.
P-508 A pilot study to identify Single Nucleotide Variants Limitations, reasons for caution: The main limitations of the study include
(SNVs) as predictors of oocyte/embryo quality in fertile women its retrospective nature, whereby IVF practices have changed considerably over
undergoing Preimplantation Genetic Testing for Monogenic the period of the study (2013-2019). The polymorphisms identified with likely
Disorders (PGT-M) significant impact, require prospective validation in other PGT-M cycles, as well
G. Kakourou1, T. Mamas1, C. Vrettou1, E. Tsorva2, S. Zacharia2, as on embryo samples of variable quality.
C. Oraiopoulou3, E. Chatzi4, E. Kourkouni5, G. Kourlaba5, Wider implications of the findings: Infertility is a complex condition.
D. Christopikou2, M. Mastrominas2, M. Moysidou3, A. Identifying prognostic factors in ART patients is confounded by multiple variables
Chatziparasidou3, I. Georgiou4, J. Traeger-Synodinos1 (parental age, stimulation and fertilization protocols, embryo transfer stage/
1 conditions etc.). PGT-M facilitates investigation of a fertile ART population,
National & Kapodistrian University of Athens, Department of Medical Genetics,
minimizing many confounding variables, and potentially facilitating the identifica-
Athens, Greece ;
2 tion of genomic biomarkers predictive of gamete/embryo quality.
Embryogenesis, Assisted Reproduction Unit, Athens, Greece ;
3 Trial registration number: This study was supported by a State Scholarships
Embryolab SA, IVF Unit, Thessaloniki, Greece ;
4 Foundation (IKY) for post-doctoral research and co-funded by national grants
Genetics and IVF Unit- Faculty of Medicine- University of Ioannina,
and the European union (ESPA 2014-2020). Trial registration number not
Department of Obstetrics and Gynecology, Ioannina, Greece ;
5 applicable
Center for Clinical Epidemiology and Outcomes Research, Cleo, Athens, Greece
Study question: Are SNVs in 26 selected genes potentially involved in preim- P-509 Applied machine learning based on time-lapse technology
plantation development, associated with oocyte and embryo quality in fertile and reproductive history data designed for pre-implantation
women undergoing PGT-M? embryo ploidy prediction.
Summary answer: SNVs in 10/26 studied genes revealed significant associ- S. Davies1, D. Christopikou1, N. Kallioras2, P. Tsakalis2, E. Tsorva1,
ations with oocyte number, fertilization rate and blastocyst formation in fertile S. Zaxaria1, K. Economou1, R. Najdecki3, E. Timotheou3, P. Tatsi3,
women undergoing PGT-M. T. Chartomatsidou3, C. Papanikopoulos1, N. Lagaros2, M.
What is known already: Genetic profiling of prospective parents has been Mastrominas1, E. Papanikolaou3
one of the approaches for identifying biomarkers of IVF success. Specific geno- 1
c/o Embryogenesis-Private Centre for Reproductive Studies, embryology,
types have been associated with IVF failure, gamete/embryo quality etc., with Maroussi- Athens, Greece ;
most studies focusing on infertile populations. Recent studies identified SNVs 2
Institute of Structural Analysis & Antiseismic Research- National Technical
(rs1801133, rs1801131 in MTHFR, and rs2305957 on chromosome 4 linked to University of Athens, School of Civil Engineering, Athens, Greece ;
INTU, SLC25A31, HSPA4L, PLK4, MFSD8, LARP1B and PGRMC2) associated with 3
Assisting Nature- Private IVF Unit, Ob/gyn, Thessaloniki, Greece
embryo quality or chromosomal status in infertile women. Additionally, preim-
plantation development may be influenced by pathways involved in follicle devel- Study question: To determine whether Artificial Intelligence (AI) algorithms
opment, meiosis, mitosis and DNA repair, for which many other genes have based on time-lapse (TLM) morphokinetic parameters and reproductive history
been implicated in human and animal studies. can be used to predict embryo ploidy status.
Study design, size, duration: DNA from fertile prospective mothers was Summary answer: Cleavage time intervals from 5-6-7-8-9-cell stage to time
genotyped using exonic NGS (QiaseqTM Targeted Custom Panel, Miseq) for 18 of morulation, along with maternal age and PGT indication enhances the pre-
genes (AIRE-AMH-AURKA-AURKB-AURKC-FSHR-HSPA4L-HUWE1-INTU-KHDC3L- diction of blastocyst ploidy.
LARP1B-MFSD8-MTHFR-PGRMC2-PLK4-SENP7-SLC25A31-WBP1) and 9 selected What is known already: Time-lapse technology allows continuous monitoring
SNVs in a further 8 genes: rs175080(MLH3), rs1799963(F2), rs6025(F5), of embryo development and has assisted in progression towards automated
rs5918(ITGB3), rs5985(F13A1), rs1805087(MTR), rs1801394(MTRR), and objective systems to assess the embryos and improve IVF treatment.
rs28756992(MLH3) and rs2305957(HSPA4L). The study (September 2017- Artificial intelligence technology, based on data and pattern exploitation, has
2019), approved by the University of Athens Bioethics Committee and National been reported as a tool to identify embryo viability and predict IVF outcome.
Authority of Assisted Reproduction, involved PGT-M cycles performed between However, studies reporting an association between morphokinetic parameters
2013-2019. Statistical analysis was performed by the Center for Clinical and embryo ploidy status are controversial and do not support the predictive
Epidemiology and Outcomes Research. value of time-lapse analysis for preimplantation genetic testing of embryo ploidy
Participants/materials, setting, methods: The study was performed at thus far.
the UoA Laboratory of Medical Genetics, and focused on fertile couples under- Study design, size, duration: Double blind, retrospective longitudinal cohort
going PGT-M. NGS was performed for 85 women undergoing 107 PGT-M-cycles. study. Embryo morphokinetic parameters of 383 blastocysts were collected
The following details were collected: maternal age, stimulation protocol, number from 193 patients undergoing preimplantation genetic testing for aneuploidy
of oocytes collected/fertilized, blastocyst formation. Maternal genotypes were (PGT-A) and structural rearrangements (PGT-SR) between November 2017
analysed in association with number of oocytes collected (No.O), fertilization and December 2019 from two independent IVF clinics.
rate (FR), percentage of blastocysts developed per MII oocyte (BL/MII) and Participants/materials, setting, methods: The morphokinetic parameters
percentage of blastocysts per 2PN embryo (BL/2PN). of blastocysts, subject to PGT-A/PGT-SR analysis with next generation sequenc-
Main results and the role of chance: A 20x coverage was achieved in all ing (NGS), were used to validate and test for ploidy prediction through a set of
exons and SNV regions investigated, identifying 121 variants. Using STATA SE, machine learning classifier algorithms. Data augmentation and pre-processing
v.13, to test for normality and associations between continuous variables and methods were applied on the sample to ensure unbiased results and optimal
different genotypes, significant associations (p<0.05) were detected for each of feature selection. Validation and testing were performed through a stratified
the studied parameters: 1) No.O: lower for minor allele (MA) of rs564533 five-fold split. The efficacy of ploidy prediction was quantified using ROC curves,
(KHDC3L), higher for TT vs. CC/TT genotype of rs7616677 (SENP7) and lower AUC and confusion matrices.

Main results and the role of chance: After exhaustive feature selection via on chromosomal constitution in single (monosomy or trisomy), double, and
methods like Pearson Correlation and Decision Tree algorithms the following complex aneuploidy (more than two different aneuploidies) group.
morphokinetic and reproductive history features were shown to contribute the Main results and the role of chance: A total of 4850 aneuploidies were
most to predicting euploidy: tPNf, t4, t7-tPNf, tM-t5, tM-t6, tM-t7, tM-t8, tM-t9, detected, whole-chromosome (3547/4850;74%) occurred more frequently
maternal age and indication of PGT (PGTSR vs PGTA for recurrent miscarriages, than segmental (1303/4850; 26%) mosaicism (P<0.01). The highest preva-
advanced maternal age or severe male factor). A model was then created based lence of whole-chromosome imbalance leading to aneuploidy was seen for
on those parameters which showed a significant AUC=0.70 from the ROC curve chromosome 14, 18, 21, 22 and X, while for segmental mosaicism was seen
analysis. The mean accuracy throughout the 5 validation folds was measured at for chromosomes 1,2,5, and 16. Mosaicism rates for these chromosomes
0.68 and the mean precision at 0.73. Out of 383 samples (150 euploid/ 233 did not statistically vary when stratified by maternal age. For whole-chro-
aneuploid) the classification results, obtained from confusion matrices were: 157 mosome mosaicism, trisomy was significantly more frequent than monosomy
true positives (TP), 76 false positives (FP), 96 true negatives (TN), 54 false (p<0.05) but for segmental mosaicism trisomy was less frequent (P<0005).
negatives (FN). The features used can be sorted according to their correlation Regarding the type of mosaic embryos, 53% (1209/2278) were found to be
strength (higher to lower) as follows: PGT indication, Maternal Age, tM-t9, tM-t7, composed of whole chromosomes, 30% (694/2278) of structural aneuploi-
tM-t8, tM-t6, tM-t5, t7-tPNf, tPNf and t4. It is observed that lower values of dies and 17% (375/2278) contained both whole-chromosomal and structural
maternal age, tM-t9, tM-t7, tM-t8, tM-t6, tM-t5 and tPNf correspond to higher aneuploidies. Of the whole-chromosome embryos, single aneuploidy was
probability for euploidy while the opposite stands for t7-tPNf and t4. significantly more frequent than complex (53%, vs 28%;p<0.001), and double
Limitations, reasons for caution: The main limitation of the current study aneuploidy (53%, vs 19%;p<0.001). Similarly, when grouped based on struc-
is the small sample size. Machine learning algorithms provide better results and tural abnormalities, single segmental aneuploidy was significantly more fre-
generalize efficiently when sufficient data is provided. quent than double (80% vs 16%; p<0.0001) or complex segmental aneuploidy
Wider implications of the findings: Machine learning model depicts specific (80% vs. 4%; p<0.001). Structural mosaic and whole-chromosome aneu-
cleavage time intervals, maternal age and PGT indication as significant predictors ploidy blastocysts with >50% aneuploid cells accounted for 11% and 14% of
of embryo ploidy. This model cannot substitute pre-implantation genetic testing analysed embryos, respectively.
for aneuploidy but can potentially be useful during the prioritization of embryos Limitations, reasons for caution: This study was retrospective, demon-
for transfer in cases where PGTA/PGTSR is not performed. strating the relative frequency of different type of mosaic embryos but not
Trial registration number: non applicable offering any direct insight into the clinical relevance of the findings. Additional
clinical data must be obtained to evaluate the clinical implication of chromosomal
P-510 Detailed investigation into the mosaic embryo karyotypes: mosaicism in mosaic embryo outcome.
multicenter data from 2280 trophectoderm biopsies obtained Wider implications of the findings: Our findings reported the prevalence
during preimplantation genetic testing cycles in IVF of the different kind of mosaicism in human blastocyst. Furthermore the study
F. Fiorentino1, F. Spinella1, A. Biricik1, A. Victor2, F. Barnes2, provides a detailed description of the prevalence, distribution and level of mosa-
C. Zouves2, A. Ruberti3, E. Cursio3, M. Sofia4, L. Corti4, V. Bianchi5, icism for each chromosome involved in mosaicism. These results contribute to
P. Viganò4, M.G. Minasi3, G. Ermanno3, M. Viotti2 the understanding of the nature and origin of mosaic embryos.
1 Trial registration number: None
Genoma Group srl, Molecular Genetics Laboratories, Rome, Italy ;
2
Zouves Foundation for Reproductive Medicine, Zouves Foundation, Foster City,
P-511 Whole genome insights into male infertility
U.S.A. ;
3
European Hospital-, Centre For Reproductive Medicine., Rome, Italy ; A. Mantzouratou1, L. Xanthopoulou2, A. Mania3, P. Serhal4,
4
San Raffaele Scientific Institute, U.O. Obstetrics and Gynecology, Milan, Italy ; J. Delhanty5, S. SenGupta5
5 1
Future for Family, Future for Family, Bologna, Italy Bournemouth University, Life and Environmental Sciences, Poole, United Kingdom ;
2
Cooper Genomics, Laboratory Director, London, United Kingdom ;
Study question: Which is the prevalent type of mosaicism affecting human 3
King’s Fertility, The Fetal Medicine Research Institute, London, United Kingdom ;
mosaic embryos? 4
CRGH, The Center for Reproductive and Genetic Health, London, United Kingdom ;
Summary answer: Single whole chromosome aneuploidy is the most preva- 5
University College London, Institute for Women’s Health- Preimplantation
lent form of mosaicism in preimplantation embryos with a majority involv- Genetics Group, London, United Kingdom
ing gains.
What is known already: Chromosomal mosaic embryos are characterized Study question: How is structural genetic variation associated with the causes
by the presence of chromosomally different cell lines within the same embryo. and prognosis of poor sperm parameters and embryonic aneuploidy using a
Mosaicism may involve whole-chromosome, segmental (or partial), complex or whole genome sequencing (WGS) approach.
a combination of such aneuploidies. We previously demonstrated that the repro- Summary answer: Males with severe sperm abnormalities present with sig-
ductive potential of mosaic embryos is affected by the complexity of and the nificantly higher deletion length and increased amount of CNVs throughout their
number of aneuploid cells present in trophectoderm (TE) biopsy. We also genome that could link to embryonic aneuploidy.
observed that the mosaicism involved specific chromosomes and that single What is known already: Poor sperm parameters are associated with embry-
(sub-chromosomal or whole-chromosome) aneuploidy was the prevalent type onic aneuploidy and linked to various polymorphic sites concentrated in the Y
of chromosomal mosaicism. However, our study involved a limited number of chromosome and several individual genes. Structural genomic variations in indi-
mosaic embryos and data available were insufficient to test this hypothesis. viduals have been linked abnormal sperm parameters. So far studies have shown
Study design, size, duration: This is a large-scale multicenter study on mosaic that male infertility is the result of a combination of several genetic and environ-
embryos to examine the patterns and prevalence of chromosome specific mosa- mental interactions but in most cases the approach was targeted to specific areas
icism in TE samples. The cohort consisted of 2280 consecutive mosaic embryos of the genome with varied results. The impact of genetic variation on sperm
collected between May2016-May2019. All embryos were cultured to blastocyst parameters and embryonic aneuploidy is not yet fully established. Whole genome
stage; TE biopsy was performed on Day-5 or Day6/7of development. TE biop- sequencing can provide a more complete genomic snapshot of these
sies underwent comprehensive chromosome screening utilizing validated next conditions.
generation sequencing (NGS). TE biopsies were classified as mosaic if they had Study design, size, duration: Couples presenting with repeated implantation
20%-80% abnormal cells. failure and/or recurrent miscarriage that went through PGT-A (2008 - 2015)
Participants/materials, setting, methods: Mosaic embryos composed of were invited to provide DNA samples for WGS.
mosaic chromosomes only were analyzed. Mosaicism was tabulated per chro- Several selection criteria were applied to the test group: i) referred for PGT-A
mosome, and chromosomal constitution and incidence of different type of >3 AR failures, ii) Normal karyotypes, iii) Female age less than 37, iv) Embryonic
mosaic embryos were also analyzed. For statistical analysis mosaic embryos were instability in their subsequent PGT-A cycles. Results were analysed according to
divided in three groups: whole-chromosome, segmental and mixed mosaicism. insemination method, sperm parameters and associated with the PGT-A embryo
In addition, whole-chromosome and segmental mosaicism were divided based results.

Participants/materials, setting, methods: From the invited individuals - processed on site with the Non-Invasive Chromosomal Screening (NICS) plat-
118 consented to WGS analysis. From the consented individuals, 36 (18 couples) form (Yikon Genomics, Lewes, DE). NICS uses multiple annealing and loop-
passed all the selection criteria. Within that cohort, there were 12 males needing ing-based amplification cycles (MALBAC) for whole-genome amplification
ICSI and poor sperm parameters and 9 males with normal sperm parameters followed by next-generation sequencing to obtain ploidy information on all 24
(IVF cohort) and DNA from these participants was sequenced. chromosomes.
WGS was performed with a short-read BGI platform. Variant calling and genomic Main results and the role of chance: The average patient age was 36.6
data analysis were performed with gold standard bioinformatic pipelines. years old. Of the 1,504 media samples tested with the NICS platform, 86 (5.7%)
Main results and the role of chance: Whole genome sequencing provides were initially found be inconclusive. 62 of the 86 inconclusive embryos were
a global view of the human genome and is a step towards a personalized thawed for overnight culture and next day repeat media collection based on
approach to healthcare. patient preference. The remaining 24 inconclusive were not resampled. All
This study has found that males with severe sperm abnormalities present with retested embryos resulted in usable ploidy data. The proportion of Euploid,
high DNA deletion length and increased amount of copy number variations Mosaic, and Aneuploid calls in the 1480 resulted embryos (original and retested)
compared to less severely affected individuals and to males with normal sperm were 22.9%, 32.6%, and 44.5%, respectively. Of the 62 retested embryos, 35.4%
parameters. Specifically, the ICSI group had significantly higher total CNV dele- were euploid, 29.0% were mosaic, and 35.5% were aneuploid. Data was further
tion length than the IVF group. In addition, the total number of CNV’s was stratified based on established Society for Assisted Reproductive Technology
significantly higher in the males with severe conditions like azoospermia com- (SART) age groups. Excluding mosaic results, 51.1%, 38.0%, 20.8%, 26.4%, and
pared with the males that presented with poor sperm parameters like oligosper- 10.5% were found in be euploid in patients under 35, 35-37, 38-40, 41-42, and
mia. T hese variations were found in several areas of the genome as well as in over 42, respectively.
the Y chromosome or genomic areas related to fertility. Limitations, reasons for caution: While the NICS platform has been pre-
Several polymorphic sites found in the same genomes could contribute to an vious validated, the proportion of mosaic calls can vary depending on thresholds
accumulated risk that is linked to poor sperm parameters and embryonic aneu- established within the Yikon pipeline. Additionally, caution should be taken when
ploidy. For example, in severely affected individuals, structural variations were generalizing results as certain clinical and embryologic practice parameters could
found in several genes associated with spermatogenic failure like AURKC, affect DNA capture and successful amplification.
CATSPER1, CEP19. Variations in genes associated with increased prostate cancer Wider implications of the findings: The NICS platform can be utilized
risk observed in some men with poor sperm parameters. This could explain the successfully in a private setting. Embryos resulting as inconclusive initially can be
highly heterogeneous associations in genetics of male infertility. successfully retested. Furthermore, the use of a non-invasive methods to identify
Limitations, reasons for caution: Ethnicity bias presented in relation to the embryo ploidy may reduce risk to the embryo, however this has yet to be
bioinformatic analysis as variation is flagged based on frequencies mainly focused determined.
in Caucasian populations, so this has been considered especially for the SNP Trial registration number: not applicable
analysis. For future work, a normal fertility control group with preimplantation
embryo results would facilitate further research. P-513 A homozygous missense mutation in an highly conserved
Wider implications of the findings: A complex picture of a polygenetic/ autosomal gene coding poly(A)-binding protein cause female
multifactorial risk is arising for certain infertility types, the results from such infertility due to oocyte maturation arrest at GV stage
targeted population studies can help in the identification and subsequently the O. Okutman1, A.S. Gürbüz2, U. Büyük3, B. Rinaldi4, S. Friant4,
prediction of genetic risk for heterogeneous conditions. This study adds to the J. Muller5, S. Viville1
prognostic value of WGS in male infertility and embryo aneuploidy. 1
Hôpitaux Universitaires de Strasbourg- Nouvel hôpital civil, Laboratoire de
Trial registration number: Not applicable diagnostic génétique- UF3472 Infertilité, Strasbourg, France ;
2
Novafertil, IVF Center, Konya, Turkey ;
P-512 A review of 1,504 autologous embryos evaluated using a 3
Hibrigen Biyotecnology, Research and Development, Gebze-Kocaeli, Turkey ;
non-invasive platform for preimplantation genetic testing for 4
Centre National de Recherche Scientifique CNRS- Université de Strasbourg,
aneuploidy at a private clinic Department of Molecular and Cellular Genetics- UMR7156, Strasbourg, France ;
M. Olcha1, M. Elzaky1, M. Jaremko1, Z. John1, B. Hensinger2, S. Lu2 5
Hôpitaux Universitaires de Strasbourg, Laboratoire de diagnostic génétique,
1
New Hope Fertility Center, Reproductive Endocrinology, New York, U.S.A. ; Strasbourg, France
2
Yikon Genomics, Genomics, Lewes, U.S.A.
Study question: Is it possible to identify the new causative genetic mutation
Study question: To determine if a non-invasive approach for preimplantation responsible of an oocyte maturation arrest in a Turkish consanguineous family
genetic testing for aneuploidy (PGT-A) can successfully assess embryo ploidy via whole exome sequencing?
status at a large-scale private practice setting Summary answer: Whole exome sequencing reveals a homozygous missense
Summary answer: Non-invasive PGT-A can identify ploidy status in a large mutation for the three affected sisters in a Turkish familiy with multiple loops of
cohort of embryos. Furthermore, this technology can also distinguish mosaic consanguinity.
embryos from aneuploid or euploid ones. What is known already: Women undergoing controlled ovarian hyper stim-
What is known already: PGT-A has been widely used around the world to ulation prior to in vitro fertilization (IVF) are treated by various protocols aimed
identify ploidy status of embryos. The knowledge obtained can help assist clini- at inducing multiple follicular growths. Approximately 20%–30% of oocytes
cians and patients with prioritization of embryos for transfer. The transfer of collected for IVF are meiotically immature at the time of oocyte retrieval. The
single euploid embryos has be shown to improve per cycle pregnancy rate, complete failure of all oocytes to mature in vivo is very rare, but known as causes
reduce miscarriage rate, and prevent transfer of embryos harboring certain of primary female infertility. Recently mutations in two genes, TUBB8 and PATL2
genetic anomalies such as Down Syndrome. The primary modality for obtaining have been identified in women with oocyte maturation arrest. However, for
DNA from blastocysts has been trophectoderm biopsy, which requires removal approximately 60% of cases, underlying genetic factors are still largely unknown.
of embryonic cells. Recently, the advent of non-invasive methods for PGT-A Study design, size, duration: We have recruited a Turkish familiy with mul-
allow embryonic DNA within spent media to be amplified for ploidy analysis. tiple loops of consanguinity comprising 3 sisters with repeating oocyte matura-
Study design, size, duration: Retrospective data analysis of 1,504 blasto- tion arrest at GV stage. Different stimulation protocols have been tried
cysts, which were evaluated using non-invasive PGT-A from August 1st 2019 to nevertheless collected oocytes failed to proceed into mature oocytes, even after
December 30th 2019. All embryos evaluated using non-invasive PGT-A were extended in vitro culture. All three sisters have normal karyotype; however their
included except patients using donor eggs. brother was diagnosed with Down syndrome.
Participants/materials, setting, methods: Patients electing to have their Participants/materials, setting, methods: Blood samples from all siblings,
embryos undergo PGT-A were offered to utilize a validated non-invasive parents and available non-affected family members were collected after obtaining
approach instead of conventional trophectoderm biopsy after appropriate signed consent form. Genomic DNA was extracted from peripheral blood using
informed consent. Spent media collected from embryos on day 5, 6, or 7 was Hibrigen DNA Extraction-Blood kit (Hibrigen, Istanbul, Turkey), according to

the manufacturer’s instructions. Exome sequencing of three affected sisters was NM_015091.3:c.1102C>T [p.(Arg368Trp)], and c.3619C>T [p.(Arg1207*)],
performed by Integragen Genomics via Illumina technology. Bioinformatics anal- each inherited from one parent. TOGARAM1 encodes a protein regulating micro-
ysis was executed using IG Constitutionel DNA pipeline V4.0. Detected variants tubule dynamics, it is highly conserved throughout evolution and che-12, its
were also scored and ranked by VaRank tool. orthologue in nematodes, is expressed in the cilium of a subset of sensory
Main results and the role of chance: For three samples, at least 4.8GB neurons.
DNA sequence were generated with >99% of the target exome was represented Mutant worms recapitulating the patients’ genotype showed a shorter cilium
with >25-fold coverage. Exome sequencing reveals a homozygous missense in sensory neurons and in vitro experiments confirmed aberrant tubulin binding,
mutation in an autosomal gene (STBG1) for all three affected sisters. The muta- indicating a causative role of TOGARAM1 variants in the pathogenesis of this
tion was confirmed and segregation has been shown in available family members novel primary ciliopathy, characterized by a spectrum of defects consistent with
via Sanger sequencing; parents, the cousin and one aunt are heterozygous the Meckel-Gruber phenotype,including microphthalmia, hydrocephalus and
whether second aunt is wild type for the substitution. The coding protein is well cleft palate.
conserved during evolution from lamprey to human. The identified mutation is The molecular identification of this new genetic disorder allowed to offer the
in RNA recognition domain, predicted as disease causing/deleterious through couple the opportunity to perform the preimplantation genetic diagnosis
different online tools. KO female mice are infertile, and could not generate (PGT-M).
mature oocytes neither in vivo nor in vitro. Functional studies are ongoing; the Limitations, reasons for caution: We cannot predict accurately the post-na-
effect of the mutation is studied on budding yeast. tal phenotype associated with biallelic TOGARAM1 mutations, because we have
Limitations, reasons for caution: Our study is limited to one family only only observed fetal cases. Further studies are warranted to better define the
and the phenotype is very rare. We still have to screen a cohort of patients clinical picture of this new severe disorder.
presenting the same phenotype. Wider implications of the findings: Our data strongly support the role of
Wider implications of the findings: Our results provide a new understand- TOGARAM1 as a novel causative gene of primary ciliopathy, which thus should
ing for the pathogenesis of oocyte maturation arrest and help us to identify be included in diagnostic gene panels for this heterogeneous group of disorders.
reason for GV arrest in our patients. It may help to define novel therapeutic Trial registration number: not applicable
approach to patients with similar symptoms.
Trial registration number: Not applicable P-515 PGT-A: Who and when? A systematic review and network
meta-analysis of RCTs
P-514 From molecular identification to PGT-M of a novel primary A. Pantou1, K. Sfakianoudis2, E. Maziotis3, P. Tsioulou3,
ciliopathy due to biallelic mutations in the TOGARAM1 gene S. Grigoriadis3, A. Rapani3, P. Giannelou2, M. Asimakopoulou3,
D. Zuccarello1, V. Morbidoni2, E. Agolini3, K.C. Slep4, L. Pannone5, K. Nikolettos4, T. Kalampokas5, G. Kokkali2, K. Pantos2, N. Vlahos6,
E. Grosso6, G. Gai6, B. Dallapiccola7, A. Novelli3, S. Martinelli8, E. M. Koutsilieris3, M. Simopoulou3
Trevisson2 1
Genesis Genoma Lab, Molecular Genetics, Athens, Greece ;
1
Clinical Genetics Unit, Department of Women’s and Children’s Health- University 2
Genesis Athens Clinic, Assisted Conception Unit, Athens, Greece ;
of Padova, Padova, Italy ; 3
National and Kapodistrian Univerisity of Athens, Physiology, Athens, Greece ;
2
Clinical Genetics Unit- Department of Women’s and Children’s Health- University 4
Assisted Reproduction Unit of Thrace “Embryokosmogenesis”, Assisted
of Padova, Istituto di Ricerca Pediatrica- IRP- Fondazione Città della Speranza, Conception Unit, Alexandroupoli, Greece ;
Padova, Italy ; 5
Aberdeen Maternity Hospital- Fertility Center, Assisted Reproduction Unit,
3
Laboratory of Medical Genetics, IRCCS-Bambino Gesù Children’s Hospital, Rome, Aberdeen, United Kingdom ;
Italy ; 6
Aretaieion University Hospital, 2nd Department of Obstetrics and Gynecology,
4
University of North Carolina, Department of Biology, Chapel Hill- NC, U.S.A. ; Athens, Greece
5
Department of Oncology and Molecular Medicine- Istituto Superiore di Sanità,
Genetics and Rare Diseases Research Division- Ospedale Pediatrico Bambino Gesù, Study question: What is the optimal practice for preimplantation genetic
Rome, Italy ; testing for aneuploidy (PGT-A) considering the patients’ age and the best day
6
Medical Genetics Unit, Città della Salute e della Scienza University Hospital, to biopsy?
Turin, Italy ; Summary answer: PGT-A employing complete chromosomal screening
7
Scientific Directorate, Bambino Gesù Children’s Hospital and Research Institute, appears to be beneficial only when performed on the basis of trophoectoderm
Rome, Italy ; biopsy, addressing predominantly women over 35 years old
8
Department of Oncology and Molecular Medicine, Istituto Superiore di Sanità, What is known already: Following literature search, two schools of thought-if
Rome, Italy not more-are emerging with regards to “when” and “how” PGT-A should be
employed. A number of prospective and retrospective studies revealed contro-
Study question: How to eliminate the risk of recurrence of a genetic condition versial results concerning the effectiveness of PGT-A cycles during fresh or frozen
with unknown molecular basis? ART cycles. On one hand, the advocators support that performing PGT-A holds
Summary answer: By identifying the causative gene, ascertained by robust remarkable promise with regards to successful embryo implantation, especially
functional studies, it is possible to offer PGT-M to the couple. when focusing on the distinct group of women of advanced maternal age. On
What is known already: Dysfunction in non-motile cilia is associated with a the other hand, no difference and even lower live birth rates have been demon-
broad spectrum of developmental disorders characterized by clinical heteroge- strated when opting for PGT-A in comparison to IVF cycles without PGT-A.
neity. Despite over one hundred genes have been associated with primary cil- Study design, size, duration: A systematic review and meta-analysis of 10
iopathies, with wide phenotypic overlap, some patients still lack a molecular published studies including women who underwent IVF-Embryo Transfer cycles
diagnosis. following complete chromosomal screening (CCS) of the preimplanta-
Study design, size, duration: Whole exome sequencing (WES) analysis, in tion embryo.
silico, in vitro and in vivo functional studies. Participants/materials, setting, methods: A systematic search of the lit-
Participants/materials, setting, methods: Unrelated young parents were erature was performed in the databases of PubMed/Medline, Embase and
referred following 2 TOP due to multiple fetal malformations, including micro- Cochrane Central Library, limited to articles published in peer-reviewed journals
cephaly, severe cleft lip and palate, microphthalmia, and brain malformations. up to June 2019. The initial search yielded 1819 studies. From the total yield,
Array-CGH on fetal DNA was normal. WES analysis was performed on DNA 215 studies were duplicates and 1501 were excluded on the grounds of not
from 2 fetuses and its parents, and in silico, in vitro and in vivo (in C. elegans) fulfilling criteria for inclusion. Following thorough full-text screening and citation
functional studies were carried out to explore the impact of finding mutations mining a total of 10 studies were included in the current meta-analysis.
on protein structure and function. Main results and the role of chance: A total of 10 Randomized Control
Main results and the role of chance: WES analysis identified in both fetuses Trials employing CCS were identified. PGT-A improved ongoing pregnancy/live
a compound heterozygous genotype in the orphan gene TOGARAM1: birth rates both in the pooled results (RR:1.29; 95%CI:1.07-1.56) and in the over

35 years old subgroup (RR:1.52; 95%CI:1.01-2.28), when compared to the con- with a major risk of reproductive failure. Our case is the first one for our national
trol group of morphological assessment for embryo selection. Interestingly, no trophoblastic disease center. It confirms the effectiveness of oocyte donation
statistically significant difference was observed in the younger than 35 years old to prevent recurrent hydatidiform moles.
subgroup (RR:1.10; 95%CI:0.81-1.48), when compared to the control group. Trial registration number: not applicable
When comparing D3 and D5 biopsy no statistically significant difference was
observed between the groups regarding ongoing pregnancy/live birth rates P-517 Analysis of segmental aneuploidy and mosaicism in the
(RR:0.87; 95% CI: 0.59-1.29). When comparing D3 biopsy to the control group human blastocysts. Is there any difference in pregnancy rates?
no statistically significant difference was observed between the groups regarding J.A. Ortiz1, B. Lledo1, R. Morales1, E. Garcia-Hernandez1, J. Ten2,
ongoing pregnancy/live birth rates (RR:1.10; 95% CI: 0.75-1.60). Interestingly, J. Llacer3, A. Bernabeu3, R. Bernabeu3
only D5 biopsy provided enhanced rates regarding ongoing pregnancy/live birth 1
Instituto Bernabeu, Biología Molecular y Genética, Alicante, Spain ;
(RR: 1.26; 95% CI: 1.04-1.53) when compared to the control group. 2
Instituto Bernabeu, Embriología, Alicante, Spain ;
Limitations, reasons for caution: The multifaceted nature of PGT-A success 3
Instituto Bernabeu, Medicina Reproductiva, Alicante, Spain
may stand as a valid limitation when unravelling results and drawing conclusions.
When reporting on what truly makes a PGT-A cycle successful we should clearly
Study question: What are the characteristics, associated factors and the clinical
distinguish between efficiency of diagnosis and successful outcome of the cycle
pregnancy of the embryos with segmental alterations?
leading to a healthy offspring.
Summary answer: The ongoing pregnancy rates of mosaic segmental embryos
Wider implications of the findings: The results of the present study indicate
are similar to those of euploid embryos, therefore, embryo transfer with seg-
that PGT-A may improve clinical outcomes and live birth rates only when per-
mental mosaicism could be considered.
formed on women aged over 35 years old and on blastocyst stage embryos.
What is known already: Chromosomal abnormalities in IVF embryos can be
However, further RCTs investigating the exact requirements to ascertain a ben-
detected by PGT-A. By using NGS or aCGH, it is possible to detect segmental
eficial effect of PGT-A should be conducted.
(subchromosomal) errors that occur when small portions of DNA are duplicated
or removed from the embryo. The origin of the segmental alterations is mainly
mitotic and occurs in the first embryonic divisions. Many of the characteristics
P-516 Pregnancy after oocyte donation in a patient with NLRP7 of these alterations have not been studied, such as: proportion of chromosomes
mutation and a history of recurrent hydatidiform molar pregnancy with segmental errors, hot spot regions, type of segmental errors, length of
F. Scheffler1, C. Cozette1, J. Massardier2, I. Touitou3, J. Gondry4, alterations, or percentage of aneuploid cells. Moreover, the effect of segmental
H. Copin1, M. Ben Khalifa1, R. Cabry-Goubet1 mosaicism in the pregnancy rate is still controversial.
1
CHU Amiens Picardie SUD, Service de médecine et biologie de la reproduction- Study design, size, duration: A total of 5286 embryos from 1822 PGT-A
cytogénétique et CECOS de Picardie, Amiens, France ; cycles were included in this study (October 2014-April 2019). The trophoecto-
2
CHU Lyon Sud, Centre Français des Maladies Trophoblastiques- Hospices Civils de derm biopsies on D5/D6 blastocysts were analysed by aCGH (46%) or NGS
Lyon- Pierre Benite, Lyon, France ; (54%). The detection of segmental alterations was validated by the analysis of
3
CHU Montpellier, Cellules souches- plasticité cellulaire- médecine régénérative et embryos from patients with structural chromosomal alterations. Embryos with
immunothérapies- INSERM- Université de Montpellier- Département de génétique ≤25% aneuploid cells were considered euploid, between 25-50% were classified
médicale- maladies rares et médecine personnalisée- CEREMAIA, Montpellier, as mosaic and aneuploid with >50%. Only euploid or mosaic embryos were
France ; transferred.
4
CHU Amiens Picardie SUD, Service de gynécologie et obstetrique, Amiens, France Participants/materials, setting, methods: The main indications for
PGT-A were advanced maternal age, abnormal sperm FISH and recurrent
Study question: Is oocyte donation the best option in case of genetic hydatid- miscarriage or implantation failure. Embryo analysis were performed using
iform moles? Agilent SurePrint G3-8x60K-CGH microarrays or Veriseq-NGS (Illumina), with
Summary answer: Oocyte donation appears to be the best option for a previous whole genome amplification. The comparison of ongoing pregnancy
normal and successful pregnancy in women with a history of recurrent complete rate between euploid embryos, segmental mosaic and full chromosome mosaic
hydatidiform moles was performed by a binary logistic regression in which maternal and paternal
What is known already: Molar pregnancies are benign trophoblastic diseases ages, embryo quality and day of biopsy were introduced as confounding factors
associated with a risk of malignant transformation. If aetiology remains mostly (SPSSv20.0).
unknown, the risk of recurrent molar pregnancy is around 1.5% after one molar Main results and the role of chance: A total of 5286 embryos were anal-
pregnancy and around 25% after 2 molar pregnancies. Genetic mutations have ysed (41.3% aneuploid and 2.6% not informative). Segmental alterations were
been described, increasing hugely this risk of trophoblastic diseases. In case of detected in 12.4% of the analysed embryos, 42.9% of these embryos presented
mutations, the probability to obtain a normal pregnancy is estimated around segmental deletions, 25.1% duplications and 32.0% both. The average size of
1.8%. In patient with pathogenic mutations in the NLRP7 gene, only 3 normal the segmental imbalances was 56.87 Mb (8 to 170 Mb). Interestingly, sites of
pregnancies and deliveries after oocyte donation have been published. chromosome breakage associated with segmental errors were not random. A
Study design, size, duration: Case report 58.2% of the segmental alterations were located near the telomeres, 7.4% in
Participants/materials, setting, methods: We report the case of a the proximity of the centromere, 4.7% in the middle of the arms and 29.7%
Caucasian 30-year-old woman whose previous five spontaneous pregnancies affect a whole arm. Analysing the factors associated to segmental errors, we
had a negative outcome: a spontaneous miscarriage and 4 complete hydatidiform observed an inverse correlation among segmental alteration and maternal age:
moles, without gestational trophoblastic neoplasia. There was no history of 15.3% in women until 35y versus 10.3% in >36y women (p<0.0001). No factors
recurrent pregnancy failures in her family. Genetic testing revealed that the of paternal origin showed any association. On the other hand, embryonic quality
patient carried two pathogenic mutations in the NLRP7 gene (Y318CfsX7 and showed a strong association with segmental errors (A: 10.3%; B14.5%; C:18.4%,
c.2982-2A>G). D:18.2%;p<0.0001). We also analyzed the clinical data of embryos with seg-
Main results and the role of chance: We proposed an oocyte donation in mental mosaicism versus embryos with full chromosome mosaicism or euploid
order to obtain a normal pregnancy. Four mature oocytes were done and two in single embryo transfer cycles. The ongoing pregnancy rate was higher for
embryos transferred on day 3. This technic enabled a complication-free, singleton segmental mosaics compared with full chromosome mosaics (42.1% vs
pregnancy that resulted in a healthy term live birth of a 2900g female, in spite 22.7%;p=0.066) and similar to cycles where euploid embryos were transferred
of a pre-eclampsia at the end of the pregnancy. The pathological placenta exam- (42.1% vs 33.9%;p=0.254).
ination was normal and we could observe a spontaneous normalization of Limitations, reasons for caution: The main limitation of this study is the
BHCG. The couple is now waiting for another oocyte donation. use of different techniques for the PGT-A. The NGS is more sensitive and specific
Limitations, reasons for caution: none to detect segmental errors than aCGH. To avoid this bias, the analysis technique
Wider implications of the findings: Women presented with mutations in (aCGH or NGS) was introduced into the statistical analysis as a confound-
the NLRP7, KHDC3L or PADI6 genes are unlikely to obtain normal pregnancies, ing factor.

Wider implications of the findings: Segmental-chromosomal alterations Summary answer: Live birth rate with single blastocyst transfer with or with-
are relatively frequent in embryos and chromosome breaks close to the telomere out PGT-A in freeze-all strategy is not superior to fresh ET in
are more likely. Maternal age and embryo quality are risk factors. Finally, ongoing normoresponders.
pregnancy rates of mosaic segmental embryos are similar to euploid embryos, What is known already: Cryopreservation of human embryos is now a
consequently the transfer of mosaic segmental embryos could be considered. routine procedure in assisted reproductive technologies for better embryo-en-
Trial registration number: Not applicable dometrium synchrony and the lower risk of ovarian hyperstimulation syndrome
at the same time resulted in live births increased. However, there are some
P-518 Embryo morphology affects euploid blastocysts concerns need to be evaluated in freeze-all strategy such as the efficacy in normal
implantation or poor responders. Aneuploidy is a leading cause of implantation failure and a
significant cause of failure in IVF cycle, PGT-A has been deemed as a promising
M. Gomez Peña1, L. Kopcow2, M. Rocha2, G. Terrado2,
approach to improve pregnancy rate but on the other hand PGT-A requires
M. Papayannis1, E. De Martino1, M. Horton2, F. Sobral2,
consideration of multipl factors
I. De Zúñiga2, A. Oubiña2, C. Bisioli1
1
Study design, size, duration: This retrospective study includes patients who
Pregna Medicina Reproductiva, ART Embryology, Buenos Aires, Argentina ;
2
underwent ICSI-ET procedures at Istanbul Yeni Yüzyil University Department
Pregna Medicina Reproductiva, Clinical department, Buenos Aires, Argentina of Reproductive Medicine&Infertility between January 2016 and December 2018.
Fresh embryo transfers (n:72) frozen-thawed embryo transfer(FET) (n:264) and
Study question: Is blastocyst euploidy the only relevant factor to predict
frozen-thawed embryo transfer after PGT-A (FET-PGT-A) (n:56) were analyzed.
implantation?
Participants/materials, setting, methods: Women aged< 42 years with
Summary answer: Good quality euploid blastocysts have a better implanta-
normal uterine cavity , normo-responders who had 5-14 oocytes at retrieval,
tion and pregnancy potential.
single blastocyst transfer, ovulation induction with gonadotropin-releasing hor-
What is known already: Euploidy has been regarded in the literature as the
mone antagonist (GnRH-a), sperm found in ejaculate, endometrial thickness ≥
only guarantee of a successful implantation and live birth. Thus, irrespective of
7 mm on the day of hCG injection in fresh ET cycles or of progesterone start
other embryological and clinical variables such as patient age, blastocyst mor-
in FET cycles ,first complete cycle in the center. Only blastocysts having normal
phology or biopsy day. This assertion, however, remains controversial.
karyotypes were transferred in PGT-A cycles.
Study design, size, duration: Retrospective comparative study of 222 elec-
Main results and the role of chance: A total of 392 couples who had fresh
tive single frozen-thawed blastocyst transfers after preimplantation genetic testing
embryo transfer or a first frozen embryo transfer with or without PGT-A after
for aneuploidy (PGT-A) via next generation sequencing (NGS). Sampling
freeze-all cycles were analyzed in the study. Fresh ET, FET and FET+PGT-A
occurred between June 2016 and September 2019.
groups were comparable with regard to number of oocytes collected (8.8±2.3,
Participants/materials, setting, methods: We compared implantation (IR),
8.7±2.9 and 9.1±2.4) and fertilized (5.9±2.3, 6.1±2.2 and 6.3±2.1) as well as
ongoing pregnancy (OPR) and miscarriage (MR) rates between:
the number of blastocysts (2.3±1.2, 2.3±0.9 and 2.4±1.3) available for the
transfer, consecutively.
• Embryo biopsy day 5 (n= 138) vs. day 6 (n= 84). Positivity of β-hCG level (>5 IU/L) was detected in 38 (52.7 %) women in
• Female age <37 yo (n= 89) vs ≥ 37 yo (n= 133) fresh ET group, 168 (63.3 %) women in FET group and 33 (58.9 %) women in
• Excellent/Good (n= 138) vs. Fair/Poor (n= 84) quality embryos. FET+PGT-A group following single blastocyst transfer (p>0.05).
All pregnancies were singletons and no severe OHSS was observed in all
Student’s t-test and Chi-squared test were used as appropriate. groups. Moderate OHSS was detected in 3 cases (4%) in fresh ET cycles and 11
Main results and the role of chance: IR (43.5% (n=64/147) vs 38.8 % cases (3.4%) in all FET cycles.
(n=33/85); p=NS), OPR (39.2% (n=49/138) vs 29.7% (n=25/85); p=NS) and Live birth rates were 24 (33.3%) for fresh ET group, 98 (37.1%) for FET and
MR (5.8% (n=8/138) vs 7.2% (n=6/84); p=NS) were similar between embryos 24 (42.8%) for FET+PGT-A group. Although it was higher in FET+PGT-A group
biopsied on D5 vs D6 respectively. compared to fresh ET group it did not reach statistical significance (p>0.05).
We also found no differences regarding patients age <37 vs ≥ 37 in IR (49.5% Limitations, reasons for caution: The limitations of our study include its
(n=45/91) vs 38.0% (52/137); p=NS), OPR (39.3% (n=7/89) vs 31.5% retrospective nature, the lack of hormonal assessments in the follicular phase
(n=7/133); p=NS) or MR (7.8% (n=35/89) vs 5.3% (n=42/133); p=NS). and imprecise allocation of patients into each group (physician’s or patient’s
Excellent and good quality blastocysts achieved better IR (49.0 (n=70/143) vs preference).Otherwise, it was an advantage that all procedures were performed
31.7% (n=27/85); p=0.01) and OPR (39.85% (n=55/138) vs 26.2% (n=22/84); by the same embryologist.
p=0.02) than fair and poor ones. MR was not significantly different between Wider implications of the findings: This was first study comparing fresh
different quality embryos (7.2% (n=10/138) vs 4.8% (n=4/84); p=NS). ET with FET with and without PGT-A in the literature.Although live birth rate
Limitations, reasons for caution: This study has biases due to its retrospec- was higher in PGT-A and FET group than fresh ET group but did not reach
tive design. The small sample size and the strong subjectivity among embryolo- statistical significance. There is a need for further prospective randomised studies
gists when grading blastocysts are additional limitations. to confirm this subject properly.
Wider implications of the findings: Although euploidy is considered as a Trial registration number: 2019/71
main predictor of reproductive success our analysis shows that embryo grading
might also play a role. Therefore diploid chromosome number alone might not P-520 Developmental incompetent preimplantation embryo: a
be sufficient to predict a successful implantation. two Societies consensus
Trial registration number: Not applicable R. Ciriminna1, A. Novelli2, L. De Santis3, M.G. Minasi4, L. Rienzi5,
R. Gualtieri6, D. Paoli7, V. Pisaturo8, C. Scarica9, D. Zuccarello10
P-519 Freeze-all cycles with or without preimplantation genetic 1
AMBRA, On behalf of the Italian Society of Embryology- Reproduction and
diagnosis for aneuploidy superior to fresh embryo transfer in Research SIERR, Palermo, Italy ;
normo-responders? 2
IRCCS-Bambino Gesù Children’s Hospital Laboratory of Medical Genetics, On
M. Karacan1, M. Keskin2, M. Kurek Eken2, M. Yanık1 behalf of the Italian Society of Human Genetics SIGU, Rome-, Italy ;
3
1
Yeni Yuzyıl University Medical Faculty, Department of Reproductive Medicine and San Raffaele Scientific Institute -Ob/Gyn IVF UNIT, On behalf of the Italian
Infertility, Istanbul, Turkey ; Society of Embryology- Reproduction and Research SIERR, Milan, Italy ;
4
2
Yeni Yuzyıl University Medical Faculty, Department of Reproductive Medicine and Center for Reproductive Medicine- European Hospital, On behalf of the Italian
Infertility, İstanbul, Turkey Society of Embryology- Reproduction and Research SIERR, Rome, Italy ;
5
GENERA centres for Reproductive Medicine, On behalf of the Italian Society of
Study question: Are freeze-all cycles with or without preimplantation genetic Embryology- Reproduction and Research SIERR, Rome, Italy ;
6
diagnosis for aneuploidy superior to fresh embryo transfer in normo- University of Naples “Federico II”- Department of Biology, On behalf of the
responders? Italian Society of Embryology- Reproduction and Research SIERR, Naples, Italy ;

7
Laboratory of Seminology - Sperm Bank “Loredana Gandini” - Sapienza” Study question: The cystic fibrosis carrier screening on sperm donors, is better
University of Rome, On behalf of the Italian Society of Embryology- Reproduction to do it through a complete analysis of the CFTR gene by NGS or by genotyping?
and Research SIERR, Rome, Italy ; Summary answer: We recommend to study the CFTR gene by NGS for the
8
International Evangelical Hospital- Reproductive Medicine, On behalf of the cystic fibrosis carrier screening on sperm donors.
Italian Society of Embryology- Reproduction and Research SIERR, Genoa, Italy ; What is known already: In accordance with the legal regulations in force in
9
Casa di Cura Villa Salaria in partnership with Institut Marques- Center for Spain and under the European Union (EU) Directive on human tissues and cells,
Reproductive Medicine, On behalf of the Italian Society of Embryology- gamete donors should be screened (tested) for autosomal recessive genes
Reproduction and Research SIERR, Rome, Italy ; known to be prevalent. Due to the high prevalence of cystic fibrosis, many
10
Clinical Genetics Units- University Hospital of Padova, On behalf of the Italian scientific societies recommend the cystic fibrosis carrier screening for sperm
Society of Human Genetics SIGU, Padova, Italy donors. However, none has specified whether the CFTR gene should be studied
by complete sequencing and non-directed analysis of variants or by using a panel
Study question: What is the definition of “developmental incompetent” pre- of targeted variants (genotyping panel).
implantation embryos, which can therefore be discarded, in the absence of a Study design, size, duration: It is a descriptive observational study, with 935
clear regulatory reference law? men aged 18-35 years, who were evaluated in the framework of a sperm dona-
Summary answer: SIERR and SIGU define as “developmental incompetent” tion programme in Spain, from January 2014 to June 2019.
a preimplantation embryo with specific characteristics related to ploidy, type of Participants/materials, setting, methods: We did the complete study of
cell division, chromosomal and molecular status. the CFTR gen by Next Generation Sequencing (NGS) in 935 caucasian sperm
What is known already: In 2010 SIERR published the first document on the donors in a Spanish private sperm bank. We have determined the frequency of
definition of “developmental incompetent embryos”, in the absence of a specific sperm donors carrying pathogenic variants of the CFTR gene and we have
indication in the Italian IVF Law 40/2004 on which embryo can be discarded at analysed the detection rate of these carriers that would have been detected by
the end of IVF process. applying the genotyping tests for CFTR gene mutations that are most commonly
Nine years later, SIERR, in collaboration with SIGU felt the need to update and used in the assisted reproduction in Spain.
implement the document, in the light of the development of new diagnostic Main results and the role of chance: Of the 935 sperm donors, 159 (17%)
capabilities related to modern technologies. The present consensus redefines were carriers of at least one pathogenic variant in the CFTR gene. The frequency,
the criteria according to which a preimplantation embryo can be defined as therefore, was approximately one in six. In total, 39 different variants were found.
developmental incompetent. Of the 39 variants found in the sperm donors, 22 variants (56,41%) would not
Study design, size, duration: Five SIERR embryologists and 2 SIGU geneti- have been detected by any of the four genotyping tests. And only four of the
cists (a clinician and a biologist), representative of their respective scientific 39 variants (10,27%) would have been detected by all of the genotyping tests.
societies worked as a committee in 6 board meetings, through 2018 and 2019, Our study shows there is great heterogeneity in the pathogenic variants of the
in order to draw up the document. The consensus was then approved by the CFTR gene that are included in the genotyping tests most commonly used in the
steering committee of the two scientific societies and made open through pub- context of assisted reproduction. Less than 50% of the variants included in the
lication on their respective websites. panels are common to all these tests. The number of carrier donors detected
Participants/materials, setting, methods: Over a hundred between sci- by genotyping would be 8.2%-51,6% from total number of carrier donors
entific articles and guidelines have been selected through systematic research of detected by NGS.
specific reference items. The most recent morphology and morphokinetics Limitations, reasons for caution: The limitations of this study is that it is
studies and the most advanced preimplantation genetic diagnosis technologies, not a population study (men had to meet exclusion criteria with excellent semen
allowed the working committee to develop standard criteria to define a devel- parameters), and second that it is a relatively homogeneous population which
opmental incompetent embryo. may not extend to other ethnic groups. In consequence, our population is biased
Main results and the role of chance: The criteria to define a developmental in this respect.
incompetent embryo have thus been established as: a) presence of a number Wider implications of the findings: The NGS presents a higher detection
of pronuclei different from two; b) no further cell division 24 hours after the rate of cystic fibrosis carrier than genotyping. There is a significant high repro-
previous observation; c) blastocyst affected by complex aneuploidy; d) blastocyst ductive risk when targeted panels are used.
affected by complete or partial monosomy in homogeneous form of an auto- We recommend to study the CFTR gene by NGS for the cystic fibrosis carrier
some; e) blastocyst affected by complete or partial trisomy in a homogeneous screening on sperm donors to reduce the risk of having offspring affected.
form of chromosomes: 1, 2, 3, 4, 5, 7, 10, 11, 12, 14, 19; f ) blastocyst affected Trial registration number: N/A
by polyploidy or haploidy; g) blastocyst affected by in uterus lethal monogenic
P-522 Effects of the Differences in Trophectoderm (TE) Biopsy
disease.
Procedures on the Results of Next-Generation Sequencing (NGS)
Limitations, reasons for caution: Regarding the genetic criteria identified
Analysis
to define a developmental incompetent embryo, it is necessary to take into
account the percentage of diagnostic error intrinsic to each technique in preim- K. Takeuchi1, Y. Kuwatsuru2, Y. Kuroki2, Y. Fukumoto2,
plantation genetic testing as well as the possible detection of chromosomal M. Tokudome2, H. Moewaki2, E. Hashimoto2, M. Watanabe2,
mosaicism, which varies according to the technique adopted. T. Iwakawa1, T. Tabira3, T. Kato4, H. Kurahashi4, M. Takeuchi3
Wider implications of the findings: This document fills a legislative gap of 1
Takeuchi Ladies Clinic / Infertility Center, Infertility, Aira-shi- Kagoshima-, Japan ;
2
Italian law while providing a useful tool to support the choice to not transfer. It Takeuchi Ladies Clinic / Infertility Center, laboratory, Aira-shi- Kagoshima-, Japan ;
3
allows the IVF centers to discontinue these embryos from further culture, reduc- Takeuchi Ladies Clinic / Infertility Center, Gynecology, Aira-shi- Kagoshima-, Japan ;
4
ing the number of cryopreserved embryos, and opens to the donation of incom- Fujita Health University, Division of Molecular Genetics. Institute for
petent embryos to scientific research. Comprehensive Medical Science, Toyoake-shi- Aichi, Japan
Study question: We examined how differences in TE biopsy procedures affect
the incidence of chromosomal abnormalities, embryo transfer rate, and embryo
P-521 Cystic Fibrosis: Complete CFTR gene analysis vs. CFTR mosaicism based on NGS analysis.
genotyping on sperm donors carrier screening Summary answer: Of 204 embryos, 53 (25.9%) were appropriate for transfer,
M. Molina Romero1, A. Yoldi Chaure1, P. Navas Bastida2, M. Gañán while 37 (18.1%) were completely normal. Embryo mosaicism was less frequent
Parra3, Á. Vaquero Argüelles1, J.L. Del Pico Sánchez1, J.P. Ramírez when more cells were biopsied.
López1, J.A. Castilla Alcalá1 What is known already: According to recent studies, results for NGS-based
1
CEIFER Biobanco, Genetic Department, Granada, Spain ; preimplantation genetic testing (PGT) and incidences of embryo mosaicism differ
2
CEIFER Biobanco, Genetic Department, Córdoba, Spain ; depending on facilities and procedures. Currently, we are performing embryo
3
CEIFER Biobanco, Genetic Department, Sevilla, Spain biopsies with a laser procedure, but this procedure has yielded a certain

percentage of mosaic embryos. Therefore, we considered it necessary to re-ex- Main results and the role of chance: No significant difference was observed
amine the effects of different TE biopsy procedures. in ploidy distribution between LA and non-LA groups in overall patients (p=0.10).
Study design, size, duration: The subjects comprised 18 patients (204 Furthermore, no significant difference was found in mosaic and aneuploid rates
embryos) who underwent NGS-based PGT for structural rearrangements between the two biopsy groups in different age spans. Divided by embryo mor-
(PGT-SR) from 2016 to 2019. In addition, 24 surplus embryos were subjected phology, there were no significant differences in ploidy distribution between the
to rebiopsy and diagnosed with NGS. The average age of patients at oocyte two biopsy groups within the same morphologic grading: [good] mosaic rate
retrieval was 35.1 ± 2.5 years. (p=0.92) and aneuploid rate (p=0.94); [median] mosaic rate (p=0.57) and aneu-
Participants/materials, setting, methods: Analysis results were examined ploid rate (p=0.66); [fair] mosaic rate (p=0.68) and aneuploid rate (p=0.48). In
retrospectively. In addition, we performed three patterns of TE biopsy: (A) the analysis of reproductive outcomes, biochemical pregnancy rate (BPR), clinical
excessive laser irradiation of cells, (B) no washing of the cells obtained by biopsy pregnancy rate (CPR), implantation rate (IR), and miscarriage rate (MR) between
during tubing, and (C) the flicking method. These results were compared with the two biopsy groups were also comparable: [BPR] 65.9% vs. 71.5% (p=0.37);
those obtained previously. [CPR] 56.0% vs. 59.2% (p=0.63); [IR] 53.8% vs. 56.2% (p=0.73); [MR] 2.2% vs.
Main results and the role of chance: The rate of obtaining a normal embryo 3.8% (p=0.5).
was 18.1% (37/204), while that of obtaining embryos appropriate for transfer Limitations, reasons for caution: The main limitation of this study is the
in accordance with ESHRE regulations was 25.9% (53/204). The incidence of a retrospective nature. The challenges of PGT-A may occur during both the manip-
mosaic pattern was 26.4% (54/204). Besides structural abnormalities, many ulation and ploidy testing, including biopsied cell number or amplification artifacts.
abnormalities were found in chromosomes 4, 15, 16, 21 and 22. Wider implications of the findings: The study showed that utilization of
Analysis of the rebiopsied embryos showed that a mosaic pattern occurred laser for TE biopsy does not affect the embryo ploidy and reproductive out-
in two samples with small numbers of collected cells in A, with no other effects comes, and routinely using laser-assisted biopsy would not increase the risk of
observed. In B, a mosaic pattern occurred in two samples, with an overall embryo damage.
increase of noise observed. In C, fewer mosaic patterns were observed in all Trial registration number: not applicable
embryos. However, the average number of cells collected with the flicking
method and the laser procedure was 8.5 and 5.1, respectively, suggesting that P-524 A randomized controlled trial comparing the two
the number of cells available for TE biopsy affected the occurrence of a mosaic trophectoderm biopsy protocols for preimplantation genetic
pattern. testing for aneuploidies (PGT-A): a prospective sibling oocyte
Limitations, reasons for caution: The sites of mosaic patterns may vary study
based on which site of the embryo is biopsied. M. Serdarogullari1, O. Coban1, F.K. Boynukalin2, Z. Yarkiner3,
Little data is available because only PGT-SR has been approved in Japan. H. Bankeroglu4, C. Simon5, A. Capalbo6
Wider implications of the findings: The flicking method seemed to produce 1
British Cyprus IVF Hospital, Embryology, Nicosia, Cyprus ;
fewer mosaic patterns. More samples remained unaffected by excessive laser 2
Bahceci Health Group, Obstetrics and Gynaecology, İstanbul, Turkey ;
irradiation, suggesting cell numbers available for TE biopsy affected the occur- 3
Cyprus Science University, Software engineering, Kyrenia, Cyprus ;
rence of mosaic patterns. In our hospital, we perform TE biopsy under conditions 4
British Cyprus IVF Hospital, Obstetrics and Gynaecology, Nicosia, Cyprus ;
where we can obtain more TE cells by extending culture periods. 5
Igenomix Foundation, Medical, Valencia, Spain ;
Igenomix Italia, Genetics, Marostica, Italy
P-523 Laser-assisted trophectoderm biopsy does not affect the Study question: Is there any difference between the two most frequently used
embryo ploidy and reproductive outcomes in following single biopsytechniques in PGT-A cycles?
euploid embryo transfer Summary answer: Embryo quality was superior in the direct trophectoderm
J.Y. Hsieh1, Y.P. Chang1, T.H. Chuang1, M.J. Lee1, H.L. Wang1, biopsy approach without cleavage stage zona opening while blastocyst biopsied
H.H. Lai1 rate was equivalent.
1
Stork Ladies Clinic, Stork fertility center, Hsinchu, Taiwan R.O.C. What is known already: Selecting euploid embryos for embryo transfer
during assisted reproductive treatment, preimplantation genetic testing for aneu-
Study question: Does laser-assisted trophectoderm biopsy affect the embryo ploidy has now been increasingly utilized worldwide. Nowadays with improving
ploidy in preimplantation genetic testing for aneuploidy (PGT-A) cycles and technology, trophectoderm biopsy (TB) application at blastocyst-stage and sub-
reproductive outcomes in following single euploid embryo transfer? sequent comprehensive genetic screening by next generation sequencing is well
Summary answer: Laser-assisted trophectoderm biopsy does not affect the applied and adopted in PGT-A cycles in clinical practice. Based on zona pellucida
embryo ploidy and following reproductive outcomes. handling where differ by the time of zona opening before biopsy, there are two
What is known already: The laser plays an important role in assisted repro- approaches for TB. However, there is not enough comparative evidence in the
ductive technology, such as assisted hatching and embryo biopsy. High energy literature indicating the superiority of one approach to another.
light is emitted by laser and the derived heat dissolves cells, thus making cell Study design, size, duration: This prospective sibling oocyte study was per-
separation easier and convenient. Previous studies indicated that degradation formed in British Cyprus IVF Hospital between May and December 2019. The
of DNA occurs at high temperature, which makes the double strand break and inclusion criteria were: ≥ 6 fertilised zygotes using fresh donor or own oocytes,
disintegrate. Laser-assisted trophectoderm (TE) biopsy is still an invasive inter- PGT-A cycles using NGS by TB. Primary outcome measure is the rate of biopsied
vention to the embryos, whether the derived thermal effect impairs the embryo blastocysts out of the number of utilized MII. Sample size was set at 1224 biopsied
genetic materials and implantation capacity is not known already. embryos to exclude a difference of 7% between the two groups (alfa 0.05, beta 0.80).
Study design, size, duration: The data was retrospectively collected from Participants/materials, setting, methods: This interim analysis includes
January to December in 2018, 1303 blastocysts were analyzed. The methods 58 cycles and 686 embryos. For each cycle, normally fertilised oocytes were
of TE biopsy were classified into laser-assisted (LA) and non laser-assisted (non randomly assigned into two groups. Embryos in group-I had zona opening with
LA) biopsy groups, and the ploidy states of embryos between the two groups laser on day 3 to make trophectoderm cells herniate through the hole before
were compared. Only patients with following single euploid embryo transfers the biopsy while group-II contains embryos without zona opening. Main second-
were included in the analysis of reproductive outcomes. ary outcome measures were blastocyst morphology at the time of biopsy, num-
Participants/materials, setting, methods: The output of laser was 400 ber of biopsied cells, time of biopsy procedure and genetic analysis outcomes.
mW within 0.2-0.24 msec. The biopsies were tested via Veriseq PGT-A (Illumina) Main results and the role of chance: In this study, 58 patients are taken
and analyzed by BlueFuse Multi. The comparison of embryo ploidies between into analysis consisting total of 686 embryos (355 embryos in Group I, 331
LA and non-LA groups was by age (<35, 35-37, 38-40, >40 years), and by embryos in group II). Between the study groups, no statically significant difference
embryo morphology: good (AA, AB, BA), median (BB), and fair (BC, AC). The in the number of 2PN (p= 0.852), blastocysts developmental rate (p=0.724),
definitions of euploid, mosaic, and aneuploid were based on aneuploid percent- total number biopsied embryos (3.41±2.30 vs 2.91±2.19, p= 0.233), average
age: [euploid] <20%, [mosaic] 20-79%, and [aneuploid] >79%. biopsy time per embryo (p= 0.223) and the biopsied cell count (p= 0.151) were

observed. Regarding to the biopsied embryo quality on day 5 and 6, the pro- If we consider the embryo MTHFR genotype, the aneuploidy rate was 22.3% in
portions of embryos in three categories (best, good, poor) were found to be 677CC blastocysts vs. 15.7% in blastocysts with at least one mutated allele
statistically significant different among study groups (p=0.044). In detail, the (p=0.464), and 17.9% in embryos 1298AA vs. 18.3% in embryos 1298AC/CC
higher proportions of best quality embryos in group II (p=0.015) and the good (p=0.988). On the other hand, the incidence of embryo mosaicism was 13.5%
quality embryos in group I (p=0.022) were observed while no statistically sig- in 677CC group vs. 5.4% in 677CT/TT group (p=0.019) and 7.5% in 1298AA
nificant differences were found regarding poor quality embryos (p=0.565). vs. 9.1% in 1298AC/CC group (p=0.817). The only significant difference was
Furthermore, between the groups the percentage of euploidy rate in analysed observed in mosaicism rate among 677C>T genotype.
embryos did not show statically significant difference (51.6 vs 53.4 %, p= 0.758). The ongoing pregnancy rate was compared between different embryo genotypes
In detail, there was not significant discrepancy in no results rate (1.3 vs 1.7 %, and no differences were found (41.9% in 677CC vs. 39.5 in 677CT/TT, p= 0.611
p=0.750), type of aneuploidies as mosaicism (6.5 vs 9.3 %, p= 0.557) and com- and 37.4% in 1298AA vs. 43.6% in 1298AC/CC, p= 0.276). Morevoer, no dif-
plex aneuploidies (24.7 vs 20.4 %, p= 0.564) among groups ferences were observed among different maternal genotypes.
Limitations, reasons for caution: This is an interim and our results should Limitations, reasons for caution: The limited sample analysed precludes
be taken as preliminary; therefore, the clinical significance of the finding needs definite conclusions. Larger sample size studies are warranted to confirm our
to be validated in the planned sample size. However, the study is already suffi- findings. Future studies focused on genes involved in embryo aneuploidy, implan-
ciently powered to exclude major differences in the primary outcome measure. tation and pregnancy development are needed.
Wider implications of the findings: This is the first study to evaluate dif- Wider implications of the findings: Our analysis suggests that neither
ferent biopsy approaches by using sibling oocyte design that could assist embry- maternal nor embryo MTHFR genotype influences the aneuploidy or ongoing
ologist while seeking more convenient approach during embryo biopsy for PGS pregnancy rate. The only correlation was observed in mosaicism rate among
if adequate evidence reached. 677C>T embryo genotypes. Further research is need to clarify the role of this
Trial registration number: non applicable genotype on the processes that lead to mosaicism in the embryo.
P-525 Do maternal and embryo MTHFR gene polymorphisms
have any influence on embryo chromosomal abnormalities and P-526 Consistent results of non-invasive pre-implantation genetic
the ongoing pregnancy rate? testing for aneuploidy (niPGT-A) of human embryos using two
G. Eva M.1, R. Morales1, H. Codina1, A. Cascales1, B. Lledo1, J.A. different techniques for chromosomal analysis
Ortiz1, J. Ten2, J. Llacer3, A. Bernabeu3, R. Bernabeu3 B. Lledo1, R. Morales1, J.A. Ortiz1, A. Rodriguez-Arnedo2, J. Ten2,
1
Instituto Bernabeu, Molecular Biology and Genetics, Alicante, Spain ; J.C. Castillo3, A. Bernabeu3, J. Llacer3, R. Bernabeu3
2 1
Instituto Bernabeu, Embriology laboratory, Alicante, Spain ; Instituto Bernabeu, Molecular Biology, Alicante, Spain ;
3 2
Instituto Bernabeu, Reproductive Medicine, Alicante, Spain Instituto Bernabeu, Embriology, Alicante, Spain ;
3
Instituto Bernabeu, Reproductive Medicine, Alicante, Spain
Study question: Is there any relationship between maternal and embryo
MTHFR polymorphisms (c.677C>T and c.1298A>C) and embryo chromosomal Study question: Are the results of niPGT-A similar when analysed with dif-
abnormalities or the ongoing pregnancy rate of euploid embryos? ferent genetic techniques?
Summary answer: Maternal and embryo MTHFR polymorphisms do not Summary answer: Consistent results (diagnosis and concordance rate) are
increase the aneuploidy rate, as well as they are not correlated with the ongoing obtained with different genetic techniques used for niPGT-A, thus suggesting
pregnancy rate. discordances might be attributable to mosaicism or DNA-contamination.
What is known already: MTHFR enzyme is involved in folic acid metabolism What is known already: The concept of niPGT-A has generated huge interest
and play a role in reproduction. Many investigations have linked point mutations in reproductive medicine. Variable success and concordance rates have been
in the MTHFR gene with embryo alterations and problems in pregnancy. The reported and discordances seem related to embryonic mosaicism, favoured
single-nucleotide-polymorphisms 677C>T and 1298A>C are the most relevant. elimination of aneuploid cells, DNA contamination and the method used for
Maternal genotype has been associated with embryo aneuploidy rate in previous analysis (amplification/detection). The niPGT-A efficacy has been restricted by
studies. Moreover, a negative correlation between the embryo genotype and technical complications associated with the low quantity and quality of the DNA,
the implantation rate has been described. However, it is still a controversial issue. presenting technical challenges for genetic analysis. It is not clear which method
The aim of our study was to investigate the effect on embryo aneuploidies and is the most appropriate. The aim of this study was to assess the accuracy of
mosaicism and the correlation with pregnancy rate. niPGT-A by comparing two different chromosomal analysis techniques.
Study design, size, duration: MTHFR genotype (677C>T and 1298A>C) was Study design, size, duration: A prospective blinded validation study was
analysed in 77 women who performed an IVF/ICSI cycle with their own oocytes performed from September 2018 to December 2019, including 302 chromo-
and also carried out a preimplantation genetic testing for aneuploidy (PGT-A) from somal analyses. Two hundred and seventy-six corresponded to trophectoderm
January 2016 until December 2018. Moreover, MTHFR genotype was analysed in biopsies (TE) from couples who attended our clinic for PGT-A and whose cor-
the 189 embryos biopsied in the PGT-A cycles from these patients. In addition, respondent spent blastocyst medium (SBM) was evaluated by two methods.
218 DNA samples from trophoectoderm biopsies belonging to a different group Eight blank-medium were evaluated for DNA contamination detection. Finally,
of patients were also genotyped for MTHFR polymorphisms. to investigate the diagnostic discrepancies: TE-aneuploid embryos were thawed
Participants/materials, setting, methods: MTHFR genotyping was deter- and re-analyzed using TE and inner cell mass (ICM) biopsies.
minated with TRF-plus Thrombosis Risk Panel (Elucigene Diagnostics). Embryo Participants/materials, setting, methods: Embryos were cultured in con-
chromosomal analysis was performed using Agilent SurePrint G3-8x60K-CGH tinuous media (Global Total LP) until D3, washed three times and cultured again
microarrays or Veriseq-NGS (Illumina), with previous whole genome amplifica- until embryo biopsy. Embryos were biopsied on D5 or D6. TE analysis was
tion. The main parameters analysed among MTHFR genotypes (both in embryos carried out using Veriseq (Illumina®). Frozen euploid embryos according to the
and in patients were the embryo aneuploidy and mosaicism rate. Additionally, TE biopsy were thawed and transferred in the subsequent cycle. SBM collection
the ongoing pregnancy rate of euploid blastocysts (n=243) was assessed. The was done following TE biopsy and samples were stored at -80ºC. SBM chromo-
differences between groups were evaluated by chi-square and binary logistic somal analysis was performed using Veriseq (Illumina®) and NICS (Yikon®).
regression and (SPSSv20.0). Main results and the role of chance: We obtained genetic results in 96.8%
Main results and the role of chance: The aneuploidy rates obtained were of TE-samples vs 90.4% in SBM using both techniques(p>0.05). The mosaicism
comparable in embryos coming from patients with homozygous normal genotype rate was higher in SBM (30.4% SBM-Yikon;28.3% SBM-Veriseq) compared to
and those embryos from women with at least one mutated allele (54.7% in TE-biopsies (14.1%) (p= p=0.013; p=0.031) regardless of the technique used.
677CC vs. 30.2% in 677CT/TT, p=0.058 and 37.8% in 1298AA vs. 42.7% in We performed comparisons between both SBM techniques showing 95.2% of
1298AC/CC, p=0.212). Furthermore, no differences were observed in mosa- consistency in the diagnosis. Regarding diagnostic concordance (euploid-euploid
icism rate (24.0% in 677CC vs. 13.8% in 677CT/TT, p=0.345 and 17.1% in vs aneuploid-aneuploid) between each SBM-technique and TE-biopsy, we obtained
1298AA vs. 17.3% in 1298AC/CC, p=0.865). 74.6% SBM-Yikon vs 72.3% SBM-Veriseq. However, when we considered embryos

biopsied on D6 these rates reached up the 92.0% and 86.5%, respectively. after morphine withdrawal. ChIP-sequencing analysis confirms a decrease of
Analysing the full chromosome concordance, the cytogenetic results were exactly H3K27me3 enrichment on the whole coverage of the X chromosome. The
the same as the TE-biopsy in 45.2% SBM-Yikon and 41.7% SBM-Veriseq. Moreover, Smchd1 gene expression was also down-regulated in in-vitro-morphine-treated
in 20.1% SBM-Yikon and 23.3% SBM-Veriseq the results were discordant only in blastocysts, preventing the XCI in early embryo. This is consistent with the
mosaicism diagnosis. The remaining were partial (22.6% SBM-Yikon vs 23.3% SBM- increase in blastocyst rate observed after morphine chronic treatment. Our
Veriseq) and complementary (4.8% SBM-Yikon vs 3.3% SBM-Veriseq) discor- results clearly prove that chronic morphine treatment prevents the XCI, modi-
dances. To identify the cause of discrepancies we reanalysed TE-aneuploid fying PRC2 and Dnmt1 gene expression in the short term and maintaining the
embryos, 55.6% of the discrepancies were due to DNA contamination (maternal repression upon Smchd1 in the long term. SMCHD1 plays a role as a key regu-
origin), 22.2% to embryo mosaicism, 11.1% to low resolution in SBM-Yikon and lator of epigenetic cellular memory induced by morphine, conferring to the cell
11.1% low resolution in both techniques. Finally, we analysed the IVF outcome, a stable activate state
the ongoing pregnancy rate was 50% for TE-euploid-SBM-Yikon-aneuploid vs Limitations, reasons for caution: To perform the in-vitro analysis.
33.3% TE-euploid-SBM-Veriseq-aneuploid. Wider implications of the findings: Morphine alters normal X chromo-
Limitations, reasons for caution: The main limitation of the study is the some-wide silencing, providing insights into the epigenetic mechanisms underlying
sample size, especially for the subgroup analysis. Larger, prospective studies are cell memory induced by morphine and establishing the bases to understand how
warranted to draw definite conclusions on the accuracy of niPGT-A in repre- environmental factors can cause epigenetic changes, which leads to health prob-
senting the genetic constitution of the whole embryo with potential to improve lems or diseases.
pregnancy outcomes. Trial registration number: DTS18/00142
Wider implications of the findings: Diagnostic concordance between
PGT-A and niPGT-A seems independent of the technique used for genetic anal- P-528 Cigarette smoke alters the expression of genes involved in
ysis. Interestingly, concordance was higher for day-six-biopsied embryos. the inflammatory response in mouse uterus
Therefore, niPGTA may be influenced by factors such as DNA contamination M.C. Budani1, E. Carletti2, G.M. Tiboni2
and embryo mosaicism. Optimization of culture conditions and medium retrieval 1
University “G. d’Annunzio” Chieti-Pescara- Italy, Department of Medicine and
constitutes potential targets to improve NIPGT-A reliability. Aging Sciences, Chieti, Italy ;
University ‘G. d’Annunzio’ Chieti-Pescara- Italy, Department of Medical- Oral and
Biotechnological Sciences, Chieti, Italy
P-527 Morphine treatment prevents the X chromosome
inactivation and maintains a long-term repression by SMCHD1 Study question: What are the effects of a whole body exposure to cigarette
epigenetic regulator. smoke on stress and toxicity pathways in the mouse uterus?
N. Subiran1, I. Muñoa-Hoyos1, M. Araola-Lasa1, I. Urizar-Arenaza1, Summary answer: The in vivo cigarette smoke exposure induced an over-ex-
M. Gianzo1, J. Irazusta1 pression of pro-inflammatory cytokines and a down-regulation of extracellular
1
University of Basque Country, Physiology, Bilbao, Spain matrix metallopeptidases in mouse uterus.
What is known already: Cigarette smoking was associated to a lower chance
Study question: To elucidate how morphine can cause stable epigenetic of clinical pregnancy and live birth rates, and a dramatic increase in the incidence
changes on X-chromosome inactivation (XCI) process that can be memorized of spontaneous miscarriages in women. The noxious effects of cigarette smoke
and persist in the long term. exposure have been studied especially in the ovaries. Research linking smoking
Summary answer: Chronic morphine treatment prevents the XCI through to uterus damage mainly relies on studies in vitro testing a single cigarette
PRC2 and Dnmt1 in the short term and Smchd1 maintains this long-term repres- smoke-derived component. Smoke components seemed to exert adverse effects
sion in mESC and blastocysts. on endometrial angiogenesis in human primary endometrial cells and in mouse
What is known already: XCI is an important epigenetic process that exem- endometrial cells as well and on uterine cells proliferation with a role in decreas-
plifies the developmentally controlled formation of silent chromatin, where one ing the endometrial thickness.
of the X chromosomes in females is inactivated for dosage compensation. The Study design, size, duration: Female C57BL/6 mice (aged 8 weeks) were in
XCI involves several steps, which implies the initiation of chromosome-wide vivo exposed to the smoke originated by 3 cigarettes once a day, 7 days/week for
silencing by XIST, the formation of a repressive compartment through Polycomb 2 months using a specific whole-body mainstream exposure system. Control mice
complexes and finally the maintenance of the stable repression by DNA meth- were placed in a restrainer and exposed to room air only for the same time period.
ylation and other epigenetic regulators in which SMCHD1 plays an important Participants/materials, setting, methods: At the end of exposure, mice
role. Although XCI is part of normal development, there is a big concern in were euthanized and the uteri collected. From these, the extracted RNA was
understanding how environmental factors can cause stable epigenetic changes used to perform a real-time PCR using an array consisting of a panel of 84 key
that lead to health problems. genes involved in stress and toxicity pathways. Representative genes with >2
Study design, size, duration: To evaluate if morphine can induce cellular fold differential expression identified in the array were validated by real-time
epigenetic memory, OCT4-reported mESCs were chronically treated with mor- PCR gene expression assays.
phine during 24h, 10-5µM (P1). After morphine removal, mESCs were seeded Main results and the role of chance: No adverse changes in body conditions
and maintained in culture for 48h (P2) and 96h (P3). To elucidate the role of and atypical behaviors were noted as result of cigarette smoke exposure. The
morphine in early embryo development, two cell- embryos stage were chron- genes that were significantly up-regulated in the gene array were as follows: reactive
ically treated with morphine for 24h and in-vitro cultured up to the blastocyst protein C (Crp, p-value < 0.05), growth arrest and DNA-damage-inducible45γ
stage in the absence of morphine. (Gadd45γ, p-value < 0.05), interferon γ (Ifnγ, p-value < 0.01), and interleukin1α
Participants/materials, setting, methods: Transcriptomic analyses and (Il1α, p-value < 0.005). Cigarette smoke was associated in the gene array with a
H3K27me3 genome wide distribution were carried out by RNA-Sequencing and down regulation of matrix metallopeptidase-9 (Mmp9, p-value < 0.05) compared
Chip-Sequencing respectively. Validations were performed by RNA-RT-qPCR to control group. The qRT-PCR analysis validated the differential expression profile
and Chip-RT-qPCR. of the representative genes deregulated at least two folds in the array.
Main results and the role of chance: Dynamic transcriptional analyses Limitations, reasons for caution: The experimental nature of the study
identified a total of 932 differentially expressed genes (DEGs) at P1 and, in the limits the capacity to extrapolate to the human field.
absence of morphine, 1196 DEGs at P2 and 2138 DEGs at P3, providing strong Wider implications of the findings: This is the first study showing the
evidence of the existence of transcriptional cellular memory induced by mor- capacity of in vivo cigarette smoke exposure in inducing an over-expression of
phine. Chronic morphine led to a down-regulation of epigenetic regulators inflammatory genes and concomitant down-expression of MMP9 in mouse
important for XCI, such as PCR2 complex and Dnmt1 in the short term. High- uterus. These data support the idea that inflammation cigarette smoke-derived
throughput screening approaches showed up Smchd1 as a key regulator of a may interfere with the tissue remodeling process responsible for the embryo
long-term epigenetic cellular memory induced by morphine. Morphine caused implantation.
a down-regulation of Smchd1 gene expression that can persist over the time Trial registration number: not applicable

P-529 To flick or to pull? The effect of trophectoderm biopsy What is known already: According to the embryonic culture protocol, the
methodology on mosaicism incidence culture medium is usually renewed on day 3 of development, although currently
L. Coll1, M. Parriego1, S. Mateo1, B. Carrasco1, S. García1, the use of a single step medium makes it unnecessary. In the past, different
M. Boada1, B. Coroleu1, A. Veiga1 techniques were used to perform AH. Nowadays in most laboratories, the use
1 of laser has been unified. There have been studies on the technique and its
Dexeus Mujer, Obstetrics- Gynecology- and Reproductive Medicine, Barcelona,
usefulness, but due to its heterogeneity, there have been no conclusive results.
Spain
In this way there are no specific data related to the heating of the dishes and the
Study question: Is mosaicism incidence related to differences in biopsy culture medium and their effect on the results.
methodology? Study design, size, duration: Three months retrospective study (October
Summary answer: Mosaicism incidence is not related neither to the number 2017- December 2017) with 190 patients of the PGT-A program. They were
of laser pulses during biopsy nor to the use of flicking/pulling methodology. divided in two post-hatching categories on day 3: AHR (assisted hatching
What is known already: Chromosomal mosaicism may be an inherent feature renewal) and AHNR (assisted hatching no renewal).
in preimplantation human embryos. However, reported incidences vary widely Participants/materials, setting, methods: The AHNR group included 94
among different IVF and PGT-A settings. Although such differences could be patients and AHR group included 96 patients. The AH was performed by laser
attributed to the use of different diagnostic methodologies and criteria, settings Octax Ò on day 3 directly on the EmbryoscopeÒ dish and the biopsy was done
with the same diagnostic platforms have observed different mosaicism incidence. at blastocyst stage. Chromosomal analysis method was NGS (Thermo Scientific,
This points to the fact that extrinsic factors can also have an impact. Moreover, INC., USA). For embryonic categorization, the ASEBIR classification was used
it has been speculated that the biopsy procedure itself and biopsied sample being A cathegory the best prognosis blastocyst and D the worst. The statistical
handling could induce the generation of artefactual mosaicism leading to an analysis was performed by Fisher’s exact test.
overestimation of such phenomenon. Main results and the role of chance: A total of 1505 embryos were
Study design, size, duration: This is a prospective observational study analysing obtained, of which 238 were biopsied in the AHNR group and 242 in the AHR
the chromosomal constitution of 484 trophectoderm biopsies from PGT-A cycles group. For the AHNR group, 105 blastocysts were euploid (44.12%) and 133
performed from May 2019 to January 2020 in a single IVF setting. Mosaicism aneuploid (55.88%). Regarding the 242 of the AHR group, 110 were euploid
incidence was evaluated in relation to different biopsy methodologies. (45.45%) and 132 aneuploid (54.55%). With reference to the the morphology
Participants/materials, setting, methods: Trophectoderm biopsies were by categories, the data for AHNR were A: 5.35%; B: 48.18%; C: 27.26%; D:
performed by flicking or pulling depending on embryo characteristics and ana- 19.21% and for AHR A: 5.97%; B: 49.61%; C: 26.25% and D: 18.17%. No sta-
lyzed by NGS. Data on the quality of the biopsied sample (with or without lysed tistical differences were found between both groups (p>0.05).
cells) was analysed with regards to biopsy methodology. Additionally, the number Thus, it seems that renewing the culture medium on day three of development
of laser pulses and the biopsy methodology (flicking or pulling) were analyzed using single step medium does not provide an improvement with respect to the
for mosaicism incidence. Time from freezing to amplification was also assessed morphology of the blastocysts, as well as regarding the percentage of euploidy.
in relation to mosaicism. Therefore, we can conclude that the single step medium is a reliable option for
Main results and the role of chance: In 73.8% of cases (357/484), ≤3 laser the PGT-A program since the AH does not modify the culture conditions.
pulses were needed to obtain a biopsy. Moreover, in 34.3% of biopsies Limitations, reasons for caution: Retrospective study and the limited num-
(166/484), the trophectoderm sample was obtained by pulling methodology vs ber of cases. Further studies are necessary to corroborate these results. In
65.7% (318/484) in which the methodology was flicking. The percentage of addition, study conditions may vary from other laboratories, for example: culture
samples showing signs of lysis was higher when flicking methodology was used conditions, the power of the laser and the medium used for embryo culture.
compared to pulling (20.8% vs 10.8%; p-value = 0.01). Therefore, results may not be comparable between laboratories.
The incidence of mosaicism did not vary among different biopsy operators Wider implications of the findings: The introduction in the laboratories
(p=0.1). In relation to biopsy methodology, similar mosaicism incidence was of the time-lapse technology with new and modern dishes as well as the use of
observed for pulling and flicking methods (13.9% vs 16.0%; p-value =0.5). The one step media, has meant a change in the working protocols. The combination
number of laser pulses applied was not related to mosaicism either (3.30±0.75 of both factors with AH requires more studies that show the possible influence
in mosaic embryos vs 3.33±0.66 in non-mosaic embryos; p-value = 0.67). on clinical outcomes.
Concerning biopsied sample storage, no differences were observed regarding Trial registration number: 1503-VLC-017-AM
the elapsed time from biopsy to amplification (10.4d ±10.4 in mosaic embryos
vs 11.0 d ±13.8; p-value=0.7). P-531 Differential endometrium DNA methylation and gene
Limitations, reasons for caution: These are preliminary results. For a deeper expression patterns underline mechanisms of reproductive failure
analysis controlling for potential confounding factors more data are needed. in women with unexplained recurrent miscarriage (RM) and
Embryologists were homogeneously trained and followed the same SOPs. As a recurrent implantation failure (RIF)
consequence, there was small variability in biopsy methodology. R. Zhang1, S.W. Hung1, X. Chen2, T.C. Li2, C.C. Wang1
Wider implications of the findings: Each PGT setting can choose its own 1
Chinese University of Hong Kong, Obstetrics and Gynecology, Hong Kong,
procedures as long as trophectoderm biopsy is performed within homogeneous Hong Kong ;
methodology and high quality standards. However, it remains to be assessed whether 2
Prince of Wales Hospital- The Chinese University of Hong Kong, ART unit-
different biopsy methodologies have an effect on embryo reproductive outcome. Obstetrics and Gynecology, Hong Kong, Hong Kong
Study question: What is the underlying mechanism cause the differential gene
P-530 Euploidy and embryo quality with single step medium expression in endometrium in women with unexplained RM and RIF, and causes
with or without renewal post assisted hatching (AH) in a PGT-A their endometrial receptivity disorder?
program using Embryoscope dishes. Summary answer: Differential endometrium DNA methylation-regulated
A. Delgado Mendive1, A. Mercader1, L. Escrich1, D. Beltran1, genes for reproductive failure were identified. RIF and RM were associated with
A. Tejera1, N. Grau1, F. Insua1, M. Nohales1, A. Mifsud1 abnormalities in endometrium cell adhesion and metabolic pathway, respectively.
1
Instituto Universitario IVI Valencia, ivf and pgd laboratory, Valencia, Spain What is known already: Unexplained RM and RIF are two major challenging
reproductive failure attribute to endometrium. Those women have normal
Study question: Is the culture medium affected by the heating of the uterus, parental karyotype and basal hormone level; negative for autoimmune
EmbryoscopeÒ dish after the AH and how could it influence embryo develop- antibodies and thromobophilia, uNK and plasma cells, and even good embryo
ment and euploidy rates? quality. Abnormal endometrium in molecular levels may contribute to the failure.
Summary answer: The results regarding euploidy and embryo quality between Our previous study of endometrium RNA sequencing (RNAseq) identified dif-
both groups were similar and AH did not seem to modify the conditions of the ferential transcriptome profiles between RM and RIF. Epigenetics can modify
culture medium. gene expression without change in gene structure. Evidences have shown that

abnormal DNA methylation modification will result in abnormal endometrial of MGC from women with DOR (Fan et al. 2019) and we have previously shown
genes expression and thereby reproductive failure. thatMGC have a distinctive epigenetic ageing profile compared with other
Study design, size, duration: This is a prospective study of DNA methylation somatic cell types (Olsen et al. under review). But whether the MGC from
and gene expression of endometrium in genomic scale. Fertile control women women with DOR differ from MGC from other women is not known.
and unexplained RM and RIF undergoing IVF treatment were recruited. 5 endo- Study design, size, duration: Multicenter cohort study based on a retro-
metrium samples in each group were included for gene discovery study. Another spective analysis of prospectively collected data (September 2016 – June 2018)
10 endometrial samples in each group were included for validation and functional and material (blood and MGC) derived from 119 healthy women undergoing
studies. All the samples were collected and analysed in 2019. IVF or ICSI treatment following controlled ovarian stimulation with a standard
Participants/materials, setting, methods: Endometrial samples were gonadotropin-releasing hormone antagonist protocol. The public fertility clinics
obtained from control fertile women, and women with RM and RIF. All samples at the Copenhagen University Hospitals in Herlev and Hvidovre, Stork IVF clinic
were collected on LH+7 in natural cycle and stored in -80oC before assays. and the fertility clinic at Skåne University Hospital included patients in the study.
Whole genome bisulfite sequencing (WGBS) was used to assess DNA methyl- Participants/materials, setting, methods: MGC were obtained from
ation and RNA sequencing (RNAseq) was used to assess transcriptome pattern women with varying ovarian reserve status (defined from age dependent
in the same endometrium samples. Differential methylated genes (DMG) were anti-Müllerian hormone (AMH) levels) by isolation from pooled follicles imme-
selected and target methylation specific-PCR (MS-PCR), RT-qPCR and immu- diately after oocyte retrieval. DNA from the MGC aggregates was extracted
nostaining were used to validate the results. and analysed with the Illumina EPIC array, at the Human Genotyping Facility
Main results and the role of chance: RNAseq identified 969 differentially Genetic Laboratory, Dept. of Internal Medicine Erasmus MC Rotterdam.
expressed genes (DEGs) between RIF and controls, 370 DEGs between RM and Subsequently, the data went through a bioinformatic analysis. All analysis included
controls, and 872 DEGs between RM and RIF. WGBS detected higher overall adjustment for chronological age.
methylation level in RIF and controls than RM. 157989 differentially methylated Main results and the role of chance: None of the epigenetic clock tools
regions (DMRs) were found in RM versus controls, 85834DMRs in RIF versus that were applied showed an association between age acceleration and ovarian
controls, and 133116 DMRs in RIF versus RM in all genome regions. If promoter reserve in either cell type. Performing a differential DNA methylation variability
region only, we found 79 differentially methylated genes (DMGs) in RIF versus analysis by comparing DOR or high ovarian reserve samples to controls (normal
controls, and 51 DMGs in RM versus controls, and 76 DMGs in RM versus RIF. ovarian reserve), differential variability between DOR and normal samples was
For RIF, most DMGs were enriched in calcium ion binding and plasma membrane observed at 4,199 CpGs in MGC, while only 447 between high and normal (FDR
mainly into cell adhesion pathway. For RM, most DMGs were enriched in protein < 0.05). Regardless of ovarian reserve, very few signals were detected in leu-
binding, integral component of membrane and mitotic cell cycle mainly into kocytes and no overlaps with the signals in MGC were found. Several genes
metabolic pathway. Based on biological and molecular functions, 4 DMGs includ- central to folliculogenesis, i.e. inhibin subunit beta B (INHBB), AMH, insulin-like
ing AQP3, AURKB, CRISP2, CRYBB2 were chosen for validation and functional growth factor 2 (IGF2), were associated with variably methylated CpGs. Gene
studies. ontology analysis of genes associated with the variable CpGs in DOR in MGC
Limitations, reasons for caution: Although we attempted to eliminate revealed an enrichment for categories related to cell-to-cell adhesion (p<0.001).
embryo factors by embryo morphology grading before transfer, molecular com- Furthermore, we found a higher number of epimutations in the MGC from
position of the embryos was still unknown. Apart from DNA methylation, other women with DOR.
epigenetic mechanisms, such as histone acetylation and methylation; and Limitations, reasons for caution: The granulosa cells were collected during
non-coding RNA, on the endometrium abnormalities are still not known. ovarian stimulation which may influence DNA methylation. We were not able
Wider implications of the findings: Important endometrium genes regu- to correct for the FSH dose. However, we have previously shown no association
lated by DNA methylation for the reproductive failure were identified. This not between FSH dose and DNA methylation profile.
only increases the understanding one of the underlying molecular mechanism Wider implications of the findings: Clarification of a potential unstable
for the conditions, but also provides potential therapeutic targets to improve methylome and epigenetic dysfunction in MGC being associated with the poor
the pregnancy outcomes in these women. ovarian reserve found in women with DOR may be important for future treat-
Trial registration number: CREC 2016.160-T ment and prevention strategies.
P-532 Identification of a unique epigenetic profile in women with
diminished ovarian reserve P-533 Preferential allelic segregation of DNA triplet repeat
K.W. Olsen1, J. Castillo-Fernandez2, A.C.H. Chan3, N.L.C. expansion on PGT-M embryos from Myotonic Dystrophy type 1
Freiesleben4, A. Zedeler5, M. Bungum6, A. Cardona7, J. Perry7, and Huntington´s disease
S. Skouby1, R. Borup3, E.R. Hoffmann3, K. Gavin2, M.L. Grøndahl1 P. Villa1, R. Quintana2, Q. Ramiro3, M. Sandra4, J.V. Martinez-
1
Copenhagen University Hospital Herlev, Department of Obstetrics and Sanchis5, M. De la Orden5, I. Moya5, T. García-Gimeno3, A. Monzo3
1
Gynaecology- Fertility Clinic, Herlev, Denmark ; La Fe University Hospital-Joint First authors, Women’s Health Area, Valencia,
2
Babraham Institute, Epigenetics Programme, Cambridge, United Kingdom ; Spain ;
3 2
Faculty of Health and Medical Sciences- University of Copenhagen, DNRF Center La Fe University Hospital-Joint First authors, Women’s Health Area, Valencia,
for Chromosome Stability- Department of Cellular and Molecular Medicine, Spain ;
3
Copenhagen, Denmark ; La Fe University Hospital, Human Reproduction Unit- PGD-Unit, Valencia, Spain ;
4 4
VivaNeo Fertility Clinics, Stork IVF Clinic A/S, Copenhagen, Denmark ; La Fe University Hospital, Department of Genetics, Valencia, Spain ;
5 5
Copenhagen University Hospital Hvidovre, Department of Obstetrics and La Fe University Hospital, Human Reproduction Unit. IVF Lab, Valencia, Spain
Gynaecology- The Fertility Clinic, Hvidovre, Denmark ;
6
Skåne University Hospital, Reproductive Medicine Centre, Malmö, Sweden ; Study question: Are there any differences in parental origin of triplet repeat
7
University of Cambridge Addenbrooke’s Hospital Cambridge, Medical Research expansion in human embryos after PGT-M from Myotonic Dystrophy type 1 or
Council Epidemiology Unit, Cambridge, United Kingdom Huntington’s Disease carriers?
Summary answer: Triplet repeat expansion mutations in DMPK and HTT
Study question: Does the epigenetic profile of mural granulosa cells (MGC) genes have a preferential gender pattern of transmission to embryo yield.
and leukocytes from women with diminished ovarian reserve (DOR) differ from What is known already: Myotonic dystrophy 1 (MD1) and Huntington’s
other women? disease (HD) are progressive autosomal dominant neurodegenerative disorders
Summary answer: MGC from human ovarian follicles have a distinctive epi- caused by a CGG or CAG repeat expansion in the DMPK or HTT genes, respec-
genetic profile in women with DOR. The same does not apply for leukocytes. tively. Preimplantation genetic testing (PGT) is a diagnostic procedure available
What is known already: DOR is defined as a reduced number of oocytes for these individuals, because they carry a higher risk of transmitting this genetic
and can be physiological due to advanced reproductive age or pathological due condition to their offspring. It has been proposed preferential expansion depend-
to premature loss of fertility. Previous studies have found an increase in apoptosis ing on the gender parental carrier.

Study design, size, duration: Information about 82 PGT-M cycles performed average. The obtaining of 3 mature oocytes were associated with 28.6% chance
from 2011 to 2019 in 14 MD1 and 18 HD couples was collected retrospectively. of having at least one healthy embryo. This probability increased significantly
MD1: 21 PGT-M cycles in 9 female-carrier and 9 cycles in 5 male-carrier couples. with the number of mature oocytes. Ten mature oocytes led to more than 60%
HD: 39 cycles in 14 female-carrier and 13 cycles in 5 male-carrier couples. chance of obtaining a healthy embryo. This probability did not exceed 69% and
Participants/materials, setting, methods: All procedures were performed plateaued from 15 metaphase II oocytes collected. In the FP group, the mean
at the Human Reproduction and Genetic Units in a tertiary University Hospital. number of mature oocytes vitrified was 5.1 ± 3.6 per FP cycle. On average, 3
PGT provided direct testing of embryos obtained after intracytoplasmic sperm cycles of ovarian stimulation were performed per patient leading to a mean of
injection, using polymerase chain reaction PCR of microsatellite markers (indirect 15 mature oocytes vitrified.
method) and by expansion amplification (direct method) as the genetic testing Limitations, reasons for caution: This study is retrospective, analyzing a
protocol to detect the risk allele and the expanded CTG/CAG repeat in 366 limited number of cycles, without possibility to determine threshold values of
day 3 embryos, 167 from MD1 and 199 from HD. oocytes that might predict the number healthy embryo.
Main results and the role of chance: The percentage of non-affected Wider implications of the findings:
embryos related to the parent-of-origin in DM1 was 35.5% (43/121) in female The present results should be confirmed by further analyzes. Patients included
carriers and 19.6% (9/46) in male carriers. PGT-M in HD resulted in 39.6% in a fresh PGT cycle usually show higher values of ovarian reserve tests when
(67/169) in female carriers and 16.7% (5/30) in male carriers. compared with FMR1 premutated carriers seeking FP. Whether the oocyte com-
In both conditions, we observed significantly higher ratio of non-affected petence differs in these groups of patients is unknown. This information might
embryos in female carriers when compared to male carriers. (38%: 110/290 vs balance our conclusion.
18.4% (14/76), respectively. Chi square test P=0.026). Trial registration number: not applicable
Of the total sample, 38 embryos (10.4%) were misdiagnosed, mainly due to
allele drop out (ADO) or amplification failure. P-535 Antral follicle responsiveness to exogenous FSH is not
Limitations, reasons for caution: This study has a retrospective design. The altered in FMR1 mutation carriers undergoing stimulation for
number of patients is far low in some groups, although the number of embryos preimplantation genetic testing.
is adequate for statistical tests. Misdiagnosed embryos has a potential to bias M. Grynberg1, M. Pinto1, A. Naouel1, V. Gallots1, A. Mayeur2,
results. C. Sonigo1
Wider implications of the findings: In both conditions, the observed non-af- 1
Hôpital Antoine Béclère, Reproductive Medicine & Fertility Preservation, Clamart,
fected embryos rate is far below of the theorical 50% expected, according to
France ;
autosomal dominant inheritance. Our results must be taken into account in order 2
Hôpital Antoine Béclère, Reproductive biology unit, Clamart, France
to offer the appropriate genetic counseling in carrier patients before PGT-M.
Study question: Is antral follicle responsiveness to exogenous FSH, assessed
by the follicular output rate (FORT), altered in FMR1-mutation carriers?
P-534 Probability of producing at least one healthy transferable Summary answer: FMR1-mutation carriers do not show alteration in antral
cleavage-stage embryo after preimplantation genetic testing for follicle responsiveness to exogenous FSH assessed by the FORT index
fragile X syndrome What is known already: Female carrying FMR1 mutation (premutation or
C. Sonigo1, M. Sadoun1, L. Hester2, J. Steffann3, A. Benoit1, full mutation) may be candidate to preimplantation genetic testing (PGT) to
V. Gallot1, N. Frydman2, M. Grynberg1 achieve a pregnancy with a healthy baby while avoiding the burden of prenatal
1
Antoine Béclère Hospital, Reproductive Medicine and fertility preservation, genetic diagnosis and the risk of pregnancy termination. Several lines of evidence
Clamart, France ; indicate that female carrying FMR1 premutation are at higher risk of diminished
2
Antoine Béclère Hospital, reproductive biology unit, Clamart, France ; ovarian reserve and further premature ovarian insufficiency. However, whether
3
Necker Enfants Malades Hospital, Genetic Unit, Paris, France this genetic profile alter the capacity of small antral follicle to respond to exog-
enous FSH during controlled ovarian stimulation remains unelucidated.
Study question: What are the chances of obtaining a healthy transferable Study design, size, duration: Monocentric retrospective observational study.
cleavage-stage embryo according to the number of mature oocytes in FMR1- Thirty-three patients carrying either FMR1 premutation (n=17) or full mutation
mutated carriers undergoing preimplantation genetic testing? (n=16) were referred to our PGT centre from 2006 to 2018.
Summary answer: Fifteen mature oocytes provide 69% chance of obtaining Participants/materials, setting, methods: Eighty-five PGT cycles for fragile
at least one healthy embryo after preimplantation genetic testing (PGT) in FMR1- X syndrome, 43 in premutated patients and 42 in fully mutated women, were
mutated carriers. analyzed. COS outcomes were compared to 85 PGT cycles performed for male
What is known already: PGT may be an option to achieve a pregnancy with translocation, matched on female age, BMI, antral follicle count, and protocol
a healthy baby for FMR1 mutation carriers. In addition, FMR1 premutation is of stimulation. FORT was determined by the ratio between the pre-ovulatory
associated with a higher risk of diminished ovarian reserve and premature ovarian follicle count (PFC, 16-22 mm) on the day of oocyte triggering × 100/ antral
insufficiency. As a consequence, oocytes vitrification may be offer to these young follicle count measured just before initiation of the stimulation.
women as a method of fertility preservation. The cryopreserved oocytes can Main results and the role of chance: By design, patients were comparable
then be used to achieve pregnancy, with or without PGT. The number of vitrified in terms of age, BMI and markers of the follicular ovarian status. Overall, after
oocytes to allow the transfer of a healthy embryo following PGT has never been similar starting dose (253±91 vs. 247±82, p=0.7 and total dose of gonadotropin
investigated. administered (2623±1071 vs. 2620±1029 IU, p=0.98), the numbers of oocytes
Study design, size, duration: This is a monocentric retrospective observa- recovered and mature oocytes (12.1±6.1 vs. 11.8±4.9, p=0.70; and 9.4±5.2 vs
tional study analyzing 33 premutated (n=17) or fully mutated (n=16) candidates 9.9±4.1; p=0.70, respectively) were not significantly different in FMR1-mutation
for PGT for fragile X syndrome between 2006 and 2018. carriers and controls. In addition, FORT index remained similar between both
In the meantime, 15 FMR1 premutated carriers underwent oocyte vitrification groups (42.6±23.9% vs 42.4±27.2%, p=0.96, respectively).
for fertility preservation (FP). Comparison of FMR1 premutation and full mutation carriers showed lower
Participants/materials, setting, methods: Eighty-five PGT cycles were values of ovarian reserve tests in permutated patients (AMH 3.5±3.2 vs. 6.2±3.6
performed for fragile X syndrome, 43 in premutated patients and 42 in fully ng/mL, p=0.0004 and AFC 18.7±9.4 vs. 22.9±8.9 follicles, p=0.03). Therefore,
mutated women. For each cycle, we estimated the number of mature oocytes significantly higher doses of total FSH were needed (2988±1184 vs. 2256±867
for obtaining at least one healthy embryo after PGT using Visual Basic for IU, p=0.002) to obtain the similar oocyte yield (12.2±6.6 vs. 11.9±5.7, p=0.8)
Applications (VBA) analysis. and number of mature oocytes (12.2±6.6 vs. 11.9±5.7, p=0.8). The FORT index
Main results and the role of chance: Overall, the total number of retrieved was significantly higher in FMR1 premutation carriers when compared to full
and mature oocytes were 12.1 ± 6.1 and 9.4 ± 5.2 respectively. Among the 7.2 mutation carriers (48.8±26.5 vs.37.4 ±19.5%, p=0.03).
± 4.6 day-3 embryos obtained, a mean number of 3.4 ± 3.0 embryos were Limitations, reasons for caution: This study is retrospective, analyzing a
morphologically eligible for biopsy, leading to 1.5 ± 1.6 healthy embryo on limited number of cycles. In addition, women included in a PGT program do not

reflect the whole population of FMR1-mutation carriers. Indeed, those with the Wider implications of the findings: Presented results suggest that addition
lowest values of ovarian reserve tests are often not eligible for PGT. of GM-CSF does not change the general percentage of euploid embryos.
Wider implications of the findings: FMR1-mutated carriers eligible for PGT Although no benefit of GM-CSF on percentage of chromosomally normal
may not require specific adjustments in their stimulation protocol. Further anal- embryos was observed in this study, there is a need for further randomized
ysis is required to confirm these data on a larger population. studies that could correlate findings with other factors.
P-536 Difference in aneuploidy rates between embryos biopsied P-537 Adopting machine learning (ML) and artificial intelligence
on day 5, 6 and 7, cultured in medium with and without (AI) methods to minimize the use of invasive biopsy techniques in
granulocyte-macrophage colony-stimulating factor (GM-CSF) embryo assessment and selection
A. Bruszczynska1, M. Rusin2, M. Fidzianska2, M. Jagiello2, N. Zaninovic1, J. Sierra2, J. Barnes3, P. Khosravi3, I. Hajirasouliha3,
A. Wozniak.1, J. Liss1,3, A. Knight1, K. Lukaszuk1,4,5 Q. Zhan1, J. Malmsten1, Z. Rosenwaks1
1
1
Invicta, Fertility and Reproductive Centre, Gdansk, Poland ; Weill Cornell Medicine, The Center for Reproductive Medicine, New York- NY, U.S.A. ;
2
2
Invicta, Fertility and Reproductive Centre, Wroclaw, Poland ; The Manhattan College, Computational Biology, New York, U.S.A. ;
3
3
University of Gdansk, Department of Medical Biology and Genetics, Gdansk, Weill Cornell Medicine, Department for Precision Medicine, New York- NY, U.S.A.
Poland ;
4 Study question: Can an AI-informed technique be developed to select
Medical University of Warsaw, Department of Gynecological Endocrinology,
embryos with a high probability of euploidy and indicate when more invasive
Warsaw, Poland ;
5 methods (PGT) should be adopted?
Medical University of Gdansk, Department of Obstetrics and Gynecological
Summary answer: Incorporating AI-ML methodology for embryo assessment
Nursing- Faculty of Health Sciences, Gdansk, Poland
results in a high-fidelity embryo ploidy prediction model that can provide guid-
Study question: Are aneuploidy rates different for embryos qualified for tro- ance when more invasive assessment techniques are required.
phectoderm biopsy on day 5, 6 or 7 of culture, in medium with or with- What is known already: Selecting the best embryo for transfer is an enduring
out GM-CSF? objective in IVF treatments. Embryo grading and time-lapse technology have
Summary answer: PGT-A results showed an increasing proportion of abnor- helped to determine key attributes of embryo quality that correlate to clinical
mal blastocysts on days 6 and 7 comparing to day 5 but not difference for cultures outcomes. The development of a noninvasive method of embryo screening as
with/without GM-CSF. an alternative to invasive and expensive PGT is vital.
What is known already: Chromosome analysis is performed to ensure that Study design, size, duration: This is a retrospective study of blastocysts
embryo transferred to the patient has a correct number of chromosomes to (N=10491) of known ploidy status with embryo morphokinetic and morphology
reduce the risk of abnormal pregnancy, miscarriage and failed implantation. annotations and associated patient clinical profiles.
Granulocyte-macrophage colony-stimulating factor has a beneficial effect on the Participants/materials, setting, methods: Patients undergoing their first
development of human embryos in assisted reproductive technology (ART). or second ICSI cycle using PGT were included. All embryos were cultured with
Study design, size, duration: This is a retrospective study performed using a time-lapse (TLM) system. Variables studied included the morphokinetic timings,
the medical records of patients who underwent IVF procedures with PGT-A TLM images of the embryos, and blastocyst grades of the ICM, trophectoderm,
between January 2017 and December 2019 at INVICTA Fertility Clinic, Poland. and expansion. Ploidy was determined by PGT methods conducted on-site.
1854 embryos cultured to a blastocyst stage were examined for aneuploidy. Main results and the role of chance: Multiple ML approaches and AI (deep
Trophectoderm biopsy was performed on day 5 of development or day 6 and learning) algorithms were implemented to determine the most expedient and
7 for slower growing embryos. Embryo culture medium was selected based on reproducible method to provide a reliable assessment of embryo ploidy. To
the clinicians and patients decision. address the role of chance, 80:20 bootstrapping methods were used to develop
Participants/materials, setting, methods: Analysis included 655 treatment ranges in the model parameters and predictions. The area under the receiver
cycles. Median patient age was 37 (interquartile range: 33-39). Embryos were operating curve (AUC) obtained from the ML model was 0.734. This model,
qualified for trophectoderm biopsy on days: 5, 6 or 7 depending on their devel- using a preset threshold, was able to detect 25% of blastocyst ploidy status with
opment. PGT-A was performed using next generation sequencing (NGS). high accuracy (95% pos. and 94% neg.) that an invasive biopsy was not necessary.
Additionally, the collected data was classified according to the type of culture For the remaining 75% of the population, the model estimates could be incor-
medium – with GM-CSF (EmbryoGen/BlastGen, Origio) or without (G1 PLUS/ porated into other clinical information to determine the acceptable level of risk
G2 PLUS, Vitrolife). to undertake an invasive biopsy. The deep learning model, trained on embryo
Main results and the role of chance: Among the 1854 analyzed embryos, images of blastocysts at 110 hours, showed an accuracy of 62.5%. The addition
584 were cultured in medium containing GM-CSF. In that group 229 embryos of embryological information improved the accuracy of the model to 70.4%.
were qualified for biopsy on day 5. 68.1% of them were euploid and 31.9% Limitations, reasons for caution: The single-center and retrospective nature
aneuploid. On day 6, material was collected from 342 embryos - 52.6% euploid. of the analysis could be a limiting factor, although the findings were validated
On day 7, among 13 examined embryos and 23.1% were euploid. The differences through a large dataset.
in proportions of euploid embryos biopsied on different days were shown to Wider implications of the findings: The implementation of AI/ML algo-
be statistically significant (p<0.001). rithms trained on a large data set of blastocysts of known ploidy suggests that
Among the 1270 embryos cultured in the standard medium (i.e., without noninvasive approaches may be used to assess embryos with a high level of
GM-CSF), on day 5, biopsy was performed on 405 embryos and 64.2% of them confidence. These techniques may also help us to identify embryos for which
were euploid. On day 6 – 55.9% were normal, on day 7 – 50.0%. This comparison PGT is recommended.
showed that with each next day, the percentage of embryos with normal PGT Trial registration number: not applicable
results decreases. The differences in proportions of euploid embryos were sta-
P-538 Pericentromeric DNA transcripts are involved in the
tistically significant (p<0.001).
formation of membraneless mitochondria-associated structures at
There also appears to be a linear trend in the decrease of euploidy rate with
the end of oocyte maturation in human..
each additional day needed to the embryo to reach qualification for biopsy
(GM-CSF: p<0.001, no-GM-CSF: p<0.001). N. Enukashvily1, A. Dobrynin1, N. Korchagina2, A. Prjibelski3,
No significant difference was observed in general percentage of correct result D. Shafranskaya4
1
between embryos cultured with or without GM-CSF (58,1% vs 59,9%). Institute of Cytology- RAS, The Non-Coding DNA Lab, St. Petersburg, Russia C.I.S. ;
Limitations, reasons for caution: The possible limitation is that embryo 2
Ava-Peter-Scandinavia Clinics, Embryology Dept, st. Petersburg, Russia C.I.S. ;
3
culture medium selection was not randomized but rather based on medical St. Petersburg State University, Center for Algorithmic Biotechnology, St.Petersburg,
indications (outcomes of previous cycles) and also on financial resources of the Russia C.I.S. ;
4
patients, as GM-CSF supplementation was more expensive for the patient. St. Petersburg State University, Biology Dept, St.Petersburg, Russia C.I.S.

Study question: Whether maternal long non-coding RNAs (lncRNAs) tran- 3

Ava-Peter – Scandinavia ART Clinic, Clinic on Nevsky prospect, St. Petersburg,
scribed from pericentromere non-coding tandemly repeated (TR) DNA are Russia C.I.S. ;
4
involved in the formation of membraneless structures in GV-MI oocytes. St. Petersburg State University, Faculty of Biology, St. Petersburg, Russia C.I.S.
Summary answer: In late preovulatory oocytes, pericentromere tandemly
repeated DNAs are transcribed. The transcripts are clustered in some mem- Study question: Whether maternal long non-coding RNAs (lncRNAs) tran-
braneless RNP structures containing helicases and associated with scribed from pericentromere non-coding tandemly repeated (TR) DNA are
mitochondria. accumulated in late maturating oocyte
What is known already: Tandemly repeated DNA (TR DNA) constitutes Summary answer: The maternal transcripts of pericentromere TR DNA are
approximately 10% of the human genome. TR DNA family includes microsat- accumulated in the late oocyte maturation. They are present in transcriptomes
ellites, minisatellites, and satellites. Classical human satellites 1, 2, 3 (HS1, HS2, of MI, MII oocytes.
HS3) underlie the pericentromere region. Pericentromeric satellites transcripts What is known already: Non-coding RNAs play an important role in mam-
play an important role in mammalian embryogenesis. Transcripts of satellite malian gameto- and embryogenesis. Human pericentromere is built on TR DNA
DNA are present in mouse embryos prior to zygotic activation. However there of “classical” satellites family - human satellite (HS) 1, 2, 3 (HS1, HS2, HS3).
are no data about pericentromeric TR RNA in human oocytes. Blocking the pericentromere TR DNA transcription leads to embryo develop-
In mammals, pericentromeric transcripts are involved in assembling mem- ment arrest in mouse. Transcripts of TR DNA are present in mouse embryos
braneless bodies. However, in mammalian oocytes membraneless bodies prior to zygotic activation in small quantities suggesting they are not only tran-
enriched in non-coding RNAs and helicases are believed to be disassembled in scribed in embryo but are accumulated in maternal ooplasm. However there
the meiosis I prophase., are no data about pericentromeric TR DNA and their transcripts in human
Study design, size, duration: GV and MI donor oocytes are not used for oocytes.
gametes banking and are usually discarded. They were donated for scientific Study design, size, duration: We analysed 8 RNA-Seq published datasets
purposes after the local ethic committee permission was granted and a donor of healthy donors MI and MII oocytes obtained by 4 independent research
informed consent was signed. groups. The biocomputational analysis results were verified by fluorescence in
The HS 2,3 transcripts distribution, their colocalisation with helicases and mito- situ hybridization (FISH). FISH was carried out on MI human preovulatory
chondria were studied in GV (n=12), MI (n=8) oocytes. The presence of HS2,3 oocytes without morphological abnormalities from healthy donors. These
transcripts in published transcriptomes of healthy donors was verified with oocytes were excluded from egg donation program and were used in the present
computational methods. study if an informed consent was signed by a donor.
Participants/materials, setting, methods: The HS 2, 3 transcripts distri- Participants/materials, setting, methods: Raw reads were quality- and
bution was studied with RNA-DNA FISH. The colocalisation of the transcripts adapter-trimmed in order to check the presence of sequences containing homol-
with helicases was estimated by immunoFISH. Double immunostaining was used ogy regions with the HS2,3 probe. Only transcripts containing highly confident
to study the spatial relationship of helicases and the import receptors Tom20 matches with HS2,3 were selected . Several polyadenylated transcripts were
of mitochondrial preprotein translocases of the outer membrane (Tom). The detected (length 300-640 bp). They were analyzed for regions of similarity,
presence of HS2,3 transcripts in published transcriptomes of healthy donors multiple sequence alignment. The same probe as in transcriptome analysis was
was verified with computational analysis of published healthy donors used as a DNA-RNA FISH probe.
transcriptomes Main results and the role of chance: All four selected transcripts belonged
Main results and the role of chance: HS2, 3 transcription increased in in to pericentromere TR DNA families. Multiple alignment of all the 4 transcripts
oocytes after the GV stage. In GV oocytes, the intranuclear DDX5 helicase was and pairwise alignment of sequences revealed that they are not identical and
adjacent to the heterochromatic ring surrounding the post-nucleolar body. In belong to different families of classical satellites. All the assembled transcripts
late GV-MI and in MI, the HS2, 3 transcripts were colocalised with the helicases shared homology (identity more than 75%) with different satellite lncRNA pre-
granules that overlapped with the oocyte mitochondria. In published transcrip- viously obtained by from heat-shocked HeLa cells. One of the transcripts we
tomes, several polyadenylated transcripts from different datasets were detected. assembled shared high degree homology (> 80%) with the boundary region
All the transcripts belonged to HS TR DNA families. The cDNAs had the typical between centromeric DNA and pericentromeric HS. Moreover, the same tran-
signatures of HS sequences. One of transcripts contained the sequence we script contained the complete HS3 sequence that we had found earlier tran-
previously identify in transcriptomes of cancer and fetal cells. scribed in some cancer cell lines, senescent lung fibroblasts and some tissues of
Thus, in late preovulatory oocytes, different HS families of TR DNA are tran- a developing human embryo. The transcripts revealed by computational analysis
scribed. The transcripts are clustered in some membraneless RNP structures were detected in MI oocytes by RNA-FISH. The transcripts were associated in
that contain helicases and are closely associated with mitochondria being there- granules scattered throughout the ooplasm.
fore similar to Balbiany bodies described previously in early mammalian oocytes. Limitations, reasons for caution: The sample size for FISH analysis in
To date no membraneless granules associated with mitochondria were described this study was small due to difficulties in obtaining human preovulatory oocytes
in oocytes at the late stages of maturation. Mitochondria associated membrane- without morphological abnormalities. Further studies, including analysis of
less coacervate bodies assembled on the base of pericentromeric transcripts transcription of HS2,3 dynamics by live-cell imaging, may help define the role
might be a place for such spatial sequestration in the same way as in early oocytes. that perform of these transcripts in oocyte maturation and early
Limitations, reasons for caution: The experiments were performed on embryogenesis.
fixed cells. The in vivo visualisation of the structures will be carried out to prove Wider implications of the findings: According to our data, transcription
the existence of the granules in a living oocyte of HS2,3 occurs in late maturing oocytes that were assumed to be transcrip-
Wider implications of the findings: Transcription of HS2,3 occurs in late tionally silent. The appearance of HS3 transcripts in the MI oocyte suggests that
maturing oocytes that were assumed to be transcriptionally silent. The existence these transcripts might be important for oocyte maturation or early
of membraneless bodies was demonstrated for the first time. The obtained embryogenesis.
results give a new perspective to study the late oogenesis. Trial registration number: not applicable
P-540 correcting a PLCζ mutation in the human germ line to
P-539 Transcription of human pericentromeric non-coding DNA overcome hereditary infertility
at the end of human oocyte maturation B. Bekaert1, A. Boel1, M. Popovic1, P. Stamatiadis1, R. Reddy
M. Dobrynin1, A. Prjibelski2, N. Korchagina3, D. Shafranskaya4, Guggilla1, S. Chuva de Sousa Lopes M. C.2, P. De Sutter1,
N. Enukashvily1 P. Coucke3, B. Heindryckx1
1
1
Institute of Cytology of Russian Academy of Sciences, Laboratory non-coding UGhent, Department of Human structure and repair, Ghent, Belgium ;
2
DNA, St. Petersburg, Russia C.I.S. ; Leiden University Medical Center, Department of Anatomy and Embryology,
2
St. Petersburg State University, .Center for Algorithmic Biotechnology, St. Leiden, The Netherlands ;
3
Petersburg, Russia C.I.S. ; UGhent, Department of Biomolecular Medicine, Ghent, Belgium

Study question: Can clustered regularly interspaced short palindromic repeats Study question: To determine the relationship between 5mC and 5hmC
(CRISPR)/Cas9 gene editing be used for correction of a single base pair substi- according to the sperm chromatin protamination, in 3 sperm populations: nor-
tution in phospholipase C zeta (PLCζ)? mally protaminated, less protaminated, and deprotaminated, in fertile and infer-
Summary answer: CRISPR/Cas9 administration during ICSI enables the cor- tile males
rection of a single base pair substitution in PLCζ, possibly by interhomologue Summary answer: High DNA 5mC level is a marker for well-protaminated
homologous recombination. spermatozoa, documenting proper spermatogenesis. 5mC/5hmC levels
What is known already: Mutations in the PLCZ1 gene, encoding for phos- are highest in properly-protaminated spermatozoa, but disrupted in
pholipase C zeta (PLCζ) -the sperm-related oocyte activating factor- can result oligoasthenozoospermia
in failed fertilization after ICSI, which can be overcome with assisted oocyte What is known already: A special role in etiology of male infertility play
activation (AOA). However, children may still inherit the disease-causing muta- epigenetic modifications, including methylation (5mC) and hydroxymethylation
tion resulting in the need for future reproductive treatment. A possible method (5hmC) of DNA. Global DNA methylation level is high in normozoospermia,
to overcome mutation transmission is gene correction with CRISPR/Cas9 during which confirms a correct spermatogenesis. Published data also indicate the link
ICSI. From previous studies using CRISPR/Cas9 in the human germline, it is still between sperm quality and the protamination state of sperm chromatin.
called into question whether gene correction predominantly occurs through Study design, size, duration: 50 patients with oligo-/oligoasthenozoosper-
interhomologue homologous recombination or the use of the added repair mia and reproductive failure (P group) vs. 30 normozoospermic controls
template. (K group). Both groups: normal karyotype (46,XY), 25-30 years old, and a lack
Study design, size, duration: We aimed to correct a failed fertilization-re- of smoking habits, any stimulants/drugs usage, and toxic work environment.
lated paternal mutation by delivering CRISPR/Cas9 components during ICSI. A Participants/materials, setting, methods: For the first time, a sequential
gRNA-Cas9 protein complex specifically designed to target the mutant allele, staining protocol was applied, which allowed to analyze the 5mC/5hmC levels
and a repair template harboring the desired base pair substitution and an addi- revealed by immunofluorescent (IF) stainings using the same spermatozoa with
tional synonymous mutation, were injected together with patient’s sperm. AOA previously determined chromatin protamination status (aniline blue staining). Also
was performed following ICSI to overcome fertilization failure. Fertilized zygotes TUNEL examination was performed to check the sperm DNA fragmentation status.
were cultured for 3-5 days. These reconstructed embryos (n=12) were thor- Main results and the role of chance: In the P group, both: the protamination
oughly analyzed to assess gene editing efficiency. level of sperm chromatin, as well as 5mC and 5hmC levels were decreased,
Participants/materials, setting, methods: We recruited one male patient when compared to the K group. Results obtained in P group also pointed out a
with a heterozygous base pair substitution in PLCZ1 (c.136-1G>C) leading to higher interindividual heterogeneity. The 5mC and 5hmC levels were diversified
failed fertilization in all oocytes after routine ICSI. Donated in vitro or in vivo in respect to the chromatin protamination status: protaminated sperm cells
matured oocytes containing clusters of smooth endoplasmic reticulum were showed the highest 5mC and 5hmC values, in both: K and P groups. A negative
targeted, next to a control group without gene editing. Targeted next-generation correlation was found between 5mC level vs. correct protamination (both groups
sequencing was used to assess correction potential and short tandem repeat of males; p<0.05). Negative correlation was also identified for 5hmC level vs.
(STR) analysis to characterize whether mutation-carrying or wild-type sperm correct protamination in the K group, in contrast to the P group, where 5hmC
gave rise to the embryo. level increased concurrently with the chromatin protamination status (p<0.05).
Main results and the role of chance: In the control experiments where no Sperm DNA fragmentation was higher in the P group vs. K group.
CRISPR/Cas9 was added, ICSI with the patient’s sperm and AOA treatment, Limitations, reasons for caution: Analyzes on bigger number of patients
resulted in a mutation distribution of approximately 50%. High specificity of the are required
designed CRISPR/Cas9 components for targeting the paternal mutant PLCZ1 allele Wider implications of the findings: Measuring of 5mC/5hmC status of
was demonstrated by the absence of unspecific insertions/deletions (indels) in sperm DNA evidences the correctness of spermatogenesis and its disruption
embryos originating from wild-type sperm (n=5). When the mutant PLCZ1 allele may be a marker of reproductive failure.
was observed, indicated by the presence of the mutant chromosome-related STR Study funding/competing interest(s)
markers, either no editing (n=3), additional mutagenesis (indels) (n=2) or correc- 2015/17/D/NZ5/03442, National Science Centre in Poland. Authors declare
tion (n=2) occurred. Remarkably, our results suggest that the correction process no competing interests.
may not require the presence of a repair template, due to the absence of the Trial registration number: na
synonymous mutation incorporated into the repair template in the DNA sequence
of the corrected embryos. In addition, for the STR markers close to the Cas9 cut P-542 Transfers of chromosomal-abnormal “mosaic” and
site only one novel (presumably the maternal) allele was present, while both “aneuploid” embryos in patients previously refused such transfers
maternal and paternal alleles were identified for STR markers further upstream at centers were embryos had been produced
and downstream from the Cas9 cut site. This suggests that the correction resulted D. Barad1, D. Albertini2, E. Molinari2, N. Gleicher1
from interhomologue homologous recombination, by which the maternal wild-type 1
Center for Human Reproduction, n/a, New York, U.S.A. ;
allele is used as a template to repair the double-strand break. 2
Center for Human Reproduction, Laboratories, New York, U.S.A.
Limitations, reasons for caution: One of the major limitations of CRISPR/
Cas9 is the occurrence of mosaicism and off-target editing, which still remains to
Study question: What are outcomes of transfers of chromosomal-abnormal
be evaluated. Moreover, the number of targeted embryos should be increased to
embryos (“mosaic” and “aneuploid”) after patients were refused such transfers
more reliably estimate the accuracy and efficiency of the CRISPR/Cas9 protocol.
by their IVF centers, where those embryos were established?
Wider implications of the findings: Our findings demonstrate that CRISPR/
Summary answer: Transfers of such embryos result in significant life birth
Cas9 may serve to correct heritable infertility mutations in the germ line. Our
rates with basically no risk for births of chromosomal-abnormal offspring.
results further confirm the occurrence of interhomologue homologous recom-
What is known already: Like earlier studies, the STAR study recently again
bination, which may lead to loss of heterozygosity. Additional studies should be
failed to demonstrate outcome benefits from chromosomal testing of embryos
undertaken to evaluate the safety/efficiency of this method prior to clinical
prior to transfer (PGT-A). Moreover, hundreds of chromosomal-normal off-
applications.
spring have been reported born following transfer of chromosomal-abnormal
embryos (“mosaic” and “aneuploid”), without even a single chromosomal-ab-
normal birth. Yet, PGT-A practice continues, and most IVF centers still refuse
P-541 Immunostaining of global 5mC and 5hmC levels of DNA in transfers of embryos, by PGT-A declared “mosaic” and/or “aneuploid.”
differentially protaminated human sperm chromatin Study design, size, duration: Prospective observational study of 50 patients
M. Olszewska1, O. Kordyl1, M. Kamieniczna1, M. Fraczek1, who moved 278 allegedly chromosomal-abnormal embryos (per PGT-A)
P. Jedrzejczak2, M. Kurpisz1 between June 2016 and October 2019 to our center after having been refused
1
Institute of Human Genetics- Polish Academy of Sciences, Poznan, Poland ; transfer of some of these embryos at the centers where those embryos had
2
Poznan University of Medical Sciences, Department of Infertility and been produced. The center’s experience preceding 2016 has been
Reproductive Endocrinology, Poznan, Poland reported before.

Participants/materials, setting, methods: PGT-A was performed by NGS involved chromosomes. This limitation may lead to a misintepretation of break-
or CGH. Consents pointed out risks of chromosomal-abnormal pregnancies. points number affecting chromosomes.
Without evidence for a 80% threshold of “aneuploid” DNA between “mosaic” Wider implications of the findings: FISH techniques aren’t capable of
and “aneuploid” biopsies, and no differences in reported outcomes, we do not identifying all complex chromosomal rearrangements, mainly those involving
differentiate between “mosaic” and “aneuploid” embryos. Transfers are recom- centromeric regions of acrocentric chromosomes, investigations of those abnor-
mended in absence of 3 or less known “survivable” chromosomal abnormalities. malities should be performed with a sensitive methods.The advent of new tech-
Patients are, however, entitled to override. Consents since 2014 oblige to early nologies allowed better appreciations of a genome complexity.WGS brought a
prenatal diagnostics and termination with aneuploidies. required information to explain the occurred genomic events.
Main results and the role of chance: 32 patients underwent transfers (age Trial registration number: NA
40.6 ± 4.1 years); 31/32 (96.9%) had at least one previous IVF cycles (total,
n=176); 20/32 (62.5 %) had at least one prior pregnancy (overall, n=40); but P-544 Comparison of aneuploidy rates between embryos obtained
only 6/32 (18.8%) prior live births. In 32 transfers, 101 embryos were placed from fresh and frozen oocytes in the same patients: Analysis more
(median 3; range 1-5), 74 embryos were mosaic, 19 aneuploid and 8 undeter- than 1000 embryos in the UAE.
mined. Three live births and two ongoing pregnancies beyond 20 weeks (total Y. Alhelou1, S. El Shawarby1, M. Fakih1, A. Hellani2,
likely live birth rate 5/32 (15.6%) were established; 4 pregnancies were miscar- N.A. Mat Adenan3, J. Ali4
ried (total pregnancy rate 28.1%), 2 concordant with PGT-A results, 1 discordant 1
Fakih IVF Fertility Center, IVF Department, Abu Dhabi, United Arab Emirates ;
(normal 46,XX - the patients had SLE), and 1 patient refused testing. 2
Viafet genomics, PGD Lab, Dubai, United Arab Emirates ;
Limitations, reasons for caution: At mean age 40.6, patients in this study are 3
University of Malaya, Obstetrics and Gynaecology, Kuala Lumpur, Malaysia ;
significantly older than in any previously reported patient population receiving “abnor- 4
University Of Malaya, Obstertrics and Gynaecology, Kuala Lumpur, Malaysia
mal” embryos. Here presented data, therefore, likely underestimate pregnancy and
live birth outcomes and overestimate miscarriage rates for younger women. Study question: Does oocyte vitrification affected the chromosomal content
Wider implications of the findings: Transfer of selected, by PGT-A desig- of the resulting embryos?
nated to be chromosomal-abnormal embryos, whether “mosaic” or “aneuploid,” Summary answer: There is no significant aneuploidy rate between the
under current reporting standards is safe, and results in delivery of significant embryos that resulted from fresh oocytes versus that resulted from frozen
chromosomal-normal offspring which never would be born if these embryos oocytes in the same patients.
had not been transferred. What is known already: Oocyte vitrification is one of the routine practices
Trial registration number: n/a in assisted reproduction. It gained more popularity after achieving high survival
rates of more than 90% and pregnancy rates comparable to fresh oocytes.
P-543 A whole genome analysis for simplified a preimplantation Therefore, there is a need to know if oocyte vitrification can affect embryo
genetic testing of a rare and a complex interchromosomal euploidy
reciprocal insertion: thorough investigations for a straightforward Study design, size, duration: Retrospective data analysis was performed on
interpretation. 1050 embryos of 84 patients under age of 35 years between November 2017
R. Zenagui1, I. Bernicot1, T. Anahory1 and December 2019. Every patient was performed an oocyte vitrification cycle
1
Montpellier Hospital, PGT depatment, Montpellier, France followed by a fresh ICSI cycle. On the same day of the fresh ICSI cycle, the
vitrified eggs were warmed and ICSI was performed. The embryos resulting
Study question: Should whole genome sequencing (WGS) be considered for from both fresh and vitrified eggs had day 5 biopsy followed by PGS..
the diagnosis of complex chromosomal abnormalities before a preimplantation Participants/materials, setting, methods: Every patient was done an
genetic testing of structural rearrangement (PGT-SR) attempt? oocyte vitrification cycle followed by a fresh ICSI cycle. On the same day of the
Summary answer: WGS should be considered to establish the complexity fresh ICSI cycle, the vitrified eggs were warmed and were done ICSI. The
of chromosomal abnormalities with accuracy before PGT-SR investigations. embryos resulted from both fresh and vitrified eggs were done day 5 biopsy
What is known already: Insertions are rare events and interchromosomal followed by Preimplantaion Genetic Screening (PGS) using Next Generation
reciprocal insertion (IRI) with four breakpoints are exceptional, with only 11 Sequencing with High Resolution (NGS HR PGS). The structural and numerical
cases reported to date. To the best of our knowledge, no preimplantation genetic chromosomal abnormalities were analysed and compared.
testing for structural rearrangement (PGT-SR) was performed to investigate the Main results and the role of chance: The biopsied embryo that resulted
management of patients with IRI abnormalities. from fresh oocytes had an aneuploidy rate of 49.2% versus 53.8% aneuploidy
Study design, size, duration: A male carrier of rare IRI 46,XY,ins(14;22) rate for embryos resulting from frozen oocytes. The p value was 0.13 which
(q11.2 ;q11.2) and his partner were directed to our center for PGT analysis after means the result is statistically insignificant. In addition, the rate of monosomies,
three spontaneous miscarriages. The patients were informed of the investigations trisomies and complex abnormalities did not show any statistical significance.
and gave their consent before participation in the study. Limitations, reasons for caution: The frozen/thawed oocytes were not
Participants/materials, setting, methods: A genome sequencing of a male recruited from the same fresh ICSI cycle. The retrospective nature of he study
carrier was conducted at the French National Research Centre for Human and the population of the study are all 35 years and younger
Genomics. Library generation was performed with TruSeq® DNA PCR-free kit. Wider implications of the findings: The study provides reassurance to young
The genome was sequenced on a HiSeq X5 following the manufacturer’s instruc- patients undergoing oocyte freezing for variance indications that the technology
tions. DNA sequences were mapped to the reference human genome sequence itself does not affect the rate of chromosomal abnormalities and aneuploidy rate.
(GRCh37). Structural variations were called using MANTA. FISH assays were As far as we know this is the first study in the UAE that investigate the aneu-
used to detect all chromosome segments involved in chromosomal rearrange- ploidy in fresh and frozen oocytes.
ment and to identify transferred normal/balanced embryos. Trial registration number: not applicable
Main results and the role of chance: Whole genome sequencing analysis
allowed to determine accurately the chromosomal breakpoint numbers between P-545 Evaluation of Severe Male Infertility in the Light of
the chromosome 14 and the chromosome 22. Against all odds, two breakpoints New Genetic Variations
were identified instead of four as expected in IRI rearrangements. The junction M. Gavaz1, B. Umay Kara1, F. Gokdagli Sagir1, B. Yuksel1,
fragments sequencing confirmed the location and the number of both break- M. Cetinkaya1, S. Kahraman1
points. Finally, the patients were informed that the chromosomal abnormality 1
Istanbul Memorial Hospital, Assisted Reproductive Technologies and Reproductive
was a reciprocal translocation instead of interchromosomal reciprocal insertion. Genetics Center, Istanbul, Turkey
Thereby, a FISH probe strategy was adapted to highlight all chromosomal imbal-
ances, PGT-SR attempt was achieved and a healthy baby was born. Study question: Is it possible to classify the severe male infertility cases and
Limitations, reasons for caution: No FISH probes could be established predict their treatment outcome/prognosis according to causative genetic
between the centromeres and the chromosomal breakpoints on either or both variation(s)?

Summary answer: The genetic variations identified in severe male infertility Study question: Does telomere length in human triploid zygotes differ between
cases were correlated to the morphological sperm findings and the outcome oocyte- and sperm-derived sets of metaphase chromosomes and is it associated
with the genetic variations. with maternal and paternal age?
What is known already: The majority (30–60%) of infertile males do not Summary answer: In human triploid zygotes, sperm-derived telomeres are
receive a clear diagnosis; therefore, they are reported as idiopathic with a strong longer than oocyte-derived telomeres; telomere length does not depend on
suspicion of genetic underpinnings. This is particularly evident in cases of infer- either maternal or paternal age.
tility and repetitive ART failures with normal semen parameters. Genome-wide What is known already: Telomeres represent complexes of short tandem
association studies in male infertility have affected several genomic regions, and DNA repeats and proteins that protect the ends of chromosomes. Telomere
all exome sequencing studies have identified coding variants associated with male shortening caused by cell divisions, exogenous factors or genetic effects leads to
infertility. WES has been successful in identifying new genes such as TEX15, chromosomal end fusion, degradation of chromosomes and cell death. Considering
MAGEB4, SUN5, SRA1 in all categories of etiologic spermatogenic dysfunction that telomere length (TL) in preimplantation embryo may be an important pre-
(quantitative, qualitative, hypothalamic-pituitary axis and ductal obstruction or dictor of its developmental potential, the question about TL regulation in game-
dysfunction). togenesis and embryogenesis is of high importance. In contrast to telomeres in
Study design, size, duration: We carried out a cohort study at Istanbul sperm which become longer with age, oocyte telomeres are short. Whether these
Memorial Hospital from December 2018 to December 2019. We included 25 gamete-specific patterns are maintained after fertilization in zygote is unclear.
patients with severe familial male infertility without identified etiology after stan- Study design, size, duration: A total of 23 triploid zygotes were obtained
dard genetic assessment (karyotype and Y chromosome microdeletion). We from 20 couples referred for in vitro fertilization during a ten month period. The
classified male infertility under 3 groups such as non-obstructive azoospermia maternal age ranged from 23 to 39 years (mean 32.04±0.8); the paternal age
(NOA) (n=7), oligoasthenoteratozoospermia (n=7), motility and morphological ranged from 23 to 45 years (mean 34.15±1.2). The study was approved by the
(n=11) defects. Successful treatment outcome was evaluated as being able to Institutional Ethics Committee; the participating couples signed informed consent.
reach ongoing pregnancy. Participants/materials, setting, methods: For metaphase preparations,
Participants/materials, setting, methods: We obtained blood samples the zygotes were treated with colchicines, hypotonic solution and fixed on glass
from all patients. Genomic DNA was extracted using the QIAampDNA Mini slides. Parental origin of chromosomes was identified immunocytochemically by
QIAcube Kit (Qiagen, France) according to the manufacturer’s instructions. weak DNA methylation and strong hydroxymethylation in the sperm-derived
Whole exome sequencing was performed by NovaSeq 6000 Series Sequencer (paternal) chromosomes versus strong DNA methylation and weak
via Illumina technology. The coverage of target sequence sequencing is not less hydroxymethylation in the oocyte-derived (maternal) chromosomes. Relative
than 99%. Patients gave their informed consent to undergo genetic testing to TL was assessed using quantitative fluorescence in situ hybridization (qFISH)
uncover the etiology of male infertility. (Telomere PNA FISH Kit/Cy3, Agilent) by dividing the telomeric fluorescence
Main results and the role of chance: Totally, 30 different previously by the subtelomeric fluorescence measured in ImageJ1.49v.
identified genes that related with male infertility were detected. The most Main results and the role of chance: Out of 23 triploid zygotes, four zygotes
common variants were detected in NANOS1, AR, CCDC40 and RSPH1 had additional maternal chromosome set and 19 zygotes had additional paternal
genes. We detected five unique variants at NANOS1 gene in 5 patients. chromosome set. According to the relative TL values, the zygotes were categorized
including a deletion of serine (p.S83del) residues that was identified as patho- into four types. In most zygotes (21 out of 23), the relative TL values were higher
genic. From five men with definitive sperm factors, two patients were iden- in the paternal chromosomes. In two zygotes, the relative TL values appeared to
tified heterozygous variations in CCDC40 gene: c.1318-14A>G which altered be higher in the maternal chromosomes. In two zygotes with additional paternal
the protein sequence and resulted in the ultrastructural defects in the micro- chromosome sets, only one out of two paternal sets had higher relative TL values
tubule structure of cilia. In another patient, a variation in the CCDC40 gene compared to the maternal one. In two zygotes with additional maternal chromo-
that caused p.Asp284His at c.850G>C was identified as disease-causing. some sets, only one out of two maternal sets had lower relative TL values com-
Different types of variants in the RSPH1 gene were detected in three patients pared to the paternal one. The Wilcoxon matched pairs signed rank test showed
with a history of parental consanguineous marriage. No positive pregnancy that the relative TL was significantly higher in the paternal chromosomes compared
was obtained in NOA patients with variations in RSPH1, ZYMND15, SEPT12, to the maternal ones (p<0.0001). A few zygotes demonstrating unusual patterns
DNAAF1, CCDC40 and NANOS1. Variation (c.849-20dup) in SPATA16 of the relative TL values seem to be the exception rather than the rule. This
gene was found in 2 patients with morphological defect phenotype. Five of observation may suggest that some cases of developmental arrest in triploid
the sixteen patients with variants associated with primary ciliary dyskinesia embryos are due to the altered TL pattern. The relative TL neither in sperm-de-
had positive pregnancy and four had negative pregnancy, and three patients rived nor in oocyte-derived chromosomes correlated with paternal (r=-0.058;
had been waiting for embryo transfer. Variants in the DNAAF5, RSPH1 and p=0.736) or maternal age (r=0.155; p=0.44), respectively.
DNAI2 genes are the most common in patients resulting in negative Limitations, reasons for caution: As normal diploid zygotes were not avail-
pregnancies. able, the study was performed on triploid ones. The developmental potential
Limitations, reasons for caution: The main limitation of our study is the of triploid human embryos is confirmed by their capacity for implantation and
small number of patient. This study should be extended to include more patients even a full-term development suggesting that basic developmental processes in
from the same family who suffer from male infertility. triploid zygotes could be similar/identical to those in normal ones.
Wider implications of the findings: Our findings identified new variations Wider implications of the findings: Our findings may suggest that after
responsible of the severe male infertility phenotype. This study provides an fertilization, TL in sperm-derived chromosomes is “reprogrammed”. Being
understanding of undefined variations involved in male spermatogenesis and hypomethylated, paternal chromosomes are prone to recombination and, thus,
progress in the diagnosis of infertility. to alternative lengthening of telomeres (ALT). ALT may be crucial for cleavage
Trial registration number: - capacity and explains the absence of correlation of TL in sperm-derived chro-
mosomes with paternal age.
P-546 Telomere length in oocyte- and sperm-derived metaphase Trial registration number: not applicable
chromosomes in human triploid zygotes
O. Efimova1, A. Pendina1, M. Krapivin1, I. Mekina2, E. Komarova2, P-547 Frequency of mitochondrial genetic variations in human
A. Tikhonov1, O. Chiryaeva1, Y. Sagurova3, V. Baranov1, cumulus cells and their association with mitochondrial function,
A. Gzgzyan2, I. Kogan2 embryo quality and BMI
1
D.O. Ott Research Institute of Obstetrics- Gynecology and Reproductology, K. Kumar1, M. Venturas2, D. Needleman2, C. Racowsky3, D. Wells4
1
Genomic Medicine, St. Petersburg, Russia C.I.S. ; University Of Oxford, Nuffield Department of Women’s & Reproductive Health,
2
D.O. Ott Research Institute of Obstetrics- Gynecology and Reproductology, Oxford, United Kingdom ;
2
Assisted Reproductive Technologies, St. Petersburg, Russia C.I.S. ; Harvard University, MCB and SEAS, Cambridge- MA, U.S.A. ;
3 3
St. Petersburg State University, Genetics and Biotechnology, St. Petersburg, Russia Harvard Medical School, Brigham & Women’s Hospital, Boston-MA, U.S.A. ;
4
C.I.S. Juno Genetics Ltd, Juno Genetics Ltd, Oxford, United Kingdom

Study question: What is the frequency of mitochondrial genetic variations in 1

Genomic Prediction Clinical Laboratory, Clinical Laboratory, North Brunswick,
human cumulus cells and are these associated with mitochondrial function, U.S.A. ;
2
embryo quality or patient BMI? Care Fertility Group, Care Fertility Department of Research and Development,
Summary answer: Neither cumulus cell mtDNA quantity nor the presence Nottingham, United Kingdom ;
3
of specific mtDNA genetic variants were associated with embryo quality. Columbia University, Department of Pediatrics, New York, U.S.A.
However, associations with patient BMI were detected.
What is known already: Cumulus cells fulfil a vital role in support of oocyte Study question: To develop a test for evaluating miscarriage risk in IVF patients
developmental competency. Cumulus cell function relies on adequate energy and preimplantation embryos.
production, which in turn depends on the quantity and genetic competence of Summary answer: Development and validation of a novel M2 saliva test for
its mitochondria. Mitochondrial DNA mutations can be inherited or they can screening patients with infertility, and the first method capable of M2 haplotyping
accumulate in somatic cells over time, potentially contributing to ageing. Such in preimplantation embryos.
mutations may be homoplasmic (affecting all mtDNA in a cell) or they may display What is known already: Thrombophilia is linked to increased risk of miscar-
varying levels of heteroplasmy (affecting a proportion of the mtDNA). Currently, riage and other placental mediated pregnancy complications (PMPC). Expressed
little is known concerning the frequency or associations between mitochondrial on the apical surface of the syncytiotrophoblast, the anticoagulant protein
genetic variation in cumulus cells and oocyte health. Annexin A5 (ANXA5), helps maintain placental vasculature. Genetic variation,
Study design, size, duration: Human cumulus cells, associated with 290 known as the M2 haplotype, in the core promoter of the ANXA5 gene reduces
oocytes from 85 IVF patients, were analyzed. Patient age, body mass index (kg/ protein levels and increases the risk for adverse pregnancy outcomes. Infertile
m2), infertility diagnosis, fertilization status and embryo development were couples are 14% more likely to be carriers than the general population. Low
recorded with respect to each cumulus complex. Mitochondrial DNA quantity dose heparin treatment is an effective strategy to combat the complications in
was measured using a validated quantitative PCR method. A subset of 119 carrier couples, but direct testing on preimplantation embryos may serve as a
samples had their entire mitochondrial genomes sequenced using a high-depth more effective alternative.
massively parallel sequencing approach. Metabolic functions were assessed Study design, size, duration: Test performance was measured by com-
non-invasively using fluorescence lifetime imaging microscopy (FLIM). paring Sanger sequencing on parental blood DNA and quantitative real-time
Participants/materials, setting, methods: Massive parallel sequencing not (q)PCR on saliva DNA, cell line 7-cell replicates, cell line 7-cell samples and
only permitted accurate detection of mutations, but also precise quantification corresponding purified DNA, trophectoderm biopsies and DNA isolated
of levels of mutations in cases of heteroplasmy. Sequence variants in the mtDNA from the corresponding embryonic stem cell line, Mendelian inheritance
were evaluated using Mitomaster and HmtVar to predict their potential impact. expectations in embryos, embryo sanger sequencing, and SNP microar-
FLIM allowed measurement of the autofluorescence of NADH and FAD+, which ray-based linkage analyses. Embryo qPCR allele dropout rates, M2 frequencies
are crucial for mitochondrial function. Multivariate generalized linear regression observed in an IVF patient population, and inheritance patterns in IVF-derived
models were used to assess correlations among variables. P<0.05 was consid- embryos.
ered statistically significant. Participants/materials, setting, methods: Initial assay development was
Main results and the role of chance: Sequencing the entire mitochondrial performed on 6 IVF patient samples. Additional data was obtained from 14 cases
genome in 119 cumulus cell samples revealed a total of 952 synonymous single of PGT where parental and embryo biopsy DNA (n=107) were available for
nucleotide variants (SNVs) with respect to the reference genome (i.e. no amino research and 55 patients ordering clinical M2 testing. M2 testing was performed
acid change) and another 626 non-synonymous SNVs (altered amino acid at Genomic Prediction Clinical Laboratory under CLIA certification and CAP
sequence). Most SNVs were homoplasmic and therefore presumed to have been accreditation.
inherited, but 21 were heteroplasmic and potentially acquired by somatic muta- Main results and the role of chance: Concordance rates between sanger
tion. Additionally, 104 homoplasmic variants detected in 63 samples were pre- sequencing and qPCR was 93% (13/14) on parental blood DNA and 100% (6/6)
dicted to be pathogenic, with a likely impact on ATP production. A significant on saliva DNA. Concordance rates between all replicates of the cell line 7-cell
association was observed between BMI and the number of non-synonymous samples were 100% (24/24). Concordance rates between 7-cell aliquots and
variants (p=0.037) and the quantity of mtDNA in cumulus cells (p<0.001). corresponding bulk DNA was 100% (20/20). The concordance rate between
Significant difference was also observed for FLIM measurements of flavin adenine trophectoderm biopsies and DNA isolated from the corresponding embryonic
dinucleotide (FAD) long lifetime in cumulus cells that had pathogenic mtDNA stem cell line was 100% (1/1). Concordance between qPCR and Sanger sequenc-
mutations (p=0.046). Grouping samples according to patients’ mitochondrial ing on trophectoderm biopsies was 100% (2/2). Trophectoderm biopsy qPCR
haplogroups revealed a significant association of nicotinamide adenine dinucle- results were 97% (104/107) concordant with expectations from Mendelian
otide (NADH) long lifetime (p=0.025) and NADH short lifetime (p=0.012) with inheritance rules and 100% (107/107) when including SNP array based linkage
haplogroup. This suggests that mitochondrial metabolism varies between hap- analyses. There was an M2 frequency rate of 18% in patients ordering clinical
logroups. Moreover, the number of detected potentially pathogenic variants testing. When one parent was a carrier, half (33/66) of the embryos were M2
were also significantly different among mitochondrial haplogroups (p=0.005). carriers. In families where both parents were carriers, 76% (16/21) of embryos
No correlations were observed with respect to cumulus cell mtDNA quantity were M2 carriers.
and patient age, infertility diagnosis, fertilization status or embryo quality. Limitations, reasons for caution: Embryo biopsy qPCR displayed a 7% allele
Limitations, reasons for caution: Although, a large number of samples drop out rate and required concurrent SNP array based linkage analysis to obtain
were analysed in this study, an expanded sample size is needed to determine accurate genotypes.
whether subtle relationships do exist among between mtDNA mutation/copy Wider implications of the findings: We have developed and validated a
number and patient/oocyte/embryo characteristics. We are continuing to pro- saliva DNA-based M2 haplotyping test for infertile patients, and a new ability to
cess more samples. perform “PGT-M2 haplotyping” for carrier couples undergoing IVF. Future work
Wider implications of the findings: No relationship was detected between will include prospective analysis of clinical outcomes following embryo selection
cumulus cell mtDNA quantity or sequence variants (including pathogenic) and and non-selection with PGT for M2 carrier status.
clinically relevant endpoints for corresponding oocytes. Nonetheless, some of Trial registration number: not applicable
the findings raise interesting biological questions, particularly regarding the inter-
play of metabolism and BMI.
Trial registration number: NOT APPLICABLE

P-548 A novel test for IVF miscarriage risk: Annexin A5 M2 REPRODUCTIVE ENDOCRINOLOGY
haplotyping in IVF patients and preimplantation embryos
B.R. Rana1, R. Zimmerman1, D. Marin1, J. Xu1, E. Messick1,
L. Tellier1, D. Egli3, S. Fishel2, N. Treff1

P-549 None of The Parameters of Bologna and Poseidon Criteria Study question: Does the activity of total Adenosine deaminase (ADAT) in
Has a Significant Impact on Pregnancy Rates the follicular fluid have a relation with ovarian reserve and clinical parameters
B. Devranoglu1, H. Bozdag2, E. Özkaya1, Ç. Yayla Abide1, linked to IVF outcomes?
E. Çöğendez1, E. Tozkır1, E. Darici1 Summary answer: Follicular fluid(FF) ADAT activity in DOR group was higher
1 and had a negative correlation with BMI,a positive correlation with FSH and no
University of Health Science- Zeynep Kamil Maternity and Childeren’s Diseases
relation with IVFoutcomes.
Health Training and Research Hospital, Department of Obstetrics and Gynecology,
What is known already: In the human reproductive tract,the function of the
istanbul, Turkey ;
2 enzymes that affect adenosine metabolism,especially ADA,have been investi-
Health Sciences University Sultan Abdülhamid Han Training and Research
gated.The activity of ADA was evaluated in the male reproductive system.
Hospital, Department of Obstetrics and Gynecology-, istanbul, Turkey
Inhibition of ADA activity increases adenosine levels and it resulted an improve-
Study question: What are predictive values of the individual parameters of ment of sperm fertilisation.In addition, it is suggested that ADA receptors might
Bologna and Poseidon criteria for clinical pregnancy and cycle cancellation in have a role in capacitation and acrosome reaction.Alteration in ADA activity
women with poor ovarian reserve or previous poor response? seemed to be related with male infertility.For the female reproductive system,that
Summary answer: These criteria, Bologna or Poseidon, did not have any women with elevated ADA activity have the highest risk for recurrent sponta-
benefit to ameliorate management to obtain a better outcome. neous abortion.To the best of our knowledge,ADA activity has not yet been
What is known already: Even several authors have attempted to define the systematically addressed in the female infertility.
diagnosis of poor ovarian response (POR), this terminology has not been clarified Study design, size, duration: This study is a retrospective cohort study. The
among international publications. According to a published systematic review of data of all individuals, who underwent their IVF/ICSI at IVF Unit of Istanbul
47 randomized controlled trials, there were 41 different POR definitions. To Research and Education Hospital from May 1st 2019 to July 31st 2019 were ret-
outline the POR definition in a standardized manner, Ferraretti et al. have pro- rospectively reviewed. Total of 106 women were eligible and enrolled in this study.
posed new definitive criteria known as “Bologna Criteria.”. A New strategy, Participants/materials, setting, methods: The women were between
POSEIDON, has been introduced to predict the prognosis of patients under- 23–45 years old, had an infertility etiology of diminished ovarian
going ovarian stimulation for IVF. reserve(DOR),tubal factor(TF),male factor(MF) or unexplained infertility.
Study design, size, duration: Seven hundred twenty-two cases were retro- Patients were excluded if they had two or more type of infertility etiologies,
spectively screened from the database and reclassified based on both Bologna diseases of the immune system, hematologic malignancies, ovarian tumors, endo-
and Poseidon criteria separately, and assisted reproductive technology (ART) crine disorders or smoking. Blood samples were drawn before oocyte retrieval.
outcomes were compared for each corresponding group. The diminished ovarian FF from the first aspirated follicle was used for ADAT measurement.Its relation
reserve was defined according to basal follicle-stimulating hormone (FSH) with ovarian reserve markers and IVF outcomes were determined.
value >10 IU/L, antral follicle count <6, or anti-Müllerian hormone <1.1 ng/ Main results and the role of chance: A total of 106 infertile women were
mL and/or a previous poor ovarian response. The presence of one of these enrolled in the study. Thirty-three of the women had DOR, 22 had TF, 21 had
criteria made us diagnose poor ovarian reserve. unexplained infertility and 30 had MF infertility.
Participants/materials, setting, methods: Poor ovarian reserve was deter- AMH level of DOR group was significantly lower than that of the other groups
mined in patients who fulfilled the above-mentioned criteria. Individualized (p<0.01). FSH level of DOR group was significantly higher than that of the other
gonadotropin dose and protocol were selected based on our institutional con- groups (p<0.05). Mean level of serum estradiol on the day of hCG administration
ventional ovarian hyperstimulation protocols defined for cases with poor ovarian (p<0.01), total number of oocytes retrieved (p<0.01) and MII oocyte count
reserve. All these cases were retrospectively screened from the database and (p<0.01) of DOR group were significantly lower than that of the other groups.
reclassified based on both Bologna and Poseidon criteria separately, and ART There was no statistically significant difference among the other groups (p>0.05).
outcomes were compared for each corresponding group. Plasma lymphocyte, monocyte and eosinophil counts were not statistically
Main results and the role of chance: The highest pregnancy rate was different between groups (p>0.05).
observed in Poseidon group 1 (10.1 % vs. 4.8 % in Poseidon group 2, 4.6 % in Activity of plasma ADAT was significantly higher than activity of FF ADAT in
Poseidon group 3, 4.3 % in Poseidon group 4) however the difference between all of the infertile groups (p<0.01). Activity of FF ADAT was higher in DOR
groups did not reach statistical significance (p > 0.05). No pregnancy was group than the others (p<0.01).There was no statistically significant difference
observed in Bologna groups which included cases fulfilled the criteria age and among the other groups (p> 0.05).
ovarian reserve or age and oocyte number, on the other hand, Bologna group The relation of ADAT activity with BMI showed a negative correlation in DOR
with oocyte number and ovarian reserve criteria had 4.7 % pregnancy rate, and group (r:-0.507). Additionally, ADAT activity of DOR group and FSH levels,
pregnancy rate was 3.7 % in group of women who fulfilled all criteria of Bologna showed a positive correlation (r:0.352).
classification. Pregnancy rates were not found to be statistically significant Limitations, reasons for caution: Study population was small and ADA
(p > 0.05). enzyme activity could be calculated as ADA1 and ADA2 rather than ADAT.
Limitations, reasons for caution: This was a retrospective study and no Wider implications of the findings: Increased ADAT activity can lead to
data available in terms ongoing and live birth rates. reduced adenosine levels, which might be resulted in disturbed fertility process.
Wider implications of the findings: Our data showed that Bologna or The activity of FF ADAT activity might be important for fertility work-up. Further
Poseidon did not have any benefit to change management to obtain better studies are needed.
outcomes but may be utilized to provide better and precise information and Trial registration number: IRB: IEAH7/2019
counseling for the patients with poor ART outcomes before starting the treat-
ment cycle. P-551 Predictive factors influencing pregnancy rate in frozen
Trial registration number: not applicable embryo transfer.
N. Hayashi1, N. Enatsu1, T. Iwasaki1, J. Otsuki2, Y. Matsumoto1,
P-550 Adenosine deaminase activity in the follicular fluid of S. Kokeguchi1, M. Shiotani1
infertile women with diminished ovarian reserve can act as a 1
Hanabusa Women’s clinic, medical office, Kobe, Japan ;
predictor of ovarian reserve 2
Okayama University Assisted Reproduction Technology Center, medical office,
G. Keskin1, B. Bacanakgil1, H.H. Kalem2, A. Köse2, A. Atıcı1, Okayama, Japan
S. Yalcınkaya1, M. Ayyildiz3
1
Health Science University Istanbul Research and Education Hospital, Department Study question: What factors influence pregnancy rate (PR) in frozen embryo
of Obstetrics and Gynecology, Istanbul, Turkey ; transfer (FET)?
2
Health Science University Istanbul Research and Education Hospital, Department Summary answer: Endometrial thickness and the position of transferred air
of Embriyology, Istanbul, Turkey ; bubbles influenced clinical pregnancy in FET cycles.
3
Health Science University Istanbul Research and Education Hospital, Department What is known already: There have been many reports evaluating single
of Biochemistry, Istanbul, Turkey factors which affect embryo transfer (ET) outcomes including embryo quality,

female age, endometrial thickness and the technique of the transfer procedure. (PCB), Polybrominateddiphenylethers (PBDE) or Dichlorodiphenyltrichloroethane
Most studies have confirmed that the PR is better in women with a thicker (DDT)/Dichlorodiphenyldichloroethylene (DDE), less is known about two exten-
endometrium than those with a thinner endometrium. Another significant vari- sively utilized EDCs: Mirex and the technical mixture of nonylphenols.
able was the position of the transferred air bubble, which is often regarded as Study design, size, duration: Consecutive patients undergoing ICSI due to
an indicator for the position of the transferred embryo. However, precisely male subfertility were included. Patients with endometriosis or polycystic ovaries
which factors and the degree to which they each affect ET outcomes is still were excluded. Human FF was collected at the Fertility Center Dortmund
unknown. between 2016 and 2017 (n=210). Only FF from the first two punctured follicles
Study design, size, duration: This study was a retrospective study, and was collected. An additional multiple-choice questionnaire was provided to
included 938 FET cycles involving single frozen-thawed good quality blastocyst identify potential sources of EDC, because the social environment, consumers
(Gardner grade ≥ 3BB) between August 2017 and January 2018 in a single behaviour and food habits may play an essential role regarding EDC uptake.
fertility clinic. Participants/materials, setting, methods: The concentration of Mirex,
Participants/materials, setting, methods: FET was performed with either five different NP isomers, DDT/DDE, six PCB and four PBDE congeners in the
a hormone-replacement cycle or natural-cycle protocol. The primary outcome FF was analysed using gas-chromatography coupled with mass-spectrometry
of this study was clinical pregnancy defined as the detection of gestational sac using selected ion monitoring. The samples were extracted and derivatized
in the uterus by using transvaginal ultrasound at 5 weeks gestational age. The before quantification of selected EDC. Standard solutions of pre-named con-
significance of several parameters including endometrial thickness, position of centrations and solvent blanks were injected with every measurement. To identify
the transferred air bubble, self-evaluation score by physicians, and the uterus a potential association between EDC-exposure, -uptake, clinical parameters and
direction at ET as predictors of clinical pregnancy were evaluated using univariate socioeconomic factors, a statistical evaluation was performed.
and multivariate analysis. Main results and the role of chance: All investigated EDCs were present
Main results and the role of chance: Among 938 ET cycles, 462 (49.3%) in every FF sample showing a broad distribution of concentrations between
were considered clinically pregnant. Among the variables, endometrial thickness individual samples. NP was present in the FF at 10.4±11.1ng/g FF [mean±sd],
was positively associated with clinical pregnancy in a linear trend. The clinically Mirex at 1.1±0.9ng/g FF, PCBs at 0.1±0.1ng/g FF, PBDEs at 0.04±0.02ng/g FF
acceptable threshold was calculated as 10.0mm. The position of the transferred and DDT/DDE at approximately 0.7±0.8ng/g FF. There was no significant cor-
air bubble and the clinical pregnancy rate showed a curvilinear relationship indi- relation between EDC concentration and the number of retrieved oocytes. With
cating that the clinical PR increased as the air bubble position got closer to 6 mm regard to clinical parameters, an increased concentration of NP, Mirex, DDT/
apart from uterine fundus, remained steady until 10 mm, and began decreasing DDE, PCB153, PBDE99, 100 and BB153 in FF samples correlated negatively
after 10 mm. The clinical PR was significantly higher in cases with a self-evaluation with the maturation rate (p<0.05). Nearly all examined EDCs including NP
rating of A, compared with ratings of B or C (50.2% vs 42.1%, p<.05). Comparing isomers and Mirex lead to a relative decrease in the number of 2PN and hence
the clinical PR based on uterus direction, there was no difference between the a significantly reduced fertilization rate (p<0.05). In contrast to the maturation
anteflexion, retroflexion, and straight groups (59.5%, 47.6%, and 42.9% respec- and fertilization rate, no significant influence of EDC concentration on clinical
tively, p = .45). Univariate analysis of predictive parameters identified endome- pregnancy rate could be observed.
trial thickness, self-evaluation score by physicians, and position of air bubbles as Regarding socioeconomic factors, no impact was observed regarding patients’
significant predictors of clinical pregnancy, of which endometrial thickness and residential area, diet, source of food products, nicotine or caffeine consume,
position of air bubbles appeared to be independently related to clinical pregnancy stays abroad as well as professional life. However, a significant correlation
(odds ratio; 1.56 and 1.34, 95% confidence interval; 1.20 – 2.03 and 1.02 –1.77 between EDC concentration and source of supply of textiles (fashion discounters
respectively, p<.05). vs. retail shop vs. fashion boutiques) was found (p<0.05), in particular for NP.
Limitations, reasons for caution: Patients with slight complication such as Limitations, reasons for caution: Limitations of this study include difficulties
leiomyoma or hydrosalpinx without surgical indication and adenomyosis were in extrapolating the findings to the general population, because no data of women
included in this study. Therefore, we could not completely exclude the influence not undergoing ICSI are available. Data regarding the exact molecular mechanism
of these factors Additionally, this study was a non-randomized, retrospective of NP and Mirex respectively, are limited.
study, and a larger prospective study would be needed to optimize ET strategy. Wider implications of the findings: EDC uptake and exposure to NP, Mirex,
Wider implications of the findings: We recommend that physicians should PCB, PBDE or DDT/DDE may adversely affect female reproductive outcome.
focus on maintaining their patients’ endometrial at the greatest possible thickness Higher EDC levels in the FF correlate negatively with the maturation and fertil-
during the ET cycle. Moreover, the transferred air bubbles, that means a position ization. Consumers´ behaviour play an essential role regarding the individual
of the transferred embryo, should be placed between 6 to 10 mm from the EDC uptake.
fundus during ET procedure. Trial registration number: NCT01385605
P-553 Aging attenuates ovarian circadian rhythm
P-552 Analysis of Nonylphenols (NPs), Mirex and selected G.L. Ding1, Z.R. Jiang1, K.X. Zou1, H.F. Huang1
endocrine disrupting chemicals (EDCs) in the follicular fluid (FF) 1
The International Peace Maternity and Child Health Hospital, Shanghai Jiao Tong
of women undergoing intracytoplasmic sperm injection (ICSI) University School of Medicine, Shanghai, China
T. Trapphoff1, A. Bullach2, S. Zühlke2, M. Spiteller2, S. Dieterle1
1
Kinderwunschzentrum Dortmund, Dortmund Fertility Centre, Dortmund, Germany ; Study question: The objective of our study was to investigate the effect of
2
Technische Universität Dortmund, Analytische Chemie und Umweltchemie INFU, aging on ovarian circadian rhythm.
Dortmund, Germany Summary answer: Subfertility in older women is partially due to ovarian
circadian dysrhythmia as a result of aging.
Study question: Are selected EDCs including Mirex and NPs present in the What is known already: The circadian clock is also known as the physiological
FF of women undergoing ICSI, and are they associated with clinical and socio- clock. In mammals, the circadian clock system is composed of coordinated and
economic parameters? synchronized cell and tissue clocks, including the central clock located in the
Summary answer: NP, Mirex and selected EDCs can be quantified in FF and suprachiasmatic nucleus (SCN) of the basal hypothalamus and the peripheral
may lead to a reduced maturation and fertilization rate in patients under- clock in the various tissues of the body.The ovarian circadian clock is a peripheral
going ART. circadian clock that is regulated by neuroendocrine signals from the SCN, playing
What is known already: EDCs are exogenous substances known to interfere an important role in the physiological process of the normal reproductive system,
with the mammalian hormone system. EDCs mostly origin from synthetic chem- such as ovulation and steroid release. However, the effect of aging on the ovarian
icals used in industry, pharmacy or agriculture. Exposure to EDCs has been shown clock has hardly been explored.
to affect female reproduction. It is known that different EDCs alter the embryonic Study design, size, duration: Human granulosa cells were obtained by fol-
development, spindle formation, chromosomal alignment or gene expression of licular aspiration from women undergoing in vitro fertilization (IVF), which were
oocytes and embryos. Besides well studied EDCs like Polychlorinatedbiphenyls divided into two groups, 34 cases of young group (under 40 years old) and 18

cases of old group (over 40 years old). Female C57BL/6 mice of 12-week-old outcomes were comparable in blastocyst embryo transfer cycles, between dif-
were determined as young group and 8-month-old as old group. Three to five ferent P level with fresh or frozen-thawed embryo transfer. The clinical outcomes
mice per group were sacrificed every fourth hour to collect liver and ovary of frozen-thawed cycles were comparable between patients with different P
tissues. level on hCG day. For patients with P ≤3.56nmol/L, we suggested fresh embryo
Participants/materials, setting, methods: The ovarian luteinized granulosa transfer. And for those p>3.56 nmol/L, we suggested blastocyst embryo transfer
cells from young and old patients were isolated. Anti-Müllerian hormone (AMH) with fresh embryo or frozen-thawed embryo transfer.
was examined. For mice, the stage of the estrous cycle was determined by vaginal Limitations, reasons for caution: The present study was a single center
smears. During proestrus, three to five mice per group were sacrificed every retrospective study, the cut-off level of progesterone only suitable for our center.
fourth hour (ZT 0, ZT 4, ZT 8, ZT 12, ZT 16, ZT 20) to collect liver and ovarian Wider implications of the findings: The present study suggest individualized
tissues. Real-time quantitative PCR was used to detect the expression of embryo transfer strategy for patients with different progesterone levels, and
clock genes. pointed out that there should not be “one for all” strategy in IVF/ICSI cycles.‘’
Main results and the role of chance: All the examined circadian clock genes Trial registration number: NOT APPLICABLE
(Clock, Bmal1, Per1, Per2, Cry1 and Rev-erbα) in human granulosa cells showed
a downward trend in expression with aging. And, the mRNA expression levels P-555 Melatonin – a new fertility wonder drug? A systematic
of the circadian clock genes were negatively correlated with age (P<0.05). Older review on the impact of melatonin on fertility
patients (≥ 40 years of age) had significantly reduced serum anti-Müllerian hor- M. Mejlhede1, U.B. Knudsen2
mone (AMH) levels. Except for Rev-erbα, all the other examined circadian clock 1
Aarhus University, Health, Aarhus, Denmark ;
genes were positively correlated with the level of AMH (P<0.05). The circadian 2
Aarhus University, Clinical Institute of Medicine, Aarhus, Denmark
rhythm in the ovaries of older mice was changed significantly, although the cir-
cadian rhythm in the livers of older mice was basically consistent with that of Study question: How does melatonin effect assisted reproductive technologies
young mice. There was no difference between the two groups in the expression (ART)-outcomes?
of the differentiation marker LHCGR and the expression of the internal refer- Summary answer: Most of the included studies showed a significant improve-
ence β-actin, suggesting that the decreased expression of clock genes was indeed ment in one or more ART-outcome but not in clinical pregnancy rate or live
related to age but was not related to a reduction in the extent of granulosa cell birth rate.
differentiation. What is known already: ART are associated with significant levels of oxidative
Limitations, reasons for caution: Our findings from human granulosa cells stress, which can have a negative impact on the pregnancy rates by modifying
suggested a difference in the expression of circadian clock genes at certain time the quality of oocytes, embryos and sperm. The anti-oxidant melatonin has been
points. But the effect of aging on the rhythm of the ovarian clock could not be investigated as a supplement in in vitro fertilization embryo-transfer (IVF-ET)
verified in human samples. So we also carried out mouse experiments in protocols to see if it can improve the ART-outcomes.
our study. Study design, size, duration: A systematic review in accordance with the
Wider implications of the findings: In our study, we found that age is closely Preferred Reporting Items for Systematic Reviews and Meta-Analyses
related to ovarian circadian clock. Subfertility in older women is partially due to (PRISMA). Systematic data-search of PubMed, Embase and Cochrane Central
ovarian circadian dysrhythmia as a result of aging. More studies will be needed Register of Studies from the earliest available online year of indexing up to
to explain mechanistically how aging affects the circadian clock of the ovary. April 2019 was conducted. Fifteen trials were included. The total number of
Trial registration number: not applicable patients was 1.781. One retrospective cohort study reported the results in
embryo transfer cycles and thereby contributing with 13.372 unique embryo
P-554 Embryo transfer strategy for patients with normal P level transfer cycles.
on hCG day: a large sample retrospective cohort study Participants/materials, setting, methods: The population consisted of
S. Yang1, L. Rong1, Z. Xiumei1, W. Lina1, W. Haiyan1, M. Caihong1, infertile women/men undergoing IVF-ET including intracytoplasmic sperm injec-
L. Ping1, Q. Jie1 tion (ICSI).
1
Peking University Third Hospital, Reproductive Medical Center, Beijing, China The intervention was melatonin in vivo/vitro.
The primary outcome was clinical pregnancy rate (CPR).
Study question: The present study was aimed to investigate the transfer strat- The secondary outcomes were live birth rate (LBR), sperm quality-parame-
egy for patients with different level of “normal” P level on hCG day. ters, fertilization rate, oocytequantity/quality, quality of embryos and implanta-
Summary answer: The clinical outcomes of fresh embryo transfer were better tion rate.
with lower P level on hCG day. Methodological quality was assessed for each study using the CHECKLIST
What is known already: The “normal” P level might also affect the clinical from the European Society of Human Reproduction and Embryology (ESHRE).
outcomes of IVF/ICSI. More and more clinicians consider the “frozen-all” strat- Main results and the role of chance: No significant changes in CPR in
egy safe and effective, but is this strategy proper for everyone and is there more relation to use of melatonin were demonstrated. The majority of studies did
individualized choice for patients? not show effect on LBR. One study showed a significant reduction in LBR.
Study design, size, duration: It is a single-center retrospective cohort study, Three studies showed a significant improvement of fertilization rate and one
and the study period was patients accepted oocyte pick-up between 1st Jan. study showed a significantly higher implantation rate. Eight studies demon-
2011 and 31th Dec. 2016. During the study period, 20,658 cycles were full-filled strated a significant rise in embryo quality. Oocyte quality and/or quantity
the inclusive and exclusive criteria, including 4915 cycles of frozen-all patients were showed to be significant improved by seven studies. The two studies,
had accepted frozen-thawed embryo transfer by the end of Jun. 2019 (first who investigated sperm quality parameters, both demonstrated a significant
frozen-thawed cycles after frozen-all in fresh cycles). There were 15,473 fresh improvement in the measured parameters. Ten studies had poor methodolog-
cycles and 4,915 frozen-thawed cycles were analyzed respectively. ical quality.
Participants/materials, setting, methods: The cycles were divided into Limitations, reasons for caution: The included studies were small and the
two groups depending on the embryo transfer strategy, fresh embryo transfer methodological quality of the studies low or very low. Therefore, there is insuf-
or frozen-all. And both groups were further divided into four subgroups depend- ficient evidence to make any conclusion on whether melatonin could influence
ing on Quartile of P level and embryo stage (D3/D5). the result of ART.
Main results and the role of chance: The clinical outcomes of fresh embryo Wider implications of the findings: A systematic review and meta-analysis
transfer refer to clinical pregnancy rate (CPR), implantation rate (IR), and live from 2013 demonstrated a pooled risk ratio of 1.21 for CPR in favour of mel-
birth rate (LBR) were decreased with increasing P level, especially when atonin. A Cochrane Systematic Review investigated the effects of use of anti-
P>3.56nmol/l, which were lower than those accepted frozen-thawed embryo oxidants including melatonin on female subfertility. The CPR improved (not
transfer (p<0.05). The patients accepted D3 embryo transfer and P≤3.56nmol/L, statistically significant) in the group receiving melatonin compared to
the fresh embryo transfer outcomes were better than frozen-thawed cycles, control-group.
including higher CPR, IR, LBR, and lower miscarriage rate (p<0.05). The clinical Trial registration number: not applicable

P-556 The transfer of an extra low quality embryo: Is it ever 3

worth it? Scientific research, São Paulo, Brazil ;
4
D. Braga1,2, A. Setti1,2, R. Ferreira3, A. Iaconelli Jr.4, E. Borges Jr.2,4 Fertility Medical Group, Clinical department, São Paulo, Brazil
1
Fertility Medical Group, Scientific research, Sao Paulo, Brazil ;
2
Study question: Is it ever worth it to add an extra embryo derived from an
immature oocyte, when an embryo derived from metaphase-II (MII) oocyte is
Scientific research, Sao Paulo, Brazil ;
3
transferred?
Fertility Medical Group, IVF laboratory, Sao Paulo, Brazil ;
4
Summary answer: The transfer of an extra embryo, derived from rescue in
Fertility Medical Group, Clinical department, Sao Paulo, Brazil
vitro maturation (rIVM) is not worth it.
What is known already: In vitro maturation (IVM) has been reported mostly
Study question: Is it ever worth it to add an extra low-quality embryo to a
in women with polycystic ovary syndrome, where, oocytes are on purpose
high-quality embryo transfer in IVF cycles?
recovery at immature stages in natural cycles or after low-dose gonadotropin
Summary answer: The transfer of an extra low-quality embryo is worth it
stimulation. Success rates after IVM appear inferior to in vivo-matured oocytes.
for patients >37 years-old. For younger women, this dramatically increases the
Unlike in scheduled IVM, in stimulated cycles, pharmacologic doses of gonado-
risk of multiple pregnancies.
trophins induce the growth of follicles, which, under natural conditions, would
What is known already: Although morphological evaluation has been the
become atretic. Ovarian stimulation leads to the retrieval of oocytes at different
main strategy applied in order to select embryos for transfer, it has been shown
nuclear stages and some may spontaneously mature in vitro. Whether rIVM of
that even aneuploid embryos are able to reach high morphological scores and
oocytes retrieved from stimulated cycles may improve the ICSI outcomes is to
vice versa. In an effort to promote singleton gestation and reduce the number
be elucidated.
of multiple pregnancies, guidelines for the limits on the number of embryos to
Study design, size, duration: Data analyzed in this historical cohort study
be transferred in IVF cycles have been published. Recommendations strict the
were obtained via chart review of 2021 patients undergoing controlled ovarian
number of embryos transferred considering patients age and treatment prog-
stimulation for ICSI cycles, between 2016 and 2018. After ovum-pickup oocytes
nosis. Information concerning the developmental stage is also provided, however,
were assessed for nuclear status. Mature oocytes that had released the first polar
less is known about the quality of transferred embryos.
body were used for ICSI. Immature oocytes, at prophase I (PI) and metaphase
Study design, size, duration: Data analyzed in this historical cohort study
I (MI) stages, were cultured. Those that spontaneously matured in vitro were
were obtained via chart review of 1249 patients undergoing intracytoplasmic
eventually injected and transferred.
sperm injection (ICSI) cycles between 2016 and 2018. For all cycles one or two
Participants/materials, setting, methods: Cycles, performed in a private
embryos were transferred on day five. The embryo morphology was assessed
university–affiliated IVF-center, were split into groups depending on the origin
16–18h post-ICSI and on the mornings of days two, three, and five of embryo
of transferred embryos: Group-Single-MII (n=454, one embryo, derived from
development and all evaluated parameters were used for the classification of
an MII-oocyte was transferred), Group-Double-MII (n=1483, two embryos,
embryos into top- or low-quality embryo.
derived from MII-oocytes were transferred, Group-Double-Mixed-MI (n=72,
Participants/materials, setting, methods: This study was performed in a
one embryo derived from an MII-oocyte and one from a MI-oocyte were trans-
private university–affiliated IVF center. Cycles were split into groups depending on
ferred, and Group-Double-Mixed-PI (n=12, one embryo derived from an MII-
the number and quality of transferred embryos. Group Top-Quality (n=871), in
oocyte and one from a PI-oocyte were transferred). Outcomes were compared
which a single top-quality embryo was transferred, Group Mixed-Quality (n=378),
among the groups.
in which one top-quality and one low-quality embryo were transferred. The impact
Main results and the role of chance: Overall 21,220 oocytes were
of the transfer of extra low-quality embryos on singleton and multiple pregnancies
retrieved, in which 15,200 (71.6%) were in the MII-stage, 1830 (8.6%) were in
rates, in different female age ranges was investigated by general mixed models.
the MI-stage, and 4,190 (19.7%) were in the PI stage. When adjusted for the age
Main results and the role of chance: The pregnancy rate was significantly
and endometrial thickness, the pregnancy rate was increased when two MII-
increased when the double embryo transfer, including one top-quality and one
derived embryos were transfer in comparison with the transfer of a single MII-
low-quality embryo, was compared with the single top-quality embryo transfer,
derived embryo, however, no significant differences were noted when a single
regardless of the age: 33.5% vs 44.6%, p=0.003, for patients <35 years-old, 27.9%
vs 36.9%, p<0.027, for patients between 35 and 37 year-old, and 25.4% vs 24.1%, MII-derived embryo transfer or a double embryo transfer, including one MII-
derived embryo and one MI- or one P1-derived embryo, was performed (18.0%,
p=0.041 for patients >37 years-old, for Top-Quality and Mixed-quality groups
39.4%, 33.3% and 15.3.0%, p<0.001, for Group-Single-MII, Group-Double-MII,
respectively. The transfer of an extra low-quality embryo also increased the mul-
Group-Double-Mixed-MI and Group-Double-Mixed-PI, respectively). The same
tiple pregnancy rates in younger patients: 0.0% vs 41.3%, p<0.001, for patients
results were observed for the implantation rate: the Group-Double-MII, pre-
<35 years-old and 27.9% vs 36.9%, p=<0.001 for patients between 35 and 37
sented the higher implantation, followed by the Group-Single-MII and Group-
year-old, for Top-Quality and Mixed-quality groups respectively. For patients >37
Double-Mixed-MI, while the Group-Double-Mixed-PI, presented the lowest rate
years-old, no differences in the multiple pregnancy rate was noted when one
(17.7%, 29.4%, 20.8% and 11.8%, p<0.001, for Group-Single-MII, Group-
(high-quality) or two (one high- and on low-quality) embryos were transferred:
Double-MII, Group-Double-Mixed-MI and Group-Double-Mixed-PI, respec-
15.3% vs 21.1%, p=0.694, for Top-Quality and Mixed-Quality groups respectively.
tively). The miscarriage rate did not differ among the groups (9.9%, 9.3%, 9.1%
The increment of one low-quality embryo increased the odds of multiple preg-
and 25.0%, p=0.911, for Group-Single-MII, Group-Double-MII, Group-Double-
nancies in 37% for patients <35 years-old (OR: 0.631, CI: 0.487 - 0.817, p<0.001)
Mixed-MI and Group-Double-Mixed-PI, respectively).
and in 44% in patients between 35 and 37 year old (OR: 0.563, CI: 0.340 – 0.932,
Limitations, reasons for caution: The embryo quality may be a source of
p=0.026). The miscarriage rates did not differ among the groups, for any age.
bias and a reason for caution.
Limitations, reasons for caution: Retrospective nature of this study and
Wider implications of the findings: These findings raise the question about
the small sample size may be a reason for caution.
the contribution of rIVM in ICSI cycles. Although previous studies demonstrated
Wider implications of the findings: The addition of an extra low-quality
that rIVM may increase the number of embryos available for transfer, the quality
embryo may increase the pregnancy rate and be valuable for older patients, in
of these embryos must be carefully evaluated, since its implantation potential
which the pregnancy chance is lower. However, for younger patients, this results
seems to be limited, especially for PI-derived embryos.
in increased multiple pregnancy chance, raising the question on whether taking
on this level of risk is worth it.
P-558 Serum sex hormone–binding globulin (SHBG) levels during
P-557 The contribution of the rescue in vitro maturation to ICSI controlled ovarian hyperstimulation as a predictor of ovarian
cycles response in women without polycystic ovary syndrome
R.C. Ferreira1, D. Braga2,3, A. Setti2,3, A. Iaconelli Jr.4, E. Borges Jr.3,4 J. Zhai1, S. Li1, Y. Zhu1, Z.J. Chen1, Y. Du1
1 1
Fertility Medical Group, IVF Laboratory, São Paulo, Brazil ; Ren Ji Hospital- School of Medicine- Shanghai Jiao Tong University, Center for
2
Fertility Medical Group, Scientific research, São Paulo, Brazil ; Reproductive Medicine, Shanghai, China

Study question: Is the serum SHBG level during controlled ovarian hyperstim- potentially involved in the etiology, prevalence, and modulation of PCOS phe-
ulation (COH) associated with ovarian response? What’s the difference between notypes. Together environmental factors with genetics contribute to the com-
polycystic ovary syndrome (PCOS) and non-PCOS patients? plicated pathology of PCOS. As a strong environmental risk factor, circadian
Summary answer: Serum SHBG levels, especially SHBG on hCG-day, could clock is essentially involved in various diseases. However, the functional role of
be recognized as a predictor of ovarian response during COH in non-PCOS, circadian clock in PCOS remains to be elaborated.
but not in PCOS participants. Study design, size, duration: We tried to elaborate the influence of altered
What is known already: Previous study reports the positive correlation of circadian clock gene expression caused by constant light or darkness exposure
serum SHBG levels with the total number of follicles in infertile women with on traits of PCOS in rats and to explore the effects of 5 different rescue treat-
tubal and/or male-factor who undergo IVF and Recent literature has also ments. We utilized HepG2 cells and KGN cells to verify the molecular mecha-
stressed the importance of SHBG measurement in the diagnosis and manage- nisms. Additionally, human leukocytes and serum of PCOS (n=20) and
ment of PCOS. However, the correlation between serum SHBG levels and non-PCOS (n=20) patients were collected to measure expressions of circadian
ovarian response in COH cycles and the difference between PCOS and non- clock genes and other target genes.
PCOS patients remain unclear and need to be investigated. Participants/materials, setting, methods: The influence of circadian dis-
Study design, size, duration: This is a prospective study. A total of 120 ruption on hallmarks of PCOS in rats and the effects of rescue treatments were
participants undergoing the first IVF cycle were recruited to our study. The serum evaluated by estrus cycle detection, ovary morphology, hormone and metabolic
SHBG levels were detected to analyze the role of SHBG in the prediction of factor measurements. And the expressions of circadian clock genes in different
ovarian response in both PCOS and non-PCOS patients. The duration of this rat models and patients were measured with quantitative PCR and western blot.
study is from August 1st, 2018 to October 1st, 2019. The roles of circadian genes in hyperinsulinemia, hyperandrogenism and apop-
Participants/materials, setting, methods: A total of 120 participants (60 tosis of ovarian granuloma cells were investigated with small interfering RNA-
non-PCOS and 60 PCOS participants) receiving GnRH-antagonist protocol were mediated knockdown and plasmids-mediated overexpression.
recruited to our study. The serum samples were collected every 2 days from Main results and the role of chance: After exposure to constant darkness,
each participant and the concentrations of serum SHBG and sex hormones were decreased brain and muscle ARNT-like protein 1 (BMAL1) promoted insulin
tested to investigate the relationship between serum SHBG levels and ovarian resistance via glucose transporter 4 (GLUT4), and decreased period (PER) 1
response during COH. Additionally, the number of follicles and the endometrium and PER2 promoted androgen excess via insulin-like growth factor-binding pro-
thickness were monitored using B-ultrasound. tein 4 (IGFBP4) and sex hormone binding globulin (SHBG) in the liver.
Main results and the role of chance: Serum SHBG concentrations Hyperinsulinemia and hyperandrogenism shared a bidirectional link promoting
decreased along the COH cycle of GnRH antagonist protocol both in PCOS aberrant expression of circadian genes and inducing apoptosis of ovarian gran-
and non-PCOS participants. Serum SHBG levels on human chorionic gonado- uloma cells. Notably, the altered expressions of circadian clock genes in dark-
tropin (hCG)-day were positively correlated to numbers of retrieved oocyte ness-treated rats matched those of PCOS patients. Furthermore, melatonin
and embryo in all participants. Furthermore, both basal and hCG-day SHBG treatment relieved the hyperinsulinemia and hyperandrogenism of dark-
levels of high responders (>15 retrieved oocytes) significantly increased com- ness-treated rats via BMAL1, PER1, and PER2. Restoring normal light/dark
pared with those of normoresponders (4-15 retrieved oocytes) in the analysis exposure for 2 weeks reversed these conditions via BMAL1.
of all participant (n=120). However, when we analyzed separately, the positive Limitations, reasons for caution: We couldn’t compare circadian genes in
correlation between serum SHBG levels and ovarian response was only observed liver or adipose from women with/without PCOS due to the scarcity of samples.
in non-PCOS participants, without any association in PCOS group. Finally, Although the adipose of rats kept in continual darkness exhibited abnormal
receiver operating characteristic (ROC) curve analysis demonstrated that serum BMAL1 expression, we did not conduct experiments to account for the
SHBG levels on hCG-day could be used as a predictor of ovarian response in unchanged expression in P-AKT pathway under the same conditions.
the non-PCOS group (P=0.0015), but not in the PCOS group (P=0.5142). Wider implications of the findings: Our findings provide a theoretical basis
Limitations, reasons for caution: This study did not investigate the rela- for illustrating the critical function of circadian clock genes, especially BMAL1,
tionship between serum SHBG level and pregnancy outcomes due to the frozen PER1, and PER2 in PCOS, which might aid the development of feasible preventive
embryo transfer protocol. Secondly, we limited the treatment protocol, age, and therapeutic strategies for PCOS in women with biorhythm disorder.
AMH and other indicators in order to control the interfering factors. Thus, the Trial registration number: not applicable
relationship between SHBG and these factors can’t be studied.
Wider implications of the findings: This study provides a means to reassess P-560 Effect of Ovarian Stimulation on IUI Outcomes - Managing
the clinic value of using serum SHBG levels, especially before COH and on the Expectations
hCG-day as an ovarian response predictor in non-PCOS patients. Moreover, it B. Wusk1, F. Roth1, V. Hungerbuehler1
also improves our understanding of the intrinsic mechanisms of folliculogenesis 1
fiore lab, Institute of Reproductive Medicine, St. Gallen, Switzerland
and the difference between PCOS and non-PCOS patients.
Study question: IUI - natural vs FSH stimulation: how much affects an individual
medication the outcome - a single-center, retrospective database analysis over
P-559 The circadian clock gene functions as a novel therapeutic 11 years.
target for constant darkness-induced insulin resistance and Summary answer: Compared to natural cycles, the application of individual
hyperandrogenism of polycystic ovary syndrome FSH regimen led to a significant increase of 32% of βHCG-positive pregnancies
S. Li1, J. Zhai1, W. Chu1, X. Geng1, Z.J. Chen1, Y. Du1 (10.3% vs 13.6%, p≤0.001).
1
Ren Ji Hospital- School of Medicine- Shanghai Jiao Tong University, Center for What is known already: A number of contributing factors during ART lead
Reproductive Medicine, Shanghai, China to the concern that children conceived by ART might be exposed to increased
health risks as compared to naturally conceived children.
Study question: What are the mechanisms underlying metabolic and repro- IUI is a simple and non-invasive technique with minimal monitoring and risks. It
ductive dysfunction caused by arrhythmic circadian clock and their involvement can be performed without expensive infrastructure with a reasonable success
in polycystic ovary syndrome (PCOS)? rate within a few cycles in most centers. IUI is undoubtly a more cost effective
Summary answer: The arrhythmic expressions of circadian clock genes due and patient-friendly strategy than ART (Pennings and Ombelet 2007).
to constant darkness induced the metabolic and reproductive hallmarks of PCOS Study design, size, duration: A single-center, retrospective database analysis
in rats. over 11 years (2008-2019). Exclusion criteria were: HIV, AID, cryopreserved
What is known already: As a complex endocrine condition characterized by sperm, leucospermia. From a total of n=7164 IUI cycles, 6148 (85.8%) were
oligo/anovulation, high androgen levels and polycystic ovaries, PCOS is closely included in the study . The average IUI attempt per couple was 2.9 (2097 couples
associated with genetic susceptibility, and environmental risk factor plays an with 6148 cycles).
important role in the expression of those genetic traits. Environmental toxins, Participants/materials, setting, methods: The study population repre-
diet and nutrition, socioeconomic status, and geography are suggested to be sented daily clinical IUI practice. Average age of the female population has been:

2
34.0+4.6 years. Administration or not of FSH has been expressed as yes or no IVIRMA Abu Dhabi, Statistical department, Abu Dhabi, United Arab Emirates ;
3
(68% vs 32%). Initial progressive motility (59.8±16), increased after gradient University of Copenhagen, Laboratory of Reproductive Biology- The Juliane Marie
density preparation by 32% (78.4±13). Centre for Women- Children and Reproduction-, Copenhagen, Denmark ;
Main results and the role of chance: From 2008-2019, our overall IUI 4
Ansh Labs, Hormonal laboratory, Webster, U.S.A.
pregnancy rate (βHCG+) was 12.6% per cycle. For natural cycles, the pregnancy
rate per heterologous insemination was 10.3% (n=201/1946), whereas for Study question: To evaluate the role of Inhibin A as a potential predictive
stimulated IUI cycles the pregnancy rate (573/4202; 13.6%) was significantly marker of oocyte maturity in ovarian stimulation cycles for IVF / ICSI
increased by 3.3% (p≤0.001). Summary answer: On the day of final oocyte maturation, Inhibin A correlates
Limitations, reasons for caution: Since we used center specific sperm better with the number of retrieved and mature oocytes as compared to E2
preparation procedures (density gradient preparation, constant temperature What is known already: Monitoring of ovarian stimulation for IVF/ICSI by
setting of 34.5°C, HEPES buffer) data comparison with other studies are limited. transvaginal ultrasound (TVUS) and measurement of E2-levels is critical for the
Wider implications of the findings: Here, we add substantial information planning of final oocyte maturation and oocyte retrieval. E2-levels are supra-
how FSH stimulation can affect the outcome of individual IUI cycles and could physiological due to multifollicular growth and do not facilitate an accurate
assist in couple counselling and planning. determination of oocyte maturity. In contrast to E2, Inhibin A levels increase
Trial registration number: not applicable when a minimum follicular size of 12-15mm is attained , the minimal follicular
size required to retrieve a mature oocyte . Therefore, the combination of Inhibin
P-561 The mental health of women with polycystic ovary A measurement and TVUS may present a more reliable predictive parameter
syndrome: a systematic review and meta-analysis of oocyte maturity as compared to E2 plus TVUS.
C. Chan1,2, X. Yin2, Y. Ji2, C. Chan, Lai Wan2 Study design, size, duration: Prospective observational study, performed
1
from September 2018 to January 2019, including data of 145 patients recorded
University of Hong Kong, Centre of Assisted Reproduction and Embryology,
at the beginning of ovarian stimulation and 136 patients on the day of final oocyte
Pokfulam, Hong Kong ;
2
maturation.
The University of Hong Kong, Department of Social Work and Social
Participants/materials, setting, methods: Patients with primary / second-
Administration, Hong Kong, China
ary infertility and an indication for IVF/ICSI, undergoing ovarian stimulation in a
Study question: How is the generalized mental health status of women GnRH-antagonist-protocol, were included. On day 2 / 3, AFC was recorded
with PCOS? and blood samples were taken.Monitoring of the stimulation was performed
Summary answer: The mental health situation of PCOS patients was quite according to clinical routine and blood samples were repeated on the day of
worrisome, which has highlighted the importance of developing psychological final oocyte maturation. For measurement of E2, Inhibin A and Inhibin B, samples
health care interventions for them. were thawed and analysed with the same batch of reagents.
What is known already: PCOS has been proposed to be associated with Main results and the role of chance: Hormonal results and ovarian stim-
several mental health problems, including somatic symptoms, anxiety, depression, ulation parameters from the IVF-treatments were available from 145 patients at
body dissatisfaction, and eating disorders, diminished sexual satisfaction, and stimulation start (day 2/3) and from 136 patients on the trigger day. 9 patients
lowered health-related quality of life, etc. did not undergo oocyte retrieval. Patient characteristics include (mean ± SD):
Study design, size, duration: A systematic review and meta-analysis of age 35.4 ± 6.5 years, BMI 28.2 ± 4.8 kg/m², infertility duration 3.9 ± 3.0 years,
published literature comparing the mental health of women with and without previous stimulations 3.1 ± 4.2. Correlations were calculated using Pearson´s
PCOS. Ten English and Chinese databases were searched up to 12/31/2018. (ρ) coefficient and probability values (p-value).
46 studies, including 30,989 participants (9,265 women with PCOS and 25,638 On the trigger day, a strong correlation (Pearson´s Coefficient) was found
controls), were qualified for review according to the inclusion criteria. between the total follicle number and Inhibin A / E2 (ρ 0.78 / 0.71) and between
Participants/materials, setting, methods: Random effect models were the number of follicles ≥ 15 mm and Inhibin A (ρ 0.72). Pearson´s Coefficient
introduced, and subgroup analysis, sensitivity test, and meta-regression were showed a strong correlation between Inhibin A and the number of retrieved
carried out to determine the source for heterogeneity among studies. and mature oocytes (ρ 0.82 / 0.77), whereas E2 had a moderate correlation
Main results and the role of chance: Twenty-eight studies reported depres- (ρ 0.69 / 0.69) respectively, for these parameters.
sion symptoms, 22 studies were on anxiety, 16 studies showed QoL status, 12 The area under the curve (AUC) for Inhibin A as a predictor for ≥ 10 mature
studies were about sexual dysfunction, five on emotional distress, four on binge oocytes was ρ 0.91 (CI95% [0.87 ; 0.96]) and ρ 0.84 (CI95% [0.7769 ; 0.9124])
eating, and four on somatization. Women with PCOS reported significantly higher for E2. The optimal predicted threshold level of Inhibin A for ≥ 10 mature
depression (SMD=0.64; 95%CI: 0.50-0.78), anxiety (SMD=0.63; 95%CI: 0.50- oocytes was 668.1 pg/mL (sensitivity=88.0%, specificity=82.0%).
0.77), lower QoL (SMD=-0.55; 95%CI: -0.71 to -0.39), and not significant sexual Limitations, reasons for caution: The limitation of the study is that the
dysfunction (SMD=-0.23; 95%CI: -0.51 to 0.04). Studies from different countries, decision, when to administer medication for final oocyte maturation, was based
adopting various diagnosis criteria, using diverse instruments, as well as in different on E2-level and follicle size, as the Inhibin A results were not available and
years, have reported heterogenetic results. Women with PCOS in China reported therefore not used as a decision tool in this setting.
a larger effect size of depression and anxiety than patients from other countries. Wider implications of the findings: As Inhibin A on the trigger day cor-
Limitations, reasons for caution: Although subgroup analysis, meta-regres- relates better as compared to E2 with the number of follicles ≥ 15 mm and the
sion, and sensitivity tests were carried out to determine the source for high number of retrieved and mature oocytes, Inhibin A in combination with ultra-
heterogeneity, only 81.63% of the heterogeneity can be explained by this study. sound monitoring of follicular development may present a promising tool to
There were still unknown sources for heterogeneity among included studies on facilitate planning of oocyte retrieval.
the depression of PCOS patients. Trial registration number: clinicaltrials.gov., number NCT03607409
Wider implications of the findings: PCOS patients suffer from depression,
anxiety, and experience a lower QoL, whereas their sexual function is not distinct P-563 Intra-Uterine Insemination + Controlled ovarian
from that of healthy control. Studies using different instruments, from different hyperstimulation versus In Vitro Fertilization in Unexplained
countries, and adopting different diagnosis criteria, reported heterogenetic Infertility: A systematic review and meta-analysis
results. PCOS patients in China reported a high level of depression and anxiety. A. Nandi1, R. Gangopadhyay2, D. White1, T. El-Toukhy1
Trial registration number: N/A 1
Guy’s and St Thomas’ Hospital NHS Trust, Assisted Conception Unit, London,
United Kingdom ;
P-562 Inhibin A - a promising predictive parameter of oocyte 2
Watford General Hospital- Hertfordshire- UK, Department of Obstetrics and
maturity in ovarian stimulation for IVF/ICSI
Gynaecology, Wembley, United Kingdom
B. Lawrenz1, L. Depret Bixio2, C. Coughlan1, C. Andersen3,
B. Kalra4, G. Savjani4, L. Melado1, H. Fatemi1, A. Kumar4 Study question: What is the relative efficacy of IUI+COH compared with IVF
1
IVIRMA Abu Dhabi, IVF department, Abu Dhabi, United Arab Emirates ; in couples with unexplained infertility?

Summary answer: Results showed a higher success rate following IVF com- Participants/materials, setting, methods: Age-dependant comparison of
pared to IUI+COH, particularly in older women, but no difference in agonist vs antagonist protocols have revealed a slight to distinct increase of
younger women. pregnancy rates in favor of agonistic stimulation.
What is known already: Unexplained infertility is a diagnosis of exclusion
when all standard fertility investigations, namely tests of ovulation, tubal patency
and semen analysis, are normal. The two most applied treatment options are
intra-uterine insemination and IVF. However, there is an ongoing debate on the age βHCG+ (%) βHCG+ (%)
n n p value
effectiveness of the treatments and the United Kingdom (UK) National Institute group agonist antagonist
for Health and Care Excellence (NICE) fertility guidelines recommend IVF treat-
ment rather than IUI after 2 years of expectant management. We sought to 26-30y 53.2 62 52.7 203 ns
conduct a systematic review of existing evidence to assess the relative efficacy 31-35y 53.2 188 49.6 470 ns
of the two treatments.
Study design, size, duration: We searched Medline, Embase, CIHNL, Pscy 36-40y 51.6 153 39.9 592 p=0.009
Info, and Cochrane Library from 1980 till November 2019. Only RCTs comparing 41-45y 42.1 19 23.4 273 p=0.07
IUI + COH (using either clomiphene citrate or injectable gonadotropins or both)
with IVF with female patients aged 18 - 43 years and diagnosed with unexplained
infertility were included for the systematic review.
Two authors reviewed citations from primary search independently and any Main results and the role of chance: The agonist protocol yields in higher
disagreement was resolved by mutual discussion and consultation with a third pregancy rates (βHCG+) in females ≥35 years as compared to the antagonist
author.Participants/materials, setting, methods: The titles and abstracts protocol, whereas in age groups ≤ 34 years no differences were observed.
were scrutinised to identify relevant articles. The full texts of all potentially Limitations, reasons for caution: The retrospective character of the analysis
relevant articles were retrieved and reviewed to identify articles, which fulfil the limits the statement of a general recommendation.
study inclusion criteria. Study characteristics including study type and setting, Wider implications of the findings: Age-related administration of GnRH
inclusion criteria, recruitment procedure, intervention and outcome data were should be confirmed by prospective, randomized, clinical studies.
extracted. All included studies were then assessed for trial quality as per Trial registration number: not applicable
Cochrane guidelines. From each primary study, data were pooled and analysed
using Revman software 5.2 versions. P-565 Randomised control trial comparing the effects of
Main results and the role of chance: In total eight RCTs were included. The myoinositol to metformin on ART outcome in women with
quality of evidence was moderate to low quality due to inconsistency across the PCOS undergoing In-vitro fertilisation (IVF) cycle
trials and imprecision. The pooled result showed that IVF was associated with a K. Rajasekaran1, N. Malhotra1
statistically significant higher live birth rate (RR 1.53, 95% CI 1.01 – 2.32, P <0.00001 1
All India Institute of medical sciences AIIMS, Department of obstetrics and
I2 = 86%) with no significant difference in multiple pregnancy rate or OHSS rate.
gynaecology, new delhi, India
Sensitivity analysis based on women’s age and a history of previous IUI or IVF
treatment showed no significant difference in the live birth rates (RR 1.01, 95% CI Study question: Does Myoinositol in comparison to metformin in infertile
0.88 -1.15, I2 = 0%, 3 RCTs) between the two treatments in treatment-naïve women PCOS women, improves the ART outcomes, oocyte quality, pregnancy rates
younger than 38 years. In women over 38 years, the live birth rates were significantly and decreases the incidence of OHSS
higher in the IVF group (RR 2.15, 95% CI 1.16 – 4.0, I2 = 42%, 1 RCT). Summary answer: Myoinositol reduces unsuitable oocytes, improves the
Limitations, reasons for caution: The main limitation encountered in this menstrual pattern, insulin resistance, hormone profile, quality of embryos, IVF
review was the considerable clinical and methodological heterogeneity present outcomes and clinical pregnancy without significant change in OHSS.
among the primary studies, including differences in the definition of unexplained What is known already: Myoinositol and metformin are insulin sensitisers.
infertility, study design and inclusion criteria as well as treatment protocols used Metformin reduces testosterone concentrations, optimises IVF outcome, reduces
for IUI+COH and IVF. risk of OHSS in PCOS but is associated with gastrointestinal side effects and
Wider implications of the findings: Our study included only direct com- lactic acidosis resulting in reduced patient’s compliance. Myoinositol improves
parisons between IUI+COH and IVF as they are the most commonly applied ovarian function, oocyte quality, LH/FSH ratio, reduces serum androgens,
treatment and showed overall higher success with IVF but no difference in young improves ovulation and fertility outcome without the side effects of metformin.
women <38 years. This will certainly enable clinicians to individualise the care Myoinositol acts at the level of insulin receptors and improves hyperinsulinemia,
to these couples. positively correlate with quality and maturity of oocytes reducing mean number
Trial registration number: not applicable of immature and degenerated oocytes, quantity of gonadotropins or units of
FSH necessary for ovarian stimulation in IVF protocols.
P-564 Agonist vs antagonist administration: is there an age-related Study design, size, duration: The present study is a randomised controlled
difference? A retrospective analysis trial conducted after the ethical committee approval, in the ART Clinic, depart-
F. Roth1, B. Wusk1, V. Hungerbuehler1 ment of Obstetrics and Gynaecology, All India Institute of Medical sciences,
1
FIORE, IVF Lab, St. Gallen, Switzerland New Delhi from March 2017 to March 2019. A total of 231 patients infertile
PCOS women undergoing IVF cycles were enrolled in the study and asses for
Study question: Agonist vs antagonist: Is there a best practice/algorithm to eligibility and 102 were randomised and allotted as 50 and 52 in group
suggest for a specific age group of the female idiopathic patient? 1(Myoinositol) and group 2(Metformin) respectively.
Summary answer: In the age group of ≥35 years, a significantly increased Participants/materials, setting, methods: Recruited patients received
pregnancy rate (βHCG+) for the agonist protocol has been observed. myoinositol 2gm twice daily(group1) and metformin 850mg twice daily(group
What is known already: Controlled ovarian hyperstimulation (COH) is a 2). Pre and post treatment clinical(menstrual pattern,BMI), hormonal profile(L-
fundamental step in ART. GnRH antagonist stimulation protocol has shorter H,FSH,Testosterone, prolactin, AMH), biochemical parameters(HOMA IR,
duration of treatment and reduced administration of gonadotropins. GnRH fasting glucose, insulin) and side effect profile assessed. After 3 months of ther-
agoinst protocol has shown to be better in folliculogenesis and pregnancy rate, apy, patients were recruited for IVF cycles involving controlled ovarian stimula-
despite its costly and time-consuming. tion, cycle monitoring, oocyte recovery, insemination of oocytes and follow up
Study design, size, duration: A single-center, retrospetive database analysis with fertilisation, cleavage, transfer of good grade cleavage embryos or blasto-
of 1538 antagonist- and 422 agonist protocols over a 10y year period (2009- cysts pregnancy outcomes and OHSS incidence
2019) has been analyzed. Female patients have been divided into four age groups: Main results and the role of chance: Myoinositol had significant
26-30 years; 31-35 years; 36-40 years and 41-45 years. Excluded were cases of: fertilisation rate(p<0.001), embryos retrieved(p<0.001), cleavage rate(p 0.008),
disovulation, endometriosis and TESE, MESA or cryopreserved sperm. grade 1 embryos(p 0.042) with no significant difference in implantation

rate(p 0.537),number of oocytes retrieved(p0.097),metaphase II oocytes(p 0.176) Main results and the role of chance: The primary objective of the trial was
and Grade1 oocyte(p 0.204). Embryo transfer cancelled was significantly high in met, as non-inferiority was established for the number of oocytes retrieved for
metformin group(p 0.035). Starting dose of FSH, total dose of FSH and duration follitropin delta compared to follitropin beta (9.3 vs 10.5; LL 95% CI 2.3). Among
of gonadotropin stimulation were comparable. patients with AMH <15 pmol/L (40.9%), the proportion of women with <4
Clinical pregnancy rate was significantly high in Myoinositol group 36.0% oocytes was similar in the two treatment groups (11.6% vs 12.3%). Among
(18/50); 95% CI 18.0(0.9,35.0) p value 0.043) compared to metformin(18% patients with AMH ≥15 pmol/L (59.1%), treatment with follitropin delta resulted
(9/50)). The spontaneous pregnancy rate were comparable, but Group 1 had in significantly (p<0.05) fewer patients with ≥15 oocytes (22.0% vs 42.0%) or
higher number of patients conceived spontaneously(13,26%) compared to ≥20 oocytes (8.0% vs 19.0%). Blastocyst transfer was performed for 79.4% and
Group 2(6,12%) (p value 0.074).The total number of IVF pregnancy rates ( p 79.7% in the follitropin delta and follitropin beta groups, respectively, and the
value 0.715), total number of IVF pregnancy rate per number of IVF cycles (p clinical pregnancy rate for these women was 31.9% and 29.8%, respectively. The
value 0.314) and the total number of IVF pregnancy rate per embryo transfer clinical pregnancy rate per started cycle was 25.3% for follitropin delta and 23.7%
(p value 0.368) were also comparable between the Groups. Incidence of OHSS for follitropin beta. The occurrence of OHSS was reduced to approximately half
was 10% (5/50) and 20% (10/50) in Group 1 and Group 2 respectively, com- with follitropin delta compared to follitropin beta, with an incidence of 11.2%
parable between both Myoinositol and Metformin Group (p value 0.001). vs 19.8% (p<0.05) for OHSS of any grade, and 7.1% vs 14.1% (p<0.05) for
Myoinositol group had increased regularity in menstrual pattern(0.001) and moderate/severe OHSS. Two patients in the follitropin beta group were hos-
improvement in fasting insulin (p 0.001),HOMA IR (p 0.001),Serum AMH(p pitalised due to OHSS for a duration of 16 and 33 days, respectively, while there
0.001) and Serum SHBG(p 0.032) thereby suggesting decreased insulin resistance. were no hospitalisations in the follitropin delta group.
Limitations, reasons for caution: Most patients with PCOS recruited in our Limitations, reasons for caution: The individualised follitropin delta dosing
study were brittle PCOS, with high LH, AMH and hyperandrogenemia with regimen based on the patient’s serum AMH and body weight was developed
sub-optimal response during ovulation induction and IUI. In IVF cycles only fresh after completion of a phase 2 trial in non-Japanese women. Based on a subse-
embryo transfer and not FET cycles were analysed. These were attributed to quent phase 2 trial conducted in Japan, the individualised dosing regimen was
be the reason for lower Pregnancy outcomes. evaluated to also be applicable to Japanese women.
Wider implications of the findings: Myoinositol in comparison to met- Wider implications of the findings: This trial confirms that the ovarian
formin in infertile PCOS women has better IVF outcome and oocyte quality, response associated with the individualised dosing of follitropin delta preserves
however to make these results more robust, a large multi-centric trial with larger pregnancy rates and results in an improved OHSS risk profile for Japanese IVF/
sample size and longer duration is necessary. ICSI patients as already observed in non-Japanese women.
Trial registration number: CTRI/2018/05/014196 Trial registration number: NCT03228680
P-566 A randomised, assessor-blind, controlled phase 3 non- P-567 Midfollicular gonadotropin dose increases do not appear to
inferiority trial assessing the efficacy and safety of individualised change the ovarian response determined by the dose selected at
follitropin delta dosing regimen in Japanese IVF/ICSI patients the start of the stimulation
O. Ishihara1, M. Kitamura2, J.C. Arce3, X. Japanese Follitropin J.C. Arce1, B. Klein2
Delta Phase 3 Study Group4 1
Ferring Pharmaceuticals, Reproductive Medicine & Maternal Health, Copenhagen,
1
Saitama Medical University, Department of Obstetrics & Gynaecology, Denmark ;
Iruma-gun- Saitama, Japan ; 2
Ferring Pharmaceuticals, Global Biometrics, Copenhagen, Denmark
2
Ferring Pharmaceuticals, Development, Tokyo, Japan ;
3
Ferring Pharmaceuticals, Reproductive Medicine & Maternal Health, Copenhagen, Study question: Can gonadotropin dose increases during stimulation lead to
Denmark ; more oocytes or blastocysts compared to maintaining a fixed dose selected
4
On behalf of the participating sites, Fertility clinics, Japan, Japan based on biomarkers and patient characteristics?
Summary answer: Gonadotropin dose increases in the midfollicular phase
Study question: To establish the efficacy and safety of controlled ovarian do not appear to alter the ovarian response or blastocyst availability defined by
stimulation with follitropin delta individualised fixed-dose regimen versus folli- the starting dose.
tropin beta conventional dosing in Japanese women. What is known already: Studies have indicated that increasing the gonado-
Summary answer: Individualised treatment with follitropin delta versus follitropin dose after the first 5 days (or later) in anticipation of low ovarian response
tropin beta resulted in non-inferiority in number of oocytes retrieved, compa- does not rectify the response (van Hooff et al, 1993; Khalef et al, 2002). As
rable clinical pregnancy rates and significant reduction of OHSS. follicular recruitment occurs in the late luteal and early follicular phase, such dose
What is known already: The follitropin delta dosing regimen stratifies patients increases during the stimulation cycle does not lead to increasing the number of
according to response potential and applies an individualised dose based on an oocytes retrieved.
ovarian biomarker (AMH) and a patient characteristic (body weight). This Study design, size, duration: Post-hoc analysis of a randomised, asses-
approach has in previous clinical trials conducted outside of Japan been demon- sor-blind, controlled trial in 1,326 women, 18-40 years, undergoing their first
strated to optimise ovarian response and reduce OHSS risk without compro- IVF/ICSI cycle. Patients randomised to follitropin alfa (GONAL-F, Merck)
mising success rates. (N=661) received a daily starting dose of 150 IU for the first five days which
Study design, size, duration: Randomised, assessor-blind, controlled trial thereafter could be adjusted by 75 IU. Patients randomised to follitropin delta
conducted in 347 Japanese women, 20-40 years (mean 34.1), undergoing their (REKOVELLE, Ferring Pharmaceuticals) (N=665) received an individualised dose
first IVF/ICSI cycle. Randomisation was stratified by AMH at screening (<15 based on AMH and body weight, and the dose was fixed throughout stimulation.
pmol/L, ≥15 pmol/L). The primary endpoint was number of oocytes retrieved Participants/materials, setting, methods: Investigators were blinded to
with a prespecified non-inferiority margin (lower limit of 95% confidence interval treatment allocation. The frequency of investigator-requested dose increases
(LL 95% CI) >-3.0 oocytes). A GnRH antagonist protocol was applied. Clinical on stimulation day 6 was similar in the two treatment groups, with 24.5% (n=162)
pregnancy was assessed 5-6 weeks after single blastocyst transfer. OHSS was in the follitropin alfa group and 22.6% (n=150) in the follitropin delta group. The
classified using Golan’s system. investigator’s requests for gonadotropin dose adjustments were only imple-
Participants/materials, setting, methods: The follitropin delta mented in the follitropin alfa group, while patients in the follitropin delta group
(REKOVELLE, Ferring Pharmaceuticals) dose was individualised based on serum remained on the assigned daily dose.
AMH and body weight, and was fixed throughout stimulation – AMH <15 Main results and the role of chance: The follicular development and serum
pmol/L: daily dose of 12 µg, AMH ≥15 pmol/L: daily dose decreasing from 0.19 endocrine profile on stimulation day 6 were comparable between the women
to 0.10 µg/kg body weight by increasing AMH (min-max 6-12 µg). Elecsys® in the follitropin alfa and follitropin delta groups for whom the investigator rec-
AMH, Roche Diagnostics was used. The follitropin beta (FOLLISTIM/ ommended to increase the dose. These women had 12.3±5.8 and 12.8±6.7
PUREGON, MSD) dose was 150 IU/day for the first five days and could there- follicles in the follitropin alfa and follitropin delta groups, respectively, of which
after be adjusted. there were 4.8±3.6 and 4.2±3.4 follicles ≥10mm. The median [interquartile

range] estradiol was 1509 [898;2264] pmol/L for follitropin alfa and 1228 pregnant women. We detected expression of 17BHSD1,3,5.7.12., star, CYP11A1,
[762;2076] pmol/L for follitropin delta, while inhibin B was 434 [263;706] pg/ CYP17A1, CYP21A1, CYP19A1,3BHSD1-2.11BHSD1-2 in the subcutaneous adi-
mL and 430 [282;637] pg/mL, respectively. Among the women with investiga- pose tissue of pregnant women but 17BHSD2 was undetectable in both groups.
tor-requested dose increases, the ovarian response and blastocyst development PCOS subcutaneous AT showed significantly higher level of CYP11A1, CYP17A1,
were comparable between the follitropin alfa patients with dose increases and CYP21A1, STAR and 11BHSD1-2 mRNA when compared with the non-PCOS
the follitropin delta patients on a fixed dose. For follitropin alfa and follitropin women (P < 0.05). There was no significant difference between
delta, respectively, these women had 7.4±5.0 versus 7.4±5.1 oocytes retrieved, CYP19A1,3BHSD1-2. 17BHSD1,3, 5.7.12. mRNA abundance between two group.
2.5±2.4 versus 2.3±2.4 blastocysts, and 1.3±1.7 versus 1.3±1.9 good-quality Limitations, reasons for caution: Although we collected 48 samples during
blastocysts. The dose increases in the follitropin alfa group logically influenced one year, the small sample size is a limitation. Nevertheless, the strength of the
the gonadotropin consumption. The mean daily gonadotropin dose in this observed differences in aged-matched and BMI-matched subjects suggests that
sub-population was 196±26 IU (14.4±1.9 µg) follitropin alfa versus 10.2±2.4 µg power was not an important issue and that a larger study would be able to detect
follitropin delta, with a mean total gonadotropin dose of 1865±607 IU (137±45 same findings.
µg) follitropin alfa versus 99±26 µg follitropin delta. Wider implications of the findings: This is the first report for gene expres-
Limitations, reasons for caution: Although the population with investiga- sion profiles in AT of PCOS mothers which opens the road for exploration
tor-requested dose increases was similar in the follitropin alfa and follitropin regarding gene expression related glucocorticoids and mineralocorticoids metab-
delta groups, it should be noted that the decision to request an increase of the olism in AT for pregnant women suffering from PCOS.
starting gonadotropin dose was per the investigator’s judgement and not based Trial registration number: enter ‘not applicable’ for non-clinical trials
on protocol-specific criteria.
Wider implications of the findings: These findings stress the importance P-569 A multicenter and multinational non-interventional study
of appropriate dose selection before starting stimulation. The data indicate that exploring the POSEIDON criteria – An analysis of 11042 cycles
increasing the dose during stimulation does not modify the ovarian response that using real-world data from Brazil, Turkey, and Vietnam.
can be obtained with a fixed-dose regimen. Maintaining a fixed-dose regimen S. Esteves1,2, H. Yarali3, L. Vuong4, F. Bento1, I. Özbek3, P. Mehtap3,
throughout treatment is convenient for both the patient and clinic staff. A.H. Le5, T. Pham5, T. Ho5, P. Humaidan2,6, C. Alviggi7
ANDROFERT, Andrology & Human Reproduction Clinic, Campinas, Brazil ;
2
Faculty of Health- Aarhus University, Department of Clinical Sciences, Aarhus,
P-568 differences between steroids gene expression in
Denmark ;
subcutaneous fat of PCOS and non-PCOS pregnant women 3
Anatolia IVF, Endocrinology, Ankara, Turkey ;
N. Emami1, A. Alizadeh1, A. Moini2, M. Shahhosseini3 4
University of Medicine and Pharmacy at Ho Chi Minh City, Department of
1
Royan Institute for Reproductive Biomedicine- ACECR- Tehran- Iran, Department Obstetrics and Gynecology, Ho Chi Minh, Vietnam ;
5
of Embryology- Reproductive Biomedicine Research Center, tehran, Iran ; My Duc Hospital, IVF Unit, Ho Chi Minh, Vietnam ;
2 6
Royan Institute for Reproductive Biomedicine- ACECR- Tehran- Iran, Department Skive Regional Hospital, Fertility Clinic, Skive, Denmark ;
7
of Endocrinology and Female Infertility- Reproductive Biomedicine Research University of Naples Federico II, Department of Neuroscience- Reproductive
Center-Breast Disease Research Center BDRC- Tehran University of Medical Science and Odontostomatology, Naples, Italy
Sciences- Tehran- Iran- De, ;
3
Royan Institute for Reproductive Biomedicine- ACECR- Tehran- Iran-, Department Study question: What is the cumulative live birth rate per started cycle (CLBR)
of Genetics- Reproductive Biomedicine Research Center- Royan Institute for in low prognosis women stratified according to the POSEIDON criteria?
Reproductive Biomedicine- ACECR- Tehran- Iran- Department of Cell and Summary answer: CLBR varies in POSEIDON groups (48% in G1, 24% in
Molecular Biology- School o, G2, 29% in G3, 10% in G4), impacted by female age and number of oocytes.
What is known already: The POSEIDON criteria have been introduced as
Study question: Are there differences between gene expression related to a new classification for low-prognosis women undergoing assisted reproductive
steroid metabolism in abdominal subcutaneous AT in PCOS and non- PCOS technology (ART). The low prognosis patient is classified into four groups accord-
pregnant women? ing to the results of ovarian reserve markers, female age, and the number of
Summary answer: PCOS SAT(subcutaneous adipose) showed significantly oocytes retrieved in previous IVF/ICSI cycles of conventional ovarian stimulation
higher level of those gene incorporated in glococorticoids and mineralocorticoids (OS). The low prognosis is supposedly caused by the impact of a decreased
but not sex steroids genes number of oocytes, which limits the number of embryos produced. This con-
What is known already: A series of steroidogenic and steroid-inactivating dition might be aggravated by advanced female age, thus negatively impacting
enzymes have been detected in human adipose tissue. The abnormal conversion the CLBR per started cycle.
of androgen precursors to both active and inactive forms at the receptor level is Study design, size, duration: This study analyzed the data of 11,042 con-
important for the pathogenic of metabolic diseases found in PCOS..Most recently, secutive infertile couples who initiated IVF/ICSI treatment in three large Fertility
it has been suggested that adipose tissue may contain the steroidogenic machinery Centers in Brazil, Turkey, and Vietnam between 2015 and 2017. Ovarian reserve
necessary for the initiation of steroid biosynthesis de novo from cholesterol. was assessed before the cycle by either anti-Mullerian hormone (AMH) or antral
Study design, size, duration: . Samples and demographic data were collected follicle count (AFC), or both. The included patients were treated using conven-
from three hospitals in Tehran, Iran. The Subjects were 45 Iranian pregnant tional OS protocols.
women underwent cesarean section; 13 PCOS (poly cystic ovarian syndrome) Participants/materials, setting, methods: Patients were stratified into five
and 32 were non–PCOS, whose samples from subcutaneous fat of abdomen groups based on female age, ovarian markers, and number of oocytes retrieved,
were taken.Diagnosis of these PCOS subjects is responsibility of the medical including the previously described low prognosis POSEIDON groups (Groups
practitioner related to Royan Institute.At the time of caesarean section, the 1-4), and a group of patients not fulfilling the criteria (Non-POSEIDON; NP).
surgeon took 3-4 g subcutaneous fat upon entering the abdomen were collected . The primary outcome was the CLBR per started cycle, including all fresh and
Participants/materials, setting, methods: Total RNA extraction and frozen-thawed embryo transfers. The secondary outcomes were the number
removal of genomic DNA from the adipose tissue were performed using the of embryos per group and the prevalence of low prognosis patients in dataset.
RNeasy lipid Mini Kit (Qiagen, cat. no: 74004).Twenty nanograms of total RNA Main results and the role of chance: The CLBR of patients fulfilling the
was used for cDNA synthesis according to manufacturer’s instruction. Messenger POSEIDON criteria was, on average, ~30% lower than NP patients. Younger
RNA quantification was performed by qRT-PCR on the Step-One RT-PCR sys- unexpected poor/suboptimal responders (G1; n=1,480) and expected poor
tem (Applied Biosystems, USA).Gene expression data were analyzed using 2− responders (G3; n=852) had a CLBR of ~48% and ~29%, respectively
ΔΔCt algorithm to calculate the 15 steroids gene mRNA level relative to the (p<0.0001). The CLBRs of older unexpected poor/suboptimal responders (G2;
level of GAPDH mRNA level. n=1,726) and expected poor responders (G4; n=1,711) were ~24% and ~10%,
Main results and the role of chance: No significant differences were found respectively (p=0.001). Overall, the CLBR of NP patients (59.2%; n=5,273) was
with respect to age and body mass index (BMI) among non-PCOS and PCOS higher than POSEIDON patients (p<0.0001). Younger (G1b; n=1,214) and older

(G2b; n=1,301) unexpected suboptimal responder (4-9 oocytes) patients had P-571 Is ovarian response related to adverse perinatal outcomes
a CLBR of ~54% and 28%, respectively, which were higher (p=0.0001) than that after fresh embryo transfer in GnRH antagonist downregulated
of unexpected poor responder (1-4 oocytes) patients (G1a: 23%, n=266; G2a: IVF/ICSI treatment cycles?
12%, n=425). The number of embryos varied among the POSEIDON groups, E. Bardhi1, C. Blockeel1, W. Cools2, S. Santos-Ribeiro3,
impacted by the number of retrieved oocytes, and were overall lower than that A. Racca1, S. Mackens1, M. De Vos1, N. Polyzos4, B. Popovic-
of NP patients (p<0.0001). POSEIDON patients represented ~50% of all Todorovic1, M. De Brucker1, H. Tournaye1, P. Drakopoulos1
treated patients (Brazil: 64%; Turkey: 55%; Vietnam: 40%), with regional differ- 1
Uz Brussel, Centrum voor Reproductieve Geneeskunde CRG, Avenue du Laerbeek
ences primarily owing to the average age of treated women. 101- 1090 Jette, Belgium ;
Limitations, reasons for caution: Retrospective study design, with different 2
Uz Brussel, Interfaculty Center Data processing & Statistics ICDS, Avenue du
practices in participating centers, might influence cycle cancellation, oocyte yield, Laerbeek 101- 1090 Jette, Belgium ;
number of embryos, and therefore, CLBR. However, our data provide the first 3
IVI-RMA Lisboa-, Instituto Valenciano de Infertilidade, Lisboa, Portugal ;
multicenter real-world evidence to validate the concept of low prognosis –in 4
Dexeus University Hospital- Salud de la Mujer Dexeus, Department of
terms of CLBR per started cycle– in POSEIDON patients. Reproductive Medicine, Barcelona-, Spain
Wider implications of the findings: The CLBR per cycle in POSEIDON
patients is affected by both female age and number of oocytes retrieved. Thus, Study question: Is there an association between ovarian response, as
in addition to aid in patient counseling, the POSEIDON criteria may be used to expressed by the number of oocytes retrieved, and adverse perinatal outcomes?
guide clinical management with a specific focus on therapeutic strategies focused Summary answer: Ovarian response is not related to adverse perinatal
on optimizing the oocyte yield. outcomes.
Trial registration number: NA What is known already: Several studies have found that singleton IVF preg-
nancies are at higher risk for adverse perinatal outcomes compared to naturally
P-570 Levothyroxine and subclinical hypothyroidism in patients conceived ones. Possible explanations for this include infertility itself, embryo
with recurrent pregnancy loss specific epigenetic modifications due to embryo culture and vascular endothelial
dysfunction associated with ovarian ageing. However, one of the most accredited
H. Yoshihara1, M. Sugiura-Ogasawara1, S. Goto1, K. Tamao1
1
hypothesis for these findings, blames the altered endometrium function resulting
Nagoya City University Graduate School of Medical Sciences, Obstetrics and from the non-physiological endocrine milieu that follows ovarian stimulation, which
Gynecology, Nagoya, Japan in turn can be related to ovarian response. Up to date, the safety of ART in terms
of obstetrical and neonatal outcomes still remains under investigation.
Study question: It was unclear whether subclinical hypothyroidism (SCH) Study design, size, duration: This was a retrospective, single-center cohort
affects subsequent live births and whether levothyroxine is effective in improving study including infertile women attending a tertiary university center from January
the live birth rate in patients with recurrent pregnancy loss (RPL). 2009 to December 2015.
Summary answer: SCH did not affect live births and levothyroxine did not Participants/materials, setting, methods: All women who underwent
improve the live birth rate. their first ovarian stimulation cycle in a GnRH antagonist protocol, with a fresh
What is known already: It has been reported that the frequency of miscar- embryo transfer that resulted in a singleton live birth, were eligible for this study.
riage was significantly higher in women with SCH as compared with women on Patients were categorized into four ovarian response groups according to the
thyroid-stimulating hormone (TSH) < = 2.5 mIU/L and that intervention reduces number of retrieved oocytes: 1–3 (category I), 4–9 (category II), 10–15 (cate-
the adverse pregnancy outcome in antithyroid peroxidase antibody (TPO)- gory III), or > 15 oocytes (category IV).
positive women with SCH. Main results and the role of chance: The overall number of patients ana-
One study showed that out of 286 patients with two or more early losses, lyzed was 964: 67 in category I, 450 in category II, 269 in category III and 178 in
19% had SCH, however, that there was no difference in the subsequent live birth category IV.
rate between patients with SCH and euthyroidism (69% and 74%), either with Neonatal weight (in grams) was comparable between all groups (3222 ± 607g
or without treatment for SCH (71% and 67%). vs. 3254 ± 537g vs. 3235 ± 575g vs. 3200 ± 622g, P=0.85) as well as the term
Study design, size, duration: An observational cohort study was conducted of delivery (38.4 ± 2.4 vs 39 ± 1.7 vs 39 ± 2.0 vs 38.6 ± 2.4 weeks, P=0.5).
at Nagoya City University Hospital between 2010 and 2019. The study included Birth weight z-scores and neonatal gender did not differ among the four oocyte
1566 pregnancies of 1120 patients with a history of 2 or more pregnancy losses. categories (P=0.9 and P=0.17, respectively). The incidence of preterm birth and
Participants/materials, setting, methods: 4D-Ultrasound, hysterosalpin- low birth weight was also comparable across the different oocyte categories
gography, chromosome analysis for both partners, antiphospholipid antibodies (P=0.127 and P=0.19, respectively). Finally, gestational diabetes, hypertension
and blood tests for TSH, free thyroxine (FT4) and diabetes mellitus were per- disorders of pregnancy, placenta previa, intra-uterine growth restriction, pre-
formed before a subsequent pregnancy. SCH was defined as having a serum mature rupture of membranes, hospitalization for threatened preterm labor,
TSH > 2.5 mIU/L with a normal level of FT4. Live birth rates were compared delivery mode, APGAR scores at 1, 5 and 10 minutes and major fetal malfor-
between SCH and euthyroid patients treated with and without levothyroxine mations did not differ significantly among ovarian response categories either.
after excluding patients with an abnormal chromosome in either partner and Multivariate regression analysis (adjusting for female age, body mass index (BMI),
those with a uterine anomaly. initial stimulation dose, type of insemination procedure), number of embryos
Main results and the role of chance: The prevalence of SCH was 15.2 % transferred and embryo stage at transfer, revealed that the number of oocytes
(170/1120). Subsequent live birth rates were 79.2 % (42/53) for the levothy- was not associated with neonatal birth weight.
roxine group, 67.6 % (73/108) for the untreated SCH group and 69.6 % Limitations, reasons for caution: This is a large observational study based
(606/871) for the euthyroid group. After excluding miscarriages with embryonic on retrospective data collection. Despite our robust methodological approach,
aneuploidy, chemical pregnancies and ectopic pregnancies, live birth rates were the presence of biases related to retrospective design cannot be excluded.
91.3 % (42/46) for the levothyroxine group, 90.1 % (73/81) for the untreated Wider implications of the findings: The data of our study are reassuring and
SCH group and 90.9 % (606/667) for the euthyroid group. The live birth rates refute potential fears of a negative impact from enhanced ovarian response on
per pregnancy were 93.5% (58/62), 88.9% (96/108) and 90.3% (833/922), neonatal outcomes. Future, larger investigations are needed to validate these results.
respectively. There was no significant difference in the live birth rate among the Trial registration number: not applicable
three groups.
Limitations, reasons for caution: The sample size of SCH patients was
relatively small. Further RCT is necessary to confirm these results. P-572 17β-oestradiol/nomegestrol pre-treatment for ovarian
Wider implications of the findings: The measurement of TSH and FT4 stimulation GnRH antagonist protocols do not affect clinical
might not be necessary for the screening of patients with RPL who have no pregnancy rates.
features of hypothyroidism. M. Samama1,2,3, R.C.P. Piscopo4, F. Ikeda2,3, V.H. Kussumoto2,
Trial registration number: not applicable A. Sartor3,5, L. Semião-Francisco3, J. Ueno2,3, Z. Jarmy_Di Bella6

1
Escola Paulista de Medicina UNIFESP-EPM- Department of Gynecology- Sao P-573 Anti-Müllerian hormone (AMH) lower reference values
Paulo- Brazil., post-doctoral research trainee Department of Gynecology, observed in a population of Indian women compared to French
Sao Paulo / SP, Brazil ; women, using an automated AMH assay
2
Gera Institute, Post-graduation, São Paulo, Brazil ; B. Mougin1, J. Kaur2, P. Bourron1, S. Bellvert3, M. Loubet3,
3
Gera Institute, Clinical, São Paulo, Brazil ; N. Mahajan2
4
Gera Institute, Post Graduation, São Paulo, Brazil ; 1
5
Biomérieux SA, Medical Affairs, Marcy l’Etoile, France ;
Gera Institute, Post-graduation, São Paulo, British Virgin Islands ; 2
6
Mother & Child Hospital, Fertility Clinic, New Delhi, India ;
Escola Paulista de Medicina UNIFESP-EPM- Department of Gynecology- Sao 3
Biomérieux SA, Clinical Affairs, Marcy l’Etoile, France
Paulo- Brazil., Professor Head of the Division of Department of General
Gynecology, São Paulo, Brazil Study question: How much do reference VIDAS® AMH values differ for
women between India and France?
Study question: Whether pre-treatment with an oral contraceptive pill with Summary answer: Significantly lower serum AMH levels were observed for
natural oestrogen, 17β-oestradiol/nomegestrol, has effects on hormonal, women aged 20 to 44 living in India compared to women living in France.
embryological and clinical outcomes for women undergoing assisted reproduc- What is known already: Serum AMH testing is routinely performed in female
tive techniques. patients for the assessment of the ovarian reserve. Several published studies
Summary answer: 17β-oestradiol/nomegestrol favor top quality embryo have reported differences in age-specific AMH reference values in various coun-
formation and was not associated with deleterious effects on biochemical and tries. Reference intervals have been previously defined for VIDAS® AMH, in
clinical pregnancy. populations living in France and in India.
What is known already: Pre-treatments with combined oral contraceptive Study design, size, duration: This study compared the age-specific reference
pill (COCP) suppress the woman’s own hormones production and could improve VIDAS® AMH values defined during dedicated studies performed in France and
ovarian response to the hormone therapy in in vitro fertilization cycles. In the in India, for the 20-44 year age range.
GnRH antagonist protocols with synthetic oestrogen combined oral contracep- Participants/materials, setting, methods: Healthy fertile women with no
tive pill pre-treatment, the rate of live birth/ongoing pregnancy was lower than endocrine or gynaecological disorders were enrolled in the studies conducted
with no pre-treatment. in India and France. Serum AMH concentrations were measured using the
Study design, size, duration: Cohort study in an assisted reproduction tech- bioMérieux VIDAS® AMH assay, and comparative analyses were done for the
niques (ART) post-graduation program setting and no pre-treatment patients 20-44 year age range, for 748 Indian and 356 French women. Descriptive sta-
set in an assisted reproductive center in Sao Paulo, Brazil, which ran from 2017 tistics and linear regression modelling after log transformation were used to
to 2019, with a total number of patients of 130. estimate the differences.
Participants/materials, setting, methods: Three groups were analyzed: Main results and the role of chance: Overall lower median AMH values
no pre-treatment patients were recruited from the ART center, patients from were observed for India with calculated differences of 1.52, 0.50, 1.59, 0.54 and
the post-graduation program received natural oestrogen combined oral contra- 0.19 ng/mL for the five age classes covering the 20-44 year age range, respec-
ceptive pill and were compared to a synthetic COCP group, whose data was tively. The equations for the linear regression models were the following: log(AM-
analyzed retrospectively as a reference population. Patients underwent pre-treat- H)=-0.0358*(age)+1.4916 for France and log(AMH)=-0.0396*(age)+1.4006 for
ment with COCP before GnRH antagonist ovarian stimulation protocol for in India. These results confirm previous data reporting lower serum AMH values
vitro fertilization (IVF). Statistical analysis was conducted using generalized linear in women living in India compared to women living in France.
models or binomial regression. Significance was defined if p<0.05. Limitations, reasons for caution: This study focused on women within the
Main results and the role of chance: Significant effects of treatment group 20-44 year age range. Despite selected inclusion criteria and quite large size of
were present in the number of embryos (no pre-treatment mean=2.3; 17β-oestra- the two study populations, more studies are necessary to further characterize
diol/nomegestrol group mean=3.41; p=0.006) and number of top-quality the lower serum AMH levels observed in Indian women, in populations repre-
embryos (day 3) (no pre-treatment group mean=1.3; 17β-oestradiol/nomegestrol senting the regional and ethnical diversity for India.
mean=2.64; p=0.031). The mean number of mature oocytes was significantly Wider implications of the findings: These results are in line with CLSI
higher in natural oestrogen COCP compared to synthetic COCP, when the analysis guideline recommendations to check local reference intervals, that particularly
was controlled for the duration of COCP treatment (17β-oestradiol/nomegestrol makes sense for AMH. They should also stimulate more studies to cover the
group mean=6.28 and ethinylestradiol/gestodene group mean=4.34; p=0.014). population diversity worldwide, and to contribute to the adoption of the appro-
However, there were no significant differences in the mean number of mature priate usage of this biomarker for management of infertile women.
oocytes between 17β-oestradiol/nomegestrol group and no pre-treatment group Trial registration number: Not applicable
(means=6.284 and 5.56, respectively; p=1). 17β-oestradiol/nomegestrol was not
statistically associated with deleterious effects on biochemical pregnancy (odds
ratio=0.32, CI 95%=0.074–1.392; p=0.129). Ethinylestradiol/gestodene lowered P-574 Influence of cross-sex hormone therapy in trans*persons on
chances of having positive biochemical pregnancy when compared to the no laboratory profile and BMI
pre-treatment group (odds ratio=0.138, CI 95%=0.028–0.694; p=0.016). K. Feil1, A.L. Zippl1, E. Bürstmayr1, B. Böttcher1,
17β-oestradiol/nomegestrol was not associated with negative clinical pregnancy K. Winkler-Crepaz2, B. Toth1
outcome (odds ratio=1.714, CI 95%=0.339–8.676; p=0.515). Mean total dose 1
Medical University of Innsbruck, Gynecological Endocrinology and Reproductive
of follicle stimulating hormone administered for each treatment group was 2349.02 Medicine, Innsbruck, Austria ;
in 17β-oestradiol/nomegestrol group, 2259.24 in no pre-tretment group and 2
Paracelsus Medical University Salzburg, Gynecology and Obstetrics, Salzburg,
2859.21 for ethinylestradiol/gestodene. Austria
Limitations, reasons for caution: There might be a possible bias of unac-
counted variables, such as the number of blastocysts transferred, considering Study question: What influence has cross-sex hormone therapy (CHT) on
that in the clinical setting in which the study took place, patients are preferentially concentration of gonadotropins, sex steroids, liver enzymes, triglyceride, hemo-
programmed to transfer at day 3, and thus possibly altering the ratio of total globin and body mass index (BMI)?
blastocyts to embryos transferred. Summary answer: Cross-sex hormone therapy seemed to be safe within the
Wider implications of the findings: Natural oestrogen COCP was not study population. Significant changes were present in trans*men for SHBG and
associated with deleterious embryological or clinical outcomes, and significantly hemoglobin.
improved the number of embryos and top-quality embryos. Therefore, 17β-oes- What is known already: For many trans*persons CHT is the milestone of a
tradiol/nomegestrol prescription might benefit IVF cycle scheduling, and possibly life as a member of the desired sex. For this purpose, male and female sex
better synchronized follicular growth and future positive clinical pregnancy with steroids are applied, which can also lead to major side effects.
cryopreserved embryos obtained from 17β-oestradiol/nomegestrol patients. Study design, size, duration: This retrospective study was performed at
Trial registration number: Not Applicable our Department between 2005 and 2017. The concentration of gonadotropins

and sex steroids as well as liver enzymes, triglycerides, hemoglobin and body P-576 Cumulative Live birth rates in advanced maternal age
mass index were analysed before and three, six, twelve and twenty-four months (AMA) patients (40-43 years old) with autologous oocytes: the
after start of CHT. Demographic data were also evaluated. predictive factors
Participants/materials, setting, methods: In total 84 patients (33 trans*- P. Oger1, L. Delaroche1, V. Barbet2, X. Guettier1, G. Kerbrat1,
women and 51 trans*men) were included in the study. Endogenous hormone V. Maget1, G. Soudré1, E. Genauzeau1, P. Meicler1, F. Lamazou1
production was suppressed using GnRH agonists. Transdermal or oral Estradiol 1
Ramsay santé Hôpital Privé de Parly 2, ART Center IFM P2, Le Chesnay, France ;
was administered to trans*women and transdermal or intramuscularly testos- 2
Capionis, biostatistics, Bordeaux, France
terone to trans*men.
Main results and the role of chance: The mean age at first visit was 20.9± Study question: What are the predictive factors of the cumulative live birth
6,58 SD years in trans*men and 32.7 ± 15,90 SD years in trans*women. LH and rates (CLBR) in women aged between 40-43 years using their autologous
FSH decreased in both sexes immediately after starting GnRHa, but not to levels oocytes?
below the detection limits. The sex steroids showed a rapid adaptation to the Summary answer: In women aged between 40-43 years old, additional vit-
reference ranges of the desired sex. SHBG decreased significantly in trans*men, rified embryos is the only predictive factor to CLBR.
while hemoglobin increased significantly to the normal male ranges. There were What is known already: In Vitro Fertilization (IVF) in AMA patients is becom-
no significant changes in BMI, but a trend towards weight gain in both sexes. ing increasingly important. In this population, the low success rates and the high
Limitations, reasons for caution: As the follow up period of 2 years is risks of miscarriages are mainly linked to the oocyte aneuploidy rates which
short, no long term safety assessments can be made so far. increase with the maternal age. Several studies showed the probability of having
Wider implications of the findings: CHT seemed safe in the study population. a healthy baby in the AMA group of patients is less than 10% per cycle when
Since there were no significant differences and no pathological increase in liver using autologous oocytes and without any preimplantation genetic testing for
enzymes, a routine check for liver enzymes does not appear to be necessary. aneuploidy (PGT-A). These low success rates question the management of AMA
Trial registration number: Not applicable patients.
Study design, size, duration: This is a single-center retrospective study. The
P-575 Polycystic Ovarian Syndrome related hyperandrogenism institutional review board approved this study and the patients’consent to the
and elite sport activity – a systematic review use of their data was prior obtained.
C. Lambalk1, E. Bos1, M. Caanen1 Participants/materials, setting, methods: All IVF cycles performed from
1
Amsterdam University Medical Centers Location VUMC, Reproductive Medicine, November 2015 to December 2018 in women between 40-43 years old using
Amsterdam, The Netherlands their own oocytes were retrospectively analyzed. The main endpoint was the
CLBR defined as a live-born delivery ≥28 weeks after the fresh or one of the
Study question: Is there a higher prevalence of PCOS among athletic women subsequent frozen embryo transfers per pick-up. A multivariate logistic regres-
compared to the general population? sion model was used to assess the influence of the clinically relevant factors.
Summary answer: Based on the information of the current literature, no The p-values <0.05 were considered to be statistically significant.
decisive evidence was found for a higher prevalence of PCOS among female Main results and the role of chance: A total of 569 IVF cycles in 316 couples
elite athletes. were included. Mean female age was 41.4 ± 0.88 and mean AMH level was 2.0
What is known already: The population of sporting women and physically ± 2.1 pg/mL. Using female age categories, the CLBR was 8.6% for 40-41 patients,
active women at a high level of sport is increasing. This may lead to the Athlete 9.6% for 41-42 patients and 4.3% for 42-43 patients ; CLBR was improved in
Triad consisting of altered bone mineral density, low energy availability and patients with supernumary vitrified embryos (OR [95%CI] = 2.78 [1.16-6.4]
menstrual dysfunction. It is suggested that Polycystic Ovarian Syndrome (PCOS) (p=0.02)). Multivariate logistic regression model did not highlight any other
might be a cause for the menstrual disorders in female athletes. There is growing predictive factors on the CLBR: neither the maternal age (p=0.20), the etiology
concern about androgen levels and top sports performance, doping control and (p=0.80), the type of infertility (p=0.83), the smoking status (p=0.21), the AMH
fair play. The PCOS condition is positioned between two main issues in the field level (p=0.54) nor the number of mature oocytes (p=0.58) influenced individ-
of female top sports as cause for menstrual irregularities related to higher endog- ually the CLBR.
enous testosterone levels. Limitations, reasons for caution: Although the main outcome was CLBR,
Study design, size, duration: A systematic literature review was performed the design was retrospective. The results of this study differed from those avail-
up to October 2019 in the following databases: PubMed, Cochrane, Embase able in the current literature.
and Sportdiscus. To be included in the review, papers had to include elite athletes Wider implications of the findings: This study provides to AMA patients
women diagnosed with PCOS by the Rotterdam Criteria performing any type and clinicians a tool for building realistic expectations and providing accurate
of sport at national or international level. Furthermore, the papers needed a individual counselling. These preliminary results call for a prospective randomized
focus on PCOS, hyperandrogenism, oligomenorrhea or amenorrhea prevalence controlled trial including higher number of cycles.
outcomes and androgen measurement. Trial registration number: IRB00010835
Participants/materials, setting, methods: Fourteen studies of an initial
536 were selected by two researchers independently. The included studies con-
sisted of 10 observational cross-sectional studies, two case-control studies P-577 Late follicular phase ovarian stimulation protocol
(observational and experimental) one prospective cohort and one retrospective without exogenous pituitary modulators
cohort review of medical reports. The results are presented by outcome cor- Y. Fu1, X. Zhu1
responding with the 3 hallmarks of PCOS and were plotted in forest plots. 1
Shanghai Ninth Hospital, Reproduction, Shanghai, China
Main results and the role of chance: PCOS and Hyperandrogenism showed
a non significant and menstrual dysfunction a significant favoured prevalence in Study question: Is it feasible to perform ovarian stimulation from late follicular
the athletes’ group. The reported prevalence of PCOS diverged from phase without exogenous pituitary modulators during in-vitro fertilization (IVF)/
1.4% to 44%. intracytoplasmic sperm injection (ICSI) treatments?
Limitations, reasons for caution: None of the studies primarily aimed at Summary answer: Late follicular phase ovarian stimulation could be per-
exploring the PCOS prevalence among elite athletes. Overall, the majority of formed without exogenous pituitary modulators.
the studies had a high risk of bias. What is known already: Gonadotropin-releasing hormone antagonist
Wider implications of the findings: Exposure to higher androgen levels in (GnRH-ant) was the most commonly-used modulator to prevent the premature
female elite sport is currently a fierce matter of debate. This discussion requires luteinizing hormone (LH) surge when ovarian stimulation was initiated in the late
higher quality data than currently available on prevalence of the most frequently follicular phase. Our team of researchers firstly introduced the usage of exog-
occurring female natural condition of PCOS among top sport women. This enous progestational agents as an alternative pituitary modulator to gonadotro-
merits well designed and powered prospective studies. pin releasing hormone agonist (GnRH-a), and GnRH-ant for the prevention of
Trial registration number: not applicable premature LH surges in ovarian stimulation. Recently, we have documented

ovarian stimulation stared in late follicular phase with a dominant follicle diameter stimulation. With continuous PSL treatment, we administrated estrogen plus
of ≥14mm before spontaneous ovulation could be performed without exoge- progesterone for 10-14days to induce withdrawal bleeding. At initiation of bleed-
nous pituitary modulators. ing, we started ovarian stimulation using 100mg clomiphene. The clomiphene
Study design, size, duration: We retrospectively screened out the data of treatment was continued up to 4weeks until their follicles reaching to 14-18mm
normal-ovulatory patients who performed ovarian stimulation in late-follicular in diameter. Then, 15,000-20,000IU hCG was injected. Oocyte retrieval was
phase with a dominant follicle diameter of ≥10 mm in the absence of exogenous performed at 36hours after hCG.
pituitary modulators (namely late stimulation (LS) protocol) from women who Main results and the role of chance: The median age of participants was
treated with IVF/ICSI for the first time with the “freeze-all” strategy from May 37 years-old (range 26-41) with normal serum AMH (median 5.58 ng/ml, range
2016 to December 2018, in our setting, and compared the reproductive results 3.00-6.46) levels and normal antral count (range 7-15). However, they had
with early-follicular phase started ovarian stimulation using progesterone increased serum FSH (median 44.6mIU/ml, range 19-95) and LH (median
protocol. 31.4mIU/ml, range 8.2-51.4) levels as well as low estrogen levels (median 27.6
Participants/materials, setting, methods: 404 participants were included: mIU/ml, range 0-52.6). Those patients showed no recent spontaneous follicle
116 subjects for the study group and 288 subjects for the control group. In the growth and long term amenorrhea (median 18.5 years, range 11-22 years). They
study group, only gonadotropin (Gn) was injected from the start of ovarian also did not respond ovarian stimulation before this procedure. Among 4
stimulation till the trigger day. In the control group, Gn and micronized proges- patients, we succeeded follicle growth in two cases. In case 1, the follicle started
terone soft capsule were added from menstrual cycle day 3. The primary out- to growth after 22−25 days of clomiphene treatment, whereas follicle growth
come was the number of mature oocytes. could induced by15-19 days of clomiphene treatment in case 2. Total of 16 and
Main results and the role of chance: The number of mature oocytes was 20 preovulatory follicles (range 3-5 and 2-7 per stimulation) were found and
9.67±5.33 in the study group and 9.38±5.15 in the control group (P=0.693). total 10 and 11 of mature oocytes were obtained in case 1 and 2, respectively.
Total Gn dose (1962.28 ±517.06 versus 1626.04 ±311.75 IU) was greater and Seven out of 10 and 7 out of 11 oocytes were fertilized and all embryos devel-
mean Gn duration (10.48±2.44 versus 8.78±1.53) was longer in the study oped to high-quality embryos (grade 1-3) based on Veeck criteria in case 1 and
group than in the control group (P<0.001). The number of follicles with 2, respectively. All high-quality embryos were cryopreserved for future transfer
diameter >14 mm was similar between the two groups, while number of fol- under hormone replacement cycles.
licles with diameter >10 mm in the study group was greater than that in the Limitations, reasons for caution: The prevalence of ROS is extremely rare,
control group (13.33±6.67 versus 11.87±6.02, p=0.02). Additionally, there was the number of participants is small. We need to complete the procedure to
no statistic difference in the number of mature oocytes between the two groups check if the pregnancy and live birth will be achieved.
(9.67±5.33 in the study group versus 9.38±5.15 in the control group, P=0.693). Wider implications of the findings: We developed a new approach for
Similarly, the two groups were comparable regarding number of oocytes successful follicle growth to allow oocytes retrieval in ROS patients by autoim-
retrieved, fertilized oocytes, cleaved embryos and good-quality embryos. No mune suppression using PSL. If we can detect the autoimmune antibodies
secondary premature LH surges were detected in all the participants. The clinical responsible for ROS, this procedure could be a more useful method for groups
pregnancy rate was comparable in the two groups (54.55% versus 56.48%, P = of ROS patients hard to be treated.
0.718). The implantation rate was similar in the two groups(36.94% vs. 37.77%, Trial registration number: UMIN000034464
P=0.829). No differences were found in the rate of biochemical pregnancy,
multiple pregnancy, ectopic pregnancy and early miscarriage (P > 0.05). P-579 Peroxiredoxin 4 protects against ovarian ageing by
Limitations, reasons for caution: The limited cases and retrospective design ameliorating D-galactose induced oxidative damage in Mice
resulted in the bias of our study. Further studies in a large sample size and Y. Meng1, X.R. Liang1
continuous follow-up are still needed to determine the long-term safety of for 1
The First Affiliated Hospital of Nanjing Medical University, State Key Laboratory
children conceived with this novel protocol. of Reproductive Medicine, The Center for Clinical Reproductive Medicine
Wider implications of the findings: Our data suggest late follicular phase
ovarian stimulation without exogenous pituitary modulators was a feasible pro- Study question: What the effect and underlying molecular mechanism dose
tocol when performing random-start ovarian stimulation. In addition, it may be Prdx4 protein play in D-galactose induced ovarian ageing?
provided as an effective alternative for patients who choose frozen-embryo Summary answer: D-galactose induced ovarian ageing is accelerated in Prdx4-/-
transfer (FET) during infertility treatment or fertility preservation. mice, which results from accelerated apoptosis in granulosa cell via oxidative
Trial registration number: not applicable stress and ER stress-related pathways.
What is known already: Prdx4, a member of the Prdx family, is an important
P-578 Development of a new infertility treatment for resistant ER-resident antioxidant in cells. As revealed by our previous study, the expression
ovary syndrome through autoimmune suppression of Prdx4 was detected in ovarian granulosa cells and closely related to ovarian
Y. Tanaka1, K. Kawamura1 function. We also discovered that it was located in ER of granular cell and
1
International University of Health and Welfare school of medicine, Department involved in regulating the ER stress. D-galactose induced ovarian aging in mice
of obstetrics and gynecology, Chiba, Japan has been extensively used to study mechanisms of diminished ovarian
reserve (DOR).
Study question: Can suppression of autoimmune status using corticosteroid Study design, size, duration: In this study, we established mice model with
hormone induce follicle growth in patients with resistant ovary syndrome (ROS)? gene prdx4 knock out (Prdx4-/-) by the CRISPR/Cas9 technology. Adult (5
Summary answer: Treatment of corticosteroid hormone, predonisolone months) wild-type and Prdx4-/- mice were both intraperitoneally injected with
(PSL) was effective for induction of follicle growth in ROS patients. D-galactose (150 mg/kg/day) daily for 6 weeks. Ovarian function, oxidative
What is known already: ROS shows hyper gonadotropin and low estrogen, damage, ER stress and granulosa cell apoptosis in the ovaries were evaluated in
similar to premature ovarian insufficiency (POI). Infertility is the common complaint the two groups.
with ROS and POI. Although no follicle is found in ovaries of POI under ultrasound, Participants/materials, setting, methods: We detected the representative
ROS patients have normal number of antral follicles which are unresponsive to estrous cyclicity during 12 consecutive days. Secondly, we calculated ovary-to-
endogenous and exogenous FSH. Presence of genetic mutation or autoimmune body weight ratio and detected the level of E2. Thirdly, the number of different
antibodies to FSH or FSH receptor was found in some ROS patients, majority of types of follicles were counted by hematoxylin and eosin (H&E) staining, and
causes are still unknown. So far only two case reports of live birth in ROS patients apoptosis in granulosa cells were tested by TUNEL. In addition, we observed
have been published by using an approach of in vitro maturation of oocytes . the expression of senescence-associated protein and oxidative stress related
Study design, size, duration: Case series study. From, April 2018 to Dec factors via immunohistochemistry. Eventually, three ER stress pathways related
2019, we enrolled four ROS patients with written informed consent after per- markers were examined by western blot.
mission of this procedure from the ethical committee of the hospital. Main results and the role of chance: The HPG axis was more disrupted
Participants/materials, setting, methods: After diagnosis of ROS, the (P<0.01), and ovarian weight was relatively lower in the Prdx4-/- mice. The
participants received 15-20mg/day PSL for 4weeks before starting ovarian numbers of atretic follicles and apoptotic granulosa cells were significantly

increased in Prdx4-/- mice (P<0.05). In addition, the Prdx4-/- mice showed 1

Cantonal Hospital of Frauenfeld, Obstetrics and Gynecology, Frauenfeld,
increased expression of oxidative damage-related factors (8-OhDG, 4-HNE, Switzerland ;
2
and NTY) and ovarian senescence-associated protein P16 (P<0.05). Moreover, Cantonal Hospital of Lucerne, Obstetrics and Gynecology, Lucerne, Switzerland ;
3
levels of the proapoptotic factors CHOP and activated caspase-12 protein gyn-zentrum, Gynecology, Lucerne, Switzerland ;
4
(P<0.01), which are involved in the endoplasmic reticulum stress pathway and VivaNeo, Reproductive Medicine, Berlin, Germany
level of cell apoptosis-related BAX protein were elevated in the ovaries of the
Prdx4- /- mice. Study question: What is the course of sex steroids, gonadotropins and AMH
Limitations, reasons for caution: This is a descriptive study in mice. More after hysterectomy and bilateral salpingectomy?
further experiments are necessary to explore the molecular mechanism for Summary answer: DHEA-S, testosterone, androstenedione, estradiol, pro-
Prdx4 on ovarian function protection. gesterone, LH, FSH, SHBG and AMH showed significantly decreased blood levels
Wider implications of the findings: This study suggests that Prdx4 shows at the morning after surgery compared to preoperative/long-term samples
protective effect on ovarian function by reducing apoptosis of ovarian granulosa What is known already: Bilateral salpingectomy in benign conditions is widely
cell via downregulating oxidative stress and ER stress-related pathways. performed to prevent ovarian cancer. Recent Meta-Analyses demonstrate no
Trial registration number: not applicable impairment of ovarian function of this procedure, even though an impaired
postoperative ovarian stromal blood flow was described. Whether a hysterec-
P-580 GnRH agonist as an adjuvant to hCG for final oocyte tomy and bilateral salpingectomy negatively affects sex steroids, gonadotropins
maturation and AMH remains to be determined.
I. Park1, H. Sun1, J. Kim1, J. Lee1, S. Shim1, K. Lee2, G. Jeon3 Study design, size, duration: Prospective cohort study (February 2013 –
1 January 2015) of 40 premenopausal women with ovary-sparing elective hyster-
Liora Fertility Center, Obstetrica & Gynecology, Busan, Korea- South ;
2 ectomy and bilateral salpingectomy for benign uterine conditions or premalignant
Mamapapa& baby OBGY, Obstetrics and Gynecology, Ulsan, Korea- South ;
3 high-grade cervical pathologies.
Haeundae paik Hospital, Obstetrics and gynecology, Busan, Korea- South
Participants/materials, setting, methods: Serum concentrations of
Study question: Does triggering final oocyte maturation with standard dosage DHEA-S, testosterone, androstenedione, estradiol, progesterone LH, FSH,
of hCG and additional GnRH agonist make better IVF outcomes? SHBG, and AMH were assessed one day before, one day after and 4-6 weeks
Summary answer: GnRH agonist as an adjuvant to hCG for final oocyte mat- after surgery in a public hospital. Postoperative sampling was performed strictly
uration increases the number of mature oocytes and improves embryos quality. at 07.30 a.m. (±1 hour).
What is known already: A single bolus of hCG has been the gold standard Patients with presurgical evidence of malignancies, suspect ovarian morphology
for triggering final oocyte maturation in ART cycles. GnRH agonist trigger has in transvaginal ultrasound, menopausal symptoms, menopausal hormone treat-
been utilized in GnRH-antagonist cycles for inducing final oocyte maturation ment, baseline FSH values >25 IU/L or baseline AMH values <0,6 pmol/l were
while reducing the risk of ovarian hyperstimulation syndrome (OHSS). GnRH excluded.
agonist trigger could help retrieving more mature oocyte. However, after GnRH Main results and the role of chance: Mean age and BMI were: 44,83 years
agonist trigger, luteal phase defect results in lower pregnancy rate in fresh transfer and 24,53 kg/m2, respectively. At the day after surgery all hormones decreased
cycle. Final oocyte maturation triggering with both standard dosage of hCG and significantly: DHEA-S (p < 0.002 mean: 4.10/2.37/3.69 mmol/L); testosterone
GnRH agonist could help getting better IVF outcomes in GnRH antagonist cycles. (p < 0.001 mean: 0.95/0.49/0.86); androstenedione (p < 0.24 mean:
Study design, size, duration: This retrospective study included women 4.87/4.18/4.26 nmol/l); estradiol (p < 0.05 mean: 553.73/309.84/475.72
undergoing IVF cycles using GnRH antagonist protocol between January 2013 pmol/L); progesterone (p < 0.13 mean: 1.96/0.98/3.41 ng/ml); LH (p < 0.3
and December 2017. A total of 4335 low and normo-responders were assessed. mean: 10.58/9.17/12.98 IU/L); FSH ( p < 0.46 mean: 11.15/10.79/16.54 IU/L);
Participants/materials, setting, methods: When at least 2 follicles reached SHBG ( p < 0.22 mean: 74.64/67.13/73.65 nmol/L); and AMH (p < 0.46 mean:
18 mm in diameter, final oocyte maturation was triggered by hCG 10000 IU 5.23/4.99/7.12 pmol/l). There was no statistically significant correlation
alone or hCG 10000 IU plus GnRH agonist (leuprolide 1.0mg). And oocyte between hormone levels and operation time, blood loss or any further variable.
aspiration was done after 36 to 38 hours later. We divided the patients into two Serum hormone changes did not correlate with age or BMI. However, by 4-6
groups by triggering method. Group A (n=3776): trigger by hCG 10000 IU. weeks after surgery, circulating hormone values had returned to preoperative
Group B (n=559): trigger by hCG 10000 IU + GnRH agonist (leuprolide 1.0 mg) levels. We conclude that this hormonal decrease reflects an impaired hormonal
Main results and the role of chance: In each group, age (36.3 vs. 36.4), regulation after surgery. These hormonal changes are transient, lasting no longer
endometrial thickness (10.1 mm vs. 9.9 mm) and the number of transferred than 4-6 weeks.
embryos (1.9 vs. 1.9) did not show significant differences. There was no signif- Limitations, reasons for caution: These findings are based on a small sample
icant difference between two groups in total number of oocytes [9.9 vs. 10.4, size. The results need to be confirmed in larger patients groups and should
P=0.054]. Oocyte recovery rate (total number of oocytes/ follicles>11mm) include cortisol serum levels to depict stress levels.
[118.7% vs. 131.0%, P<0.01], the number of mature oocytes [6.1 vs. 6.4, Wider implications of the findings: These data reveal a postoperative
P=0.032] and the number of 2PN [6.7 vs. 7.1, P=0.029] were significantly higher dysregulation of the pituitary-gonad axis. It should be further explored how long
in Group B than in Group A. Group B had higher embryo quality score [75.6 vs. these changes last and what kind of surgeries are involved. This might be import-
85.0, P<0.01] than Group A. Ongoing pregnancy rates were similar between ant knowledge for the initiation of hormonal therapies after surgery.
two groups [43.9% (1535/3494) vs. 44.3% (203/458), P=0.190]. In both group, Trial registration number: Switzerland: EKNZ
there was no case of severe OHSS.
Limitations, reasons for caution: This is retrospective study. The sample P-582 Recombinant-FSH or human-menopausal-gonadotrophin
size of each group shows difference between two groups. Randomized control during ovarian stimulation resulted in similar mean euploid
study will be needed to confirm our conclusions. blastocyst rates per cohort of inseminated oocytes: an
Wider implications of the findings: Triggering final oocyte maturation with observational study on 961 patients.
standard dosage of hCG and additional GnRH-agonist increases the number of
S. Colamaria1, D. Cimadomo1, A. Vaiarelli1, M. Giuliani1,
mature oocyte and 2PN. It also improves embryo quality. Routine use of dual
C. Argento1, G. Bruno1, S. Ferrero1, G. Fabozzi1, R. Mazzilli1,
trigger with hCG and GnRH agonist could be a good option to improve IVF
A. Conforti2, C. Alviggi2, M. Schimberni3, N. Ubaldi4, L. Rienzi1,
outcomes without increasing OHSS or treatment burden.
F.M. Ubaldi1
1
Clinica Valle Giulia, Genera centers for reproductive medicine, Rome, Italy ;
2
University of Naples- Federico II, Department of Neuroscience- Reproductive
P-581 Negative short-term effect of hysterectomy and bilateral Science and Odontostomatology, Naples, Italy ;
salpingectomy on sex steroids, gonadotropins and AMH 3
Clinica Valle Giulia, Bioroma, Rome, Italy ;
4
J. Hutmacher1, V. Uerlings2, A. Günthert3, J.M. Weiss4 Catholic University of the Sacred Heart, Obstetrics and Gynecology, Rome, Italy

Study question: Do the type (recombinant-FSH or human-menopausal-go- P-583 GnRH-agonist and urinary-human-chorionic-gonadotrophin
nadotrophin [hMG]) and dose of gonadotrophin adopted for controlled-ovari- trigger of final oocytes’ maturation result in similar oocytes’
an-stimulation affect the euploid-blastocyst-rate per cohort of metaphase-II competence: an observational study on 2725 cycles with
oocytes (m-EBR per MII)? aneuploidy testing
Summary answer: The type and total dose of gonadotrophins do not affect C. Petriglia1, D. Cimadomo1, A. Vaiarelli1, S. Colamaria1,
the m-EBR per MII. M. Giuliani1, G. Bruno1, R. Maggiulli1, D. Soscia1, M.G. Amendola1,
What is known already: Controlled-ovarian-stimulation (COS) is one of the G. Nastri1, F. Innocenti1, G. Fabozzi1, N. Ubaldi1, F.M. Ubaldi1,
cornerstones of IVF. Its purpose is to obtain an adequate response in terms of L. Rienzi1
oocytes’ number and quality to improve treatments’ efficacy and efficiency by 1
Clinica Valle GIulia, Genera center for reproductive medicine, Rome, Italy
obtaining several competent embryos. Although some data suggested that
recombinant-FSH and hMG for COS in long agonist protocols perform similarly, Study question: Does the trigger of final oocytes’ maturation (GnRH-agonist
the evidence is limited in antagonist protocols, i.e. the most commonly used at versus urinary-human-chorionic-gonadotrophin [u-hCG]) affect their compe-
present. Therefore, the decision on which gonadotrophins should be used for tence during IVF cycles with preimplantation-genetic-testing-for-aneuploidies
COS is still uncertain, especially in patients at their first COS (naïve) and/or in (PGT-A)?
freeze-all strategies. Summary answer: GnRH-agonist and u-hCG involve similar maturation rates
Study design, size, duration: Observational study including all naïve patients per oocyte-pick-up (OPU) and mean euploid blastocyst rates per cohort of
(n=961, mean-maternal-age:39.2±3.5yr, mean-AMH:2.2±1.7ng/ml, mean- metaphase-II oocytes (m-EBR per MII).
BMI:21.5±2.5) indicated for trophectoderm biopsy-based preimplantation-ge- What is known already: The conventional trigger of final oocytes’ maturation
netic-testing-for-aneuploidies (PGT-A) in the period 2013-2018. All patients after controlled ovarian stimulation (COS) is hCG. Nevertheless, standing the
underwent a GnRH-antagonist protocol. The kind of gonadotrophin was chosen growing application of GnRH antagonist protocols for COS, GnRH-agonist trig-
depending on patients’ compliance to the type of administration (pen for recom- ger has been introduced to almost eradicate the incidence of ovarian hyperstim-
binant-FSH versus syringe for hMG) and gynecologists’ judgment. The dose was ulation syndrome (OHSS) in patients at high risk. While it is clear that this strategy
established according to patients’ characteristics. The primary outcome was the affects the window of implantation when conventional luteal phase support is
m-EBR per MII in the two groups. used, no evidence has been instead produced regarding its effect on oocytes’
Participants/materials, setting, methods: To achieve 80% power (α=0.05) competence. Lastly, it is still uncertain whether GnRH-agonist trigger might be
to rule-out a 5%-difference in the primary outcome we required 792 and 158 efficiently adopted in an unselected population of patients candidate to freeze-all.
naïve patients in the recombinant-FSH and hMG groups, respectively, assuming Study design, size, duration: Observational study including 2725 OPU for
that 1/5 would use hMG. Only completed PGT-A cycles (live-birth achieved or PGT-A performed between 2013 and 2018 at a private IVF center (maternal
no euploid embryos produced/left) with all MII-oocytes undergoing ICSI were age:39.5±3.3yr, AMH:2.1±1.8ng/ml, BMI:21.7±2.7). Also cycles without MII,
included. Secondary outcomes were: mean number of oocytes, blastocysts and zygotes or blastocysts were included. The primary outcome was the m-EBR per
euploid blastocysts, maturation and blastocyst rates, and cumulative-live-birth-de- MII in the cycles adopting GnRH-agonist (n=1294) versus u-hCG (n=1431). The
livery-rate (CLBdR) per cycle. main secondary outcome was the mean maturation rate per OPU (MII/COCs).
Main results and the role of chance: 773(80.4%) and 188(19.6%) patients All outcomes were adjusted for patients’/cycles’ putative confounders in gen-
used recombinant-FSH and hMG, respectively. The groups were similar for eralized-linear-models and multivariate logistic-regression-analyses
maternal/paternal age, AMH, BMI, sperm factor, cause of infertility, and trig- Participants/materials, setting, methods: All patients had regular men-
ger of ovulation. The dose and duration of COS were lower with recombi- strual cycles. Different gonadotrophins and doses were used in antagonist COS
nant-FSH (2447±813IU versus 2672±871IU,p<0.01;9.7±1.7days versus protocols (recombinant-FSH,n=1048, 38.5%; recombinant-FSH+LH-activi-
10.0±1.7day,p=0.02), nevertheless the number of cumulus-oocytes-com- ty,n=1065, 39.1%; hMG,n=282, 10.3%; biosimilar-FSH,n=90, 3.3%; corifollitro-
plexes (COCs) was higher (n=8185,10.6±6.5 versus n=1709,9.1±5.7,p<0.01). pin-alpha+recombinant-FSH/hMG,n=240, 8.8%) according to patients’
The maturation rates per COCs (MII/COC) were similar (74%±19% versus characteristics and gynaecologists’ judgement. When ≥2 follicles reached a
72%±21%,p=0.3), therefore also the number of MII was higher with recom- diameter ≥17–18mm, the trigger was performed 35hr before OPU with a sub-
binant-FSH (n=5827,7.5±4.4 versus n=1213,6.4±4.3,p<0.01). More blasto- cutaneous bolus of GnRH-agonist (Buserelin:0.5ml) or intramuscular injection
cysts were biopsied with recombinant-FSH (n=1926,2.5±2.2 versus of u-hCG (10,000IU). Denudation, ICSI, blastocyst biopsy, comprehensive-chro-
n=402,2.1±2.1,p=0.02), although the blastulation rate per MII was similar mosome-testing, and vitrified-warmed euploid blastocyst transfers were con-
(33%±25% versus 33%±28%;p=0.5). The number of euploid blastocysts ducted after OPU.
was slightly higher with recombinant-FSH (n=897,1.2±1.5 versus n=186, Main results and the role of chance: The adoption of the GnRH-agonist
1.0±1.5,p=0.05), while the m-EBR per blastocysts were comparable trigger increased during the observational period (~10% to ~60%), especially in
(34%±37% versus 30%±37%,p=0.1). Lastly, both the m-EBR per MII patients collecting >10 cumulus-oocyte-complexes (COCs) (~30% to ~80%).
(15%±20% versus 14%±20%,p=0.14) and the CLBdR (n=251/773,32.5% This reduced the incidence of patients referring symptoms of a moderate-severe
versus n=50/188,26.6%;p=0.1) were similar. Once corrected for maternal OHSS from ~2% to none, with no impact on the competence of the retrieved
age, AMH and sperm factor, neither the kind nor the dose of gonadotrophins oocytes. Overall, GnRH-agonist trigger was adopted in younger patients
was associated with the m-EBR per MII in a generalized-linear-model (p=0.2 (38.8±3.4yr) who collected more COCs (n=17593, 13.6±7.4 per OPU) than
and p=0.5). Once corrected for maternal age, number of oocytes and sperm u-hCG (40.2±3.1yr; n=10552 COCs, 7.4±4.6 per OPU). Nevertheless, the
factor, neither the kind nor the dose of gonadotrophins was associated with trigger adopted, also when adjusted per maternal age and number of COCs in
the CLBdR in a multivariate logistic-regression-analysis (p=0.2 and p=0.7). a generalized-linear-model, did not associate with the mean maturation rate per
Limitations, reasons for caution: The study was powered to exclude a OPU (p=0.1). A similar statistical approach outlined that the mean fertilization
5%-difference in the primary outcome, therefore secondary outcomes are obser- rate per MII, the mean blastulation rate per cohort of zygotes, and the mean-EBR
vational. A cost-effectiveness analysis is required. The choice of the gonadotro- per cohort of biopsied blastocysts were also independent of the trigger adopted
phin was based on patients’ compliance with the administration method and (p=0.5, p=1.0, and p=0.3, respectively). Not even the primary outcome under
gynecologist’s judgment, thus randomized-controlled-trials investigating the investigation (i.e. m-EBR per MII) was affected by the trigger used (p=0.83). At
CLBdR per cycle are still required. last, the cumulative-live-birth-delivery-rate (CLBdR) per completed cycle (i.e.
Wider implications of the findings: Recombinant-FSH in naïve patients ≥1 live-birth achieved or no euploid embryos left/produced) was independent
undergoing blastocyst culture, PGT-A and vitrified-warmed euploid embryo from the trigger adopted (Odds-Ratio: 1.0,95%CI 0.85-1.31, adjusted-p=0.6) in
transfer(s) is suitable as first-line COS approach. Indeed, it may result in larger a multivariate-logistic-regression-analyses corrected for confounders.
cohorts of oocytes of a similar competence as hMG, but involving a higher Limitations, reasons for caution: This is a retrospective study. No cost-
patients’ compliance (lower dose, shorter COS and a well-accepted type of effectiveness analysis was conducted. GnRH-agonist trigger was performed
administration). independently from LH levels, although this might identify specific subpopulations
Trial registration number: None of patients with a suboptimal response. Lastly, properly-conducted

Randomized-Controlled-Trials are still eagerly requested, especially in antagonist QUICKI and Irisin in addition to GTT may play a pivotal role in institution of
COS protocols. lifestyle changes that may help in delaying the progression to type 2 DM.
Wider implications of the findings: Throughout the years the adoption of Trial registration number: not applicable
GnRH-agonist trigger after antagonist COS protocols has sharply increased for
cycles planned for PGT-A with blastocyst culture, biopsy, and freeze-all. This did
P-585 Prokineticin receptor 2 (PROKR2) mutations in functional
not impact oocytes’ competence, while almost eradicating OHSS. U-hCG trigger
hypothalamic amenorrhoea
is still used in ~50% of the patients collecting ≤10 COCs.
Trial registration number: None A. Tranoulis1, D. Mavrogianni2, A. Soldatou3, A. Pampanos2,
T. Leoutsakou4, P. Drakakis5, D. Loutradis6, L. Michala7
P-584 Role of Homeostasis model assessment (HOMA2-IR), 1
Guy’s and St Thmas’ NHS Foundation Trust- King’s College, Obstetrics and
Quantitative insulin sensitivity check index (QUICKI) and Irisin Gynaecology, London, United Kingdom ;
in diagnosing insulin resistance among young girls with Polycystic 2
Alexandra Hospital- Athens University Medical School, Molecular Biology Unit-
ovarian syndrome Division of Human Reproduction, Athens, Greece ;
3
H. Sagili1, B. Sharan1 ’P&A Kyriakou’ Childran’s Hospital- National and Kapodistrian University of
1 Athens- School of Medicine, Second Department of Paediatrics, Athens, Greece ;
JIPMER, Obstetrics and Gynaecology, Puducherry, India 4
Alexandra Hospital- National and Kapodistrian University of Athens, Department
Study question: Do HOMA2-IR, QUICKI and Irisin diagnose additional cases of Molecular Biology, Athens, Greece ;
5
of insulin resistance in young girls with Polycystic ovarian syndrome (PCOS) in Alexandra Hospital-Athens University Medical School, Division of Human
comparison with 75 gm OGTT? Reproduction- IVF Unit- First Department of Obstetrics and Gynaecology, Athens,
Summary answer: HOMA2-IR, QUICKI and Irisin appear to detect additional Greece ;
6
cases of insulin resistance in young girls with PCOS in comparison with 75 Alexandra Hospital- Athens University Medical School, Division of Human
gm OGTT. Reproduction- IVF Unit- First Depertment of Obstetrics and Gynaecology, Athens,
What is known already: Young girls with PCOS have a predilection to Greece ;
7
become insulin resistant with an increased risk of impaired glucose tolerance Alexandra Hospital- National and Kapodistrian University of Athens- School of
(IGT) and type 2 Diabetes Mellitus (DM). Because impaired glucose tolerance Medicine, First Deparment of Obstetrics and Gynaecology, Athens, Greece
is often asymptomatic, screening for insulin resistance (IR) is recommended. The
gold standard method of assessing Insulin resistance namely Euglycemic hyper- Study question: To identify any possible mutations in prokineticin recptor 2
Insulinemic clamp technique is expensive and not practical. Currently 75 gm oral (PROKR2) gene in subjects with functional hypothalamic amenorrhoea (FHA).
Glucose tolerance test (OGTT) is being offered as a screening method for insulin Summary answer: PROKR2 mutation screening in 41 women with FHA
resistance but might not identify all cases of IR. The role of other surrogate resulted in the identification of eight novel different mutations in five patients.
markers for diagnosis of IR is not yet established. What is known already: Although the proximate cause of FHA is the abnor-
Study design, size, duration: This cross sectional study was carried out over mal gonadotropic-releasing hormone (GnRH) secretion, it is seemingly more
a period of two years.82 young girls aged 15-19 years -41 young girls with PCOS than an isolated desynchronisation of the GnRH axis. There is fair evidence to
were compared to 41 age matched controls who had presented with minor support the association of FHA with behavioural, emotional, cognitive and psy-
gynaecological complaints such as dysmenorrhea, white discharge per vaginum, chological factors. However, it remains uncertain, whether this variability is also
lower abdominal pain etc. Excusion criteria were thyroid dysfunction,hyperpro- attributable to a genetic predisposition. The PROK2 and PROKR2 singling path-
lactinaemia and treatment with Insulin sensitizers. Consecutive sampling was way is implicated in the cause of idiopatic hypogonadotropic hypogonadism
carried out. (IHH) and Kallmann’s syndrome. Two heterozygous PROKR2 mutations (c.254
Participants/materials, setting, methods: Girls recruited from G>A and c.518 T>G) have also been identified amongst patients with FHA. The
Gynaecology outpatient department of a tertiary care hospital underwent testing latter were found to be loss-of-function mutations.
for Fasting- Serum Irisin, Insulin; Blood Glucose and second hour Serum Insulin; Study design, size, duration: This is a prospective, single-centre study carried
Blood glucose after 75 gm glucose. HOMA 2-IR and QUICKI were calculated out at a tertiary referral clinic for gynaecological endocrinology and paediatric
using computer software and Serum Irisin was measured by ELISA. Cut offs for & adolescent gynaecology, spanning the period January 2016 to June 2019. We
HOMA2-IR, QUICKI and Irisin were defined based on population based studies analysed the coding sequence of PROKR2 in 84 women: 41 with FHA, 23 with
as >2.5, 0.383+0.007 for nonobese / 0.331+ 0.010 for obese and 15.43 ng/ml IHH, and 20 healthy controls respectively.
respectively. Participants/materials, setting, methods: Genomic DNA was extracted
Main results and the role of chance: Almost half the cases were obese, from peripheral blood samples using PureLink Genomic DNA Kits for purification
another 22% were overweight in our study and the body mass index between of genomic DNA. PCR was performed using primers designed to amplify the
cases and controls was statistically significant (p<0.0001).78% of the cases had two coding exons and the exon-intron boundaries. The amplified PCR products
all three features of PCOS according to Rotterdam’s criteria. Infrequent men- were sequenced using the ABI Prism 3130 Genetic Analyzer, Applied Biosystems.
strual cycles and clinical and/or biochemical signs of hyperandrogenism were Variants were validated using the sequencing Analysis v 5.2 software.
seen in 14.7% and infrequent cycles and polycystic ovaries were present in 7.3%. Main results and the role of chance: Eight novel PROKR2 mutations were
51% of cases had impaired glucose tolerance on 75 gm OGTT and there were identified in five of the 41 women with FHA, either in heterozygous state (two
no cases of overt diabetes. 43.9% cases had IR based on HOMA2-IR compared cases) or in homozygous or compound heterozygous state (three cases): 241
to 21% controls (p=0.03). 34% cases compared to 17% controls had IR based c.G>C, c.296 G>T, c.375 A>G, c.404 A>G, c.404 C>G, c.410 C>A, c.421
on QUICKI (p=0.038).19% cases compared to 9.7% controls had IR based on G>A, and c.475 G>T. Furthermore, three novel PROKR2 heterozygous muta-
fasting serum Irisin (p=0.211). Overall 21 cases were diagnosed to be insulin tions were identified in three of 23 women with IHH: c.246 C>T, c.252 C>T
resistant based on 75 gm OGTT ; 18 additional cases (43.9%) were diagnosed and c.259 G>C. Reproductive phenotypes ranged from absent to partial puberty
as insulin resistant based on a panel of markers (HOMA2-IR ,QUICKI and fasting to complete reversal of gonadotropin-releasing hormone deficiency following
serum Irisin ). 11 and 8 cases (26.8% and 19.5%) were diagnosed based on only discontinuation of treatment.No mutations were found in the cohort of 20
high HOMA2-IR and QUICKI values respectively. Similarly, 3 cases (7.3%) were controls with normal menstrual cycles.
diagnosed as insulin resistant based on only high fasting serum Irisin values . Limitations, reasons for caution: Further research to ascertain the func-
Limitations, reasons for caution: The study was done on a small sample tional impact of these variants is warranted. Differences in the genetic back-
and as a case-control study for the findings to be applicable to a larger population. grounds of various ethnic populations should also be considered.
Further, follow up of these subjects may yield additional insight into the progres- Wider implications of the findings: Identification of gene mutations under-
sion of insulin resistance and type 2 DM in young girls with PCOS. lying FHA may lead to new pathophysiology insights, improved diagnostics, and
Wider implications of the findings: As young girls with PCOS are at high novel treatment approaches.
risk of developing IGT, early identification of IR by a panel of HOMA2-IR, Trial registration number: not applicable

P-586 Impact of gonadotropin genetics profile and ovarian reserve from clearly correlating with IVF-ICSI pregnancies. On the other hand, a marginal
on controlled ovarian stimulation outcomes effect on ovarian stimulation and anthropometric characteristics of women
A. Rebecchi1, E. Papaleo1, M. Mignini Renzini2, C. Alviggi3, seems to emerge. Further biggest studies are needed.
A. Marino, Alessandro4, C. Brigante5, A. Conforti3, M. Molgora1, R. Trial registration number: not applicable
Iemmello6, I. Carbone7, L. Quaranta1, A. Allegra4
1
San Raffaele Hospital, Obstetrics and Gynecology, Milan, Italy ; P-587 Body Mass Index stratified Anti-Müllerian hormone
2
Biogenesi Reproductive Medicine Center - Istituti Clinici Zucchi Monza, obstetric thresholds for diagnosing Polycystic Ovary Syndrome
and gynecology, Monza, Italy ; X. Liu1, Y. Guo1
3
University of Naples- Federico II, Department of Neuroscience- Reproductive 1
The First Affiliated Hospital of Zhengzhou University, Reproductive Medicine
Science and Odontostomatology, Naples, Italy ; Center, Zhengzhou, China
4
ANDROS Day Surgery Clinic, Reproductive Medicine Unit, Palermo, Italy ;
5
Biogenesi Reproductive Medicine Centre- Istituti Clinici Zucchi, Reproductive Study question: Should Anti-Müllerian hormone (AMH) threshold for diag-
Medicine, Monza, Italy ; nosing Polycystic Ovary Syndrome (PCOS) be stratified according to different
6
Ospedale Maggiore Policlinico- Mangiagalli and Regina Elena, Infertility Unit, Body Mass Index (BMI)?
Milan, Italy ; Summary answer: PCOS patients with lower BMI had higher serum AMH
7
Ospedale Maggiore Policlinico- Mangiagalli and Regina Elena, Obstetrics and levels and AMHR Ⅱ mRNA expression in granulosa cells and we proposed BMI
Gynecology, Milan, Italy stratified AMH thresholds.
What is known already: Serum AMH levels are higher among patients with
Study question: o assess the difference between expected, through standard- PCOS than healthy women and PCOM (Polycystic Ovary Morphology) women
ized-3D AFC, and retrieved oocytes in relation to gonadotropin genetic profile with normal ovulation. While AMH has been reported to be positively or
after standard COS with 150 UI/day r-FSH. non-significantly related to BMI in infertile women without PCOS, in women
Summary answer: In good prognosis women, specific genetic variants seem with PCOS, AMH appears to be negatively correlated with BMI levels. AMH is
to partially influence ovarian stimulation, despite no relevant impact on pregnancy believed to be a surrogate indicator of the antral follicle count (AFC) for diag-
rates was observed. nosing PCOS. However, the heterogeneity of PCOS patients has seldom been
What is known already: Among fertility treatment, controlled ovarian stim- considered when calculating cut-off values.
ulation represents the key step, with the main aim to obtain a certain number Study design, size, duration: This was a retrospective cohort study based
of competent oocytes that will enable the best probability of achieving a live on 4347 infertile women under 35 years old treated in a reproductive medicine
birth. The ovarian response to FSH in individuals is difficult to predict. Indeed, centre of a university-affiliated hospital from February 2016 to May 2018.
women with similar demographic, anthropometric and gonadotropin profiles Granulosa cells(GCs) were collected from 22 patients undergoing transvaginal
may have different response to r-FSH due to the “sensitivity” of follicles to oocyte retrieval after controlled ovarian hyperstimulation with agonist protocol
exogenous FSH itself. Several studies demonstrated that such “sensitivity” could from December 2019 to January 2020.
be influenced by specific genotype characteristics. However, the evidence in this Participants/materials, setting, methods: The study population included
sense seems to be controversial and inconclusive. 1220 patients with PCOS, 1386 patients with PCOM and 1741 normal controls.
Study design, size, duration: We performed a multicenter, longitudinal, We divided them into normal-weight groups and overweight groups separately
prospective, interventional, cohort pilot-study, enrolling 119 women attending based on a BMI threshold of 24 kg/m2 according to a guideline based on the
four clinical Centers of medically assisted reproduction from August 2016 to Chinese population. RNA samples were extracted from GCs from 13 PCOS
November 2018. patients and 9 normal controls. Expression of AMHR Ⅱ mRNA were evaluated
Participants/materials, setting, methods: Infertile normo-gonadotropin by RT-qPCR.
patients, aged between 34 and 39, at their first COS, with normal ovarian reserve Main results and the role of chance: Regardless of normal weight or
(AFC between 8 and 16) measured with 3D automated ultrasonography. Blood overweight, PCOS patients tend to have higher AMH levels (P<0.001) and
samples were collected for gonadotropin genetic polymorphisms assessment slightly higher AMHR Ⅱ expression in GCs (Normal weight: 2.69±2.59 vs
and hormone assay. Stimulation protocol consisted of standard 150 IU/day 1.04±1.01, P=0.088; Overweight: 1.78±1.22 vs 0.82±0.42, P=0.116) than con-
r-FSH for the whole stimulation period starting from basal assessment, GnRH trol groups. In the PCOS group, normal-weight women had higher serum AMH
antagonist starting from day 5-6 of stimulation, and trigger when at least two levels than overweight women (P<0.001) and slightly higher AMHR Ⅱ expres-
follicles reached >16 mm diameter. sion(P=0.133,0.180, respectively), while in the control groups, there was no
Main results and the role of chance: 106 patients completed the stimulation such difference. The cut-off level for AMH for diagnosing PCOS was 4.77 ng/
protocol. The mean basal 3D follicular count was 13.4, the mean of follicles > 16 ml in the normal weight group and 4.15 ng/ml in the overweight group with
mm at induction was 5.8 and the mean number of retrieved oocytes was 8.4. No sensitivity of 81.8% and 85.8%, specificity of 90.5% and 84.2%, respectively.
significant correlation was found between the number of retrieved oocytes and Limitations, reasons for caution: The current GC samples collected from
different genotypes for each polymorphism. However, after 5 days of stimulation, patients were limited and the expression pattern of AMHRⅡ may be affected by
we observed a significant impact of AG genotype for FSHR p.N0680S on the the medication used during COH, thus, further experiments including larger
percentage of observed follicles ≥10 mm compared with genotypes AA (p = 0.04) sample size and validation with the animal model are needed.
and GG (p = 0.005). Nonetheless, at day of trigger, no significant differences were Wider implications of the findings: This study agreed with the former
found in the number of follicles ≥16 mm. This may be due to a fastest development study that AMH levels of patients with PCOS were affected by their BMI. Our
of follicles in the first part of stimulation for women carrying AG genotype. study suggested that BMI should be taken into consideration when assessing
LHCGR2 GG genotype leads to a shorter stimulation compared with genotypes endocrine indicators of women with PCOS, such as AMH, in both clinical practice
AA (p = 0.008) and GA (p = 0.006). The same result was obtained for homozygous and basic research.
GG genotype for the FSHB-211 polymorphism compared to the genotype GT Trial registration number: 81571409
(p = 0.04). Significant correlation was found between BMI and heterozygous gen-
otype of the FSHR p.N0680S (OR= 1.28 [1.09-1.52] p=0.004). We obtained
P-588 The activated mTOR-S6K signaling pathway increases DNA
36.8% of ongoing pregnancies per started cycle.
damage in follicles of women with polycystic ovary syndrome
Limitations, reasons for caution: Strict inclusion criteria limited the enrol-
ment of subjects planned before the study. Thus, the main findings of our study G. Jing1, S. Cai2, G. Lin3
1
could be underpowered. However, this is the first prospective study in which Reproductive and Genetic Hospital of CITIC-Xiangya, Reproduction Medicine
polymorphisms were associated to AFC assessed by a reliable, standardized Center, Changsha, China ;
2
automated method together with centralized AMH assessment. Reproductive and Genetic Hospital of CITIC-Xiangya, Reproductive Medicine
Wider implications of the findings: In normo-responder patients, the Center, Changsha, China ;
3
impact of pharmacogenomics in the personalization of treatment protocol is far Central South University, School of Basic Medical Science, Changsha, China

Study question: Does the follicular microenvironment of polycystic ovary Summary answer: Clinical pregnancy and implantation rates were significantly
syndrome (PCOS) change by regulating the expression of mammalian rapamycin increased in natural cycle (NC) as compared to hormone replacement ther-
(mTOR) pathway targets? apy (HRT).
Summary answer: The mTOR-S6K signaling, by controlling DNA damage What is known already: The gold standard protocols in FET cycles are HRT,
response and genome stability of granulosa cells and oocytes, maintains follicular true or modified NC. When comparing live birth and clinical pregnancy rates
microenvironment of female ovarian. (CPR), the superiority of one regimen over another has not been demonstrated.
What is known already: Recent researches suggest that mTOR pathway, NC can only be performed in women with regular ovulatory menstrual cycles,
which was found to be especially active in PCOS, is considered to have close while HRT cycles can be used in women with regular and irregular menstrual
relationship with the pathogenesis of PCOS. In previous studies, we determined cycles. An increased early pregnancy loss has been reported in previous studies
the role of mTOR by use of conditional knockout mice model. mTOR-dependent in HRT compared to NC, which could be related to the higher proportion of
pathways in primordial or growing oocytes differentially affected downstream patients with polycystic ovary syndrome (PCOS) or the presence of bias linked
processes including follicular development, sex-specific identity of early granulosa to possible embryo aneuploidy.
cells, maintenance of oocyte genome integrity, oocyte gene expression, meiosis, Study design, size, duration: This single center retrospective cohort study
and preimplantation developmental competence. However, the underlying mech- included a total of 802 single/double euploid FET cycles between March 2017
anism between mTOR pathway and pathophysiological changes of PCOS and October 2019. Trophectoderm biopsy samples were subjected to Next
remains largely undetermined. Generation Sequencing to test the ploidy state. Vitrification and warming were
Study design, size, duration: In this study, patients with PCOS (n = 4) performed using the Cryotop method (Kitazato, Biopharma). The CPR, implan-
diagnosed according to the 2003 Rotterdam standard underwent partial or tation rate (IR) and early pregnancy loss (EPL) were evaluated in euploid frozen
wedge resection of the ovary. The ovarian tissues used as control were obtained embryo transfers between true NC and HRT.
from female (n=3) who underwent ovariectomy for fertility preservation. Participants/materials, setting, methods: The following patient charac-
Samples were fixed and analysed by immunohistochemistry to determine mTOR teristics were analyzed: age, Anti Müllerian hormone (AMH), body mass index
pathway activation and expression of markers reflecting DNA damage, cell (BMI), primary or secondary infertility, duration of infertility, uterine abnormal-
apoptosis and proliferation. Mtor-conditional knockout mice was performed to ities (diagnosed by ultrasound), endometrial thickness, quality of the transferred
examine the underlying molecular mechanism. embryo/s, difficulty of the transfer (requirement of additional instrumentation),
Participants/materials, setting, methods: Ovarian tissues of PCOS day of biopsy and single (SET) or double (DET) embryo transfer. PCOS patients
patients and Mtor- GcKO mice were used as study models. Ovarian tissues were were excluded. The primary aim was to evaluate whether the clinical pregnancy
fixed and analysed histologically. Immunostaining for: 1) the expression of MTOR and implantation were affected by the cycle regimen.
and the activated form of its major downstream effectors—phosphorylated Main results and the role of chance: The mean patient age was 34.02±5.10
RPS6KB1, phosphorylated ribosomal protein S6 (pRPS6), phosphorylated vs 33.34±5.52 years in NC and HRT. The FET was performed in a NC (n=294,
EIF4EBP1. 2) γH2AX as a marker of DNA damage; 3) CASPASE3 as a marker 36.6%) or HRT cycle (n=508, 63.4%). Clinical pregnancy rate was significantly
of cells apoptosis; 4) KI67 as a marker of cells proliferation; higher in NC; 66.3% vs 57.1% in HRT (p=0.029) while EPL rate was significantly
Main results and the role of chance: We found that nuclear expression of lower in NC; 1.26% vs 5.05% (p=0.025), respectively. Also, the implantation
p-RPS6 level was increased in oocytes of primary and secondary follicles in PCOS rate was significantly higher in NC as compared to HRT (64.6%±46.81 vs
patients compared with control group. In the PCOS group, γH2AX expression 57.6%±47.93; p=0.046).
in oocytes and its surrounding granulosa cells of non-growing and primary fol- Patient characteristics (age, AMH, BMI, duration of infertility, uterine abnormal-
licles significantly increased (p<0.05). Expression of KI67 in granulosa cells of ities, endometrial thickness, embryo quality, embryo transfer difficulty, biopsy
growing follicles showed no significant differences between any groups (p>0.05). day and SET/DET) were similar between both groups compared.
The results of immunohistochemistry analysis showed that the levels of the Multivariate logistic regression model was performed with all significant values
active forms of phosphorylated MTOR (p-MTOR) and its downstream effectors of the univariate model (p<0.20), to check the factors predicting the clinical
were regularly expressed in oocytes and granulosa cells at different stages of pregnancy outcome. Following characteristics showed a significant (p<0.05)
folliculogenesis in mice. A major downstream effector of MTOR activation, effect on CPR: AMH: OR: 0.889 [0.795-0.995], p=0.039, age: OR: 1.36 [1.001-
p-RPS6, was gradually increased in different stages of oocyte as follicle devel- 1.071], p=0.04, transfer of a top-quality blastocyst: OR: 7.445 [3.552-15.604],
opment and strongly expressed in cumulus cells and pre-antral granulosa cells p=0.001, ET difficulty [Difficult vs Easy]: OR: 0.575 [0.347-0.954], p=0.032 and
but barely detectable in mural granulosa cells. The results of Western blot cycle regimen [NC vs HRT]: OR: 1.482 [1.035-2.121], p=0.031.
showed that BMP15 was significantly decreased in oocytes collected from sec- Limitations, reasons for caution: A prospective randomized study with
ondary follicles in Mtor- GcKO mice. Immunofluorescence staining of γH2AX large sample size would have minimized potential limitations as compared to this
revealed more DNA double-strand breaks (DSBs) in Mtor-GcKO oocytes of retrospective design.
the early- stage growing follicles. Wider implications of the findings: Patients with regular menstrual cycles
Limitations, reasons for caution: Due to the limitation by PCOS female should be offered a natural FET cycle to achieve better outcomes in terms of
tissue acquisition, the ovarian tissue detected in this study was obtained from clinical pregnancy and implantation rates.
the patients who need to perform ovarian wedge resection. Additionally, the Trial registration number: NA
underlying molecular link between mTOR pathway and DNA damage repair
should be further evaluated. P-590 Should we measure serum LH levels at the initiation of
Wider implications of the findings: These results reveal an important function ovarian stimulation in a GnRH antagonist downregulated
of the mTOR-S6K signaling in DNA damage response and suggest a mechanism IVF/ICSI cycle?
related to genome stability in the follicular microenvironment of PCOS patients. F. Di Guardo1, A. Racca1, B. Popovic-Todorovic1, S. De Rijdt1,
Our findings may subsequently provide valuable resources for developing targeted G. Verheyen1, H. Tournaye1, M. De Vos1, C. Blockeel1
interventions aimed at improving follicle recruitment in vivo for PCOS patients. 1
Universitair Ziekenhuis Brussel- Vrije Universiteit Brussel, Centre for Reproductive
Medicine, Brussels, Belgium
P-589 True Natural Cycle outweighs Hormone Replacement Study question: Does the serum LH level at the onset of ovarian stimulation
Therapy in euploid frozen embryo transfer (OS) in a GnRH antagonist cycle have an impact on reproductive outcomes?
A. Bayram1, N. De Munck1, I. El Khatib1, A. Arnanz1, A. Abdala1, Summary answer: The serum LH level at the start of GnRH antagonist cycles
A. El Damen1, L. Melado1, B. Lawrenz1, H. Fatemi1 has no impact on live birth rates (LBRs) nor cumulative live birth rates (CLBRs).
1
IVIRMA Middle East Fertility Clinic LLC, IVF Lab, Abu Dhabi, United Arab Emirates What is known already: Previous studies suggested that elevated LH levels
in the early follicular phase could be associated with lower pregnancy rate.
Study question: Does the cycle regimen used for endometrial preparation Introducing a GnRH antagonist from Day1 onwards instead of Day 6, has been
affects the outcomes in euploid frozen embryo transfer (FET) cycles? shown to suppress high LH levels at the start of OS for intrauterine insemination

in polycystic ovarian syndrome (PCOS) patients. A comparison between the Participants/materials, setting, methods: 150 participants with polycystic
administration of GnRH antagonist on Day 1 versus Day 6 of stimulation in ovary syndrome conducted in this study,The data collection was performed by
regular IVF cycles has also been performed, which demonstrated no significant using of the three structural questionnaire including demographic questionnaire,
differences neither in follicular development nor in the oocytes maturity, how- Eating Disorder Questioner(EDE-Q) and Health-Related Quality of Life
ever, so far, the effect on reproductive outcomes remains unknown. Questionnaire (PCOSQ).
Study design, size, duration: This was a retrospective, single-centre cohort Main results and the role of chance: Prevalence of eating disorder was
study including all women (aged 18-43 years) undergoing their first ovarian stim- 26.3%. Body mass index(BMI) was higher in women with eating disor-
ulation for IVF/ICSI in a tertiary referral university hospital between 2009 and der(P=0.001). Also results showed significant correlation between EDE-Q score
2018. A total of 8329 hCG-triggered GnRH antagonist cycles were included. and BMI(P=0.000). Regarding to the relationship between EDE-Q score and
Cycles with hormonal pre-treatment, IVM, managed natural cycles, oocyte dona- quality of life, the results indicated that the quality of life decreased as the EDE-Q
tion, and PGT were excluded from the analysis. score increased(P=0.000). On the other hand, BMI and quality of life were also
Participants/materials, setting, methods: LH serum levels at the initiation inversely correlated(P=.033).
of the cycle were studied in all included patients. Cycles were divided into two Limitations, reasons for caution: no limitation
groups based on a LH threshold level of 10 UI/L. The LH grouping was repeated Wider implications of the findings: Clinicians should be aware of diagnosis
using a LH threshold level of 15 UI/L in order to obtain cycles with very-high and management of eating disorder in infertile women specially PCOS patients
basal LH level. The primary outcome was CLBR. The secondary outcome to prevent its complications.
was LBR. Trial registration number: IR.SSU.SPH.REC.1397.149
Main results and the role of chance: General population baseline charac-
teristics were average female age (33.6 ± 5 years), average basal LH (5.6 ± 3.1 P-592 Ovulatory women with polycystic ovaries and their clinical
UI/L) and mean number of oocytes retrieved (9 ± 6.1). The population was outcome following individualised ovarian stimulation based on
divided into a group of 7758 cycles with basal LH<10 IU/L and a group of 571 AMH and body weight.
cycles with basal LH ≥10 IU/L. The multivariate analysis adjusting for potential H. Visnova1, E. Papaleo2, F. Sánchez Martin3, K. Koziol4, B. Klein5,
confounders did not show any difference between the two groups in terms of B. Mannaerts6
CLBR (31.8% vs. 30.9%, p value=0.678) nor LBR (23.1% vs. 21.4%, 1
IVF CUBE, Reproductive Medicine, Prague 6, Czech Republic ;
p value=0.339). The analysis was repeated with the basal LH threshold level set 2
San Raffaele hospital, Obstetrics and Gynaecology Dept, Milan, Italy ;
at 15 IU/L, resulting in a group of 8216 cycles (LH<15 IU/L) and 113 cycles 3
Ginemed, Fertility Clinic, Sevilla, Spain ;
(LH ≥ 15 IU/L). Although LBR was comparable (23% vs. 23%, p value=0.997), 4
Novum Warsaw, Fertility Clinic, Warsaw, Poland ;
CLBR in cycles with basal LH ≥ 15 IU/L was significantly higher compared to 5
Ferring Pharmaceuticals, Global Biometrics, Copenhagen, Denmark ;
those in cycles with LH<15 IU/L (31,7% vs. 41,7%, p=0.05). Multivariable regres- 6
Ferring Pharmaceuticals, Reproductive Medicine and Maternal Health,
sion analysis showed that lower age, a diagnosis of Rotterdam PCOS and higher
Copenhagen, Denmark
number of cryopreserved embryos were significantly associated with CLBR
(adjusted OR = 0.94, 95% CI=0.92-0.94, p value <0.001; 1.25, 95% CI=1.08-
Study question: How does the efficacy and safety of individualised fixed dosing
1.46, p value=0.02; 1.29, 95% CI=1.26-1.32, p value <0.001 respectively).
regimen compare to conventional dosing for ovarian stimulation in patients with
Limitations, reasons for caution: In spite of the large dataset and the rig-
polycystic ovaries?
orous methodological approach, the presence of biases related to the retro-
Summary answer: The ongoing pregnancy rates were similar but the incidence
spective design of the study cannot be excluded.
of early moderate/severe OHSS and/or preventive interventions for OHSS
Wider implications of the findings: Based on our data, the assessment of
was three times lower following individualised stimulation.
basal LH levels at the initiation of an hCG-triggered GnRH antagonist cycle could
What is known already: Individualised dosing of follitropin delta targets a
be omitted.
mean of 11 oocytes (range 8-14 oocytes) and has proven to optimize ovarian
response resulting in similar efficacy and improved safety compared to conven-
tional ovarian stimulation. The safety improvement is positively associated with
P-591 The relation of eating disorder and polycystic ovarian serum AMH levels, which is a biomarker of polycystic ovarian morphology.
syndrome in female Study design, size, duration: Retrospective comparative analysis of a subset
M. Bazrafkan1, M. Amiri2, A. Nadjarzadeh3 of 153 women with polycystic ovaries with AMH > 35 pmol/l. Following ovarian
1
Reproductive Biotechnology Research Center- ACECR, Avicenna Research Institute, stimulation, women with <25 follicles ≥12 mm were triggered using recombinant
tehran, Iran ; hCG and were eligible for fresh blastocyst transfer. Women with ≥25 follicles
2
Shahid Sadoughi University of Medical Sciences- Faculty of Health, Nutrition, ≥12 mm were either triggered with GnRHa for subsequent frozen embryo trans-
Yazd, Iran ; fer or had their cycle cancelled. Frozen embryo transfers up to one year after
3
Shahid Sadoughi University of Medical Sciences-Faculty of Health, Nutrition, the stimulation cycle were included in the analysis.
Yazd, Iran Participants/materials, setting, methods: Women aged 18-40 years were
randomised for ovarian stimulation to either an individualised fixed-dose of
Study question: The aim of this study was to clarify the prevalence of eating follitropin delta based on AMH and body weight (n=78) or to 150 IU follitropin
disorder and relation with BMI and quality of life in women with PCOS. alfa (n=75) which daily dose could be adjusted after 5 days of stimulation. Only
Summary answer: Women with PCOS had a higher BMI and eating disorder. single blastocyst transfers were performed with the exception of one double
Quality of life scores decrease by the eating disorder and body mass index blastocyst transfer in the follitropin alfa group. Continuous endpoints were eval-
increase. uated using t-tests. Binary endpoints were evaluated using chi-square tests.
What is known already: Polycystic ovary syndrome(PCOS) is an endocrine Main results and the role of chance: Women with polycystic ovaries had
disorder in women, that can lead to infertility. Eating disorder has been suggested the same characteristics in both treatment groups with an overall age of 31.8
as one of the leading causes of obesity that is a main risk factor to PCOS. Also, years, body weight of 62.8 kg, and AMH of 49.2 pmol/l. At the end of stimu-
women with PCOS had lower quality of life due to obesity and overweight that lation the number of follicles (≥12 mm) was 12.1 ±7.0 and 18.3 ±7.0 with
are the largest contributor to poor quality of life. On the other hand, obesity increased progesterone (>3.18 nmol/l) in 27% and 67% in women treated with
and weight loss is major concern in this women that lead to depression and individualised follitropin delta and conventional follitropin alfa, respectively. The
lower quality of life. overall mean (SD) number of oocytes was 9.3 ±6.7 and 17.9 ±8.7 respectively
Study design, size, duration: This cross sectional study was conducted in and there were slightly more women with blastocyst transfer (74.4% vs. 68.0%)
Avicenna Research Institute by 150 participants with polycystic ovary syndrome. and slightly less women with at least one cryopreserved blastocyst (61.5% vs
The data collection was performed by using of the three structural questionnaire 76.0%) in the follitropin delta group. The ongoing pregnancy rate per started
including demographic questionnaire, Eating Disorder Questioner(EDE-Q) and stimulation cycle following fresh blastocyst transfer was 28.2% and 24.0%, respec-
Health-Related Quality of Life Questionnaire (PCOSQ). tively. The incidence of all cases of OHSS was three times higher in the follotropin

1
alfa group i.e. 5.1% vs 16.0% (P=0.025) as well as the combined early moderate/ Rabin Medical Center-Beilinson Hospital, IVF and Infertility Unit, Petach Tikva,
severe OHSS and/or preventive interventions for early OHSS i.e. 7.7% with Israel ;
2
individualised follitropin delta and 26.7% with conventional follitropin alfa Tel Aviv University, Electrical engineering, tel aviv, Israel
(P=0.001).
Limitations, reasons for caution: This retrospective analysis includes ovu- Study question: Is there a machine learning algorithm that can predict oocyte
latory women with polycystic ovaries defined as serum AMH > 35 pmol/l. maturation rate in GnRH antagonist cycles using baseline and treatment
Anovulatory women may have different efficacy and safety profiles and should characteristics
be studied separately. Summary answer: Machine learning algorithms can predict oocyte maturation
Wider implications of the findings: The results demonstrate that individ- rate using simple parameters. We demonstrated an accuracy rate of 75% in
ualised follitropin delta treatment provides an improved balance between efficacy predicting high oocyte maturation rate
and safety in potential high responders, as it normalizes the ovarian response What is known already: GnRH antagonist cycles may be associated with
and decreases the incidence of OHSS and preventive interventions of OHSS, heterogeneous follicular development that may result in lower oocyte maturation
with less embryo transfer cancellations. rate. Oocyte maturation is affected by different baseline and treatment param-
Trial registration number: NCT01956110 eters, and has a key effect on treatment outcome. As far as we know, a prediction
model for oocyte maturation rate made by machine learning and neural network
algorithms was not described yet.
P-593 Effect of AMH, age and history of previous pregnancy on
Study design, size, duration: A retrospective cohort study of 462 women
the number of total and mature oocytes obtained in a donation
aged ≤ 38 who underwent their first IVF treatment using a GnRH antagonist
programme
protocol in one tertiary hospital. All were treated with a flexible antagonist
V. Pataia1, M. Wolska1, J. Wang2, N. Macklon2, K. Ahuja2 protocol. Median maturation rate was approximately 80%,. Treatment param-
1
London Egg Bank, London Egg Bank, London, United Kingdom ; eters of cycles with high oocyte maturation rate (≥80%, n=236) were compared
2
London Women’s Clinic, London Women’s Clinic, London, United Kingdom to cycles with low oocyte maturation rate (<80%, n=226). Patients that their
oocytes were fertilized by insemination only without ICSI were excluded.
Study question: In an oocyte donation programme, does Anti-Mullerian hor- Participants/materials, setting, methods: We used XGBoost algorithm that
mone (AMH), age and/or history of previous pregnancy predict the number of fits the training data using decision trees, and can also rate the factors according to
total and mature oocytes obtained? their influence on the prediction. We assigned a weight to each class (positive/
Summary answer: AMH correlated to the number of total and mature negative), to overcome potential biases due to the unbalanced nature of data. For
oocytes obtained from oocyte donors, but age and history of previous pregnancy the machine learning training phase, 80% of the cohort was randomly selected. We
did not. have then used the rest of the samples as a test-set, to evaluate our model’s accuracy.
What is known already: A relationship between age, AMH and ovarian Main results and the role of chance: The following data was retrieved from
stimulation response is well established in the subfertile population. However, medical records: patient’s age, gravity, parity, BMI, Infertility cause and duration,
the value of these markers in healthy oocyte donors remains to be clarified. gonadotropins doses number of oocyte retrieved, treatment parameters on the
Additionally, the effect of previous donor pregnancy on oocyte donation out- day of GnRH antagonist initiation including estradiol and progesterone levels,
comes remains largely unexplored. maximal follicle diameter and number of follicles, GnRH antagonist treatment
Study design, size, duration: Retrospective cohort analysis of 223 oocyte duration, and treatment parameters on trigger day including number of follicles,
donors. Donors were aged 18-35, registered in accordance with the Human estradiol and progesterone levels.
Fertilisation and Embryology Authority (HFEA) guidance, and underwent their The test-set samples’ parameters were fed into the trained model and the pre-
first donation cycle between January 2018 and May 2019. Donors were divided dicted outcome was compared to the actual outcome. On the randomly picked
into 6 age groups: 18-20 (n=28), 21-23 (n=60), 24-26 (n=39), 27-29 (n=45), samples used to evaluate our model, we demonstrated an accuracy rate of 75%
30-32 (n=27), 33-35 (n=24). Donors were also divided according to their repro- in predicting oocyte maturation rate in GnRH antagonist cycles. The most pre-
ductive history as previously pregnant (n=65) or never pregnant (n=158). dictive parameters arranged by descending importance order were: estradiol
Participants/materials, setting, methods: Kruskal-Wallis test was applied level on trigger day, estradiol level on antagonist initiation day, average gonado-
to compare the relationship between study parameters and outcomes between tropins units per day, progesterone level on trigger day, patients’ weight, number
age groups. The respective associations within each age group were investigated of oocytes retrieved, patients’ height, number of follicles on GnRH antagonist
using Spearman’s rank correlation. Mann-Whitney U test was performed to initiation day, patients’ age and infertility duration.
assess the effect of previous pregnancy on study parameters and outcomes. Limitations, reasons for caution: Retrospective design and lack of infor-
Main results and the role of chance: No significant differences were mation regarding follicle synchronization.
obtained in mean AMH levels, total oocytes or total mature oocytes between Wider implications of the findings: A state-of-the-art machine learning algo-
donor age groups (p>0.05). Moreover, a history of previous pregnancy had no rithm presented promising ability to predict oocyte maturation rate using simple
significant impact on study parameters (p>0.05). AMH was however positively parameters. We now plan to expand the cohort and evaluate the model in
correlated with the number of mature oocytes obtained (p≤0.05), and the larger numbers
strength of association increased with age. Similar findings applied to the total Trial registration number: not applicable
number of oocytes obtained, particularly in donors ≥27 years old.
Limitations, reasons for caution: This is a small study and a higher number P-595 Progestin Primed Ovarian Stimulation is an effective oral
of donors included would allow clinical outcomes to be assessed. All donors alternative for Antagonist Protocol in patients undergoing assisted
accepted on the donation programme had a minimum AMH level of 12pmol/L reproductive techniques: a retrospective study.
and/or antral follicle count of 12. Thus, our results may not be replicated in the J.P. Caetano1, L. Calazans1, L. Amorim1, E. Xavier1, A.L. Campos2,
general population. B. Coimbra3, R. Marinho1
Wider implications of the findings: In a cohort of healthy oocyte donors 1
Pro-Criar Medicina Reprodutiva, Medical, Belo Horizonte, Brazil ;
under 35 years-old meeting standard ovarian reserve criteria for acceptance, a 2
Pro-Criar Medicina Reprodutiva, Embriology, Belo Horizonte, Brazil ;
similar number of total/mature oocytes is likely to be obtained irrespective of 3
Hospital das Clíncias - Universidade Federal de Minas Gerais, Medical Residency,
age or history of previous pregnancy. Belo Horizonte, Brazil
Study question: Are Clinical Pregnancy Rates in Progestin Primed Ovarian
Stimulation noninferior to Antagonist Protocol in patients undergoing assisted
P-594 Prediction of oocyte maturation rate in antagonist flexible reproductive techniques (ART)?
IVF protocol using a novel machine learning algorithm Summary answer: Our results showed noninferiority of Clinical Pregnancy
O. Houri1, A. Wertheimer1, Y. Gil2, S. Danieli-Gruber1, G. Oron1, Rates in Progestin Primed Ovarian Stimulation compared to Antagonist Protocol
O. Sapir1, S. Yoel1, A. Ben Haroush1 with blastocyst transfer and freeze-all strategy.

What is known already: The blockade exerted by progestin in ovulation accounting for only 2 % of total serum P4. Importantly, salivary P4 contains free
induction involves blocking the hypothalamic-pituitary-ovary axis, changing the progesterone, only.
frequency and amplitude of GnRH pulses (La Marca et al., 2019). Early LH peak Moreover, salivary P4 fluctuates during the natural cycle, whether this is the
suppression occurs despite high oestradiol concentration (Kuang et al., 2015). case in different ART cycles is unknown until now.
In the current stage of ART development, in which embryo freezing is an increas- Study design, size, duration: An observational study including 20 patients.
ingly common practice and provides good results, new options for controlling Ten IVF patients treated with GnRH-antagonist protocol and 10 patients under-
the LH peak can be considered (Kuang et al., 2015). The use of progestins has going Hormone Replacement Therapy Frozen Embryo Transfer (HRT-FET) were
aroused interest in this regard, and the possible negative effect on the endome- included from October 2018 to April 2019. Salivary P4 was measured over a
trium is no longer a concern (Yu et al., 2018). 12-hour period from 7 a.m. to 7 p.m. on blastocyst transfer day and in HRT-FET
Study design, size, duration: A retrospective study included all ovarian patients on 6th day of vaginal progesterone treatment. Furthermore, serum P4
punctures performed at Pró-Criar Medicina Reprodutiva, Belo Horizonte, Minas was measured the same day between 11 a.m. and 1 p.m.
Gerais, Brazil, between May 2018 and May 2019 using a GnRH antagonist ana- Participants/materials, setting, methods: Patients were included from a
logue or oral progestins to block the LH peak in IVF/intra-cytoplasmic sperm Danish public fertility clinic. Saliva samples were frozen upon collection and P4
injection (ICSI) cycles for infertility treatment. A total of 266 IVF/ICSI cycles levels were measured using enzyme immunoassay kits (Salimetrics™) at a sep-
performed were analysed and 222 cycles (83.5%) were included in the study, arate clinic. Immunoassay measurements were conducted precisely according
112 in the Progestin group and 110 in the Antagonist Group. to assay specifications. Reactions were conducted in hormonespecific anti-
Participants/materials, setting, methods: Exclusion criteria were age older body-coated microtitre plates. Samples and controls were incubated simultane-
than 42 years, fresh embryo transfer, transfer at the cleavage stage (D2/D3), ously with competitive horseradish-peroxidase-bound P4. Competitive reactions
preimplantation genetic screening, cryopreservation of oocytes, shared donation were detected using the peroxidase substrate tetramethylbenzidine (TMB) via
and cycles without embryo transfer at the time of analysis. The primary outcome optical density readings at 450 nm.
evaluated was the clinical pregnancy rate at the first embryo transfer. The sec- Main results and the role of chance: The 12-hour salivary P4 profiles in
ondary outcomes were the mean MII oocytes retrieved, fertilization rate, blas- IVF patients showed substantial fluctuations on the blastocyst transfer day; how-
tocyst formation rate, mean duration of stimulation and mean dose of ever, no time related pattern was discovered. The percentage variation in IVF
gonadotropins. cycles ranged from 49 to 351%, with levels ranging from 202 pg/ml to 23953
Main results and the role of chance: The primary outcome of our study, pg/ml. One IVF patient had a mean salivary P4 level of 355 ±97 pg/ml which
Clinical Pregnancy Rate at the first embryo (Blastocyst) transfer, was 58.4% in the correlated to a low mean serum P4 level of 8.5 ng/ml. The corpus luteum (CL)
progestin group and 54.9% in the antagonist group (P = 0,735), a finding consistent deficiency was found despite the fact that the patient had 19 follicles and 12
with most studies published to date using different progestins. The mean number oocytes retrieved; this patient did not conceive.
of retrieved oocytes was 11 in the antagonist group and 9 oocytes in the progestin In HRT-FET cycles, similar fluctuations of salivary P4 levels were seen despite
group (P = 0.009). The fertilization rate was 80% for both groups (P=0.935). The the absence of a CL and the fact that P4 derived from vaginal P4 application,
rate of blastocyst formation per cycle was 50% in the antagonist group and 55.6% only. The percentage variation ranged from 99 to 395 %, and salivary P4 levels
in the progestin group (P = 0.106). Stimulation lasted a mean of 10 days in the from 8 to 21044 pg/ml. In contrast, the percentage variation in serum P4 ranged
two analysed groups and did not vary with patient age in either group. The gonad- from 11 to 47 %, only.
otropin dose used was higher in the antagonist group (2025 IU) than in the pro- In IVF as well as in HRT-FET cycles the ratio of salivary P4 to serum P4 varied
gestin group (1950 IU) (P = 0.057). In addition, the blockade was effective: there significantly between patients. This individual variation was also previously
was only one case of spontaneous ovulation, which corresponded to less than 1% described during natural cycle.
of the cycles, an incidence compatible with the 0.34 to 8% risk described in the Limitations, reasons for caution: This study included a limited number of
literature for failure to control the LH peak in antagonist protocol cycles. patients, and seven out of 160 salivary P4 levels were very high. Until now it is
Limitations, reasons for caution: This was not a randomized controlled unclear whether these high measurements are caused by contamination of the
trial, and the choice of protocol depended on the attending physician, yet sta- saliva samples, due to non-compliance with collection instructions, or a true high
tistical analysis showed that the studied populations were similar, with similar salivary P4.
mean age, BMI, duration of infertility and distribution of causes of infertility Wider implications of the findings: The analysis of salivary P4 may be a new
between the groups. patient friendly alternative to serum sampling for luteal phase monitoring in ART.
Wider implications of the findings: The use of progestins to block the LH However, future studies should explore optimal cut-off levels of salivary P4 levels
peak has the advantages of ease of oral administration and lower cost compared during luteal phase in ART. In the long term a home test could be developed.
to GnRH antagonists. It allows flexible ovulation monitoring, therefore more Trial registration number: ClinicalTrial.gov no.: NCT03725904
comfortable for the patient. The freeze-all strategy allows the embryos to be
transferred into a more physiological uterine environment. P-597 Cumulative live birth rate with recombinant follicle
Trial registration number: not applicable stimulating hormone biosimilar: a multicenter study with over
than 7,000 cycles.
P-596 Salivary progesterone - a new diagnostic tool for luteal E. Arbo1, B. Paul2, C. Avril3, S. Hamamah4, J.L. Pouly5, M.
phase monitoring in Assisted Reproductive Technology (ART) Grynberg6, B. Salle7
B. Alsbjerg1, D. Sakkas2, B. French2, H.O. Elbaek1, B.B. Povlsen1, 1
Medical Director- Gedeon Richter France, Medical Affairs Department, Paris,
R.J. Laursen1, M. Alper2, P. Humaidan1 France ;
2
1
Regional Hospital Viborg - Skive Sundhedshus, The Fertility Clinic- Skive Regional Centre Hospitalier Universitaire de Nantes- CRTI- UMR 1064- INSERM-
Hospital, Skive, Denmark ; Université de Nantes- ITUN- CHU de Nantes, Department of Biology and
2
Boston IVF, Boston IVF, Waltham MA, U.S.A. Reproductive Medecine-, Nantes, France ;
3
Clinique Mathilde, AMP Mathilde, Rouen, France ;
4
Study question: Can salivary progesterone (P4) measurement be used as an Arnaud de Villeneuve Hospital-CHU Montpellier - INSERM U 1203- Hôpital Saint
alternative and more patient-friendly diagnostic tool for luteal phase monitoring Eloi- 34295, ART-PGD Department, Montpellier, France ;
5
during ART? CHU de Clermont-Ferrand- 63003, Gynécologie Obstétrique et Reproduction
Summary answer: Luteal phase deficiency was detected by constant low Humaine, Clermont-Ferrand, France ;
6
salivary P4 in IVF. Hôpital Antoine Béclère- Inserm U1185 Université Paris-Sud- Université Paris
What is known already: The reproductive outcome of ART depends on P4 Saclay- 94276 Le Kremlin Bicêtre- France. Inserm U1133 Université Paris
levels during the luteal phase. Traditionally, P4 levels are measured in serum, Diderot- 75013 Paris- France., Department of Reproductive Medicine and Fertility
which is cumbersome for patients due to constant venopunctures. Furthermore, Preservation-, Paris, ;
7
when P4 level is measured in serum it is total P4; meaning, both the part tightly Hôpital Femme Mère Enfant Bron- 69677, Human Reproduction Department,
bound to proteins and the free bioactive portion. Free P4 is the active part Lyon, France

Study question: In routine clinical practice, is the cumulative live birth rate Study design, size, duration: This is a retrospective cohort study. None
(cLBR) followed by IVF/ICSI using Bemfola®, a biosimilar of follitropin alfa, interventions were applied to the patients. The main outcome measures are as
comparable to the originator? followed: Lipid metabolic index, including total cholesterol (TCHO), triglycerides
Summary answer: This real-world data confirms the clinical effectiveness of (TG), high density lipoprotein cholesterol (HDL-C) and LDL-C. Basal sex hor-
Bemfola® for controlled ovarian stimulation (COS) for IVF/ICSI. mone, including follicle stimulation hormone (FSH), luteinizing hormone (LH),
What is known already: Although randomised controlled phase 3 clinical progesterone(P), estradiol (E2) and AMH. IVF index, including AFC, no.RO,
trials on selected populations demonstrate the efficacy of new drugs to allow maturation rate (MR), fertilization rate (FR) and embryo formation rate (EFR).
approval for routine use, once drugs have been introduced into clinical practice, Participants/materials, setting, methods: The participants were PCOS
real world data (RWD) may assess their actual effectiveness. As in the case of patients with clomiphene resistance undergoing natural cycles from January 1,
Bemfola®, a follitropin alfa biosimilar approved on the basis or its proven efficacy 2015 to November 30, 2019. All the data were from clinical records.
related to the number of oocytes retrieved, RWD may assess additional param- Main results and the role of chance: 219 patients were eventually included
eters of clinical interest such as live birth rates following IVF/ICSI. Indeed, RWD in this study. Scatter plots and fitting curves showed that TCHO and LDL-C had
reflect the actual effectiveness in an unselected infertile population across mul- a tendency to increase first and then decrease with no.RO, while HDL-C had a
tiple indications, protocols and clinical strategies. tendency to decrease first and then increase with no.RO. The correlation
Study design, size, duration: National, multicenter, retrospective, cohort between BMI, TG and no. RO was not statistically significant. Cutoff values of
study conducted in 17 public and private ART centers. Data from all cycles of TCHO (5.89mmol/L), HDL-C (1.71mmol/L) and LDL-C (4.09mmol/L) were
women who underwent COS with a follitropin alfa, between January 2016 and identified through fitting curves. Among four BMI groups (low: <18.5Kg/m2,
February 2017 including follow-up period up to December 2017 were collected. normal: 18.5~24Kg/m2, overweight: 24~28Kg/m2 and obesity: ≥28Kg/m2),
The main assessment criterion was cumulative live birth rate (cLBR) including only AMH and four lipid metabolic index showed statistically significant differ-
fresh and frozen-thawed embryo transfers per initiated cycle. ences, while all IVF index and other basal sex hormone did not. Next, cutoff
Participants/materials, setting, methods: Data from 9,162 COS cycles values were used separately to divide each BMI group into two subgroups. The
were extracted. The present descriptive analysis includes 7,448 cycles for which results showed that significant differences were found in AMH, AFC and no.RO
COS was performed with a follitropin alfa without any association with other between LDL-C subgroups (BMI: normal), P and AFC between TCHO sub-
gonadotropin. From these, 2,478 cycles were performed with Bemfola® and groups (BMI: normal), E2 between LDL-C subgroups (BMI: overweight) and P
4,970 with another follitropin alfa (90% corresponding to the originator between LDL-C subgroups (BMI: obesity). No statistically significant differences
Gonal-f®). cLBR in both groups are displayed with their 95% confidence interval in MR, FR and EFR were observed in all subgroups comparisons.
(95%CI). Limitations, reasons for caution: The sample size of subgroups higher than
Main results and the role of chance: Mean female age were 34.1±4.7 and TCHO, HDL-C and LDL-C cutoff values is still limited, and this part of the sample
33.9±4.7 years and mean BMI were 23.9±4.7 and 23.8±4.6 kg/m² in the size should be further expanded.
Bemfola® and other follitropin alfa groups, respectively. The medians of AMH Wider implications of the findings: LDL-C may be a better predictor of
serum levels were 2.3 ng/mL (interquartile interval:1.3-3.8) and 2.6 ng/mL no.RO than BMI in natural cycles.
(interquartile interval: 1.4-4.7) and the proportion of women presenting an AMH Trial registration number: ChiCTR-ONC-17011861
serum level > 5.2 ng/mL were 14.8% and 21.5%, in the Bemfola® and other
follitropin alfa groups. The most prescribed starting dose of follitropin alfa were P-599 Replacing antagonist by clomiphene citrate to prevent
between 150-225 IU in both groups. Oocyte retrieval resulted in 7.1±5.7 and premature ovulation in IVF-ET: is it possible?
7.9±5.5 mature oocytes in the Bemfola® and in the other follitropin alfa groups, S. Ekmekjian2, H. Maroun1, C. Boyadjian1, T. Tadros1,
respectively. cLBR per cycle were 20.0% (95% CI: 18.4%-21.5%) in the Bemfola® I. Aboujaoude1
group and 20.8% (95% CI: 19.7%-21.9%) in the other follitropin alfa group. cLBR 1
Aboujaoude Hospital-, Center for Reproductive Medicine-, jaleldib, Lebanon ;
in the COS cycles that were followed by at least one embryo transfer during the 2
Lebanese University faculty of sciences II, Reproduction biology lab, Fanar,
follow-up period were 21.2% (95% CI:19.5-22.8) in the Bemfola® group and
Lebanon
21.6% (95% CI:20.5-22.8) in the other follitropin alfa group.
Limitations, reasons for caution: Although systematic biases cannot be
Study question: Do 150 mg of clomiphene citrate (CC) capable of replacing
excluded, detailed analyses revealed that the apparent biases seem to favor the
the classical antagonists in controlled ovarian stimulation (COS) to prevent pre-
originator follitropin alpha, suggesting that Bemfola® had been preferentially
mature ovulation (PO)?
prescribed to poorer prognosis patients. The comparator follitropin alpha group
Summary answer: Our data indicated that CC is as efficient as antagonist
included another follitropin alpha biosimilar, but too few cases to impact
in IVF-ET.
results (10%).
What is known already: Since the introduction of antagonists in the 90th, the
Wider implications of the findings: This is the largest multicentric cohort
antagonist protocol is becoming the most used protocol in COS for IVF-ET
describing cLBR with the use of a follitropin alfa biosimilar in a non-selected pop-
patients. However, due to their relatively high cost, infertility specialists in devel-
ulation. This real-world evidence demonstrates that the use of Bemfola® is an
oping countries try to find a more affordable molecule to avoid premature the
effective option for COS, with clinical results comparable with other follitropin alfa.
ovulation during COS. New larges studies showed that CC could be used to
prevent PO in mild stimulation using 50 mg daily. No studies to our knowledge
used CC in standard IVF-ET protocols. Therefore, we decided to do a pilot study
P-598 The Effects of Lipid Metabolism and Obesity on the in order to replace antagonists with CC 150 mg per day until triggering.
Number of Retrieved Oocytes in Polycystic Ovary Syndrome Study design, size, duration: We studied 161 patients who entered our
Undergoing Natural Cycles IVF-ET program. All patients started COS on day 3 of the menstrual cycle using
T. Liu1, J. Yan1 HMG or recombinant FSH.
1
Beijing University Third Hospital, Center for Reproductive Medicine, Beijing, China Participants/materials, setting, methods: Patients were sorted as follows:
group 1 Ganirelix (n=73) underwent the classic antagonist protocol where
Study question: To assess the effects of lipid metabolism and body mass index Ganirelix was introduced at day 6 of the COS; group 2: CC (n=88) where we
(BMI) on the number of retrieved oocytes (no.RO) in polycystic ovary syndrome replaced the antagonist by CC 150 mg. Single or double triggering for final oocyte
(PCOS) undergoing natural cycles. maturation was performed using HCG 5000 IU and/or Decapeptyl 0.3 mg when
Summary answer: The increase of serum low density lipoprotein cholesterol at least 3 follicles reached 17mm in diameter. Oocyte retrieval was conducted
(LDL-C) can improve anti-Mullerian hormone (AMH) level, antral follicle counts 35 to 37 hours after triggering.
(AFC) and no.RO when BMI is normal. Main results and the role of chance: As shown in the Table1, whereas
What is known already: The no.RO can be affected by various factors in patients and COS characteristics were similar in both groups, we observed no
controlled ovarian hyperstimulation (COH) cycles, in which BMI and lipid metab- statistical differences in terms of number of oocytes retrieved, empty follicles
olism are very important to predict ovarian response. cases and pregnancy rates.

ANT CC(n=73) ANT Org(n=88) P
Ages (years) 28.1 ± 4.71 30.3 ± 4.39 (19-35) 0.0032*

Duration of infertility (years) 7.3 ± 4.034 (0.5-19) 6.8 ± 4.348 NS
IVF-ET Indications • Male (%) 68.5 65.5 NS
• IUI failure (%) 12 13.5 NS
• Tubal (%) 11.5 11 NS
• Endometriosis (%) 8 9 NS
Duration of COS (days) 8.4 ± 1.44 8.6 ± 1.89 NS
E2 day of trigger (pg/ml) 2825.5 ± 2065 1870.4 ± 1414.60 0.01
Progesterone day of trigger >1.5 ng/ml (%) 21.1 16.7 NS
Total dose of Gonadotropins (IU) 2108.6 ± 988.15 1954.5 ± 737.72 NS
No. of total oocytes 9.8 11.3 NS
MII oocytes (%) 69.3 77 NS
Empty follicles (%) 4.1 2.3 NS
No. of embryos transferred 2.7 2.5 NS
Clinical pregnancy rate / transfer (%) 38.8 24.7 NS
Ongoing pregnancy / transfer (%) 31.3 23.5 NS
Miscarriage rate (%) 19.2 5 NS
38.831.319.2 Study design, size, duration: A prospective, randomised controlled trial was
Limitations, reasons for caution: The present study could not address the carried out from 1st January 2019 to 31st October 2019 at a tertiary infertility
exact mechanism of action of the CC on the hypothalamic-pituitary axis to centre in India. 50 patients fulfilling the criteria of Group 3 and Group 4 of
prevent PO neither discriminating possible outcome differences using lower dose POSEIDON classification were included in the study. Patients with endocrine
of CC due to limited sample size. Moreover, potential side effects of CC where disorders (thyroid, prolactin), endometrioma, history of surgery on the ovaries,
not taken into consideration in this study. sensitivity to testosterone gel, male factor infertility and deranged liver and renal
Wider implications of the findings: Our data indicate that clomiphene function tests were excluded.
seems to be as efficient as the antagonists, cheaper, and easier to use in order Participants/materials, setting, methods: Enrolled patients were ran-
to prevent premature ovulation in IVF-ET. Further randomized controlled stud- domised into two groups of 25 patients each, one group was pretreated (TTG
ies, including larger populations, are needed to confirm and expand present group) with transdermal testosterone gel ,12.5 mg/day from day 6th of previous
findings. cycle to day 2nd of stimulation cycle while patients in other group took DHEA
Trial registration number: not applicable tablet,75 mg/day for a total duration of three months (DHEA group) before
stimulation with GnRH antagonist fixed protocol followed by fresh Day 3 transfer
P-600 Transdermal testosterone vs. oral dehydroepiandrosterone Main results and the role of chance: The baseline characteristics of the
(DHEA) pre-treatment in improving IVF outcomes in diminished two groups were comparable. The primary outcome measures were the number
ovarian reserve patients (POSEIDON group 3 and 4): a of oocytes retrieved and number of grade A embryos formed (according to
randomised controlled trial Istanbul consensus). The secondary outcome measures were implantation rate,
clinical pregnancy rate, miscarriage rate and ongoing pregnancy rate. The mean
N. Sharma1, K.D. Nayar1, S. Gupta2, M. Singh1, R. Bhattacharya1,
number of oocytes retrieved in TTG group was 5±1.4 which was significantly
G. Kant1, R. Gahlot1, K.D. Nayar1
1
higher than DHEA group-3.3±1.7, (p<0.001). The mean number of Grade A
Akanksha IVF Centre- Mata Chanan Devi Hospital, Reproductive Medicine, New embryos were also significantly higher (4.28±0.88 vs 2.85±0.63, p<0.001) in
Delhi, India ;
2
TTG group. The TTG group had higher implantation rate (28% vs 20%, p= 0.49),
Akanksha IVF Centre- Mata Chanan Devi Hospital, Reproductive Medicine, New clinical pregnancy rate (32% vs 20%, p = 0.41), ongoing pregnancy rate (32% vs
Delhi, India 16 %, p= 0.38) and lower miscarriage rate (0% vs 20%, p=0.38), however, these
differences were not statistically significant.
Study question: To compare the efficacy of transdermal testosterone with
Limitations, reasons for caution: The study was done at a single centre
oral dehydroepiandrosterone in improving IVF outcomes using GnRH antagonist
with small sample size, replication with more subjects and in different centers is
protocol in POSEIDON group 3 and 4 patients.
needed. Also, cost effectiveness of either drug was not assessed.
Summary answer: Patients receiving pre-treatment with testosterone gel had
Wider implications of the findings: Pre-treatment with testosterone gel in
higher mean number of oocytes retrieved and grade A embryos as compared
DOR patients improves ovarian response to stimulation and results in higher
to the patients receiving oral DHEA
number of oocytes retrieved and good quality embryos resulting in improved
What is known already: Diminished ovarian reserve (DOR) is associated
clinical pregnancy rates. Transdermal testosterone is advantageous because of
with suboptimal ovarian response, higher cycle cancellation rate and lower clinical
better bioavailability, easy application, patient friendly and less adverse effects.
pregnancy rate following IVF cycles. Various treatment regimens have been
Trial registration number: MCDH/2019/28
devised for management of such patients and use of adjuvants in the form of
oral or transdermal androgen is one of them. Androgens improves follicular
response to gonadotropin stimulation as well as increase FSH receptor expres- P-601 The role of midkine, a heparin-binding growth factor, in
sion in granulosa cells, in turn leading to better oocyte yield and pregnancy rate. human follicle development and oocyte maturation
Aim was to compare the effect of transdermal testosterone gel with oral DHEA S.E. Pors1, S.G. Kristensen1, D. Nikiforov1, L. La Cour Poulsen2,
on the ART outcome in DOR patients(POSEIDON Group 3 and 4) A. Sanchez3, I. Pla Parda3, C. Yding Andersen1

1
Rigshospitalet, Laboratory of Reproductive Biology, Copenhagen, Denmark ; Study question: To compare the efficacy of letrozole and clomiphene citrate
2
Zealand University Hospital, Zealand Fertility Clinic-, Køge, Denmark ; for ovulation induction in infertile women with PCOS for IUI cycles in Indian
3
Lund University, Department of Translational Medicine, Lund, Sweden population
Summary answer: Letrozole favors monofollicular development and has a
Study question: Does midkine, a heparin-binding growth factor found in follicle better endometrial response when compared to CC though no difference in the
fluid, affect human follicle development and oocyte maturation? clinical pregnancy and multiple pregnancy rate
Summary answer: Midkine affect human follicle development by augmenting What is known already: Clomiphene citrate is used for ovulation induction
oocyte maturation in vitro and its concentration increases in intrafollicular fluid in PCOS. Clomiphene resistance occurs in 15-20% of the patients. PCOS women
during the mid-cycle surge of gonadotropins. in India have a higher prevalence of insulin resistance(~75%) and are more likely
What is known already: In vitro maturation (IVM) of immature oocytes to have CC resistance. CC also has a negative effect on the cervical mucus and
without exogenous FSH stimulation has attracted much interest in recent years endometrium. Treatment with CC is thus associated with a discrepancy between
and immature oocytes from small antral follicles (SAF) is a new way of augment- ovulation and conception rates. Letrozole, an aromatase inhibitor used for ovu-
ing the fertility potential of women undergoing fertility treatment by having lation induction has no adverse effects on the endometrium and the cervical
ovarian tissue cryopreserved. However, regulation of human oocyte maturation mucus. It can thus act as an effective alternative to CC in Indian women
is highly complex and dynamic and far from fully elucidated. The growth factor with PCOS
midkine, acting via the NOTCH signalling pathway, has recently been identified Study design, size, duration: A prospective clinical trial on 120 infertile
in human follicular fluid (FF) and may represent an important factor for follicle patients with PCOS diagnosed according to Rotterdam criteria was carried out
and oocyte development. at a tertiary care infertility centre in India from January 2019 – October 2019.
Study design, size, duration: Follicle fluid (n=50) from SAF, immature Participants/materials, setting, methods: One hundred and twenty
oocytes (n=891) and tissue were obtained from ovaries from women undergoing women with PCOS were divided into two groups-61patients with Letrozole,
ovarian tissue cryopreservation for fertility preservation. Small antral follicles 2.5 mg/day and 59 patients with clomiphene citrate 100 mg/day from day 3-7
with different diameters were isolated from surplus medulla tissue (n=183). In of the menstrual cycle. Follicular monitoring was done and 10,000 IU of HCG
addition, FF from preovulatory follicles were aspirated from women undergoing was administered when the largest follicle was ≥18 mm. Intra-uterine insemina-
fertility treatment at different time points after final follicle maturation was tion was done 36-40 hours after HCG administration. 400mg micronized pro-
induced (n=25). All women had given written consent for use of the material gesterone was given intra-vaginally for 15 days as luteal phase support.
for research purposes. Main results and the role of chance: Baseline characteristics in both letro-
Participants/materials, setting, methods: Concentration of midkine in FF zole and CC group were comparable. Mean number of follicles ≥ 18mm on the
was analysed by proteomics and ELISA. Microarray data on gene expression day of hCG administration was 1.13±0.53 in letrozole group and 2.6±1.15 in
from SAF were obtained and analysed. Immunohistochemistry using antibodies the CC group (p <0.0001). Mean endometrial thickness on the day of hCG
against midkine and Notch2 were performed. Immature oocytes, collected after administration was 8.21±0.86 in letrozole group and 7.35±0.99 in CC group (p
ovarian tissue cryopreservation, were randomly allocated for IVM with or with- <0.0001). Ovulation rate in letrozole group was 77.04% and in CC group was
out midkine in the media (1 ug/ml). From the same patient, two FF containing 59.32%, which was not statistically significant (p= 0.05). Clinical pregnancy rate
immature oocytes from SAF were obtained. FF levels of midkine were associated was 14.75%(9/61) with letrozole group and 13.56%(8/59)with CC group, how-
to the IVM outcome. ever this difference was not statistically significant (p>0.05). Mutiple pregnancy
Main results and the role of chance: FF samples from preovulatory follicles rate was higher with CC group (2.5% (2/8)vs. 1.11%(1/9) , p=0.57) but not
were collected at five time points after ovulation trigger: 0, 12, 17, 32 and 36 h, statistically significant
respectively. Expression of midkine protein was significantly higher at 0,12 and Limitations, reasons for caution: The study was done at a single centre
17 h compared to 32 and 36 after the ovulation trigger. When measuring the with small sample size. Replication with more subjects and multiple centres
midkine content with ELISA the concentration of midkine was high at all time- is needed
points measured after administration of the ovulation trigger and only decreased Wider implications of the findings: Letrozole leads to more monofollicular
significantly at 32 h. In SAF midkine was also present in concentration ranging development and better endometrial response compared to CC. Hyperinsulinemia,
from 190-2300 ng/ml. The highest levels of gene expression of midkine was which is frequently associated with PCOS, is one of the causes for CC resistance.
present in SAF (<9 mm) compared to larger follicles (>9 mm). Preliminary results The prevalence of insulin resistance in PCOS is approximately 75%.Thus, letrozole
of the immunohistochemical investigations indicate that midkine and its potential has an important role as first line treatment for PCOS patients.
receptor, NOTCH2, both are present in granulosa and cumulus. In addition, Trial registration number: MCDH/2019/45
midkine significantly increased the maturation rate of immature oocytes (without
midkine: 27%; with midkine: 34%, p=0.04). The concentration of midkine was P-603 Drug free In Vitro Activation for woman with very advanced
compared using ELISA between FF from SAF from the same patient. Follicles maternal age
that contained an oocyte that matured contained higher concentration of mid-
L. Valkova1, N. Magunska2, B. Petkova3, T. Milachich3,
kine as compared to the follicle in which the oocyte failed to mature (n=16
K. Kawamura4, A. Shterev2
follicles from 8 patients).
1
Limitations, reasons for caution: The dynamics in the preovulatory follicles SAGBAL Dr Shterev EOOD, IVF Unit, Sofia, Bulgaria ;
2
were investigated in FF from follicles obtained in stimulated cycles and the use SAGBAL Dr Shterev EOOD, Department of Obstetrics and Gynecology- IVF Unit,
of ovulation trigger may have affected results. Sofia, Bulgaria ;
3
Wider implications of the findings: These results indicate that midkine is SAGBAL Dr Shterev EOOD, IVF Unit, Sofia, Bulgaria ;
4
a new growth factor, important for human follicular development both during Advanced Reproductive Medicine Research Center- International University of
ovulation and in folliculogenesis. Further investigations on the processes taking Health and Welfare School of Medicine, Department of Obstetrics and
place in the ovary, will increase the possibility for optimizing and developing Gynecology, Narita, Japan
future treatments especially for women with a low ovarian reserve.
Trial registration number: Not applicable Study question: What is the effect of drug-free In Vitro Activation (IVA) for
woman with very advanced maternal age (vAMA)?
Summary answer: Drug free IVA had positive effect through decreasing the
P-602 Comparison of letrozole versus clomiphene citrate (CC)
levels of FSH, LH, E2. Good quality oocytes were retrieved and top quality
for ovulation induction in infertile women with polycystic ovary
embryos were obtained.
syndrome (PCOS) in Indian population: A prospective clinical trial
What is known already: There are studies demonstrating good results in
E. Gupta1, K.D. Nayar1, S. Gupta1, M. Singh1, R. Bhattacharya1, patients with premature ovarian insufficiency (POI) using drug- free IVA. The
R. Gahlot1, G. Kant1, K.D. Nayar1 mechanism involves suppression of Hippo signaling pathway leading to secondary
1
Akanksha IVF Centre- Mata Chanan Devi Hospital, Reproductive Medicine, New follicle growth. Women with diminished ovarian reserve (DOR) as well as
Delhi, India patients with AMA and vAMA have very low number of follicles and oocytes

even after applying of ovarian hyperstimulation. Their chance for pregnancy and and also for different subgroups according to the BELRAP (Belgian Register for
live birth is minimal but number of patients with AMA or vAMA who wish to Assisted Procreation) criteria and to confounding factors selected by the LR.
have children with autologous oocytes increases constantly. Main results and the role of chance: 2216 patients were studied in all. 233
Study design, size, duration: Case report. From March to November 2019, patients constituted the freeze all (FA) group and 1983 the control (C) group
after refusing the use of donor oocytes to patients with vAMA were proposed (population 1). Patients selected according to BELRAP criteria (less than 36 years
application of drug-free IVA. Detailed information for risks and chances were old and first IVF trial) were divided as follows: 124 patients in the FA group with
provided to the women and one woman signed necessary informed consents. 1241 in the C group (population 2). For these two groups, the CLBR was respec-
The procedure was performed after permission of the Ethics Research tively 50.2% vs 58.1% P=0.021 for population 1 and 53.2% vs 63.3% P=0.023
Committee of the hospital. for population 2. LR revealed the following confounding variables: age, rank of
Participants/materials, setting, methods: To a 48 years-old woman with the attempt, tobacco, number of oocytes retrieved, number of embryos
primary infertility drug-free IVA was performed by taking pieces from both ova- obtained, and the date of oocytes retrieval before 2011 and after 2011. Once
ries under laparoscopic surgery. The cortices were dissected into small cubes these confounding variables were excluded, the FA and the C group were
(2x2x2 mm) that were auto-transplanted at the same surgery. The ovarian pieces restricted to respectively 109 and 770 patients. The CLBR stays in favour of the
were placed into peritoneal pockets of the mesosalpinx and of the remaining control group: 70.1% vs 55.9% P=0.03. The time to become pregnant is equally
ovaries followed by closure using sutures. Ovarian stimulation and fertilization in favour of the C group with a median of 5 days against 61 days for the FA group.
with donor sperm of the retrieved oocytes were performed after manipulation. Limitations, reasons for caution: Its a retrospective study carried over a
Main results and the role of chance: The hormone levels before and after long Long period of time with changes in the freezing techniques and in our
drug-free IVA were compared. They were: FSH – 50.8 vs 8.3 IU/L, LH - 38.1 transfer politics during the study. We also have an heterogeneous FA group with
vs 6.1 IU/L and E2 – 79.4 vs 56.9 pg/ml, respectively. Before manipulation the mostly hy- responders (75%).
hormonal ovarian stimulation was impossible because of high levels of gonad- Wider implications of the findings: The CLBR is significantly lower in the
otropins. After surgery, we succeeded to make hormonal stimulation three times. FA group compared to the control group (excellent rate of 70.1%) witch is the
In each cycle we achieved: follicle growth, high quality oocytes and embryos contrary of what is found in recent studies.
using donor sperm. After these three stimulations, we obtained six oocytes and But CLBR of 55.9% is still a very good rate for the FA population as its the best
five embryos. Embryo transfer (ET) was performed two times: the first one on found in the littérature.
day 2 with two embryos and second one on day 3 with one embryo. All embryos Trial registration number: not appilcable
were with top quality for the corresponding day, but pregnancy was not achieved.
One of possible reasons for the negative result could be obstruction of the P-605 Letrozole did not improve endometrium receptivity in
implantation by 3 myomas in the uterus of the patient. Laparoscopy for elimi- non-ovulation PCOS women, one RCT study for endometrium
nation of the myomas is forthcoming. Until the appropriate moment for the ET preparation before thawed embryo transfer in non-ovulation
we decided to generate embryos in different stimulation cycles with subsequent PCOS women.
cryopreservation by vitrification. We have already vitrified on day 3 two top C. Wang1, Q. Mai1, C. Zhou1
quality embryos from the third stimulation. 1
the first affiliated hospital of Sun Yat-sen University, repeoductive medical center,
Limitations, reasons for caution: The procedure has to be finished and is
Guangzhou, China
important if the pregnancy and live birth will be achieved. In contrast to positive
effect of drug-free IVA on improvement of oocyte quality, it cannot fix the issue
Study question: Does letrozole can improve the endometrium receptivity in
of aneuploidy. Additional genetic tests before implantation or after pregnancy
women with PCOS who underwent thawed embryo transfer (TET)?
could be performed.
Summary answer: Letrozole did not improve endometrium receptivity and
Wider implications of the findings: There are data about children born
clinical pregnancy rate in non-ovulation PCOS women.
after drug-free IVA in young patients. If the results are good also for patient with
What is known already: Polycystic Ovarian Syndrome (PCOS) is the most
AMA and vAMA this could be approved as useful method for this hard for
common endocrine disorder in infertile women. For non-ovulation PCOS
treatment steadily growing groups of patients.
women, hormone replace treatments (HRT) are always used for the endome-
trium preparation before thawed embryo transfer. In international PCOS guide-
line, Letrozole has been advised as the first line controlled ovarian stimulation
P-604 Does the Freeze-all strategy improves the cumulative live medicine. Letrozole is not only be used as COH medicine but also could be used
birth rate and the time to become pregnant in IVF cycles? as endometrium reparation before thawed embryo transfer in non-ovulation
A. Delvigne1, J. Vandromme2, A. Larbuisson1, S. Johnson1 PCOS women. However, there is no evidence show the effect of letrozole on
1
CHC Montlégia, ART center, Liège, Belgium ; endometrium receptivity and clinical pregnancy rate in women with PCOS who
2
CHU Saint-Pierre, Gynecology, Brussels, Belgium undergoing TET.
Study design, size, duration: One RCT study (ChiCTR1800014746)
Study question: Does the Freeze-all strategy improves the cumulative live involved PCOS women undergoing TET at Reproductive Medicine Center of
birth rate in IVF cycles? The First Affiliated Hospital of Sun Yat-sen University from 2017 to 2019. The
Summary answer: The CLBR is significantly lower in the FA group. sample size was calculated based on previous pregnancy rate of letrozole COH
Nevertheless, the CLBR in the FA group remains superior to that observed in and HRT endometrium preparation in PCOS women underwent TET.
previous studies. Considering the mean pregnancy, 5% type I error and 20% type II error, 130
What is known already: Elective freezing of all good quality embryos and patients were required in each group and 150 patients were studied in each group.
transfer in subsequent cycles, named as the freeze-all strategy (FAS) is widely Participants/materials, setting, methods: 296 non-ovulation PCOS
used for ovarian hyperstimulation syndrome (OHSS) prevention. The benefits women were involved in our RCT. 148 women were randomized selected into
of the FAS on live birth rates among high responders have been shown by many HRT endometrium preparation group and 148 women were radonmized
studies. Similarly, the benefits of frozen embryo transfer (FRET) compared to selected into letrozole COH endometrium preparation group.In letrozole COH
fresh embryo transfer (FET) has been demonstrated to prevent preterm birth endometrium preparation group,10 cycles were canceled. In HRT endometrium
and small for gestational age. Consequently, why should we limit the FAS to high preparation group, 7 cycles were canceled. SPSS ver. 20 was used for dada
responders rather than extend it to all? analysis. P value of less than 0.05 was considered statistically significant with
Study design, size, duration: A retrospective and monocentric study was Fisher corrections.
conducted between January 2008 and January 2018 comparing the cumulative Main results and the role of chance: Total 283 TET cycles were evaluated,
live birth rates (CLBR) of patients having undergone FAS to those using FET and 140 for letrozole COH endometrium preparation group (group A) and 143 for
having at least one frozen embryo during the same period. HRT endometrium preparation group (group B). The mean age in group A and
Participants/materials, setting, methods: Logistic regression (LR) was group B were 30.09 and 29.61 years (p=0.853). The base line of basic FSH, BMI,
used to identify confounding variables. Analysis were made for the entire cohort the number of oocytes and embryo transfer number were no significant

difference between two groups. The endometrium thickness on the day endo- P-607 Factors in first complete in vitro fertilisation cycle which
metrium transformation was significant higher in Group A.(10.28±1.94mm VS predict success in second complete cycle
9.21±1.59mm; P<0.001) The E2 level on embryo transfer day were significantly X. Zhang1, T. Dineen1, A. Kovacs1, R. Mihart1, J. O’Callaghan1,
lower in Group A (88.6pg/ml VS 181pg/ml; P<0.01). The progesterone level J. Culligan1, A. Tocado1, N. Daly1, M. Walsh1, D. Florio1, D.
on embryo transfer day were significantly higher in Group A . (23.22±12.39 VS McAuliffe1, J. Waterstone2
12.59±6.53; P<0.001) The average number of clinical visiting times was 3.1±0.89 1
Waterstone Clinic, Laboratory, Cork, Ireland ;
in Group A and 2.2±0.8 in Group B. (P<0.05) Clinical pregnancy rate were 2
Waterstone Clinic, Clinic, Cork, Ireland
54.7% in Group A and 60.3% in Group B. (P=0.63) The endometrium grew
more slowly in Group A. In Group A, the average day for endometrium thickness Study question: To determine the factors in first complete in vitro fertilisation
reaching 7mm and above was about 4.5 days before the day of endometrium (IVF) cycle which predict live birth (LB) in second complete IVF cycle.
transformation. However, the endometrium thickness in Group B can reach Summary answer: The patients that had LB, miscarriage or had frozen
7mm before 7 days before the day of endometrium transformation. embryos from first complete IVF cycle had a greater likelihood of achieving LB
Limitations, reasons for caution: It is a single center RCT study and the life in second cycle.
birth rate of two groups are not available right now. Our research did not What is known already: Individualised prognostics are valuable for both
evaluate the mechanism according to the endometrium receptivity in Letrozole patients and practitioners. A better understanding of the factors that determine
COH endometrium preparation group. the chance of success in IVF can help patients choose the correct fertility treat-
Wider implications of the findings: For non-ovulation PCOS women, ment options.
Letrozole COH endometrium preparation protocol can provide comparable Studies have shown that treatment response in the first cycle is reproducible.
clinical pregnancy rate. However, HRT endometrium preparation protocol could The number of aspirated oocytes is a predictor for subsequent treatment out-
be more convenient due to less number of clinical visiting times and stable high come. However, the majority of studies have considered only the outcome of
clinical pregnancy rate. It is still beyond studies about endometrium receptivity the fresh IVF transfer without the inclusion of frozen embryo transfer results.
in Letrozole COH patients. Study design, size, duration: This cohort study includes all women (Median
Trial registration number: ChiCTR1800014746 age = 36.0, 19.6% ≥ 40years) undergoing IVF treatment with homologous
oocytes in a single IVF unit who had a first (n= 4180) and a second (n=1838)
P-606 Effect of body mass index on cumulative live birth rate egg collection between 2008 and 2017. The outcome of all embryo transfers
following in vitro fertilization (fresh and frozen) from both egg collections was analysed.
L. Zhou1 Participants/materials, setting, methods: A complete cycle is defined as
1
Shenzhen Zhongshan Urology Hospital, Fertility Center, Shenzhen-Guangdong, all fresh and frozen/thawed embryo transfers derived from the same oocyte
China aspiration. LB rates of the 1st and 2nd complete cycles were calculated.
Multivariate logistic regression was used to identify prognostic factors in the first
Study question: Is it beneficial to the cumulative live birth rate of in vitro complete cycle with regard to the outcome of the second complete cycle, such
fertilization by achieving a particular body mass index prior to infertility as the number of retrieved oocytes, number of good quality embryos, blastocyst
treatment? cryopreservation and pregnancy outcome.
Summary answer: BMI does not affect the cumulative live birth rate of IVF, Main results and the role of chance: In all women LB rate was 38.0%
but it does elevate the miscarriage rate in the fresh cycles. (n=4180) for the first complete cycle, significantly higher than for the second
What is known already: It is critical to evaluate the impact of body mass complete cycle 34.0% (n=1838, p=0.003).
index on the cumulative live birth rate following in vitro fertilization, as achieving In the second complete cycle LB rate was 30.1% (364/1202) in women that
a lower BMI prior to infertility treatment is often recommended for women with did not achieve any pregnancy in the first complete cycle in comparison with
overweight and obesity. It is important to extrapolate the true benefits of weight 44.2% (156/353, OR=1.5) in women with LB and 37.1% (105/283, OR=1.3)
loss on CLBR from this study. in women with miscarriage. Moreover, embryo development patterns in the first
Study design, size, duration: This is a retrospective cohort study. A total complete cycle demonstrated a significant association with LB rate in the second
of 4166 women aged 20-40 years old with first conventional stimulation complete cycle. A significantly higher LB rate in the second complete cycle was
across different BMIs at our institution from 2012 to 2016 were stratifified found for women who had frozen blastocyst(s) in the previous cycle (OR=1.4,
into cohorts. BMI was defifined according to World Health Organization P=0.019) compared to those who had not.
guidelines. The LB rate of the 2nd complete cycle significantly increased with the number
Participants/materials, setting, methods: The primary outcome was live of oocytes retrieved in the first complete cycle. However, this association was
birth as measured by cumulative live birth rate per initiated cycle. Secondary only significant in the poor ovarian response group (number of oocytes less than
outcomes included clinical pregnancy rate, miscarriage rate and live birth rate 4) after adjusting for the factors of womens’ ages and of having frozen
independent in fresh and cryopreserved cycles. blastocyst(s).
Main results and the role of chance: There were 3384 first fresh cycles Limitations, reasons for caution: This is a cohort analysis based on retro-
and 1929 subsequent frozen embryo transfer cycles. There were 3645 live births, spective data collection. A significant number of patients did not undergo a
for an overall cumulative live birth rate of 87.49% per initiated cycle. The clinical second cycle, which may potentially bias the results.
pregnancy rate and cumulative live birth rate decreased with increasing BMI, but Wider implications of the findings: These results allow us to provide
there was no statistical significance. The miscarriage rate was lower in under- patients with more accurate advice when they have a consultation after their
weight (BMI < 18.5 kg/m2 ) and normal weight cohorts (18.5 <BMI < 24.9 kg/ first complete cycle.
m2, over weight cohort (BMI>25 kg/m2 ) of patients had a relatively higher rate Trial registration number: not applicable
in fresh cycles(P=0.041), but there was no statistical significance in all linked
cryopreserved cycles.
P-608 Optimization of oocyte retrieval rate in women undergoing
Limitations, reasons for caution: Our conclusions are limited by the ret-
ovarian stimulation for in-vitro fertilization using hCG or GnRH
rospective nature of the study, and the single-center design also weakens the
agonist for triggering final oocyte maturation
universality of our observations. It is possible that patients with a failed initial
transfer and subsequent weight loss, followed by a successful transfer, were not J. Bosdou1, E. Deligeoroglou1, O. Pavlidi1, A. Makedos1, A. Mitsoli1,
captured within this study’s time frame. G. Lainas1, T. Lainas2, I. Sfontouris2, M. Malandri1, T. Tarlatzi1,
Wider implications of the findings: BMI does not affect the cumulative live D. Sabbaidou1, A. Chatzimeletiou1, L. Zepiridis1, G. Grimbizis1,
birth rate of IVF, but it does elevate the miscarriage rate in the fresh cycle. E. Kolibianakis1
1
Therefore, it is recommended to delay embryo transfer to achieve a lower BMI Aristotle University of Thessaloniki- Medical School-, 1st Dept. of Ob/Gyn- Unit
may be beneficial to assisted reproductive outcomes. for Human Reproduction-, Thessaloniki, Greece ;
Trial registration number: no 2

Study question: At which follicular diameter is the ratio of oocytes retrieved Summary answer: In the COS cycles using letrozole, early monitoring serum
to the number of follicles > certain diameter on the day of triggering final oocyte estradiol (E2) is an independent predicting factor for supraphysiologic level of
maturation (oocyte retrieval rate) optimised? serum E2 on hCG day.
Summary answer: The smallest mean follicular diameter, leading to a lower What is known already: Supraphysiologic serum E2 levels (≥400 pg/ml) as
95% confidence-interval(CI) of oocyte retrieval rate ≥100%, is 15mm for hCG a consequence of ovarian stimulation may promote the estrogen-dependent
and 16mm for GnRH-agonist triggering. tumor growth or recurrence. Therefore, use of aromatase inhibitors is recom-
What is known already: Oocyte retrieval rate is calculated by dividing the mended to reduce the potential effects of elevated serum E2 levels in patients
number of oocytes retrieved to the number of follicles equal or greater than a with estrogen-dependent cancers during ovarian stimulation. Previous studies
specific mean follicular diameter (i.e. ≥11mm, ≥12mm, etc.), assessed in two that have shown an effective lowering of peak estrogen levels and feasible results
perpendicular planes on the day of triggering. Ideally, the smallest mean follicular of COS using the letrozole protocol have used a fixed dose of letrozole (5 mg/
diameter leading to a lower 95%CI of oocyte retrieval rate ≥100% should be day). However, a part of patients has supraphysiologic levels of estrogen asso-
used for this purpose. In this case, the number of oocytes retrieved would be ciated with ovarian stimulation despite the administration of letrozole.
expected to be ≥ to the number of follicles above this follicular diameter. Study design, size, duration: From January 2009 to December 2019, patients
Although this is an essential information for patient consultation, no such data with estrogen-dependent cancer who underwent COS with antagonist protocol
is currently available. using a fixed dose of letrozole (5 mg/ day) to keep estrogen levels low were
Study design, size, duration: A retrospective study was performed between included in this study. Cycles for fertility preservation and general in vitro fertil-
01/2018 and 12/2019, including 413 in-vitro fertilization (IVF) cycles (306 ization cycles were all included. Subjects are consisted of 74 breast cancer
women). Ovarian stimulation was performed with a fixed dose of recombinant patients, 5 endometrial cancer patients and 2 other estrogen-dependent cancer
gonadotrophin and gonadotrophin releasing hormone (GnRH) antagonist, while patients. A total of 94 COS cycles were retrospectively analyzed.
triggering was carried out with human-chorionic-gonadotrophin(hCG) or GnRH- Participants/materials, setting, methods: Administration of letrozole was
agonist, when ≥14 follicles ≥11mm in diameter were present. Thirty-six hours started on the second or third day of the menstrual cycle. Ovarian stimulation
following final oocyte maturation, oocyte pick-up(OPU) was carried out by began with a dose of 150-450 IU/day on the same day or third day of letrozole
puncturing all follicles that could be aspirated. Follicular flushing was not administration and was continued until the hCG day. Dose of FSH was adjusted
performed. according to the patient’s age, anti-Mullerian hormone (AMH), and body mass
Participants/materials, setting, methods: Follicular development was index (BMI). Early monitoring serum E2 was measured in all patients on the 4-6th
assessed by recording the mean diameter of each follicle ≥10mm by 2-dimen- day of stimulation.
sional ultrasound. On the day of triggering, the number of follicles ≥ to a specific Main results and the role of chance: Supraphysiologic levels of serum E2
diameter was calculated for all diameters recorded, leading to multiple oocyte was found in 20.2% of the patients who underwent COS with a fixed dose of
retrieval rates for each cycle. The primary outcome measure was the identifica- letrozole (5 mg/ day) administration. Subjects were classified into two groups
tion of the smallest follicular diameter on the day of triggering, with which the according to the serum E2 level on hCG day, physiologic E2 group (≤400 pg/
lower 95%CI of the calculated oocyte retrieval rate is ≥100%. ml) and supraphysiologic E2 group (>400 pg/ml). Mean age, BMI, AMH, basal
Main results and the role of chance: The mean ± standard deviation (SD) serum LH/ FSH/ E2, total dose and duration of letrozole administered, duration
age of the patients included was 38.7 (±5.2) years, the mean BMI was 25.8 (±5.5) of stimulation, total dose of gonadotropins administered were not different
kg/m2, the mean basal FSH levels were 7.8 (±3.1) IU, the mean AMH levels between the two groups. However, early monitoring serum E2 level was signifi-
were 4.5 (±6.1) ng/mL and the mean number of antral follicle count was 11.8 cantly higher in the supraphysiologic E2 group (68.9±49.3 vs. 119.8±78.1,
(±9.5). At OPU, the mean number (95% CI) of oocytes retrieved was 6.8 (6.1- p=0.001). Early monitoring serum E2 is associated with the occurrence of sup-
7.5). In 299 out of 413 cycles (72.4%), hCG was used for triggering final oocyte raphysiologic elevation of serum E2 on hCG day (adjusted odds ratio [aOR] =
maturation, whereas in 114 cycles (27.6%), this was performed with GnRH 1.013; 95% confidence interval [CI], 1.003-1.022). The best cut-off for early
agonist. The smallest follicular diameter on the day of triggering final oocyte monitoring serum E2 to predict the occurrence of supraphysiologic estrogen
maturation with hCG, at which the lower 95% CI of oocyte retrieval rate was levels was 86.5 pg/ml (sensitivity was 73.7% and specificity was 73.3%). The
≥100%, was 15 mm (mean oocyte retrieval rate: 133.7%, 95% CI: 109.8%- mean area under a receiver operating characteristic (ROC) curve (AUC) was
157.6%). The smallest follicular diameter on the day of triggering final oocyte 0.731 (p=0.002 compared with 0.5).
maturation with GnRH agonist, at which the lower 95% CI of oocyte retrieval Limitations, reasons for caution: Limitations of this study included the
rate was ≥100%, was 16 mm (mean oocyte retrieval rate: 144.2%, 95% CI: retrospective design and the small sample size. Prospective study will be needed
107.6%-180.8%). to confirm that uptitration of letrozole is effective to prevent supraphysiologic
Limitations, reasons for caution: The results obtained can be generalized levels of estrogen.
only if similar practices to the current study are used for evaluating follicular Wider implications of the findings: To prevent supraphysiologic elevation
development and performing oocyte retrieval. For instance, whether the results of serum E2, uptitration of letrozole should be considered when early monitoring
of the current study are also valid for cycles in which follicular flushing is per- serum E2 level is more than 86.5 pg/ml during COS with letrozole in patient
formed needs to be assessed. with estrogen-dependent cancer.
Wider implications of the findings: The findings of the present study allow Trial registration number: Not applicable
for precise prediction of the number of oocyte retrieved depending on the
triggering signal. In this way, patient consultation can be facilitated, especially in P-610 Severe OHSS after GnRH-agonist triggering is not
cases where an oocyte target has been set, such as preimplantation genetic eliminated if hCG is co-administered at triggering and/or during
testing or oocyte donation cycles. the luteal phase: a systematic review and meta-analysis
Trial registration number: not required P. Ioannidou1, J. Bosdou1, G. Lainas2, T. Lainas2, N. Michaleli1,
G. Grimbizis1, E. Kolibianakis1
P-609 Predictor for supraphysiologic serum estradiol elevation on 1
Aristotle University of Thessaloniki- Medical School, 1st Dept. of Ob/Gyn- Unit
hCG day of controlled ovarian stimulation (COS) using letrozole for Human Reproduction, Thessaloniki, Greece ;
and gonadotropins in women with estrogen-dependent cancer 2
S.J. Han1, M. Lee1, S.W. Kim1, H. Kim1, J.Y. Lee2, S.Y. Ku1, C.S.
Suh1, S.H. Kim1 Study question: Is severe ovarian-hyperstimulation-syndrome (OHSS) after
1
Seoul National University Hospital, Obstetrics and Gynecology, Seoul, Korea- South ; triggering with gonadotrophin-releasing-hormone agonist (GnRH-agonist) elim-
2
Hamchoon Women’s Clinic, Obstetrics and gynecology, Seoul, Korea- South inated if human chorionic gonadotropin (hCG) is co-administered at triggering
and/or during the luteal phase?
Study question: Are there predicting factors for supraphysiologic estrogen Summary answer: Severe OHSS, although low in incidence, is not eliminated,
elevation during COS with administration of letrozole in patients with estro- when hCG is administered either concomitantly with GnRH-agonist (dual-
gen-dependent cancer? triggering) and/or during the luteal phase.

What is known already: Replacement of hCG by GnRH-agonist has What is known already: Published evidence suggests that embryo develop-
decreased or even eliminated the incidence of severe OHSS. Concomitantly, ment and LBR are inferior following IVM compared to COS in patients with and
however, the probability of pregnancy after fresh transfer and standard luteal without PCOS. Although PCOS phenotype has emerged as an important factor
phase support is decreased. This has been managed by freezing all embryos and influencing COS and IVM treatment outcomes, assignment of PCOS phenotypes
deferring transfer to a future replacement cycle. Alternatively, in an effort to is lacking in patients enrolled in previous studies. PCOS women can be catego-
perform a fresh transfer, support of the luteal phase with hCG administered rized into the following phenotypes: A= hyperandrogenism + ovulatory dys-
either concomitantly with GnRH-agonist (dual-triggering) and/or during the function + polycystic ovaries, B= hyperandrogenism + ovulatory dysfunction,
luteal phase has been advocated. The potential of both strategies, however, in C= hyperandrogenism + polycystic ovaries and D= ovulatory dysfunction +
eliminating severe OHSS has not been yet systematically assessed. polycystic ovaries. Clinical outcomes of non-hCG triggered IVM and COS have
Study design, size, duration: Literature search was performed in MEDLINE not previously been compared among similar patient phenotypes.
and CENTRAL until 12/2019, in order to identify studies evaluating the incidence Study design, size, duration: This was a retrospective, single-centre cohort
of severe OHSS following GnRH-agonist triggering in high-risk women. The inci- study including 716 cycles in unique patients between 18 and 36 years of age
dence of severe OHSS was assessed separately for studies in which hCG was who underwent either non-hCG triggered IVM (30h) after a short HP-hMG
administered either concomitantly with GnRH-agonist(dual-triggering) and/or course (189 patients) or COS (527 patients) followed by ICSI and fresh or frozen
during the luteal phase, aiming to perform a fresh transfer and for studies in which embryo transfer between January 2010 and December 2016. First rank of trial
no support was administered during the luteal phase and all embryos were frozen. were included.
Participants/materials, setting, methods: Twenty-nine studies (n=2269 Participants/materials, setting, methods: Predicted high responders with
patients), published between 2000 and 2019, were included in this systematic PCO-like morphology (PCOM) and/or levels of AMH ≥3.5 μg/l, or PCOS
review and meta-analysis. Pooling of severe OHSS incidence in the included studies according to 2003 extended Rotterdam criteria, ≤37 years of age and scheduled
is reported as a weighted average, by fitting the logistic regression model without for ICSI were included. Patient characteristics and IVM or COS treatment cycle
covariate but an intercept, or by fitting the logistic-normal random-effects model data were collected. The primary objective was CLBR, defined as the rate of
without covariates but random intercepts. The confidence intervals are based on liveborns resulting from the transfer of all fresh and cryopreserved embryos
score (Wilson) or exact binomial (Clopper-Pearson) procedures. from the same cycle. Secondary outcome was embryo quality.
Main results and the role of chance: Triggering of final oocyte maturation Main results and the role of chance: IVM patients had higher BMI (24.8 vs
was performed with GnRH-agonist alone in 24 studies, whereas in five studies 24.0 kg/m2, p=0.03), higher basal antral follicle count (AFC (42.6 vs 26.6,
a combination of GnRH-agonist and hCG was used for this purpose. p<0.001)), higher AMH (10.0 vs 6.4 µg/l, p<0.001) and were younger (28.3 vs
The pooled mean number of oocytes retrieved in the high-risk for OHSS women 29.7y, p<0.001). Patients undergoing IVM had more often PCOS-A (19.7%
included in these studies was 18.2 (95% CI: 15.0-21.5). vs 8.3%) and PCOS-D (65.4% vs 24.3%), a similar incidence of PCOS-C (4.3%
The pooled incidence of severe OHSS in high risk women triggered by GnRH- vs 7.6%) and less often PCOM (10.3% vs 59.8%, overall p<0.001). IVM cycles
agonist who did not receive any form of luteal phase support was 0% (95% CI: yielded more oocytes (17.8 vs 13.9, p<0.001), lower maturation rates (51,7%
0.0-0.0, 15 studies, 1327 women). vs 77.8%, p<0.001) and lower utilization rate per fertilized oocyte (51.0% vs
The pooled incidence of severe OHSS in high risk women triggered by GnRH- 54.9%, p<0.05). After IVM, patients had more often transfer of vitrified/warmed
agonist in whom hCG was added to standard luteal phase support was 0% (95% cleavage stage embryos, whereas fresh transfer of blastocysts was more often
CI: 0.0-1.0, eight studies, 557 women). performed following COS. Overall, CLBR was 40% after IVM and 59% after
The pooled incidence of severe OHSS in high risk women triggered by a COS (p<0.001). There was no difference in CLBR between IVM and COS in
combination of GnRH agonist and hCG (dual triggering), who received only patients with PCOS-A (43% vs 33%, p=0.45) whereas CLBR was higher (p<0.05)
estradiol and progesterone was 1% (95% CI: 0.0-3.0, three studies, 282 women). following COS compared to IVM in patients with PCOS-D (65% vs 41%) and
The pooled incidence of severe OHSS in high risk women triggered by a PCOM (59% vs 30%).
combination of GnRH agonist and hCG (dual triggering) in whom hCG was in Limitations, reasons for caution: This is a large observational study based
addition administered to standard luteal phase support was 1% (95% CI: 0.0-5.0, on retrospective data collection. Despite our robust methodological approach,
two studies, 103 women). the presence of bias related to the retrospective design cannot be excluded.
Limitations, reasons for caution: Despite the absence of statistical hetero- The low number of patients with PCOS-C impedes the validity of results in
geneity, clinical heterogeneity was present among studies regarding ovarian women with PCOS-C.
stimulation protocol, triggering signal, severe OHSS definition as well as the Wider implications of the findings: The current non-hCG triggered IVM
criteria used to define high risk probability for OHSS. Moreover, the number of system is an equally efficient mild-approach alternative for COS in patients with
patients/studies analysed in certain groups was limited. PCOS phenotype A. Further development of IVM systems could enhance the
Wider implications of the findings: Although the incidence of severe OHSS potential of IVM in a broader patient population.
after GnRH-agonist triggering in high-risk women was low in all strategies exam- Trial registration number: not applicable
ined, its elimination (upper 95% CI = 0) was only present in studies where no
hCG was added. Thus, for elimination of severe OHSS, following GnRH-agonist P-612 The impact of endometrial thickness change between the
triggering, addition of hCG is not recommended. day of ovum pick and embryo transfer on pregnancy outcome
Trial registration number: not required during in vitro fertilization cycle
M. Jeong1, M.K. Chung2, G.K. Lee2, H.J. Jeong1, H.S. Lee1,
P-611 Cumulative live birth rates of oocyte in-vitro maturation M.H. Kim1, J.W. Kim1, E.B. Noe1
and controlled ovarian stimulation in infertile patients with 1
Seoul Rachel Fertility Center, Medical Department, Seoul, Korea- South ;
polycystic ovary syndrome 2
Seoul Rachel Fertility Center, Laboratory, Seoul, Korea- South
I. Segers1, S. Santos-Ribeiro1,2, L. Mostinckx1, H. Tournaye1,
G. Verheyen1, M. De Vos1,3 Study question: Is there any difference in pregnancy outcome according to
1
UZ Brussel, Centre for Reproductive Medicine CRG, Jette- Brussels, Belgium ; the change of endometrial thickness in the patients undergoing in vitro fertiliza-
2
IVI-RMA Lisboa, Reproductive medicine, Lisbon, Portugal ; tion-embryo transfer (IVF-ET) cycle?
3
Vrije Universiteit Brussel, Follicle Biology Laboratory, Jette-Brussels, Belgium Summary answer: Clinical and ongoing pregnancy rates were significantly
higher when endometrial thickness did not increase on embryo transfer (ET)
Study question: What is the clinical outcome of in-vitro maturation of oocytes day compared with ovum pickup (OPU) day.
(IVM) compared with controlled ovarian stimulation (COS) in polycystic ovary What is known already: Many studies have reported the importance of
syndrome (PCOS) patients undergoing ICSI? endometrial thickness on pregnancy outcome during in-vitro fertilization (IVF)
Summary answer: Overall, cumulative live birth rate (CLBR) was lower after cycle, most of them discovered that thin endometrium adversely affects preg-
IVM. However, in patients with hyperandrogenic PCOS phenotype A, CLBR nancy outcome. Previous studies have mostly measured the endometrial thick-
after IVM and COS was similar. ness on the human chorionic gonadotropin (hCG) trigger day in fresh IVF cycle

and examined its effect on pregnancy outcome. It is known that the endometrial What is known already: In IVF cycles, it is estimated that about 40% of the
condition is changeable not only in natural menstrual cycle but also in IVF cycle, human oocytes are capable of growing to the blastocyst stage, whereas others
yet few studies have examined the impact of endometrial thickness change within arrest at different early stages of development. It is commonly thought that much
the IVF cycle. of human embryonic developmental potential is determined before fertilization
Study design, size, duration: A retrospective study was conducted using by the content of the oocyte (maternal-effect genes). Infertile patients commonly
data from a total of 397 fresh IVF cycles undertaken at a single fertility center. show individual variability in embryo developmental rate, ranging from high to
The study period went from January 2017 to November 2018. Only cases in total developmental failure in multiple treatment cycles. The genetic determinant
which endometrial thickness on hCG trigger day, OPU day, ET day were all of this phenotype is unknown and this study aims at exploring the genetic aeti-
measured were included. Pre-implantation genetic test cycles or oocyte donation ology of extreme phenotypes of preimplantation embryonic arrest
cycles were excluded. Study design, size, duration: Ten women (five with consanguinity history,
Participants/materials, setting, methods: Each endometrial thickness of four being the offspring of first-cousin marriage) with a history of recurrent
hCG trigger day, OPU day, and ET day were examined and its impact on preg- embryo developmental failure in multiple IVF cycles were recruited at Istanbul
nancy outcome was analyzed. Subsequently, endometrial changing pattern such Memorial Hospital (Dec 2018-Nov 2019). These were all young women (average
as increase or decrease between the days were individually examined. Clinical age 32.3 y), with good ovarian reserve (average AMH 3.6) and with several failed
and cycle characteristics, and pregnancy outcomes were compared according IVF cycles (average 4.4, range 3-8), following the retrieval of high number of
to the endometrial thickness change around OPU day. mature oocytes per procedure (average 11.2).
Main results and the role of chance: The endometrial thickness of hCG Participants/materials, setting, methods: Whole-exome sequence was
trigger day or ET day was not significantly related to the pregnancy outcome. performed by Agilent SureSelect whole-exome capture and Illumina sequencing
The endometrial thickness of OPU day was significantly higher in the technology. Variant calling against the reference genome GRCh38 was done
implanted cycles than in the non-implanted cycles (9.29mm vs. 8.76mm, using Freebayes. Variants were annotated with a custom script that integrates
p <0.001), and showed the same tendency for cases with clinical pregnancy information from Ensembl99 with publicly available manually curated lists of
(9.28mm vs. 8.84mm, p=0.002) and ongoing pregnancy (9.40mm vs. 8.82mm, genes associated with embryonic development, miscarriages, lethality, cell cycle.
p <0.001). When analyzing the relationship between endometrial thickness The code is available on gitHub. The Hospital Ethical Committee approved the
changes and pregnancy outcome, only the impact of change between OPU study and data were treated anonymously.
and ET day on pregnancy outcome was identified. Baseline and cycle char- Main results and the role of chance: Variant calling identified on average
acteristics were not significantly different according to the changing pattern 436k high quality variants per samples. According to Ensembl classification 2.8%
of endometrial thickness between OPU and ET day. Interestingly, the rate of are expected to have high (0.25%) or moderate (2.56%) disruptive impact in the
implantation, clinical pregnancy and ongoing pregnancy were all significantly gene product. Variants were filtered on a per-individual base using three criteria.
higher in the ‘non-increased’ group in which endometrial thickness were equal First, alleles with frequency >0.05% in the 1000 Genomes and gnomeAD refer-
or decreased from OPU to ET day, compared with ‘increased’ group where ence populations were filtered out. Second, variants were ranked by severity
endometrial thickness were thicker on ET day. After adjusting the transferred (as estimated by Ensembl) according to their Sequence Ontology classification,
number of good quality embryos, clinical pregnancy (aOR=1.583, 95% CI: and only variant in the last percentile were retained. Finally, variants were
1.018-2.464) and ongoing pregnancy (aOR=1.734, 95% CI: 1.101-2.728) assigned a score that correlates with the gene relevance in early embryonic
rates were still significantly higher in the ‘non-increased’ group than in the development and only variants in the last fifth percentile were retained.
‘increased’ group. Overall, 62 unique variants in 54 genes were retained after filtering, all involved
Limitations, reasons for caution: Limitations of this study included the in lethal embryonic pathways. All retained variants have high impact and half of
retrospective design and the heterogeneity of the subjects, including such as them is a stop gain. Each sample carry on average 8.6 (2.7 s.d.) potentially
infertility diagnosis or ovarian stimulation method which may affect the pregnancy detrimental variants. Of particular relevance seven samples share stop gain and
outcome in IVF cycle. Therefore, prospective studies are needed before gener- splice donor mutations at four sites in three exones of the MTCH2 gene, a
alizing the results. nuclear-encoded transporters localized in the inner mitochondrial membrane,
Wider implications of the findings: Though we have generally measured involved in the energy and lipid metabolism, never reported associated to pre-
endometrial thickness on hCG day and considered that thickness to be crucial implantation embryo arrest.
factor for IVF outcome, this study result suggest that endometrial thickness on Limitations, reasons for caution: Functional genomics studies and validation
OPU day and its change until ET day may be more important clues for predicting in an independent cohort of patients with preimplantation embryo arrest phe-
pregnancy outcome. notype and of different ethnicity is required to corroborate these findings. The
Trial registration number: not applicable generation of polygenic models will also further contribute increasing discovery
rate and to develop more general and powerful predictive models for this
P-613 Genomics analysis of maternal exomes reveals new phenotype.
candidate genes and pathways for the diagnosis and prediction of Wider implications of the findings: We have identified MTCH2 and
recurrent preimplantation embryo arrest in IVF cycles several new candidate genes as the plausible cause of female infertility char-
S. Kahraman1, F. Sagir1, B. Yuksel1, M. Cetinkaya1, M. Gavaz1, acterized by early embryonic arrest. Such discoveries not only contribute to
B. Kara1, M. Yesil1, G. Akar1, S. Buonaiuto2, G. Damaggio2, the recognition of novel gene function/pathways in reproduction but also help
M. Fabiani3, A. De Marino4, V. Colonna2, C. Simon5, A. Capalbo4 the development of genetic diagnosis and suggest possible therapeutic targets
1 for infertility.
Istanbul Memorial Hospital-, ART and Reproductive Genetics Center, Istanbul,
Turkey ;
2
National Research Council CNR, Institute of Genetics and Biophysics, Naples,
P-614 NLRP3 inflammasome analysis in grannulose-cumulus cells
Italy ;
3 from low ovarian reserve patients
Igenomix Italy, Genetics and Biophysic, Marostica, Italy ;
4
Igenomix Italy, Genetics and Biophysics, Marostica, Italy ; C. Andres1, L. Rancan2, C. Cordero1, R. Pandolfi3, M. Saladino3,
5
Valencia University and INCLIVA, Department of Obstetrics and Gynecology, C. Rodriguez1, B. Linillos4, V. Elena2, J. Tresguerres4, C. Susana1
1
Valencia, Spain Clinica tambre, IVF Laboratory, Madrid, Spain ;
2
Complutense University, Biochemistry and molecular biology, Madrid, Spain ;
Study question: Is genomic analysis of maternal exomes capable of identifying 3
Clinica tambre, Andrology Laboratory, Madrid, Spain ;
new target genes to improve infertility diagnosis and support the development 4
Complutense University, Physiology Department, Madrid, Spain
of new treatment strategies?
Summary answer: Exome analysis of woman with recurrent embryo arrest Study question: Is the inflammasome NLRP3, together with other inflamma-
successfully identify genomic variants lethal at the embryonic stage increasing tory markers, activated on low ovarian reserve patients and is this correlated
diagnostic yield toward unexplained infertility with the number of follicles retrieved?

Summary answer: Inflammasome activation was observed in patients with What is known already: Recent studies cue at the association of diminished
low ovarian reserve (LOR) in comparison to donors with normal response. It is ovarian reserve with diabetes mellitus. Reduced ovarian reserve in diabetes might
not correlated with follicle number. be associated with decreased oocyte developmental competence leading to
What is known already: LOR patients are women, within the reproductive poor IVF outcome. Attenuation of ovarian gap junction protein (connexin-43)
age group, which are characterized by having a decreased ovarian follicular pool. involves decreased follicular development and oocyte growth retardation.
They present abnormal but not post-menopausal ovarian reserve test results. However, there is little information whether diabetes targets connexin-43 to
There is a wide-range of origins that can lead to this pathology. alter follicular growth. Anti –mullerian hormone(AMH) and Antral follicular
Oxidative stress associated with chronic low-grade inflammation has emerged count(AFC) are the most important factors predicting ovarian stimulation
as a plausible key contributor to the pathogenesis. The NLRP3 inflammasome can response based on retrieved oocyte number. Hence, we have tried to find out
start an inflammatory form of cell death and trigger the release of proinflammatory the effect of diabetes on ovarian stimulation response in these women under-
cytokines through caspase activation. Although it has been associated to different going IVF.
diseases, an association with low ovarian reserve has not been described to date. Study design, size, duration: This is a prospective cohort study in women
Study design, size, duration: Prospective clinical observational study conducted (n=415) with UI in age group between 25 and 35 years, conducted between
at Clinica Tambre in collaboration with the University Complutense of Madrid. A August 2017 and December 2019 at Institute of Reproductive Medicine,Kolkata.
total of 16 donors and 9 patients participated in the study. Stimulation protocols Patients have been divided to diabetic(n=216) controlled with either tablet
were the same for all patients (FSHr and triggering with GnRH analogues). Collection Metformin or insulin and non-diabetic based on ADA guidelines. All patients are
of oocyte-granulose complexes (GCs) was performed during oocyte retrieval. GCs stimulated using recombinant FSH with individualized dosing and GnRH antag-
were processed according to the procedure described by Ferrero F, et al. 2012 onist (flexible)protocol .Fresh or frozen embryo transfer has been done
throughout 2019. All patients signed the specific informed constent. within 1 year.
Participants/materials, setting, methods: mRNA expression of NLRP3, Participants/materials, setting, methods: All patients have been evaluated
IL-1β, IL-1 receptor, caspase 1, 12 and 18, and serpin were analyzed in the CGs for HbA1C, AMH & AFC. All diabetic patient either taking tablet Metformin
complexes as well as the number of follicles retrieved to evaluate correlation and/or insulin with HbA1C <7 have been included in the study group. After
with gene expression. mRNA was measured by qRT-PCR. The 2-∆∆CT method ovum pickup, follicular fluid of each patient has been evaluated by immunoblot-
was used for relative gene expression changes calculation. Non parametric tests ting for connexin-43 expression in between granulosa cells. Routinely 2 blastocyst
were used to identify significant differences between groups and spearmen cor- have been transferred.
relation to asses correlation. Two tailed Student’s t- test is used to evaluate significance of differences.p
Main results and the role of chance: Increased mRNA expression was value <0.05 is considered to be significant.
observed in LOR patients compared to donors (p=0.022). Accordingly, LOR Main results and the role of chance: Baseline parameters (Age,obesity)
patients had increased expression of IL-1B and its corresponding receptor in are comparable between two groups .AMH ( 1.69 ± 0.9 vs 2.97±0.3, p<0.05)
comparison to donors (p=0.0003 and p=0.0001 respectively). In the same man- & AFC (8±1.9 vs 10.4±2, p<0.05) are significantly less in diabetic patient than
ner, caspase 1, 12 and 18 were also significantly increased in our group of patients non-diabetic group indicating low ovarian reserve.Total gonadotropin dose
in comparison to the donor expression (p<0.0001, p=0.0008 and p=0.0003 requirement is significantly higher in diabetic group (2873±75.08 vs 2497±80.15,
respectively). Finally, pathway’s inhibitor expression: SERPIN B9 was significantly p<0.05). Oocyte recovery /patient is also lower, but not significant (7±1.5 vs
higher in donors (p=0.0002). 9±1.7). M II oocyte percentage is significantly lower in diabetic group (73±2.1
When analyzing the correlation with the oocytes retrieved, we observed no vs 80±1.5, p<0.05). Connexin-43 expression is also significantly lower in diabetic
significant correlation between the number of oocytes obtained with the expres- group obtained by immunoblot technique. Fertilization rate (62.16±5.7 vs
sion of NLRP3 and Serpin (p= 0.2977 and p=0.7158, respectively). In contrast, 73.5±2.9), cleavage rate (87.7±6.1 vs 91.8±5.3) and blastocyst development
we did observe a negative significant strong correlation in inflammation markers rates (58.6±3.9 vs 65.9±6.2) are non-significantly less in diabetic group. But
IL-18 (p=0,0069, r -0,6583) , IL-1B (p=0,0009, r-0,8244) and IL-1R (p=0,0009, clinical pregnancy rate is significantly low (56.7 vs 64.1, p<0.05) in diabetic group
r-0,824) and a significant negative mild correlation in caspase 1 (p=0,0102, after blastocyst transfer either in fresh or frozen cycle. Duration of diabetes has
r-0,5357 ) and 12 (p=0,0488, r-0,5616). negative correlation (- 0.403) with number of M II oocyte retieved.
Our results show higher NLRP3 expression in patients with low ovarian Limitations, reasons for caution: It is an observational study with limited
reserve compared to donors suggesting an activation of the pathway and its number of patients. Exact cause or mechanism of decreased response is not
possible involvement in the pathogenesis. Additionally, there seems to be a revealed by this study. Before wide application of this observation, other unde-
negative correlation in between the number of oocytes retrieved and inflam- tected cause of diminished ovarian response should be searched for and study
matory markers. with larger sample size is warranted.
Limitations, reasons for caution: Due to the low number of patients who Wider implications of the findings: Diabetic women with infertility should
participated in our study, in order to confirm our results, further analysis is be counseled regarding low ovarian reserve and decreased ovarian response
required. irrespective of age. Diabetic women are associated with increased gonadotropin
Wider implications of the findings: Finding out the causes behind idiopathic requirement and poor IVF outcome.
LOR will give us a hint for a better approach to the treatment or ways to improve Trial registration number: not applicable
IVF outcomes.
Trial registration number: Not aplicable P-616 Can we detect a biologically relevant quantity of Anti-
Mullerian Hormone (AMH) in human hair samples?
P-615 Diabetic but not women with normal metabolic phenotype This abstract
S. Sawarkar 1
hasRodríguez
, M.Á. been chosen
2
for the press
, L. Garcia-Calvo 2
programme
, A. Carbajal2
,
with unexplained infertility are in risk for decreased ovarian B. Ring1, Y.C. Vives3, M.H. Duvison4, A. Crisci1, C.O. Marieta5,
response : A prospesctive population based cohort study and will not be made
M. Jimenez6, S. Munne1, M. Bejar2
public until the abstract is
S. Kalapahar1, S. Sharma1, R. Chattopadhyay1, S. Parvin1, presented.
1
MedAnswers Inc., Research Development & Analytics, San Pedro, U.S.A. ;
S. Behera1, S. Ghosh1, P. Chakraborty1, B. Chakravorty1 2
Universitat Autònoma de Barcelona, Dpt. Animal Health and Anatomy Veterinary
1
Institute of Reproductive Medicine, Assisted Reproductive Medicine, KOLKATA, Faculty, Barcelona, Spain ;
3
India Complejo hospitalario Ruber Juan Bravo, Unidad de Reproducción Asistida,
Madrid, Spain ;
Study question: Does diabetes confer a differential ovarian response and IVF 4
Ginemed, Reproduccion asistida en Clinicas, Seville, Spain ;
outcome through diminished connexin-43 expression in women with unexplained 5
CER Santander, Medicina de la reproducción, Cantabria, Spain ;
infertility(UI) undergoing ovarian stimulation? 6
Sinae, Research, Seville, Spain
Summary answer: Ovarian reserve and ovarian response to stimulation is
lower in diabetic patient along with decreased connexin-43 expressions in com- Study question: Can we detect a biologically relevant quantity of Anti-
parison to non-diabetic women with unexplained infertility. Mullerian Hormone (AMH) in human hair samples?

Summary answer: AMH can be detected in human hair samples, and levels What is known already: Folliculogenesis is a strictly regulated process, requir-
of AMH in hair are correlative to maternal age. ing interactions of androgenic, estrogenic and gonadotropin signalling. EDs can
What is known already: AMH is a product of granulosa cells of the preantral affect this process and, thus, reproduction by creating hormonal imbalances
and small antral follicles in women. Hence, AMH is often used as a biomarker through different and sometimes not well-established molecular mechanisms.
in assessing fertility. Typically, circulating levels of AMH are tested using blood Few in vitro evidences have associated several phthalates esters and BPA to antral
samples obtained invasively through median cubital vein punctures. Hormone follicle growth inhibition. Urinary EDs are generally considered markers of expo-
concentrations in hair may serve as a comparable and possibly superior means sure, but they may not reflect accurately in vivo exposure of oocytes.
of assaying hormone levels accrued over longer periods of time. Detection of Study design, size, duration: A multicentre study involving six fertility centres
steroid hormones in hair has been used in psychoneuroendocrinological studies across Italy (Milan, Turin, Rome, Naples, Bari, Catania) was conducted between
in human and companion, farm and wild animals. This study represents the first 2018 and 2019, aiming to prospectively recruit at least 120 women attending
quantification of AMH levels in hair in humans. fertility treatments. EDs testing was carried out in a centralised Laboratory
Study design, size, duration: The study design was prospective in nature. (Seminology Laboratory “Loredana Gandini”- “Sapienza”, University of Rome)
A total of (n=152) human female participants between the ages of 18-65 years with a standardized collection protocol and a unique disposable material provider.
were included in the study over a period of 10 months (recruitment ongoing). Participants/materials, setting, methods: The designated sites recruited
Participants/materials, setting, methods: Sample collection was per- 122 women undergoing ART living in their respective geographical area for at
formed in a clinical setting. Blood and hair samples were collected from patients least 3 years, aged ≤42 years, with normal ovarian reserve (AMH≥1 ng/ml and
by nurses. Hair follicles are not required. A doctor or a clinical technician per- FSH≤12 IU/L) and no active ovarian pathologies including PCOS. Subjects com-
formed the ultrasound for measuring the antral follicle count (AFC). Biologically pleted a questionnaire investigating potential sources of contamination. Blood
active AMH was extracted from hair using a proprietary method. AMH presence and Follicular Fluid samples were taken for EDs measurement (MBP, MBzP, MEHP,
in hair extract was confirmed using Western Blotting. AMH was measured in MEHHP, MEOXP and BPA) using liquid chromatography tandem mass spec-
plasma and serum by ELISA. trometry (LC-MS/MS).
Main results and the role of chance: AMH was successfully detected in Main results and the role of chance: The main EDs found in BS and FF
samples (n=152) via western blots on denatured gel with bands at 70kDa. An were MBP (median BS: 8.96 ng/ml, FF 6.43 ng/ml), MEHP (median BS: 9.16 ng/
average level of 9.37 pg/ml (95%CI 6.77-12) was detected in hair and 3.68 ml, FF 7.68 ng/ml) and BPA (median BS: 1.89 ng/ml, FF 1.86 ng/ml) which were
ng/ml (95%CI 2.79-4.56) in serum in age-group <25 yrs. This is in contrast above the limit of detection in both biological matrices in 97%, 77% and 25.4%
to the age group >39 years, within which a mean of 3.02 pg/ml (95%CI of subjects, respectively. A significant correlation between serum and follicular
2.19-3.85) AMH detected in hair and 0.92 ng/ml (95%CI 0.43-1.41) in serum concentration was present for MBP and BPA (Spearman’s ρ: MBP 0.567,
samples. AMH measured in hair correlated with age more strongly than plasma p<0.001; BPA 0.682, p<0.001). Multivariate analyses showed a significant inter-
AMH (p-value =1.26 x10-5 (hair), p-value 0.088 (serum)). AMH levels action between serum and follicular MBP from the geographical area and the
in hair also strongly correlated with antral follicle count (AFC). daily use of plastic food packaging (PFP) (p<0.001, partial η2=0.093). In partic-
Limitations, reasons for caution: Hair is a medium that can accumulate ular, post-hoc univariate analyses detected that serum MBP concentration was
biomarkers over several weeks, while serum is an acute matrix representing only significantly associated with geographical area (p<0.001, adj. mean 13.56 ng/
current levels. Range of detection of AMH in hair was wide within individuals ml, 14.40 ng/ml, 7.61 ng/ml, South, Centre and North Italy respectively) and
from a similar age cohort. AFC testing was included in the study laterally and negatively with home PFP (p=0.004), suggesting that different sources of expo-
has limited data points. sure than PFP may impact more on serum MBP levels. Moreover, follicular but
Wider implications of the findings: We have a novel method of detecting not serum MBP was associated with irregular cycles prior controlled ovarian
AMH in a longitudinal matrix (hair) that could be a more appropriate represen- stimulation (p=0.019). No association was detected between EDs and eating
tation of hormone levels compared to acute matrices like serum or saliva. habits, living in the area of hazardous sites, work contact with potential toxicants
Moreover, our method is also the only truly non-invasive method for testing and other clinical features.
fertility hormones. Limitations, reasons for caution: Despite several potential environmental
Trial registration number: Not Applicable sources of EDs contamination, only a few of them were associated to EDs
concentration in both serum and follicular fluid, suggesting other routes of expo-
P-617 Endocrine disruptors in the serum and follicular fluid of sure and justifying the need for more prospective investigations in larger sam-
a cohort of Italian women undergoing Assisted Reproduction ple size.
Techniques Wider implications of the findings: This study represents the first Italian
biomonitoring of plastic EDs in FF. The finding of MBP and MEHP in follicular
L. De Santis1,10, F. Pallotti2,10, A.P. Dima2,10, R. Ciriminna3,10, R.
fluid confirms the few previous reports available from other countries. This
Gualtieri4,10, V. Pisaturo5,10, M.G. Minasi6,10, L.F. Rienzi7,10, C.
investigation may prove to be the basis for future prospective evaluation to
Scarica8,10, A. Lenzi2,10, F. Botrè9,10, D. Paoli2,10, On behalf of
correlate follicular EDs concentration and oocyte quality.
SIERR(10)[segreteria@sierr.it]
1
San Raffaele Scientific Institute, Ob/Gyn IVF UNIT, Milan, Italy ;
2
“Sapienza” University of Rome, Laboratory of Seminology- Sperm Bank
P-618 IL-6/sIL-6R Increases COX-2 Expression and PGE2
“Loredana Gandini”, Rome, Italy ;
3 Production in a Human Granulosa-Lutein SVOG Cell Line Via a
Centro AMBRA, Fertility Unit, Palermo, Italy ;
4 JAK2/STAT3/SOCS3 Signaling Pathway
University of Naples “Federico II”, Department of Biology, Naples, Italy ;
5
International Evangelical Hospital, Reproductive Medicine, Genoa, Italy ; L. Saijiao1, H.M. Chang2, J.H. Wang2, J. Yang1, P.C.K. Leung2
6 1
European Hospital, Centre for Reproductive Medicine, Rome, Italy ; Renmin Hospital of Wuhan University, Department of Reproductive Medicine
7
Laboratory GENERA, Centres for Reproductive Medicine, Rome, Italy ; Center, Wuhan, China ;
8 2
Casa di cura Villa Salaria in Partnership with Institut Marques, Centres for BC Children’s Hospital Research Institute- University of British Columbia-,
Reproductive Medicine, Rome, Italy ; Department of Obstetrics and Gynaecology, Vancouver, Canada
9
“Sapienza” University of Rome, Department of Experimental Medicine, Rome, Italy ;
10
SIERR, Italian Society of Embryology Reproduction and Research, Italy Study question: The functional role of IL-6 in the regulation of ovarian
Prostaglandin E2 (PGE2) in human granulosa cells is controversial and the detailed
Study question: Are human oocytes exposed to common plastic Endocrine molecular mechanisms remain to be elucidated
Disruptors (EDs) present in the follicular fluid (FF) and blood serum (BS) before Summary answer: The IL-6 trans-signaling-induced activation of JAK2/
retrieval? STAT3/SOCS3 and subsequently increase COX-2 expression and PGE2 pro-
Summary answer: Monobutyl-phthalate (MBP), monoethylhexyl-phthalate duction in hGL cells.
(MEHP) and Bisphenol A (BPA) were detected in FF and BS and associated to What is known already: As a critical paracrine regulator of multiple repro-
geographical area, source of exposure and clinical features. ductive functions, the cytokine interleukin-6 (IL-6) is expressed in human

granulosa cells and detected in follicular fluid. At present, the functional role of TS-specific complication such as cardiac events and hypertension and neonatal
IL-6 in the regulation of ovarian Prostaglandin E2 (PGE2) is controversial. outcomes.
Moreover, the detailed molecular mechanisms by which IL-6 regulates the pro- Main results and the role of chance: Seventy-nine women with TS had a
duction of PGE2 in human granulosa cells remain to be elucidated. Up to now, total 136 pregnancies of which 37% were spontaneous conceptions and 63%
no reports were related to the effect of the IL-6 trans-signaling pathway on PGE2 were achieved with ovum donation (OD). The live birth rate was 58.1% and
production and need further investigation. miscarriage rate 36%, and this was comparable in spontaneous and ovum dona-
Study design, size, duration: Primary human granulosa-lutein (hGL) cells tion pregnancies. There were no significant differences in the prevalence of
and a non-tumorigenic immortalized human granulosa-lutein cell line, SVOG, gestational hypertension, preeclampsia and gestational diabetes between the
were used for this study. Dose, time-course, inhibitors, and siRNA were per- two groups. No case of acute cardiovascular morbidity such as aortic dissection
formed in this experiment. has been identified. There were 5 cases (16%) of vertical transmission of TS to
Participants/materials, setting, methods: Primary human granulosa-lutein daughters in those with spontaneous conception.
(hGL) cells and a non-tumorigenic immortalized human granulosa-lutein cell line, Limitations, reasons for caution: This is a retrospective study reliant on
SVOG, were used for this study. The quantitative PCR and westerns blot were participant recall.
used to measure the mRNA, proteins, and phosphorylation of proteins expres- Wider implications of the findings: The higher rate of miscarriage has been
sion; immunofluorescence staining was used to measure the translocation of previously documented but the comparable rates between those undergoing
STAT3; the small interfering RNA transfection was used to knockdown the OD and spontaneous conception is reassuring. Whilst maternal cardiac morbidity
STAT3 and SOCS3 RNA, and ELISA was used to measure the prostaglandin E2 has been reported, in our cohort there were no acute events identified. The
production. 16% vertical transmission exceeds that previously recorded in the literature.
Main results and the role of chance: The IL-6 trans-signaling using the Trial registration number: n/a
combined addition of IL-6 and soluble IL-6 receptor (sIL-6R) induced COX-2
expression and PGE2 production in hGL cells. Additionally, IL-6/sIL-6R activated P-620 miR-424 suppresses the proliferation and promotes the
the phosphorylation of Janus activated kinase 2 ( JAK2) and signal transducer apoptosis of human ovarian granulosa cells by targeting Apelin and
and activator of transcription 3 (STAT3), induced the STAT3 nuclear transloca- APJ expression
tion, which was inhibited by inhibitors AG490 ( JAK2 inhibitor) and C188-9 J. Du1, X. Lin1, R. Wu1, Z. Gao2, Y. Du1, Y. Liao1, J. Wu1, W. Ke1,
(STAT3 inhibitor), as well as siRNA-mediated knockdown of STAT3. Moreover, S. Quan3
the IL-6 trans-signaling-induced activation of JAK2/STAT3 unregulated the 1
Boai Hospital of Zhongshan, Reproductive Medical Center, Zhongshan, China ;
expression of suppressor of cytokine signaling 3 (SOCS3), which, in turn, neg- 2
Boai Hospital of Zhongshan, Department of Gynecology, Zhongshan, China ;
atively regulated the JAK/STAT3 signaling pathway by suppressing the activity 3
Nanfang Hospital- Southern Medical University, Reproductive Medical Center-
of STAT3 and the subsequent COX-2 expression and PGE2 production in
Department of Obstetrics and Gynecology, Guangzhou, China
hGL cells.
Limitations, reasons for caution: we did not study the interacting effect of
Study question: This study aims to investigated the roles of miR-424 in mod-
IL-6/sIL-6R with gonadotropins because these hGL cells have been exposed to
ulating Apelin expression and GC functions of Polycystic ovary syndrome
the high doses of gonadotropins during the clinical treatment cycles, which cause
(PCOS) .
the cellular expression levels of gonadotropin receptors and the cellular response
Summary answer: miR-424 suppresses the proliferation and promotes the
to gonadotropin could be relatively low.
apoptosis of human ovarian granulosa cells by directly targeting and inhibiting
Wider implications of the findings: Our findings shed light on the cellular
Apelin and APJ expression.
and molecular mechanisms by which the IL-6/sIL-6R-induced JAK2/STAT3/
What is known already: Polycystic ovary syndrome (PCOS) development
SOCS3 signaling pathway modulates the synthesis of PGE2 in the human ovary.
is mediated by alteration of Apelin signaling in ovarian granulosa cells (GCs).
Trial registration number: The Canadian Institutes of Health Research
Study design, size, duration: We aimed to investigate the roles of miR-424
Foundation Scheme Grant (#143317); the National Natural Science Foundation
in regulating Apelin and APJ receptor expression in the context of PCOS, as well
of China Grant (# 81701412); the Nature Science Foundation of Hubei Province
as their involvement in granulosa cell proliferation and apoptosis.The follicular
Grant (# 2018CFB491)
fluid and serum samples used for analysis were collected from patients with
PCOS who were registered at the Reproductive Medical Center of Boai Hospital
P-619 Pregnancy outcomes in Turner syndrome of Zhongshan between Jun 1st, 2015 and Dec 31st, 2018. Healthy volunteers
E. Burt1, M. Davies1, E. Yasmin1, A. Cameron- Pimblett1, were included as the control group.
G. Conway1 Participants/materials, setting, methods: miRNA expression in GCs
1
University College London Hospital, Reproductive Medicine Unit, London, United were altered by transfection with specific mimics and inhibitors. Apelin concen-
Kingdom tration was determined by ELISA, and gene and miR-424 expression were ana-
lyzed by quantitative RT-PCR. Protein abundances were measured by western
Study question: Assessing pregnancy outcomes in women with Turner syn- blotting. Association of miR-424 with genomic sequences were validated by
drome (TS) dual-luciferase reporter assay. Apelin gene expression was promoted by LV-003
Summary answer: Initial results show an overall high miscarriage rate, no plasmid-mediated cell transfection. GC proliferation was analyzed by MTS
acute cardiovascular morbidity and a 16% vertical transmission of TS. method, and its apoptosis were measured by flow cytometry.
What is known already: Turner Syndrome (TS) affects 1:2500 females and Main results and the role of chance: Apelin concentration was increased
is caused by the partial or complete loss of one X chromosome. About 80% in serum and follicular fluid from PCOS patients, accompanied with accelerated
of women with TS experience primary amenorrhea and therefore the only APJ (Apelin receptor) expression and suppressed miR-424 expression in GCs.
option for fertility treatment is ovum donation (OD). The remaining 20% may miR-424 mimics suppressed Apelin and APJ expression in KGN cells by targeting
have the opportunity for a spontaneous pregnancy. Pregnancy in women with Apelin and APJ 3’-UTR regions, which was recovered by miR-424 inhibitors.
TS has been associated with excess obstetric risk such as miscarriage and miR-424 inhibited KGN cell proliferation and cell cycle progression by down-reg-
hypertension. Estimates for maternal mortality have varied between 0 and 2% ulating Cyclin-D/E expression. Moreover, miR-424 promoted KGN cell apoptosis
in TS mainly due to the risk of aortic dissection but until now there was no by elevating truncated Caspase-3 level. The regulation of KGN cell proliferation
UK data. and apoptosis by miR-424 was mediated by directly suppressing Apelin gene
Study design, size, duration: Retrospective single centre cross-sec- expression, but not the inhibition of Apelin peptide activity.
tional study Limitations, reasons for caution: We use luteinized granulosa cells and
Participants/materials, setting, methods: Over 850 women with TS have KGN cell lines, so whether there is the same effect on non luteinized granulosa
attended the adult TS clinic at University College London Hospital and those cells needs to be discussed.
who had achieved pregnancy were identified. Telephone interviews were con- Wider implications of the findings: We first revealed that miR-424 expres-
ducted to collect data regarding; mode of conception, mode of delivery, sion was greatly down-regulated in the ovarian granulosa cells from PCOS

patients, but Apelin and APJ expression showed opposite alterations, miR-424 P-622 The entire range of trigger-day endometrial thickness
could directly target the 3’UTR regions of both the Apelin and APJ gene in KGN values in fresh in vitro fertilization cycles is in direct independent
cell line. correlation with the live birth rate
Trial registration number: not applicable Y. Shufaro1, M. Simeonov2, Y. Lande2, A. Ben-Haroush3, G. Oron3,
E. Shlush2, E. Altman3, A. Wertheimer2, T. Shochat4, O. Sapir3
P-621 PRP in recurrent implantation failure, hope or hype !? 1
Beilinson Women’s Hospital- Rabin Medical Center- and the Felsenstein MRC- Tel-
A Prospective randomized controlled study Aviv University, Infertility and IVF Unit, Petach-Tikva, Israel ;
K.A. Ibrahim Rageh1, A. Barakat1, A. Mohamed2 2
Beilinson Women’s Hospital- Rabin Medical Center, Infertility and IVF Unit,
1
Al-Baraka fertility hospital, Ivf, Manama, Bahrain ; Petach-Tikva, Israel ;
3
2
Al-Baraka fertility hospital, IVF lab, Manama, Bahrain Beilinson Women’s Hospital- Rabin Medical Center- Tel-Aviv University, Infertility
and IVF Unit, Petach-Tikva, Israel ;
4
Study question: PRP in recurrent implantation failure, hope or hype !? Beilinson Women’s Hospital- Rabin Medical Center, Research Authority,
Petach-Tikva, Israel
• We designed this study to evaluate the effectiveness of endometrial perfu-
sion of PRP in improvement of pregnancy rate in RIF patients Study question: What is the association of the entire range of the trigger-day
endometrial thickness (EMT) with the live birth rate (LBR) following in vitro
Summary answer: The PRP significantly improved the pregnancy rate and fertilization (IVF) and fresh embryo-transfer (ET) ?
may be a new hope for management of recurrent implantation failure; which is Summary answer: The entire range of EMT is in direct correlation with LBR
one of the nightmares in IVF-fresh-ET cycles, even after adjusting for age and ovarian response to
What is known already: The endometrial receptivity have been accepted to stimulation.
be major limiting factors in the establishment of pregnancy. In spite of improved What is known already: The association between EMT and LBR in IVF cycles
almost all aspects of IVF: ovarian stimulation, embryo culture and transfer, the with fresh ET is unclear. Some studies reported a positive association of EMT
pregnancy rates still not satisfactory. The bottleneck is the process of implantation. with pregnancy rates but in other studies such a correlation did not exist. There
Recurrent implantation failure (RIF) is one of the nightmares in reproductive are also conflicting reports regarding the impact of an increased EMT (≥ 13 mm)
medicine and despite several strategies that have been described for manage- on treatment outcome.
ment; there is no universal agreement yet. Study design, size, duration: A cohort study of all IVF cycles with fresh day-3
Recently, PRPis described to promote endometrial growth and receptivity, ET in patients age ≤42 in a single centre. LBR was calculated for all EMT values,
PRP has been investigated as a therapeutic approach for several medical disor- stratified into 5 groups (≤6,7-9,10-12,13-15,and ≥16 mm), overall and within sub-
ders, but its use in IVF is still limited. groups of patient age (≤35,36-40,41-42 years) and ovarian response(1-7,8-15,>16oo-
Study design, size, duration: Study design; cytes). Univariate analysis and multivariate logistic regression model were used for
prospective randomized controlled study adjusting for various independent variables. General linear models were generated
sample size ; to compare continuous variables between EMT, age and oocyte groups.
150 participants Participants/materials, setting, methods: 5133 IVF cycles, performed in
duration; 2343 female patients age 42 or younger, in a single center between 2009-2017
from July 2018 to March 2019. were included. Fresh ET was performed on day 3 in all included cycles.
Participants/materials, setting, methods: 150 infertile women with history Main results and the role of chance: LBRs were as follows: 11.22%
of RIF gave their consent to be included in this study- with age below 40 yrs, (35/312) in cycles with EMT ≤6mm, 17.98% (280/2114) in cycles with EMT
body mass index (BMI) below 30 kg/m2- from July 2018 to March 2019.Divided 7-9mm, 23.44% (476/2031) in cycles with EMT 10-12mm, 25.62% (144/562)
into 2 comparable groups; all underwent antagonist protocol, In the study group, in cycles with EMT 13-15mm, and 34.21% (39/114) in cycles with EMT ≥16 mm
intrauterine infusion of 0.5 to 1 ml of PRP was performed 48 hrs before blas- (P <0.001). Similar findings were observed on further sub-group analysis accord-
tocyst transfer, pregnancy tests were done 12 days after ET. ing to patient age and ovarian response. The entire range of trigger-day EMT
Main results and the role of chance: The result of our study revealed that values in fresh cycles was found to be in direct correlation with the LBR, even
endometrial perfusion of platelet-rich plasma (PRP) significantly improved the after adjusting for confounders of age and ovarian response to stimulation. This
pregnancy rate and may be a new hope inthose patents with a history of recur- observation was confirmed also by a multivariate logistic regression analysis in
rent implantation failure (RIF). Out of 75 participants in each group, 32 got which the EMT was found to be a significant independent predictor of LBR
pregnant (43%) in the study group after endometrial PRP infusion, compared to controlling for various confounders. Excessively thick endometrium (≥13mm)
11 pregnant participants (15%) in the control group. was not detrimental, but rather beneficial, for treatment outcome.
The main strengths of our study include: (1) this is by far the first study discussing Limitations, reasons for caution: Retrospective study, proving that EMT is
this topic in our country, as our hospital is the leader IVF center here in Bahrain. an important independent prognostic factor for a favourable ART outcome. The
(2)The low risk of bias between study and control group as both of them were findings of the study are relevant only for cycles in which fresh ET is performed.
statistically comparable. (3) We exclude those patients with hematological and This study does not address the question if interventions to increase the EMT
immunological disorders, hormonal disorders, chromosomal and genetic abnor- are beneficial.
malities and uterine abnormalities (acquired or congenital) as confirmed by HSG Wider implications of the findings: The pre-trigger EMT is in significant
and U/S to limit additional factors that may affect the results of the study. (4) independent correlation with the live birth rate. We suggest that stimulation
All Blastocyst transfers were performed under ultrasound guidance by only one cycles should be planned and managed to achieve maximal endometrial prolif-
expert gynecologist with infertility fellowship. (5) The use of individual patient eration, and that fresh ET can be performed at high EMT values without endan-
data for direct comparison between both groups. (6) The consistency between gering the outcome of the cycle.
our results and those of the previous trials in the literature. Trial registration number: NA
Limitations, reasons for caution: as we are living in small country so, sample
size may need to be increased in the further coming clinical trials, also at time P-623 Aberrant BMP15/HIF-1α/SCF signaling pathway in human
of ET we select them phenotypically according to the embryo scoring, not based granulosa cells is involved in the PCOS related abnormal follicular
on genetic basis as we are usually not doing PGT as a routine; development
Wider implications of the findings: RIF is hard to be managed, and if all J. Tan1
available treatments fail, then PRP can add value. PRP that contains several 1
Jiangxi Provincial Maternal and Child Health Hospital, Reproductive Medicine
growth factors and cytokines may improve endometrial receptivity and implan- Center, Nanchang, China
tation. PRP is collected from autologous blood sample, so in comparison to
G-CSF, PRP is more accessible and affordable Study question: To explore the related regulatory mechanisms of stem cell
Trial registration number: NCT04085783 factor (SCF) expression in PCOS related abnormal follicular development.

Summary answer: BMP15 induce SCF expression by up-regulating HIF-1α in regulation of adipogenesis and adipocyte functions including lipid metabo-
human GCs, aberrance of this signaling pathway might be involved in PCOS lism in PCOS.
related abnormal follicular development. Study design, size, duration: 17 women with PCOS and 24 normal women
What is known already: Our previous studies have found that stem cell who underwent laparoscopic operation due to tubal factors were recruited. We
factor (SCF), which is an important granulosa cells (GCs) derived growth factor collected omental adipose tissue samples (approximately 1 cm3size) during the
in follicular development, is significantly reduced in PCOS related abnormal surgery and analyzed gene expression level.
follicles. However, the related regulator mechanisms of SCF expression in PCOS Participants/materials, setting, methods: Primary human preadipocytes,
are still uncompleted clear. The present study is to further investigate the regu- isolated from omental adipose tissue, were cultured in vitro.
latory mechanism of SCF expression in human GCs, which can help us to better Dihydrotestosterone-treated cells were used as study models. The regulation
understand the abnormal development of PCOS related follicles of adipogenesis was confirmed by Western blot and Oil Red O staining after
Study design, size, duration: The present study includes human serums, androgen stimulation.
follicular fluids (FFs) and GCs were collected from 69 PCOS patients and 74 Main results and the role of chance: PCOS women had increased serum
normal ovulatory patients during the IVF therapy, respectively. Human granulosa concentrations of testosterone (P = 0.001) and higher HOMA-IR levels (P =
cell line (KGN) was also used in this study. 0.04), with increased expression of androgen receptor (AR) in adipose tissue.
Participants/materials, setting, methods: ELISA was used to evaluate In vitro studies indicated that androgens, via the AR, enhanced lipogenesis in
the concentrations of SCF and BMP15 in FFs and serums. KGN cells were human primary preadipocytes. DHT-treated cells produced more lipid droplets
treated with recombinant human BMP15 alone or in combination with BMP on Day8 in vitro. Dose-response experiments performed in human preadipocyte
type I receptor inhibitors and transfected with hypoxia inducible factor-1α cultures showed that expression of adipogenic transcription factors CCAAT/
siRNA before detecting the target genes and proteins by RT-PCR and Western enhancer-binding protein alpha (C/EBPα) and peroxisome proliferator-activated
blot, respectively. The rates of metaphase II, oocyte, fertilization, embryo receptor γ (PPARγ) increased after androgen exposure.
cleavage and good quality embryo between PCOS group and non-PCOS group Limitations, reasons for caution: The mechanism of adipocyte dysfunction
were analyzed. and metabolic disorder mediated by androgen in PCOS should be further
Main results and the role of chance: The results showed that the rate evaluated.
of MII oocyte and 2PN fertilization was significantly lower in PCOS group than Wider implications of the findings: Our results suggested that adipose tissue
those in non-PCOS group, though PCOS patients retrieved much more was an important site linking androgen and metabolism in PCOS. PCOS is an
oocytes. No difference was observed for the cleavage rate and high-quality androgen excess disorder associated with adipose tissue disturbances. For obese
embryo rate between these two groups. Furthermore, the concentration of patients with hyperandrogenism, exercise accompanied with anti-androgenic
BMP15 in FF and the concentration of SCF in serum and FF were also signifi- therapy could be efficient than those only accept lifestyle interventions to achieve
cantly lower in PCOS patients. Interestingly, we found a weakened expression weight loss.
of HIF-1α and SCF in GCs from PCOS patients when compared with the Trial registration number: not applicable
non-PCOS patients. The expression of HIF-1α and SCF was significantly
increased in KGN cells after treating cells with rhBMP15, However, this pro-
motion effects of BMP15 on HIF-1α and SCF expression were obviously P-626 Vitamin D supplementation prior to initiate IVF: a
abolished by co-treatment with DM. Moreover, knock down of HIF-1α expres- randomized controlled study
sion in KGN cells by transfecting cells with HIF-1α siRNA significantly reduced M. Reschini1, V. Sarais2, S. Ferrari1, G.C. Cermisoni2, A. Paffoni3,
the expression of SCF in human GCs, in spite of activating BMP15 signaling S. Signorelli2, E. Papaleo2, E. Somigliana1, P. Viganò2
pathway. 1
Fondazione IRCCS Ca’ Granda - Ospedale Maggiore Policlinico, Infertility Unit,
Limitations, reasons for caution: The present results were obtained from Milano, Italy ;
human GCs and KGN cell lines, which might not be fully representative of the 2
IRCCS San Raffaele, Centro Scienze Natalità, Milan, Italy ;
true human PCOS environment. 3
ASST Lariana, Infertility Unit, Cantù, Italy
Wider implications of the findings: This study showed a significantly reduc-
tion of BMP15, HIF-1α and SCF in PCOS related abnormal follicles, and demon- Study question: Does vitamin D supplementation improve pregnancy rate in
strated that BMP15 could induce SCF expression by up-regulating HIF-1α women undergoing IVF?
expression in human GCs. The aberrance of this signaling pathway might be Summary answer: In IVF cycles, Vitamin D supplementation does not increase
involved in the PCOS related abnormal follicular development. the chances of pregnancy.
Trial registration number: not applicable What is known already: Vitamin D plays an important role in human physi-
ology and pathology and vitamin D insufficiency has been shown to be associated
P-624 The ovarian yield – number of oocytes per antral follicular with several diseases. There is also growing evidence supporting a role of vitamin
count (AFC) score and anti-Mullerian hormone (AMH) levels – D in reproductive health, especially in female fertility. In particular, IVF success
does not predict ART outcome. was shown to be higher in women with appropriate reserves of vitamin D.
However, a causal relation has not been established and evidence from RCTs
“Abstract withdrawn by the authors”
exploring the possible benefits of vitamin D supplementation is sparse,
under-powered and inconsistent.
P-625 Hyperandrogenism regulates adipocytes expandasion and Study design, size, duration: A multicenter randomized double blinded
differentiation in polycystic ovary syndrome placebo controlled study was conducted to investigate the possible benefits of
Y. Qian1, D. Yang1 vitamin D supplementation. Women referring for IVF in two ART Centres from
1
Sun Yat-sen Memorial Hospital, Reproductive medicine center, Guangzhou, China October 2018 to January 2019 were considered for study entry. The main
inclusion criterion was a serum vitamin D level <30 ng/ml. Eligible women
Study question: What is the effect of androgen excess on adipocyte differ- received either Vitamin D (600.000 IU) or placebo in a single oral administration
entiation in PCOS? between 2 and 12 weeks before to oocyte retrival.
Summary answer: Androgens, via the androgen receptors, exert direct effects Participants/materials, setting, methods: Inclusion criteria of patients
on adipocyte differentiation and may leads to the metabolic disorder in were: indication to IVF, age 18-39 years, ≤2 previous oocytes retrievals and BMI
PCOS women. between 18 and 25 kg/m2. Poor responders according to Bologna criteria were
What is known already: Polycystic ovary syndrome (PCOS) women are excluded as well as patients using surgically-retrieved spermatozoa. Both partic-
at risk for developing metabolic syndrome, impaired glucose tolerance and ipants and physicians were blinded to the allocation. The main outcome was
cardiovascular disease. Androgen excess is a common feature among women cumulative pregnancy rate per retrieval. The analysis was by intention to treat.
with PCOS and has been suggested as associated with adipose tissue dis- Main results and the role of chance: Overall, 738 patients were initially
turbance. We hypothesized that androgens play an important role in selected. Eighty-eight (12%) were subsequently excluded because their serum

vitamin D was ≥30 ng/ml and 20 (3%) declined participation after being initially to disparities between intervention including dose, content, population charac-
selected. The remaining 630 women were randomized, 308 received Vitamin D teristics and measured outcomes.
and 322 were treated with placebo. 21 (7%) and 22 (7%) dropped-out prior to Whilst most authors focus on identifying whether a health benefit of PUFA
initiate the cycle, respectively. The total number of women obtaining a clinical supplementation exists, potential risks must also be explored. None of the
pregnancy in treated and control women was 123 (40%) and 131 (41%), respec- included studies explored miscarriage rates, congenital anomalies or obstetric
tively (p=0.85). Women achieving ongoing pregnancies / live births were 100 outcomes such as birth weight and prematurity.
(32%) and 111 (34%), respectively (p=0.59). The Relative Risk (RR) of clinical Importantly, a complex link between genetic variation and lipid metabolism
and ongoing pregnancy / live births were 0.98 (95%CI: 0.81 – 1.19, p=0.85) and exists, thus challenging the comparison between studies, which included different
0.94 (95%CI: 0.76 – 1.17, p=0.59), respectively. When focusing on IVF cycles demographic groups. Gene-diet interactions must be considered but are difficult
characteristics, no main differences emerged. In particular, duration of hyper- to evaluate.
stimulation (p=0.91), total dosage of gonadotropins administered (p=0.98), Overall, there were distinct differences between population groups,
number of retrieved oocytes (p=0.22), IU of gonadotropins per retrieved study design and measured outcomes. Studies showing a benefit were of low
oocyte (p=0.55) and fertilization rate (p=0.33) did not differ. quality and only six of the included studies assessed live birth rates as an
Limitations, reasons for caution: The mode of administration of vitamin outcome.
D may be inappropriate for improving IVF outcomes. One may claim that an Limitations, reasons for caution: The main limitation was the disparity
earlier initiation of the supplementation (5-6 months prior to initiate the cycle) between study methodologies. Assessment of PUFA intake varied between stud-
and a daily administration could be effective. ies; some assessed dietary intake via validated food questionnaires whilst others
Wider implications of the findings: The routine administration of vitamin measured serum fatty acid or follicular fluid fatty acid concentrations. Comparison
D prior to initiate IVF in order to increase the probability of pregnancy is not between studies was also hindered by different ART protocols used in
recommended. institutions.
Trial registration number: EUDRACT 2015-004233-27 Wider implications of the findings: Although the ovarian micro-environ-
ment is an important contributor to conception and embryo development, there
P-627 Impact of polyunsaturated fatty acid supplementation on is a lack of high quality research to support dietary PUFA supplementation in
assisted reproductive technology (ART) outcomes: a systematic women undergoing ART. In conclusion,a need exists for well-designed RCTs to
review facilitate the understanding of PUFA supplementation in women undergoing ART.
E. Bayar1, S. Saso1, N. Galazis2, B. Jones1, T. Bracewell-Milnes3, Trial registration number: N/A
M. Chawla4, Y. Ahmed-Salim1, J. Ben Nagi5
P-628 A typical temperature patterns as an aid to identify
1
Imperial College NHS Trust, Obstetrics & Gynaecology, London, United Kingdom ; infertility issues and miscarriage risk
2
Northwick Park NHS Trust, Obstetrics & Gynaecology, London, United Kingdom ;
3 M. Karoshi1, B. Hurst, S2, R. Milnes3, T. Knowles4, A. Pirrie5
Chelsea and Westminster NHS Hospital, Obstetrics & Gynaecology, London,
1
United Kingdom ; Royal Free Hospital and UCL Medical School, Gynaecology, London, United
4
North Middlesex University Hospital, Obstetrics & Gynaecology, London, United Kingdom ;
2
Kingdom ; Carolinas Medical Center, Department of Assisted Reproduction, Charlotte, U.S.A. ;
3
5
The Centre for Reproductive and Genetic Health, Fertility, London, United Fertility Focus Limited, Research and Development, Warwick, United Kingdom ;
4
Kingdom University of Bristol, Faculty of Medical and Veterinary Sciences, Bristol, United
Kingdom ;
5
Study question: This review explores the role of dietary polyunsaturated fatty Fertility Focus Limited, Research & Development, Warwick, United Kingdom
acid (PUFA) supplementation in women undergoing assisted reproduction tech-
nology (ART) in order to guide recommendations for clinical practice. Study question: To determine if ovulatory abnormalities and risk of miscar-
Summary answer: There is a need for greater quality randomised control riage were associated with atypical Patterns of vaginal core body temperature
trials (RCTs) to facilitate an in depth understanding of how PUFA supplementa- (CBT) measurements from the OvuSense system.
tion may impact ART success. Summary answer: Identified atypical OvuSense (OS) CBT Patterns are more
What is known already: Modifiable risk factors are important targets for likely to occur with “infertility related diagnoses” including PCOS, PCOS with
improving reproductive health and often represent the first interventional step in regular cycles, and elevated miscarriage risk.
maximising the chances of conception. Over recent years, diet and nutritional What is known already: Three novel, atypical CBT Patterns published pre-
supplementations have received increasing attention. Women undergoing ART viously, confirmed in updated population of 18,679 ovulatory cycles: (a) “Crash
are frequently targeted with readily available over the counter supplementations To Baseline” = first nightly averaged CBT falls by >0.2 degrees Celsius (0C) to
in an attempt to boost conception chances. Fatty acids play an important role in lowest cycle CBT point (baseline), (b) “False Start” = rise of >0.10C did not
basic cellular function and animal studies have suggested a beneficial role on oocyte result in ovulation but instead a return to baseline CBT followed by ovulation
quality and embryogenesis. This has led to the suggestion that PUFA supplemen- two or more days later in the cycle, (c) “Crash After Ovulation” = final
tation may improve reproductive outcomes in women undergoing ART. CBT >0.20C lower than the post ovulatory peak CBT. It is likely OS patterns
Study design, size, duration: The primary outcomes assessed included closely reflect progesterone changes, hence cycle-related hormonal abnormal-
pregnancy, live birth, implantation and miscarriage rates. PRISMA guidelines were ities may be associated with atypical patterns.
used to conduct this review. An electronic database search was performed using Study design, size, duration: Retrospective, longitudinal, comparative,
MEDLINE, EMBASE and the Cochrane Library to identify articles published from observational study. Participants used OS vaginally at night to monitor CBT when
January 1978 to 2019. Abstracts were screened for suitability following which, not menstruating. The total study population (TSP) was 18,679 ovulatory cycles
full text articles were reviewed in detail. Additional studies identified through from 8,653 OS users recorded between March 2016 and December 2019. A
screening review articles were also considered. detailed medical questionnaire was then issued to study participants and the
Participants/materials, setting, methods: A total of 102 studies were answers from 375 respondents accounting for 1,491 of the TSP cycles was used
identified through searching of databases. A further 12 studies were retrieved for further assessment.
through manual searching and review papers. 114 papers were screened, with Participants/materials, setting, methods: TSP used to confirm prevalence
90 exclusions, leaving 24 studies for full-text review. Of these, nine studies were of cycle Patterns (a)-(c); and questionnaire assessed per respondent for each
then excluded (five papers did not address the proposed research question, following existing ‘Diagnosis’:
three were review articles and one study was on-going). Overall 15 studies were
eligible for inclusion in our systematic review. 1. Any infertility related diagnosis
Main results and the role of chance: In total, 15 studies met the inclusion 2. PCOS
criteria for this review, four of which were RCTs. Although the overall aim was 3. PCOS and regular cycles
shared between the included RCTs, meta-analysis could not be performed due 4. Previous miscarriage = gravida >0, number miscarriages >0.

Odds Ratio (OR) calculated as (a/b)/(c/d) for each Pattern + Diagnosis and without CS using Chi-square test. All confounding variables were adjusted
combination together with their 95% confidence interval: a. Positive Diagnosis for using multivariate logistic regression, based on any significant differences
(+D), Pattern >1 cycle for respondent (+P); b. -D+P; c. +D-P; d. -D-P. between the two groups generating adjusted odds ratios (aOR).
Main results and the role of chance: The prevalence of each Diagnosis in Main results and the role of chance: We identified 9,096,788 pregnancies
the total respondent population of 375 was: 217 (57.9%) with 1. Any infertility during the study period. CS complicated 135 pregnancies at a rate of 1-2 cases
related diagnosis; 156 (41.6%) with 2. PCOS; 53 (14.1%) with 3. PCOS and per 100,000 births. CS subjects were more likely to be older (p<0.001), obese
regular cycles (i.e. 34.0% of those with Diagnosis of PCOS report regular cycles); (p<0.001), have private insurance (p<0.001), chronic hypertension (24.4% ver-
145 (38.7%) with 4. Previous miscarriage. Note: respondents can potentially sus 1.8%, p<0.001), and pre-gestational diabetes (7.4% versus 0.9%, p<0.001).
have more than one Diagnosis and Pattern. The maternal mortality rate was 0.7% and 0.007% in CS and control groups,
respectively. Preeclampsia was higher in CS compared to controls after con-
(a) “Crash To Baseline” 14.0% of TSP, and 263 cycles from 164 respondents. trolling for baseline risk factors, including pre-existing hypertension (aOR 2.20,
OR: 1.44 for 1 (CI 0.95-2.18); 1.69 for 2 (1.12-2.56); 15.71 for 3 (7.90- 95% CI 1.18-4.41). Operative vaginal delivery and blood transfusion rates were
31.23); and 6.17 for 4 (3.59-10.63). high in CS compared to controls after controlling for confounding factors, aOR
(b) “False Start” 9.1% of TSP, and 202 cycles from 133 respondents. OR: 6.49 (95% CI 4.50-9.38) and aOR 3.09 (95% CI 1.35-7.07), respectively. The
1.69 for 1 (CI 1.12-2.56); 2.84 for 2 (1.83-4.39); 9.52 for 3 (4.80-18.87); rates of preterm delivery (8.9% versus 7.2%) and gestational diabetes (8.1%
and 6.99 for 4 (3.89-12.57). versus 5.8%) were not statistically different between CS and control groups,
(c) “Crash After Ovulation” 11.4% of TSP, and 216 cycles from 128 respon- aOR 0.82 (95% CI 0.45-1.56) and aOR 0.82 (95% CI 0.44-1.56), respectively.
dents. OR: 1.10 for 1 (CI 0.71-1.70); 1.61 for 2 (1.04-2.47); 1.45 for 3 Limitations, reasons for caution: This retrospective analysis utilizes an
(0.80-2.62); and 7.04 for 4 (3.82-12.99). administrative database, with its inherent limitations. Significant medical history
or adverse pregnancy outcomes may be more often reported in patients with
more significant conditions or outcomes
These results indicate strong associations between reported Diagnosis and
Wider implications of the findings: CS patients often begin pregnancies
the atypical CBT Patterns identified using the OS system.
with maladies making them at risk for complications. These patients might benefit
The high OR for each Pattern associated with miscarriage merits further
from prevention methods for preeclampsia, and increased surveillance to
investigation, as the cohort has a low pregnancy rate.
decrease maternal morbidity and mortality. Certain aflictions which were
Limitations, reasons for caution: The authors note the co-existence of
increased in case series were not found to be elevated in CS in this study
Pattern + Diagnosis is not strictly “predictive”, as each Diagnosis is by definition
historic. The population is by definition biased to one or more Diagnosis as over
57% of respondents report having been trying to conceive for a year or more
P-630 Early pregnancy loss in patients with polycystic ovary
prior to use.
syndrome: in vitro maturation of oocytes versus controlled
Wider implications of the findings: Results suggest that atypical CBT
ovarian stimulation
Patterns may aid diagnosis, and in particular elevated risk of miscarriage. It should
be noted that the absence of an existing Diagnosis does not necessarily render L. Mostinckx1, S. Mackens1, P. Drakopoulos1, I. Segers1,
the results with positive Patterns “false”, and the existence of a Pattern could S. Santos-Ribeiro2, B. Popovic-Todorovic1, H. Tournaye1,
anyway indicate investigation for ovulatory abnormalities. C. Blockeel1, M. De Vos1
Trial registration number: Atrium Health IRB File #03-19-16E 1
Universitair Ziekenhuis Brussel, Centre for Reproductive Medicine, Brussels,
Belgium ;
2
IVI-RMA Lisboa, Gynecology/Reproductive Medicine, Lisbon, Portugal
P-629 The Effect of Cushing’s syndrome on Pregnancy
Complication Rates: Analysis of More than 9 Million Deliveries Study question: Is early pregnancy loss (EPL) increased in PCOS patients
H. Baghlaf1, A. Badeghiesh2, E. Suarthana2, M. Dahan2 undergoing ICSI following in-vitro maturation of oocytes (IVM) when compared
1
with controlled ovarian stimulation (COS)?
Toronto, Obstetrics and Gynecology, Toronto, Canada ;
2
Summary answer: PCOS patients who achieved a pregnancy following IVM-
McGill University, Obstetrics and Gynecology, Montreal, Canada
ICSI were more at risk for EPL compared to their counterparts who were preg-
nant after COS-ICSI.
Study question: What are maternal and fetal morbidities that increase with What is known already: Although some studies have reported comparable
Cushing’s syndrome (CS)? miscarriage rates after IVM and COS in women with PCOS, others have reported
Summary answer: CS increases risks of pre-eclampsia, blood transfusion, increased miscarriage rates after IVM. Nevertheless, apart from their limited
and operative vaginal deliveries, but unexpectedly not preterm deliveries and sample size, many of these studies were performed using hCG-triggered IVM
gestational diabetes compared to the control group. protocols, and patient or treatment characteristics that may contribute to EPL
What is known already: Cushing’s syndrome (CS) rarely complicates preg- were not considered. Therefore, we aimed to compare EPL after non-hCG
nancy. This may be due to the induced infertility via alterations in gonadotropins triggered IVM and COS following fresh or frozen embryo transfer in patients
and androgen. CS is believed to increases maternal and fetal morbidities, including with a specific phenotypical diagnosis of PCOS.
preeclampsia, gestational diabetes, preterm delivery and intrauterine growth Study design, size, duration: This was a retrospective, single-centre cohort
restriction. This knowledge is based solely on case reports and small case series. study including 800 pregnant infertile women between 18 and 36 years of age
A recent systematic review and meta-analysis confirmed the above findings and with PCOS as defined by the extended Rotterdam criteria who underwent either
it included only case reports and case series. Studies with control groups are IVM or COS between January 2010 and December 2017.
lacking in the literature and this study was conducted to compare pregnancies Participants/materials, setting, methods: Non-hCG triggered IVM (28-
complicated with CS to pregnancies in general population 40h) after a short HP-hMG course, or conventional COS, followed by ICSI and
Study design, size, duration: We conducted a retrospective popula- fresh or frozen embryo transfer (ET). Only the outcome of the first ET resulting
tion-based cohort study utilizing data from the Health Care Cost and Utilization in a biochemical pregnancy was included in the analysis. The main outcome
Project-Nationwide Inpatient Sample database (HCUP-NIS) over 11 years from measure was EPL (biochemical pregnancy loss (BPL) + miscarriage) before ten
2004 to 2014. We created a cohort of all deliveries between 2004 and 2014 weeks’ gestation. Relevant patient characteristics were analysed in order to
inclusively. Within this group, all deliveries to women with CS were identified as develop a multivariate logistic model for independent risk factors of EPL.
part of the study group (n=135), and the remaining deliveries were categorized Main results and the role of chance: Outcomes in 329 IVM patients were
as non-CS births and comprised the reference group (n=9,096,653). compared with those of 471 COS patients. Rank of ET was comparable (0.84
Participants/materials, setting, methods: Analysis was performed to ± 1.23 vs. 1.04 ± 1.64 previous ETs respectively in the IVM and COS groups).
identify the prevalence of pregnant women with CS over the study duration. Pregnant IVM patients had higher AMH levels (11.5 ± 8.1 ng/mL vs. 7.2 ± 4.1
Demographic and clinical characteristics were compared between women with ng/mL, p< 0.001) compared to pregnant COS patients. Hyperandrogenic PCOS

phenotypes were more common among IVM patients (59.9% vs. 48.2%, the studies associating ovarian response to aneuploidy, only Haaf et al reported
p= 0.001). Women who were pregnant after COS had previously suffered EPL an increased aneuploidy rate, although statistical limitations were observed.
more often compared to IVM patients (28% vs. 17.6%, p= 0.003). Pregnancies All other 6 studies reported similar results, concluding that ovarian response
after IVM were more often obtained after frozen ET (62.9%) compared to those did not alter the euploidy rate or the total number of euploid embryos.
after COS (35.2%, p< 0.001). Transfer of a cleavage stage embryo was more Moreover, a case can be made towards better results with an increasing number
common in pregnant IVM patients (62.6%) compared to COS patients (32.1%, of biopsied embryos.
p< 0.001). The BPL rate (13.1% after IVM vs. 9.3% after COS, p=0.09) was Limitations, reasons for caution: Only one RCT has been performed. Even
similar, but IVM patients had a higher miscarriage rate before ten weeks’ gestation though the quality of the rest of the studies is high, bias associated to retrospec-
(23.4% vs. 14.6%, p=0.002). In a multivariate logistic model allowing adjustment tive studies cannot be ruled-out. Studies presented a large diversity in the assess-
for relevant confounders, IVM was the only independent factor (adjusted OR ment of embryonic aneuploidy, and, given the great heterogeneity between
1.62, 95% CI 1.12-2.33) associated with increased odds of EPL. studies, a meta-analysis could not be performed.
Limitations, reasons for caution: This is a large observational study based Wider implications of the findings: Due to the fact that the number of
on retrospective data collection. Despite our robust methodological approach, euploid embryos available for transfer increases as the number of oocytes
the presence of bias related to the retrospective design cannot be excluded. obtained does, considering the absolute number of euploid embryos seems
Wider implications of the findings: The observation of an increased risk more relevant than the proportion of euploid/aneuploid embryos, especially
of EPL following IVM warrants further research that should result in improved when cumulative LBR outcome is ascertained as the optimal outcome.
IVM laboratory protocols conducive to enhanced embryo quality and improved Trial registration number: CRD42019120803
developmental potential.
Trial registration number: not applicable P-632 Poor ovarian reserve and response are associated with
interleukin 10 levels in women undergoing in-vitro fertilization
P-631 Association of ovarian stimulation with embryonic A.M. Fabregat Reolid1, M. Hortal1, B. Lledó1, J.A. Ortiz1,
aneuploidy in in vitro fertilization (IVF) cycles: a narrative A. Bernabeu2, J. Llácer2, R. Bernabeu2
systematic review 1
Instituto Bernabeu, Biotech, Alicante, Spain ;
J. Rodriguez-Purata1, M.J. Gomez-Cuesta1, E. Cervantes-Bravo1 2
Instituto Bernabeu, Reproductive Medicine, Alicante, Spain
1
CdelaF, Clinica de la Fertilidad, Cuajimalpa de Morelos, Mexico
Study question: Are cytokines related to ovarian reserve and response in ART?
Study question: Is there an association of controlled ovarian stimulation (COS) Summary answer: Interleukin-10 is strongly and positively related to ovarian
with embryonic aneuploidy in IVF cycles coupled with preimplantation genetic reserve and response parameters, providing a potential new tool in the prediction
testing (PGT)? of controlled ovarian stimulation outcome
Summary answer: Although even natural cycles are associated with aneu- What is known already: Previous studies reported differences in the levels
ploidy, more robust stimulations are associated with aneuploidy. Nevertheless, of IL-2, Il-6, IL-8, IL-10 and VEGF in follicular fluid between young patients with
a higher response is associated with more euploid embryos. low ovarian response and normoresponder women. Cytokine levels have also
What is known already: According to recent literature, the impact of COS been analyzed in women with low ovarian reserve who have Hodgkin lymphoma,
on the rate of embryo aneuploidy in patients without the negative effect of age obtaining a negative correlation between AMH, SIL-2R, IL-6 and IL-8 levels.
acting as a confounding factor is still a subject of lively debate. Moreover, embryo Although there seems to be some evidence about the possible effect of the
aneuploidy is present even in unstimulated cycles, suggesting that, either this is immune system on ovarian function and implantation, the role it plays in the
the real incidence in human beings, or there are factors other than COS related success of ART remains unknown. Our aim was to investigate the effect of
with the IVF procedure which may increase this incidence in comparison with in cytokines in ovarian reserve and response
vivo fertilization. A systematic review of the available evidence is needed to Study design, size, duration: One hundred and twenty-six patients were
provide a succinct analysis of the most relevant findings to date. included in a retrospective study between February 2016 and November 2018.
Study design, size, duration: The study adhered to the PRISMA guidelines. Cytokines IL-2, IL-4, IL- 6, IL-8, IL-10, VEGF, IFNγ, TNFα, IL-1α, IL-1β, MCP-1
A systematic search for studies was performed at MEDLINE, ClinicalTrials.gov, and EGF were measured previously to the ovarian stimulation cycle. Patients
PubMed, Embase, CINAHL and Cochrane Library. . Searches were coordinated with abnormal results of karyotype and/or FMR1 gene or with any other factor
by an expert librarian and a statistician in March’19. Search updates were con- that could alter the ovarian reserve or response were excluded from the study
ducted in July’19. The PICO model was used to select the study population. The Participants/materials, setting, methods: To measure the levels of the
search retrieved 73 citations, and 15 were eligible for analysis (4805 cycles). different cytokines, a sandwich immunoassay with specific antibodies for the
Average quality Newcastle-Ottawa scale (NOS) score was 8. cytokines IL-2, IL-4, IL-6, IL-8, IL-10, VEGF, IFNγ, TNFα, IL -1α, IL-1β, MCP-1
Participants/materials, setting, methods: Women/couples who under- and EGF were used. The statistical analysis was performed with the Software
went COS for an IVF cycle to genetically analyze her/their embryos through Statistical Product and Service Solutions, version 20.0 (SPSS, Chicago, IL, EE.UU.)
PGT were included. There were no restrictions in the amplification method, or Main results and the role of chance: We found that the most related
the platform used to analyze the amplified DNA. Included studies were addi- cytokine with the different ovarian reserve and response markers is interleu-
tionally subclassified according to the protocol of stimulation. Within subgroups, kin-10. It significantly correlates with AMH (p=0.011), antral follicle count
special attention was put to oocytes retrieved, average and total number of (p<0.001), total oocytes (p<0.001) and MII retrieved (p=0.019). Statistically
euploid embryos, and proportion of euploid/aneuploid embryos. When possi- significant relationships had also been found between antral follicle count and
ble, an intention-to-treat (ITT) analysis was performed. IFN- γ (p=0.036), total oocytes with IL-6 and TNF- α (p=0.012 in both cases)
Main results and the role of chance: A clear direct correlation between and MII retrieved with IL-6 and TNF- α (p=0.043 and p=0.014, respectively).
COS and embryonic aneuploidy was not found. Studies have exposed that Through multivariate analysis, we obtained a model able to predict 49.9% of the
absence of COS does not rule out the occurrence of aneuploidy. It is important variability in the number of total oocytes recovered using only 3 variables: age,
to consider that when only analyzing patients that reach biopsy-stage or had antral follicle count and interleukin 10 levels. Statistical differences were observed
at least one embryo biopsied, we are excluding patients who are at the higher when we compare patients with interleukin 10 below or above the threshold of
risk of producing an aneuploid embryo: older patients or with low-quality 0.5 pg/ml in the mean number of total oocytes (6.52 versus 11.05; p<0.001)
oocytes/embryos. Although there is evidence to believe that fewer oocytes and MII retrieved (4.45 versus 8.05; p<0.001). Moreover, in women with inter-
retrieved render better quality under similar dose, more robust studies have leukin 10 below 0.5 pg/ml, the dose of gonadotropins and the number of total
proved that aneuploidy rate did not increase with ovarian response or gonad- oocytes recovered is not associated (p=0.495). Therefore, a higher dose of
otropin dosage. Particularly, no differences were found when using an agonist gonadotropins is not related to a better ovarian response in these patients
or antagonist protocol. Nevertheless, FSH-only stimulations influenced aneu- Limitations, reasons for caution: The retrospective study design and the
ploidy rates under a long-agonist, but not under an antagonist protocol. Trigger sample size could be a limitation. The study was performed in patients with
medication (hCG vs GnRH-agonist) was not predictive of aneuploidy. Among suspected implantation failure or endometriosis. Despite the strong relationship

of interleukin 10 with ovarian reserve and response, we cannot determine its of grade A fresh embryos (0,06 Vs. 0,2 p0,045), greater rate of fresh embryo
role in the ovary, beyond its immunomodulatory function transfer (38,8%Vs.60,4% p0,030) and lower embryo freezing rate (30,6%Vs.
Wider implications of the findings: Interleukin 10 level is lower in patients 14,6% p0,041).
with low ovarian reserve and response compared to normoresponder patients. Limitations, reasons for caution: In these types of studies the investigator
Moreover, if the interleukin 10 level is below 0.5 pg/ml, tailoring the treatment may incur in some manipulation about the exposure, as each subject acts as their
might not improve the outcome. In conclusion, interleukin 10 could be used as own control. The degree of certainty also decreases since between pre-test and
a biomarker to predict the ovarian response post-test there could have been other circumstances that alter the results. Also
Trial registration number: Not applicable the population size is small.
Wider implications of the findings: dFSH might be useful in
selected patients at risk of high response to stimulation in which we seek to
P-633 Could follitropin delta individualized dose, reduce the
minimize risks but still guaranteeing good results. Lower E2 levels, greater rate
overall ovarian hyperstimulation syndrome risk (OHSS)?
of fresh embryo transfer and lower freezing rate were obtained in high
A. VAZQUEZ SARANDESES1, L. De la Fuente Bitaine1, responders
L. Marqueta Marquez2, M. Abad1, M. Carrera Roig1 Trial registration number: 123
1
Hospital Universitario 12 de Octubre, reproductive medicine, madrid, Spain ;
2
Hopital Universitario 12 de Octubre, Reproductive Medicine, Madrid, Spain P-634 Transrectal ultrasound-guided oocyte retrieval and a
successful IVF treatment outcome in a patient with vaginal
Study question: Does the individualization of the starting dose of follitropin agenesis.
delta reduce the OHSS risk in selected patients at risk of high response to D. Makhadiyeva1, A. Ibragimov1
stimulation 1
Ecomed clinic, Reproductive medicine, Nur-Sultan, Kazakhstan
Summary answer: Individualized dosing with follitropin delta (d-FSH) signifi-
cantly reduced the risk of OHSS, allows more women to undergo fresh embryo Study question: To report a case of a transrectal approach for an oocyte
and those embryos had better quality retrieval procedure in a patient with vaginal agenesis receiving in-vitro fertilization
What is known already: OHSS is an iatrogenic and potentially life-threat- (IVF) treatment.
ening complication of ovarian stimulation. It mostly occurs in patients at risk Summary answer: Transrectal ultrasound(US)-guided oocyte retrieval could
of presenting a high response to stimulation, “high responders”: antral follicle be a valuable approach to achieve IVF pregnancy with a surrogate mother in a
count (AFC), high serum levels of anti-müllerian hormone (AMH) or number patient with complete vaginal agenesis.
of oocytes retrieved greater than 15. Managing each woman’s risk of OHSS What is known already: Despite of inability to child bear in the majority of
while maximizing her potential for a successful outcome constitutes one of patients with vaginal agenesis due to absence of normal reproductive tract,
the major clinical challenges in IVF treatment. Delta follitropin is administered medically assisted reproduction is possible. Ovaries are usually present and
according to an individualized dosing algorithm based on serum AMH levels patient’s oocytes could be harvested and used in IVF. The challenge lies in oocyte
and body weight, that seeks to administrate the most accurate dose to each retrieval procedure with complete absence of vagina. Abdominal US-guided
patient. oocyte retrieval including percutaneous transvesical, transurethral transvesical
Study design, size, duration: January 2017-December 2019, we included and transabdominal-transperitoneal have been previously reported as well as
women aged 18–40 with regular ovulatory cycles and diagnosed with tubal infer- laparoscopy-guided oocyte retrieval in a patient with Müllerian agenesis. As of
tility, unexplained infertility, endometriosis stages I/II or with partners with male 2019 there have not been any published reports of transrectal approach in
factor infertility, that underwent a previous cycle using recombinant follicle-stim- oocyte retrieval in humans.
ulating hormone(rFSH) and met one of the criteria for high responders. We Study design, size, duration: Case report of a patient with vaginal agenesis
performed a study with intrasubject measurements, to evaluate the changes in who undergone IVF treatment for primary infertility at a private reproductive
the response after each controlled ovarian stimulation(COS):1stcycle with rFSH medicine clinic.
Vs. 2ndcycle with d-FSH Participants/materials, setting, methods: A 35-year old patient with a
Participants/materials, setting, methods: We collected data following history of vaginal agenesis and hysterectomy presented with primary infertility.
a pre-post design and we compared the differences between each patient She was considered for transrectal US-guided oocyte retrieval and IVF with a
both cycles. T-student and χ2 test were used to analyze quantitative and standard controlled ovarian stimulation with recombinant follicle stimulating
categorical variables respectively. Baseline characteristics and stimulation hormone and luteinizing hormone covered by gonadotropin-releasing hormone
parameters were measured: age, BMI, duration of infertility, length of stim- (GnRH) antagonist. When follicles were >15mm in mean diameter recombinant
ulation, antral follicle count( AFC), dose of rFSH and dFSH, pre-ovulatory human chorionic gonadotropin (hCG) was administered to retrieve oocytes 36
follicle count(PFC), E2 serum level previous to trigger, follicular output rate(- hours later. A 22-year old woman served as a surrogate mother.
FORT), oocytes retrieved, metaphaseII oocytes, cleaved embryos, and endo- Main results and the role of chance: Following two short protocols of an
metrium thickness. ovarian stimulation and special preoperative bowel preparation with cleansing
Main results and the role of chance: A total of 50 women (100 cycles) enema 5 oocytes (3 in the first cycle and 2 in the second cycle) were recovered
were included. We present the following values as mean± standard deviation: transrectally under general anaestesia and subjected to IVF. There were no imme-
Age 32,9 (SD±), BMI 24,7 (4,2), AFC 20,8 (6,2), duration of infertility 34,7 diate or delayed postoperative complications. Transrectal approach for oocyte
months (19,1). Hormones were measured at cycle day 2-3 before starting each recovery was performed by an experienced urology and reproductive medicine
stimulation: AMH 6,4 (3,8). The mean individualized starting dose of rFSH was specialist who had been operating transrectally urological patients in the past.
149,3IU (45,6) whereas for dFSH was 7,7IU (1,9). Normal fertilization was confirmed in 4 oocytes the next day. There were
We observed significant difference between the length of the stimulation being four (two in each IVF cycle) 6-9 cell Grade I embryos vitrified on day three
shorter the rFSH one (9,4Vs.10,2 p0,031) but no difference was found when postoocyte retrieval procedure. A few months later frozen-thawed embryo
comparing the use of dual trigger: Triptorelin plus Human chorionic gonadotropin transfer of two embryos was performed following endometrial preparation with
(11 Vs.15 p0,360). We found no significant difference according to antagonist oestrogen in a healthy surrogate mother. A singleton intrauterine clinical preg-
administration days (5,4Vs.5,04 days p0,134), endometrial thickness (9,2Vs9,6 nancy was confirmed four weeks later. Hormonal supplementation was contin-
p0,603), PFC (10,1Vs.9,54 p0,512), FORT (53,7%Vs.48,7% p0,616), punctured ued until antenatal booking appointment. The pregnancy was uncomplicated
follicles (15,1Vs.13,7 p0,153), retrieved oocytes (11,4Vs.10,2 p0,246), meta- and a healthy baby boy was delivered at term in August 2019.
phase II oocytes (8,1Vs.7,1 p0,193), total cleaved embryos (1,9Vs.1,8 p0,193). Limitations, reasons for caution: a unique case of a rare condition
No significant difference was observed between groups regarding cumulative Wider implications of the findings: Transrectal approach may be useful in
pregnancy rate (34,4% Vs. 13,1%, p0,08). a select number of patients with vaginal agenesis who could not be eligible for
However, we observed significant differences when comparing the E2 serum previously reported abdominal methods of oocyte retrieval.
levels (2600ng/mlVs.1867ng/ml, p0,031) significantly lower with dFSH, number Trial registration number: not applicable

P-635 Premature ovarian insufficiency patients with viable 1

American Hospital, Women’s Health Center, Istanbul, Turkey ;
embryos derived from autologous oocytes through repeated 2
American Hospital, Asisted Reproduction Unit, Istanbul, Turkey ;
oocyte retrievals could obtain reasonable cumulative pregnancy 3
American Hospital, Assisted Reproduction Unit, Istanbul, Turkey ;
outcomes following frozen-embryo transfer 4
Koc University, Assisted Reproduction Unit, Istanbul, Turkey
X. Zhu1, Y. Fu2
1
Study question: Should follicle aspirations in natural IVF cycles be performed
Shanghai Ninth People’s Hospital, Department of Assisted Reproduction,
by only experienced clinicians?
Shanghai, China ;
2
Summary answer: Junior physicians are as good as their seniors are and even
Shanghai First Maternity and Infant Hospital, Department of Assisted
more tenacious in retrieving oocytes that develop into transferrable embryos
Reproduction, Shanghai, China
in natural IVF cycles.
What is known already: Inexperienced clinicians may fail to retrieve the one
Study question: To estimate cumulative pregnancy outcomes following fro-
and only oocyte available during natural cycle ICVF treatment.
zen-embryo transfer (FET) in premature ovarian insufficiency (POI) patients
Study design, size, duration: A retrospective analysis of 226 oocyte pick-up
undergoing in-vitro fertilization (IVF)/ intracytoplasmic sperm injection (ICSI)
(OPU) procedures performed for natural IVF cycles between 2016-2018. Data
treatments with autologous oocytes.
regarding female age, time spent for oocyte retrieval, physician performing the
Summary answer: Cumulative pregnancy outcomes following FET were rea-
procedure, oocyte maturity, fertilization, embryo transfer and pregnancy were
sonable for POI patients using autologous oocytes who could obtain viable
extracted from the electronic database.
embryos through repeated oocyte retrievals.
Participants/materials, setting, methods: Physicians were categorized
What is known already: Women with POI are often discouraged from using
based on their years of experience as senior (≥10 years, n=6), intermediate
autologous oocytes because of the high cancellation rate during ovarian stimu-
(1-10 years, n=4) and junior (<1 year, n=6). The time spent for oocyte retrieval
lation, the small number of available oocytes, the potential for poor oocyte
was calculated from the entrance and withdrawal of the aspiration needle
quality, and the large economic burden in IVF cycles. Thus, oocyte donation is
through the vaginal fornix. All OPU procedures were performed using a double
recommended to increase the chances of pregnancy in this population, which is
lumen 17G needle and 180 mmHg aspiration pressure, under general anesthesia.
still limited or even prohibited because of cultural, religious, or ethical issues in
Student’s t test and chi-square test were used where appropriate.
some countries. Additionally, some POI patients have a strong desire to be
Main results and the role of chance: Out of 226 OPU procedures, a
genetically linked to their offspring and insist upon pursuing IVF with their own
single oocyte was harvested in 179 (79.2%) and the oocyte was in M-II stage
eggs, despite the slim chances of success.
in 156 (69%). The mean female age was 40.1 ± 4.6 years (27-46). The mean
Study design, size, duration: This was a retrospective case-control study con-
duration of the procedure was 586.6 ± 270.7 seconds (126-1288) and it was
ducted in Department of Assisted Reproduction of the Ninth People’s Hospital
affiliated with Shanghai JiaoTong University School of Medicine between January shorter (537.2 ±249.1 versus 777.7 ± 269.1 seconds, p<.001) when the oocyte
could be retrieved. The rate of successful retrieval and the mean time to
2012 and June 2019. Only patients undergoing IVF/ICSI treatments with a freeze-all
retrieve the oocyte were similar among physicians irrespective of their expe-
strategy were screened, and 103 POI patients were matched with 515 normal
rience however, senior physicians terminated the procedure earlier when no
controls in terms of the same number of viable embryos at the same embryonic age.
oocyte could be obtained. The fertilization rate was 52.6% (82/156). Neither
Participants/materials, setting, methods: The number of viable embryos
the rate of fertilized oocytes nor transferrable embryos were related with the
was defined as the sum of all embryos obtained through repeated oocyte retriev-
experience of physicians. Among 66 women who had embryo transfer, 10
als in POI patients, while it referred to embryos obtained from the first oocyte
(15.1%) were pregnant.
retrieval cycle at our centre for the control group. The embryonic age in the
Limitations, reasons for caution: This is a retrospective analysis from a
control group was considered as the age of women when the first viable embryo
single center and does not intend to confirm any causality.
was obtained. The main outcome was cumulative clinical pregnancy rate (CCPR)
Wider implications of the findings: Junior physicians are on a par with their
following FET per patient.
seniors in retrieving oocytes in natural IVF cycles.
Main results and the role of chance: Patients with POI and normal ovarian
reserve had a comparable CCPR of 62.14% (64/103) and 65.24% (336/515),
respectively (P = 0.547), and no statistical difference was found in the cumulative
live-birth rate (CLBR) between the 2 groups (43.69% vs. 53.01%, P=0.084). Based
on binary logistic regression, the CCPR and CLBR showed no association with the P-637 Herbal medicine in women undergoing in vitro fertilization/
group (POI or normal ovarian reserve), whereas the numbers of embryos per intracytoplasmic sperm injection: A systematic review and meta-
transfer and the sum of all viable embryos per patient were positively associated analysis
with the CCPR and CLBR. The increase in the age of viable embryos was negatively J.Y. Jo1, C.Y. Kwon2, B. Lee3, S.H. Lee4
related to the CLBR (OR, 0.944; 95% CI, 0.904–0.986; P=0.01 for the unadjusted 1
Conmaul Hospital of Korean Medicine, Korean Obstetrics and Gynecology, Seoul,
model, and OR, 0.944; 95% CI, 0.903–0.986; P=0.009 for the adjusted model), Korea- South ;
while it had no relation to the CCPR. The clinical pregnancy rate (CPR) per FET 2
Kyung Hee University, Clinical Korean Medicine- Graduate School, Seoul,
cycle was 38.17% for the study group and 52.1% for the control group, while the Korea- South ;
CPRs per OPU cycle in the 2 groups were 11.25% and 69.9%, respectively; both 3
Korea Institute of Oriental Medicine, Clinical Medicine Division, Daejeon,
were statistically different (P < 0.05). Moreover, POI patients had a lower implan- Korea- South ;
tation rate (27.8% vs. 37.94%) and higher early miscarriage rate per transfer 4
Kyung Hee University, Pediatrics of Korean Medicine, Seoul, Korea- South
(26.76% vs. 15%) than patients in the control group (P<0.05).
Limitations, reasons for caution: Only POI patients from whom viable Study question: Does herbal medicine (HM) improve the live birth rate (LBR)
embryos could be obtained were included, and the sample size was small. In addi- and clinical pregnancy rate (CPR) in infertile women undergoing in vitro fertil-
tion, the primary outcome was calculated by the number of patients in our study ization (IVF)/intracytoplasmic sperm injection (ICSI)?
instead of the number of oocyte retrieval and FET cycles in other published articles. Summary answer: Our analysis finds a benefit of HM in women undergoing
Wider implications of the findings: Our data may provide valuable information IVF/ICSI.
for POI patients who insist on using their own oocytes, and the method of repeated What is known already: Combination of IVF and HM has widely used in
oocyte retrievals and FET may be beneficial for the treatment in POI patients. Asian countries. A previous systematic review suggested that combination of
Trial registration number: Not applicable IVF and CHM used in the included trials improve IVF success, however, high risk
of bias observed with the trials and there were no studies that reported live
birth rate (LBR).
P-636 Is successful oocyte retrieval dependent on physician Study design, size, duration: A systematic review and meta-analysis of
experience in natural IVF cycles? randomized controlled trials (RCTs) (until September 2019) that evaluated the
S. Ertas1, M. Ateskan2, O. Bulduk3, I. Keles4, B. Balaban3, K. Yakin4 effects of HM as an adjunct to IVF was conducted.

Participants/materials, setting, methods: Studies were considered for number of viable embryo and high-quality embryo. The progesterone level on
inclusion if they were RCTs; included infertile women who were undergoing trigger day was negatively associated with the number of viable embryo (B=-
IVF/ICSI; compared the effectiveness of HM administrated immediately before, 0.513, P<0.001) and high-quality embryo (B=-0.492, P<0.001) after adjusting
during and/or after IVF with placebo, no treatment, or other active treatments; female age and the number of mature oocyte.
and reported at least one of the pregnancy outcomes including LBR, ongoing Limitations, reasons for caution: As a retrospective study, our analysis
pregnancy rate, and CPR. depended on previously recorded data. The conclusion is limited to achieve one
Main results and the role of chance: From a total of 43 RCTs involving live birth per ovarian stimulation cycle.
4316 participants, HM was more likely to increase LBR (5 studies; risk ratio (RR) Wider implications of the findings: Although late-follicular phase elevated
1.34, 95% confidence intervals (CI): 1.05 to 1.72, P = 0.02) and CPR (35 studies; serum progesterone does not affect the possibility to achieve one live birth per
RR 1.38, 95% CI: 1.29 to 1.49, P < 0.00001) than not receiving adjunctive ovarian stimulation cycle, it may adversely affect the total live birth per ovarian
treatment. The CPR in HM treatment group was also improved when compared stimulation cycle because it was associated with a decrease in number of viable
with that of the placebo group (5 studies; RR 1.85, 95% CI: 1.42 to 2.42, P < embryo and high-quality embryo.
0.00001). However, compared with active controls, HM did not significantly Trial registration number: N/A
improved CPR (RR 1.19, 95% CI: 0.71 to 2.01, P = 0.51). Sensitivity analyses
restricting to RCTs with high quality did not influence the results: HM improved P-639 Segmentation (elective freeze all) results in higher clinical
LBR (2 studies; RR 1.86, 95% CI 1.24 to 2.80, I2=0%) and CPR (30 studies; RR pregnancy rates in women with high AMH (> 30 pmol/L)
1.42, 95% CI 1.30 to 1.54, I2=0%), compared to no adjunctive treatment. HM F. Roebuck1, P. Ambrose2, L. Munn2, T. Hamilton2, C. Noble1,
also increased CPR (2 studies; RR 1.77, 95% CI 1.19 to 2.63, I2=0%), compared J. Lyon3, M. Gaudoin4, R. Fleming3
to placebo. Reported adverse events such as miscarriage, ectopic pregnancy 1
GCRM Fertility, Embryology, Glasgow, United Kingdom ;
rate and incidence of ovarian hyperstimulation syndrome were not significantly 2
GCRM Fertility, Nursing, Glasgow, United Kingdom ;
different between the HM and control group. 3
GCRM Fertility, Biochemistry, Glasgow, United Kingdom ;
Limitations, reasons for caution: Given poor reporting and methodolog- 4
GCRM Fertility, Medical, Glasgow, United Kingdom
ical weaknesses of the existing studies, large-scale, long-term RCTs with rig-
orous methodological input are needed to clarify the role of HM in this Study question: To determine if segmentation in high responder women
population. resulted in higher clinical pregnancy rates (CPR) in the first frozen embryo trans-
Wider implications of the findings: There is promising evidence from the fer than with fresh transfer.
currently available RCTs to judge the effectiveness and safety of HM on preg- Summary answer: The CPR was higher in the first frozen embryo transfer
nancy or childbirth outcomes in women undergoing IVF. (FET) compared to the fresh transfer in women with high AMH levels (P < 0.001).
Trial registration number: not applicable What is known already: Pregnancy rates appear to be lower in GnRH-
antagonist-controlled cycles, possibly because of greater endometrial sensitivity
P-638 Late-follicular phase elevated serum progesterone is to circulating progesterone (Bosch et al, Fertil Steril, 2015).
associated with embryo quality but not cumulative live birth in Segmentation is probably not beneficial in ovulatory women with normal AMH
IVF/ICSI levels (Vuong et al, NEJM, 2018) but might be beneficial in anovulatory women,
especially as this latter group has an increased risk of ovarian hyperstimulation
C. Minghui1, G. Jun1, L. Lu1, M. Qingyun1, X. Yanwen1, C. Zhou1
syndrome with a fresh embryo transfer (Roque et al, Hum Reprod. Update, 2019).
1
the First Affiliated Hospital of Sun Yat-sen University, Reproductive Medicine Study design, size, duration: Historical data from our centre showed a very
Centre, Guangzhou, China modest CPR with fresh embryo transfer in women with high AMH using GnRH-
antagonist control but substantially higher CPR with their subsequent frozen transfer.
Study question: Is late-follicular phase elevated serum progesterone (LFEP) From January 2019 we prospectively elected to culture all such patients to
associated with cumulative live birth when freeze-all strategy was adopted in blastocyst and vitrified all suitable embryos for a subsequent frozen embryo
patients with LFEP during IVF/ICSI? transfer to determine if the subsequent first frozen transfer was superior to the
Summary answer: Late-follicular phase elevated serum progesterone is not historical fresh transfer outcomes.
associated with cumulative live birth when freeze-all strategy was adopted in Participants/materials, setting, methods: All patients with AMH ≥ 30
patients with LFEP during IVF/ICSI. pmol/L received Menopur (Ferring Pharmaceuticals) using GnRH-antagonist
What is known already: Late-follicular phase elevated serum progesterone control. The primary outcome was CPR in the women having a GnRH-trigger,
during IVF/ICSI detriment endometrium receptivity and impede embryo implan- undergoing segmentation and having their first frozen embryo transfer from
tation. It is unclear whether late-follicular phase elevated serum progesterone January – October, 2019 (N = 81, Group-Segn). This was compared with
still disadvantage cumulative live birth when freeze-all strategy was adopted in the historical outcomes of the same subgroup of women in the preceding
patients with late-follicular phase elevated serum progesterone during IVF/ICSI. 2 years (but with HCG triggering) and a fresh embryo transfer (N = 158,
Study design, size, duration: This is a retrospective cohort study; A total Group-Hist).
of 6076 ovarian stimulation cycles and 5461 women who underwent IVF/ICSI Main results and the role of chance: There was no difference in age (Group-
treatment in our hospital from January 2016 to December 2017 were identified Hist = 34.1 ± 4.5 years, Group-Segn = 34.6 ± 3.7 years, P = 0.39), BMI (25.7 ±
and reviewed. 7.0 vs. 26.5 ± 6.4, P = 0.39) or AMH (48.5 ± 21.0 vs. 46.5 ± 18.8, + P = 0.47).
Participants/materials, setting, methods: All the embryos were freezed Group-Segn stimulated for longer (9.4 ± 2.2 days vs. 8.5 ± 0.7, P < 0.001),
in cycles with progesterone levels on trigger day higher than 1.50 ng/ml. received more FSH (1402 ± 324 vs. 1275 ± 106, P < 0.001) and more eggs were
Cumulative live birth rate was defined as the first live birth per ovarian stimulation retrieved (16.6 ± 8.4 vs. 13.6 ± 6.8, P = 0.003). Group-Segn got more 2PN
cycle including fresh and frozen cycles and evaluated by group. Multivariable embryos (10.1 ± 6.0 vs. 8.5 ± 5.3, P = 0.035) but there was no difference in the
linear regression and binary logistic regression was used to assess the association mean number of embryos transferred (1.1 ± 0.3 vs. 1.2 ± 0.8, P = 0.28). Group-
between progesterone levels on trigger day and embryo quality and cumulative Segn had a higher implantation rate (IR, 48% vs. 26%, P < 0.001) and higher CPR
live birth after adjusting for confounding factors. (heartbeat on scan at 8 weeks’ gestation, 52% vs. 27%, P < 0.001).
Main results and the role of chance: The cumulative live birth rate in cycles This was a prospective study comparing an additional intervention with his-
using GnRH agonist protocol was 52.7%, 58.3%, 59.9%, 49.2% and 50.0% torical data. The groups appeared to be evenly matched morphometrically
(P=0.008); the CLBR in cycles using GnRH antagonist protocol was 38.5%, although, because there was little fear of ovarian hyperstimulation syndrome
51.2%, 51.7%, 62.9% and 37.5% (P<0.001) with progesterone level on trigger when using a GnRH-agonist trigger, Group-Segn were stimulated for longer and
day ≤0.50, 0.51–1.0, 1.01-1.50, 1.51-2.00 and ≥2.01 ng/ml respectively. received more FSH.
Progesterone level on trigger day was not associated with cumulative live birth Limitations, reasons for caution: We cannot as yet determine if the cumu-
(OR 0.946, 95%CI 0.773-1.158, P=0.589) after adjusting female age, thickness lative pregnancy rate/live birth rate from a single fresh treatment cycle will be
of endometrium on the hCG trigger day, ovarian stimulation protocol, the equivalent.

Wider implications of the findings: In our study population, segmentation Study question: To investigate the use of follitropin delta, a human cell line
conferred benefit in terms of IR and ongoing CPR compared with a fresh embryo derived recombinant human Follicle Stimulating Hormone with individualised
transfer. Our study population was older than other widely-cited studies so it is dosing regimen, in routine clinical practice.
imperative that individual clinics examine their internal data to determine if a Summary answer: The use of follitropin delta, in routine care setting with a
segmentation approach might benefit their patients. broad and varied patient population, shows results aligned with the ESTHER-1
Trial registration number: N/A registration trial (NCT01956110).
What is known already: When compared to Chinese-hamster ovary (CHO)
P-640 Ovarian sensitivity index -a novel marker of ovarian derived follitropin alfa, the individualised dosing of follitropin delta resulted in
responsiveness in IVF cycles ESTHER-1 in: similar pregnancy rates; less frequent measures to prevent ovarian
C. JANDIAL1, S. Malik1, V. Bhatia1, V. Prakash1, S. Talwar1 hyperstimulation syndrome (OHSS); more women with a target ovarian response
1
southend Fertility and IVF - NEW DELHI, IVF, NEW DELHI, India (8–14 oocytes); fewer poor responses with anti-Müllerian Hormone
AMH<15pmol/L; fewer excessive responses with AMH≥15pmol/L. For the
Study question: To validate the use of OSI as measure of ovarian response follitropin delta arm, mean total dose was 90±25.3mcg, mean stimulation was
during IVF cycles by correlating it with age, AMH ,AFC , total dose of gonado- 8.9±1.9days, mean number of oocytes was 10.0±5.6; 43.3% with 8-14 oocytes,
tropins and quality of embryos 8.0±4.3 with AMH<15pmol/L (44.1%), 11.6±5.9 with AMH≥15pmol/L
Summary answer: OSI can be used as dynamic marker for ovarian reserve, a guide (55.9%). The clinical pregnancy rate was 34.9%, and the OHSS rate (all, any
for gonadotropin dose in future IVF cycles and predictor of good quality embryos grade) was 3.5%.
What is known already: In Controlled Ovarian Stimulation, different women Study design, size, duration: Interim analysis including 390 patients of a
respond differently to similar doses of gonadotropins . Preliminary studies have prospective multicentre non-interventional study using data collected from rou-
suggested that a threshold level of gonadotropins may exist and no more com- tine clinical practice performed across 38 sites in 10 countries. Women naïve to
petent oocytes are obtained if exceeded. Recent studies, on the contrary, indi- in vitro fertilisation (IVF) or intracytoplasmic sperm injection (ICSI) were included
cate that high ovarian response to gonadotropins is not so detrimental. The key in this study. Women who did not achieve pregnancy could continue with the
point is , not the gonadotropin dose or ovarian response alone, but a combina- same treatment for up to three cycles. The study is still recruiting; 1300 patients
tion of these two , is important .This is the concept addressed in the ovarian are anticipated over a period of 3 years (2018-2020).
sensitivity index. High OSI means, more oocytes are retrieved with fewer gonad- Participants/materials, setting, methods: Women are prescribed folli-
otropin doses and offers best pregnancy outcomes tropin delta with an individualised daily dose using an approved algorithm based
Study design, size, duration: This is a retrospective, single centre study on the patient’s Body weight (BW) and serum AMH levels measured by Elecsys®
which included 256 women over a period of one year. AMH plus (Roche Diagnostics). The decision to treat patients with follitropin
Participants/materials, setting, methods: Retrospective analysis which delta is independent from the decision to enrol the patient into the study. During
included 256 women with age < 42 years , no previous history of ovarian surgery, the observation period, the investigators collected data from the routine practice.
chemoradiation and had no endocrine disorders. Baseline scan was done to Main results and the role of chance: This interim evaluation included 390
determine the AFC. Subjects were treated with either long agonist / antagonist patients with mean (SD) age 33.3±4.7 years, BW 67.3±12.7 kg, and AMH
protocol as per clinical and biochemical markers. OPU was done 36 hours after 20.4±15.2 pmol/L. The total dose determined with the algorithm was 102.6±33.5
trigger injection. Ovarian sensitivity Index was calculated and corelated with age, mcg, mean treatment duration was 9.9±2.3 days. Both Gonadotropin-releasing
AMH, AFC, Total dose of Gonadotropins and top quality embryos hormone (GnRH) antagonist (84.9%) or GnRH agonist (15.1%) protocols were
Main results and the role of chance: Out of 256 women enrolled in the used. Cycle cancellation occurred in 19 patients prior to oocyte retrieval, mainly
study with a mean age of 32 years,175(68.36%) were a case of primary subfer- due to poor ovarian response. Final oocyte maturation occurred in 377 patients:
tility with an average duration of 4 years.208 women received antagonist pro- 82.2% with human chorionic gonadotropin (hCG), 13.8% with GnRH agonist and
tocol .Mean number of oocytes retrieved were 8.Statistical analysis as per 4.0% with dual trigger. Per patient who received triggering of final follicular matu-
Pearson’s Correlation Coefficient showed a negative correlation of OSI with ration, mean number of collected oocytes was 10.5±6.2: 9.7% with <4 oocytes,
age and positive correlation with AMH and AFC (p value-0.0000).Positive cor- 25.3% with 4-7 oocytes, 40.4% with 8-14 oocytes, 15.9% with 15-19 oocytes and
relation (0.8162,p value 0.000) was observed with number of oocytes recovered 8.6% with ≥20 oocytes. Mean number of retrieved oocytes was 7.7±4.1 with
and top quality embryos (0.4 and 0.3,p values 0.0000 and 0.0000 respectively. AMH <15 pmol/L (43.3%), 12.7±6.7 with AMH ≥15 pmol/L (56.6%). The major-
Further analysis of data revealed that as OSI increases the mean number of top ity of patients had embryo transfer with fresh embryos (84.4%) on day 5 (54.7%).
quality embryos increases Per started cycle, 32.3% had a positive hCG test and 27.6% had a clinical pregnancy.
Limitations, reasons for caution: A retrospective analysis, single centre, In total, 4.1% of OHSS cases (all, any grade) were reported.
smaller group and includes patients with different protocols Limitations, reasons for caution: This interim evaluation including 390
Wider implications of the findings: In failed IVF cycles, management of patients, who started and finished (with an ovum pick-up or with cycle cancel-
future cycle can be guided by OSI. Patients with higher values should receive lation) a controlled ovarian stimulation treatment for IVF or ICSI, provides results
same dose of FSH in future cycles while those with low OSI should receive higher from the first treatment cycle. The whole data-set (1300 patients) should confirm
dose of FSH. OSI can be used as a predictor of good quality embryos. these findings and allow further exploration of patients’ outcomes.
Trial registration number: NOT APPLICABLE Wider implications of the findings: Individualised follitropin delta treatment
of a broad patient population, as presented in the real-world clinical setting,
P-641 Prospective multi-centre non-interventional study to reflects a variety in the treatment approaches. It also confirms that this new
evaluate the application of individualized follitropin delta dosing regimen based on AMH levels and BW remains predictable and allows
treatment in routine clinical practice. targeting the desired ovarian response for naïve patients.
C. Blockeel1, P. Wijngaard-Boom2, G. Griesinger3, P. Larsson4, Trial registration number: NCT03393780
Y.L. Yip Sonderegger5, F. Beligotti6, S. Riviere6
1
Universitair Ziekenhuis Brussel, Centre for Reproductive Medicine, Jette- Brussels, P-642 Clinical relevance of serum progesterone level on the day
Belgium ; before frozen embryo transfer when using a combined luteal
2
Erasmus Medical Center, Division of Reproductive Medicine- Department of phase support.
Obstetrics and Gynecology, Rotterdam, The Netherlands ; N. Ramos Seixas1, P. Pirtea1, A. Benammar1, D. De Ziegler1,
3
University Hospital of Schleswig-Holstein, Department of Gynecological R. Frydman1, M. Poulain1, J.M. Ayoubi1
Endocrinology and Reproductive Medicine, Luebeck, Germany ; 1
Hôpital Foch, Service de Gynécologie et Obstetrique, Paris, France
4
Ferring Pharmaceuticals, Biometrics - Global Clinical R&D, Copenhagen,
Denmark ; Study question: To determine serum progesterone levels (SPL) with a com-
5
Ferring Pharmaceuticals, Global Market Access & Pricing, Saint-Prex, Switzerland ; bined administration (subcutaneous and vaginal progesterone) and its impact on
6
Ferring Pharmaceuticals, Global Medical Affairs, Saint-Prex, Switzerland pregnancy outcomes in frozen embryo transfer (FET).

Summary answer: We reported that the association of vaginal and subcuta- Summary answer: The accuracy of a standard immunoassay is maintained in
neous progesterone for luteal phase support (LPS) allows achieving optimal the P4-concentration range seen in IVF-patients. Furthermore, serum P4-levels
progesterone levels before FET for most patients. are stable after 2.5 years of cryo-conservation.
What is known already: ART outcomes following FET depends on proges- What is known already: Several studies both in fresh IVF-cycles and frozen
terone support efficiency. As previously reported, low SPL on the day of embryo embryo transfer cycles have demonstrated the importance of luteal P4-levels on
transfer is associated with lower outcomes in FET. Different SPL thresholds have the chance of live birth. Serum P4-levels are easily obtained using standard immu-
been described as having an impact on pregnancy rates. While Labarta reported noassays. However, consideration is required when a standard assay validated for
decreased implantation rate for SPL under 9.2ng/ml after vaginal administration, measurements in a physiological P4-range in a normal population (i.e. during the
Yovich found a lower implantation rate once SPL was lower than 15.7ng/ml or natural cycle) is used in an “abnormal” population (IVF-patients) with supra-phys-
above 31ng/ml with a pessary vaginal regimen. Alur-Gupta reported lower preg- iological P4-values. Furthermore, in large clinical trials study samples are cryo-con-
nancy rates with intramuscular progesterone LPS when SPL were under 15ng/ served for several years before analysis. Current knowledge about the long-term
ml. No study has described SPL after combined subcutaneous and vaginal pro- stability of P4 in human serum is rather incomplete and, in part, contradictory.
gesterone administration. Study design, size, duration: Validation study of laboratory factors with a
Study design, size, duration: Retrospective study in a public academic ART possible impact on the interpretation of luteal P4-levels following ovarian stim-
center between February 2019 and November 2019 including all single fro- ulation: First, a test of the precision of P4-measurements in supra-physiological
zen-thawed blastocyst transfers for women up to 43 years of age. We included P4-ranges using a standard immunoassay (Siemens Immulite 2000XPi). Secondly,
231 FET cycles for 198 patients for data analysis. an assessment of the possible changes in serum P4-levels following cryo-con-
Participants/materials, setting, methods: Patients included had a normal servation at -80°C during a 2.5-year period. Serum samples from the luteal phase
uterine cavity. They received hormonal replacement therapy (HRT) with a were obtained from 602 IVF patients to assess the P4-range following ovarian
step-up estradiol (E2) oral doses protocol, starting with 4 mg/day, reaching 8mg/ hyperstimulation.
day on day 9. Participants/materials, setting, methods: Serum samples were pooled in
After achieving appropriate endometrium thickness (>7mm), daily subcutaneous three pools to reach a P4-concentration of roughly 125, 300 and 600 nmol/l,
progesterone 25 mg and 800mg vaginal progesterone were introduced. FET was respectively. Following mixture, 10 serum samples from each pool were collected
performed after 5 days of progesterone administration. SPL was controlled and frozen. During ten successive days, one sample from each pool was analyzed
one- or two-days prior FET. Statistical analysis was performed using the two-sam- and the coefficients of variation (CV) were calculated. Another 60 luteal serum
ple Student t-test. samples were analyzed at baseline, after 9 months and after 2.5 years to assess
Main results and the role of chance: We included 231 FET cycles for 198 the long-term stability of P4-concentrations in serum.
patients for data analysis. Main results showed a mean SPL of 34.8ng/ml, on the Main results and the role of chance: The 602 serum samples for
day before FET. The lowest value of SPL was 7.4ng/ml and the highest was P4-measurement from the early or mid-luteal phase ranged from 19 nmol/l to
145.7ng/ml. With this combined protocol for LPS, only 1% of patients had 1224 nmol/l, thus up to x20 times the levels seen during the natural cycle. The
progesterone levels under 9.2ng/ml, when they were 25% in Labarta’s study inter-assay and intra-assay precision for the Siemens Immulite stated by the
with vaginal protocol support. This indicates that the combination of subcuta- supplier based on measurements within the normal range of a menstrual cycle
neous and vaginal progesterone as luteal phase support is efficient for this is <7%. The CV values for the sample pools (125 nmol/l, 300 nmol/l and 600
population. nmol/l) were 7%, 5% and 4%, respectively. In conclusion, the experimental runs
We obtained a clinical pregnancy rate (CPR), defined as the presence of an of test samples showed that the precision of the used immunoassay is maintained
embryo with cardiac activity of 29%. The miscarriage rate (clinical pregnancy in the P4-concentration ranges seen in IVF-patients following ovarian stimulation.
loss before 12 weeks) was 18%. The ongoing pregnancy rate (OPR), (CPR after Thus, luteal P4-levels in IVF-patients can be assessed by standard immunoassay
12 weeks) was 24.7% (OPR/FET). quantification with sufficient accuracy and without the risk of misclassifica-
The mean SPL was significantly higher in the group with an OPR in comparison tion bias.
with those in the miscarriage group (41.1 ±30.9 vs. 23.3 ±14.5, p<0.01; CI 95%). To evaluate the effect of storage on the serum P4-levels, a test run of 60 serum
And the OPR was significantly higher for patients presenting a SPL above 22.6ng/ samples was performed. The test runs showed comparable levels of P4 in study
Ml compared to the ones underneath (19.0% vs. 30.4%, p<0.05; CI 95%), even samples compared to the baseline results. The CV-value for P4-measurements
though there was no statistical difference in terms of: age, E2 levels, endometrial performed at baseline and the first re-run (9 months) was 3.4±2.9% and 2,7 ±2.4%
thickness or embryo morphology between the two groups. for the second re-run (2.5 years). Thus, the changes in P4-concentrations remained
Limitations, reasons for caution: An important limitation of this study is within the expected variation for the immunoassay.
that, as a retrospective and observational study, we couldn’t compare the serum Limitations, reasons for caution: The results are based only on measure-
progesterone levels to a reference population having a different luteal phase ments performed by Siemens Immulite 2000XPI. However, automated immu-
support regimen. noassay analyzers are comparable in performance and precision, and therefore
Wider implications of the findings: These findings support the value of a the results are expected to cover other immunoassay analyzers as well.
combined progesterone regimen for LPS in FET, providing optimal levels of Wider implications of the findings: The precision of a standard immuno-
progesterone. Further studies with a larger population and a control group assay is maintained in supra-physiological P4-levels in IVF-patients and can be
should be performed in order to confirm our hypothesis. used in clinical and study set-ups without the risk of misclassification bias. Short-
Trial registration number: Not applicable term cryo-conservation does not change serum P4-levels. Study samples can
therefore be frozen and stored until analysis without compromising P4 test
P-643 Validation study of the stability of serum progesterone results.
following cryo-conservation and the performance of standard Trial registration number: NCT02129998
immunoassay for quantifying supra-physiological serum
progesterone levels following ovarian hyperstimulation P-644 Outcome of in vitro oocyte maturation (IVM) in patients
L. Thomsen1, U. Kesmodel2, P. Humaidan1 with PCOS: does patient phenotype have an impact?
1
Department of Clinical Medicine- Aarhus University- Denmark., The fertility Clinic- M. De Vos1,2, S. Mackens1, S. Pareyn1, P. Drakopoulos1, T. Deckers3,
Skive Regional Hospital- Denmark., Skive, Denmark ; L. Mostinckx1, I. Segers1, S. Santos-Ribeiro4, M. Camus1,
2
Department of Clinical Medicine- Aalborg University- Denmark., The fertility G. Verheyen1, H. Tournaye1, C. Blockeel1
1
Unit- Aalborg University Hospital- Denmark., Aalborg, Denmark Universitair Ziekenhuis Brussel, Centre for Reproductive Medicine, Brussel,
Belgium ;
Study question: Is standard immunoassays for progesterone (P4) measure- 2
Vrije Universiteit Brussel, Follicle Biology Laboratory FOBI, Brussels, Belgium ;
ments accurate in IVF-patients with supra-physiological P4-levels? Furthermore, 3
AZ Sint-Lucas, Gynaecologie-Verloskunde, Brugge, Belgium ;
are serum P4-levels stable over time when cryo-conservation is used? 4
IVI-RMA Lisboa, Gynecology/Reproductive Medicine, Lisbon, Portugal

Study question: Does the phenotype of patients with polycystic ovary syn- after ART (n=121) or natural conception (n=326), within 24 hours of birth by
drome (PCOS) affect clinical outcomes of assisted reproductive technology a single caregiver who was blind to the mode of conception.
(ART) following in vitro oocyte maturation (IVM)? Participants/materials, setting, methods: Gestational age, birth weight,
Summary answer: Cumulative live birth rates (CLBR) after IVM are signifi- length and head circumference were recorded for all newborns. In female infants,
cantly higher in patients with a hyperandrogenic PCOS phenotype compared to distance from anus to the anterior clitoris (AGDAC) and to the posterior four-
those with a normo-androgenic phenotype. chette (AGDAF) were recorded. In male infants, distance from the center of the
What is known already: Hyperandrogenic PCOS phenotype has been anus to the posterior base of the scrotum (AGDAS) and to anterior base of the
reported to confer a lower CLBR after controlled ovarian stimulation (COS) penis (AGDAP) were measured.
and IVF/ICSI when compared to patients with a PCO-like ovarian morphology Main results and the role of chance: The mean age of mothers was 34.9
or with a normo-androgenic phenotype. Whether there is an influence of the years and the mean gestational age at the time of delivery was 267 days. ART
different PCOS phenotypes on success rates of IVM remains unclear. cohort was older, more likely to be nulliparous and delivered by cesarean section
Study design, size, duration: This is a single-centre, retrospective cohort at an earlier gestational week. AGDAS of male infants was approximately twice
study including 320 unique PCOS patients performing their first IVM cycle as long as AGDAF of female infants (17.6±5.0 vs 9.1±3.6 mm, p<0.001). On
between April 2014 and January 2018 in a tertiary referral hospital. average, female infants conceived by ART had shorter AGDAF (8.8±3.6 mm)
Participants/materials, setting, methods: Baseline patient characteristics than those conceived naturally (9.1±3.6 mm) but the difference was not signif-
and IVM treatment cycle data were collected. PCOS was diagnosed according icant (p >0.05). AGDAC were comparable for both groups (27.7 ± 7.1 vs
to the Rotterdam criteria. The primary outcome was cumulative live birth rate 27.4 ± 6.3 mm, p>0.05). In male infants, no significant difference was seen
(CLBR), defined as the rate of deliveries resulting from the transfer of all fresh between ART and natural conception groups in terms of AGDAS (17.4±4.6 vs
and cryopreserved embryos from the same IVM cycle. The secondary outcome 17.7±5.2 mm) and AGDAP (38.0 ± 6.7 vs 37.5 ± 6.6 mm, p >0.05 for both).
was live birth rate following the first IVM embryo transfer. Gestational age, weight, length and head circumference at birth were the stron-
Main results and the role of chance: Half of the patients (n=160) presented gest correlates of anogenital measurements. When adjusted for these covariates,
with a hyperandrogenic PCOS phenotype. These patients had a higher BMI mode of conception was not associated with differences in any of the anogenital
(27.4±5.4 kg/m2 vs 23.3±4.4 kg/m2, p<0.001) and used more often metformin measurements.
(31.2% vs 11.2%, p<0.001) in comparison with the normo-androgenic ones. Limitations, reasons for caution: A single center study with relatively small
Significantly more cumulus oocyte complexes were retrieved in the hyperan- sample size, it is not possible to rule out confounding by diet or endocrine-dis-
drogenic group (24.9±18.3 vs 19.8±13.5, p= 0.003) and, after IVM, these rupting chemicals or drugs other than progesterone that women might have
patients had more mature oocytes (11.7±9.0 vs 9.1±6.9, p=0.002). Following taken during pregnancy
the first embryo transfer, the positive hCG rate and LBR were comparable in Wider implications of the findings: Preliminary findings are reassuring for
the two patient groups (44.2% vs 36.7%, p=0.21 and 23.2% vs 26.6%, p=0.52, women undergoing ART but they need to be confirmed by future studies with
respectively). In hyperandrogenic patients, early pregnancy loss was more com- larger sample sizes
mon (20.3% vs 10.2%, p= 0.02). The CLBR (when at least one embryo was Trial registration number: not applicable
available for transfer) was not significantly different following univariate analysis
(42.7% vs 41.4%, p= 0.82). However, when using a multivariable logistic regres- P-646 Does quantity equal quality? – a morphokinetic assessment
sion model to account for confounding factors, the PCOS phenotype appeared of embryos obtained from young women with decrease ovarian
to be significantly correlated with CLBR, with more favorable results in the response to stimulation.
hyperandrogenic group (OR 2.24, CI 1.06-4.77, p= 0.03). N. Schachter - Safrai1, G. Karavani1, Y. Kantor2, I. Har- Vardi3,
Limitations, reasons for caution: These data should be interpreted with A. Buxboim2, Z. Shoham4, Y. Or4, A. Ben-Meir1
caution in the light of the retrospective nature of the study and the possibility 1
Hadassah Hebrew University Hospital, Obstetrics and Gynecology, Jerusalem,
of unmeasured confounding.
Israel ;
Wider implications of the findings: Infertile hyperandrogenic PCOS 2
Institute of Life Sciences- The Hebrew University of Jerusalem, Alexander Grass
patients undergoing IVM achieve a higher CLBR than their normo-androgenic
Center for Bioengineering- School of Computer Science and Engineering, Jerusalem,
counterparts. This is in contrast with previously reported outcomes following
Israel ;
COS and ART in which hyperandrogenic PCOS patients performed significantly 3
Soroka University Medical Center, The IVF Unit - Department of Obstetrics and
worse. Our data suggest that proper patient selection is of utmost importance
Gynecology, Beer-Sheva, Israel ;
in an IVM program. 4
Kaplan Hospital, Department of Obstetrics and Gynecology, Rehovot, Israel
Study question: Is there an association between oocyte quantity and quality
P-645 Anogenital distance in newborn infants conceived by in young women exhibiting decreased ovarian response?
assisted reproduction and natural conception Summary answer: The quantitative decrease in ovarian response is associated
S. Ercin1, S. Ertas2, O. Dundar3, O. Oktem4, K. Yakin2 with reduced oocyte quality, reflected by a slower developmental rate and lower
1
American Hospital, Neonatology, istanbul, Turkey ; implantation and pregnancy rates.
2
American Hospital, Women’s Heath Center, Istanbul, Turkey ; What is known already: It is well known there is a physiologic decline in both
3
American Hospital, Women’s Health Center, Istanbul, Turkey ; oocyte quantity and quality attributed to aging. However, it is unclear if the two
4
Koc University, Women’s Heath Center, Istanbul, Turkey are related when a premature decline in oocyte quantity is observed. While
most of the literature on this subject includes a heterogenous population and
Study question: Does anogenital distance (AGD) differ in newborn infants demonstrates conflicting results regarding embryo quality and pregnancy rate,
conceived through assisted reproduction technologies (ART) compared with a recent study included only women younger than 38 years with low ovarian
those conceived naturally response and demonstrated no difference in embryonic development and live
Summary answer: AGD measurements in infants conceived by ART are not birth rate .There is only scarce data regarding oocyte quality in poor responders
different from those of infants conceived naturally using a time lapse microscopy (TLM).
What is known already: AGD is an anthropometric measurement sensitive Study design, size, duration: The study included a retrospective assessment
to changes in the in-utero hormonal environment during the early life of fetus. of morphokinetic parameters performed by TLM from five medical centers
Alterations in AGD are associated with anomalies in neonates and reproductive between January 2013 and December 2017.The developmental process and
dysfunction in adults kinetics of 519 embryos obtained from the study group, referred as the “
Study design, size, duration: A prospective descriptive cohort study was decreased ovarian response” (DOR) group was compared to 3633 embryos of
performed on randomly selected singleton neonates (n = 447) born at a single the control group ,known as the “normal ovarian response” (NOR) group.
center between September 2017 and April 2019. Anthropometric and anogenital Participants/materials, setting, methods: The study included patients
measurements were performed in 247 male and 200 female newborns born younger than 38 years who underwent controlled ovarian stimulation (COS)

with consecutive aspiration of either 5 oocytes or less - the DOR group, or 6 moderate correlation with clinical pregnancy (r = −0.618, p<0.001). FF-AMH
oocytes or more- the NOR group. A comparison between the morphokinetic pregnancy threshold was >1.75 ng/ml. Receiver operating characteristic (ROC)
parameters, implantation and pregnancy rates of the two groups was made with analysis showed a FF-AMH sensitivity of 73.1% in predicting CPR and a specificity
additional subgroup analysis according to the implantation status. Logistic regres- of 85.3 %. The area under curve (ROCAUC) was 0.715 (p < 0.0001).
sion was conducted to assess the association between morphokinetic parameters Limitations, reasons for caution: The sample size was relatively small and
and implantation rate does not reflect a wide variety of clinically seen cases such as women with ovarian
Main results and the role of chance: Implantation and clinical pregnancy cyst (> 3 cm in diameter), polycystic ovarian syndrome, endometriosis, or those
rates were significantly lower in the DOR group compared to the NOR group with a history of previous ovarian surgery or endocrine disorders.
(31.6% vs. 44.5% and 37.7% vs. 51.5%, respectively; p<0.05). Embryos from the Wider implications of the findings: This study demonstrated that FF AMH
DOR group reached the morphokinetic milestones later than embryos obtained levels were significantly lower in fertilized oocytes than in non-fertilized oocytes.
from NOR patients. Implanted embryos in the DOR group developed faster FF-AMH is an adequate predictor of clinical pregnancy after ICSI; However
than embryos which failed to implant, however manifested a protracted course further studies on a larger group of patients are needed to correlate its relation
compared with implanted embryos from the NOR group. In a multivariate anal- with ongoing pregnancy.
ysis - normal ovarian response, age and tSB were associated with implantation. Trial registration number: not applicable
Limitations, reasons for caution: The two patients’ groups were not
homogenous in their basic characteristics. A higher rate of blastocyst culture P-648 Does thyroid autoimmunity and more specifically
and blastocysts transfer was demonstrated in the NOR patients. Important anti-thyroglobulin antibodies affect IVF/ICSI outcome?
information regarding the maximal dose of gonadotropins obtained, previous V. Uvin1, M. De Brucker1, P. Drakopoulos1, K. Poppe2,
IVF response and ovarian reserve testing was lacking. H. Tournaye1, D. Unuane3
Wider implications of the findings: Decreased oocyte quality manifested 1
UZ Brussel, Centre of reproductive medicine, Brussels, Belgium ;
by a multi-staged delayed preimplantation development in the DOR group may 2
Centre Hospitalier Universitaire Saint-Pierre, Endocrinology, Brussels, Belgium ;
highlight another pathophysiology for decreased oocyte quality not related to 3
UZ Brussel, Endocrinology, Brussels, Belgium
age. Decreased ovarian response to COS and morphokinetic parameters may
assist in predicting pregnancy.
Study question: Does the thyroid autoimmune (TAI) status and especially
anti-thyroglobulin antibodies affect IVF/ICSI outcomes of euthyroid patients?
Summary answer: Thyroid autoimmune status and more specifically anti-thy-
P-647 Follicular fluid anti-mullerian hormone: a predictive marker roglobulin antibodies do not affect the reproductive outcomes of euthyroid
of fertilization, implantation and clinical pregnancy in patients patients undergoing IVF/ICSI.
with unexplained infertility What is known already: The impact of TAI on fertility outcomes of euthyroid
A. Abdelmeged1, O. Ahmed1, A. Kotb1, E. Elgindy1, H. Baha1 women undergoing IVF/ICSI has been controversial. In literature, in addition,
1
Faculty of Medicine, Gynacology and Obstetrics, El Menia, Egypt TAI is predominately defined by elevated levels of thyreoperoxydase antibodies
(TPO-Ab). The impact of anti-thyroglobulin antibodies (TG-Ab) on fertility
Study question: Would follicular fluid anti-mullerian hormone (FF-AMH) outcome after IVF/ICSI remains unclear.
measured values predict fertilization, implantation and clinical pregnancy in Study design, size, duration: This was a retrospective study including all
patients with unexplained infertility undergoing ICSI? euthyroid patients who underwent their first IVF/ ICSI cycle in a 6 month period
Summary answer: FF-AMH level could be associated with fertilization rate at the center of reproductive medicine of the University Hospital Brussels. All
(FR), implantation rate (IR), and clinical pregnancy rate (CPR) in patients with patients were screened for both TPO-Ab and TG-Ab.
unexplained infertility undergoing ICSI. Participants/materials, setting, methods: Patients were categorized to
What is known already: In humans, AMH is considered a reliable marker for four groups based on TAI status: TAI negative, only TG-Ab positive, only
ovarian reserve, as well as for ovarian response to gonadotropin stimulation. TPO-Ab positive and both TG- and TPO-Ab positive. The main outcome was
AMH probably has a role in the regulation of follicle selection and maturation. fresh live birth rates (LBR). Secondary endpoints were positive human chorionic
FF-AMH inhibits the initiation of human primordial follicle growth and prevents gonadotropin (hCG), early pregnancy loss and cumulative LBR. Cumulative LBR
multiple selections of a dominant follicle; by reducing their sensitivity to follicle was defined as the delivery of at least one live-born infant (>24 weeks of ges-
stimulating hormone (FSH). Previous studies suggested that FF-AMH might be tation) in the fresh or in the subsequent frozen-thawed cycles.
a better predictor of ovarian responses to controlled ovarian hyperstimulation Main results and the role of chance: In total, 464 patients were included
than traditional parameters such as age, FSH, estradiol and inhibin-B. in the analysis: 402 (86.6%) in the TAI negative group, 17 (3.7%) in the TG-Ab
Study design, size, duration: This is a single-arm uncontrolled clinical trial that group, 15 (3.2%) in the TPO-Ab group and 30 (6.5%) in the TPO/TG-Ab group.
was conducted between June 2018 and 2019. The study protocol was approved Baseline characteristics such as age, body mass index, anti-Müllerian hormone
by the Institutional Ethics committee. Sixty women with the diagnosis of unex- (AMH) and antral follicle count (AFC) were similar between the four groups.
plained infertility who received controlled ovarian stimulation and ICSI treatment Only thyroid-stimulating hormone (TSH) differed significantly [1.6 (0.7) vs. 2.4
were enrolled. The inclusion criteria for women were age ≤ 38 years, normal (0.9) vs. 1.4 (0.8) vs. 1.8 (0.9), p value=0.005)]. Fresh LBR did not differ signifi-
ovulatory cycles and a body mass index (BMI) ranging from 18 to 28 kg/m2. cantly between the four groups [131/402 (32.6%) vs. 3/17 (17.7%) vs. 6/15
Participants/materials, setting, methods: This study was conducted at (40%) vs. 8/30 (26.7%), p value=0.5)]. Similarly, positive hCG and early preg-
Minia Infertility center, Minia University. Follicular fluids (FF) were aspirated under nancy loss rates were comparable between the four groups, while cumulative
transvaginal ultrasound guidance. The FF from the single dominant follicle of LBR did not show a statistically significant difference either [164/402 (40.8%)
each patient (which is aspirated first) was separated. FF was stored at - 80 °C vs. 6/17(35.3%) vs. 6/15 (40%) vs. 11/30 (36.7%), p value =0.94]. Multivariate
until assayed. AMH levels were measured by using an ultrasensitive enzyme- regression analysis indicated that after adjustment for relevant confounders the
linked immunosorbent assay. Based on the median of cycles measured values; TAI status was not significantly associated with either fresh or cumulative LBR
women were categorized as low and high FF-AMH groups. (p value = 0.24).
Main results and the role of chance: The primary outcome of this study was Limitations, reasons for caution: This is a large observational study based
fertilization rate, implantation rate, blastocyst development, embryo quality, chem- on retrospective data collection. Despite our robust methodological approach,
ical pregnancy and clinical pregnancy. Low FF-AMH group, compared to high the presence of bias related to the retrospective design cannot be excluded.
FF-AMH group, showed a significant improvement with regards to percentage of Furthermore, the rarity of TG-Ab may preclude firm conclusions.
top-quality oocytes (67.1±24.3% vs. 49.6 ± 30.3%, p = 0.014), FR (83.9±20.9% Wider implications of the findings: Our study adds further to the growing
vs. 72.4 ± 21.4%, p = 0.021), IR (57.7% vs. 16.7%, p = 0.001) and CPR (57.57% evidence that TAI (and especially TG-Ab) do not seem to affect the reproductive
vs. 16.67%, p < 0.0001). None of the remaining variables were significantly different outcomes of euthyroid patients undergoing IVF/ICSI. These results should be
between the two studied groups (p > 0.05). Further, FF-AMH value had a significant confirmed in future prospective studies.
low correlation with follicular fluid estradiol (FF E2) (r = −0.409, p <0.001) and Trial registration number: not applicable

P-649 FF-MAS oxysterol related with hedgehog signaling pathway E. Baldi1, S. Marchiani2, L. Tamburrino1, F. Benini3, M. Pallecchi4,
on folliculogenesis C. Bignozzi3, A. Conforti5, C. Alviggi5, L. Vignozzi4, G. Danza4,
S. ZIRH1, E. Bahador Zirh1, L. Karakoc Sokmensuer1, G. Bozdag2, S. Pellegrini3
S. Muftuoglu1 1
University of Florence, Department of Experimental and Clinical Medicine,
1
Hacettepe University- School of Medicine, Histology and Embryology, Ankara, Florence, Italy ;
2
Turkey ; Azienda Universitario Ospedaliera Careggi, Department Materno Infantile,
2
Hacettepe University- School of Medicine, Obstetrics and Gynecology, Ankara, Turkey Florence, Italy ;
3
Centro Procreazione Assistita Demetra, none, Florence, Italy ;
4
Study question: Does FF-MAS affect folliculogenesis via hedgehog cell signaling University of Florence, Department of Experimental Clinical and Biomedical
pathway? Sciences, Florence, Italy ;
5
Summary answer: The main effect of FF-MAS on folliculogenesis manifested University of Naples- Federico II, Department of Neuroscienze- Reproductive
by granulosa cell(GC) proliferation, is associated with the hedgehog pathway Science and Odontostomatology, Naples, Italy
(HH) proteins SMO and GLI1.
What is known already: HH is one of the main cell signaling mechanisms Study question: Are steroid hormone levels in single follicular fluid related to
effective on cell proliferation. It exerts its biological effects through signaling ovarian stimulation (rFSH or rFSH+rLH) and to Assisted Reproduction Technique
cascade that activated by smoothened(SMO) receptor consequently regulated outcomes of the corresponding oocyte?
glioma-associated oncogene(GLI) transcription factors. Follicular fluid–meiosis Summary answer: FF steroid levels were found different between the two
activating sterol(FF-MAS), which is a steroid derivative and present in follicular protocols of ovarian stimulation. FF Progesterone levels predicts the attainment
fluid, is involved in folliculogenesis. GC is critically important cell involved in of fertilization in the FSH group.
follicular development during the ovarian cycle. Polycystic ovary syndrome(P- What is known already: Conflicting results have been reported in studies
COS) defined as follicular development failure, lag of granulosa cell proliferation, concerning the relationship between steroid levels in FF and oocyte maturity or
and cystic follicles as a result, is one of the common endocrine disorders of today. Assisted Reproduction Technique (ART) outcomes. Controversy may depend
Study design, size, duration: Primary culture of granulosa cells from both PCOS on the fact that most of these studies did not consider the destiny of individual
(n=10) and healthy (male factor infertility)(n=10) woman undergoing IVF, were oocytes and utilize immunoassay to measure steroid levels, which is affected by
collected since 2018. The human immortalised granulosa cell line (hGL5, abm, cross-reactivity and shows poor specificity and sensitivity.
Canada) were also used beside primary cell culture. Each GCs divide into four Study design, size, duration: We enrolled in a prospective cohort study
groups individually: FF-MAS, FF-MAS+cyclopamine, only cyclopamine and vehicle 41 couples undergoing ICSI cycles from November 2016 to August 2018. In
used for FF-MAS and cyclopamine. Cyclopamine was used for inhibiton of HH. 13 cases follicle stimulation was performed with recombinant r–FSH alone and
Participants/materials, setting, methods: Granulosa cells of women in 28 women, with previous poor ovarian responsiveness, with r-FSH plus
undergoing IVF, have isolated from follicular fluid at oocyte pick up in Hacettepe r-LH. 316 oocytes and the corresponding collected follicular fluids (FF) were
University Infertility Center. GCs (PCOS and control (male factor[MF] infertility)) included in the study. The destiny of each oocyte was followed during the
were cultivated at least 3 passages after the isolation. ED50 dose of FF-MAS entire cycle.
was maintained by WST assay. Hedgehog pathway mollecules(SMO, GLI1) were Participants/materials, setting, methods: Levels of six steroid hormones
screened on granulosa cells from each group by using immunocytochemistry (progesterone (P), 17-OH-Progesterone (17-OH-P), Androstenedione (A),
(immunofluorescence and immunoperoxidase) and cyto-ELISA. Mean fluores- Testosterone (T), Estradiol (E1) and Estrone (E2)) were evaluated in the single
cence intensities evaluated in all micrographs by ImageJ 1.52K. FF by High Performance Liquid Chromatography coupled with tandem Mass
Main results and the role of chance: The ED50 dose of FF-MAS was Spectrometry (HPLC/MS-MS). Such levels were related to ovarian stimulation
determined as 10µM FF-MAS for 48h according to WST assays. Results were protocol and oocyte maturity, fertilization and quality of blastocysts. Clinical
evaluated according to measurement of cell proliferation and viability in response pregnancy, due to the limited number of achieved pregnancies of our cohort,
to different doses of FF-MAS. Tests were performed in GCs of PCOS, MF and was evaluated as explorative outcome.
HGL5. Experiments carried out with 10µM FF-MAS for 48h according to max- Main results and the role of chance: Our study demonstrates that FF steroid
imum non-toxic dose and duration. All the GCs were assesed in terms of GLI1 levels vary among different follicles collected from the same woman and that the
and SMO by both IF, ICC. Hedgehog pathway was slightly inhibited with cyclo- two ovarian stimulation protocols lead to differences in the steroid composition
pamine binding to signal transducer receptor SMO via canonical pathway. Indirect of FF, as 17-OH-P, A, E2 and E1 were significantly higher in FSH+LH group. In the
IF is widely considered for SMO-GLI1 proteins in all groups of granulosa cells latter group, P, 17-OH-P and E2 were more expressed in FF of follicles which yield
screening with/without FF-MAS and cyclopamine. a mature oocyte than in immature ones (median values (ng/ml): P, 7352.7
The expression of GLI1 and SMO showed different flourescence intensities [IQR:4252.2-11378.2] vs. 4270.2 [IQR: 2466.4-8064.9]; 17-OH-P, 991.2 [IQR:
between groups. GLI1 and SMO were mostly expressed in all FF-MAS groups. 596.2-1620.4] vs. 571.4 [IQR: 352.6-1072.3]; E2, 396.0 [IQR: 238.5-551.4] vs.
The mean fluorescence intensities were compared in all groups by ImageJ analysis. 245.9 [IQR: 165.3-385.6], p<0.01) and 17-OH-P was higher in FF of follicles which
A significant difference were obtained between quantitative reading of intensities yield fertilized oocytes respect to non-fertilized oocytes (median values (ng/ml):
in FF-MAS and FF-MAS+cyclopamine groups. Results of immunoperoxidase cyto- 1076.6 [IQR: 663.7-1694.6] vs. 811.9 [IQR:362.3-1385.7], p<0.05). Moreover,
ELISA for GLI1 and SMO expressions of FF-MAS groups confirmed the IF analysis. P FF levels of the oocytes which achieved clinical pregnancy were significantly
According to our findings, the highest expression was observed in FF-MAS higher respect to non-pregnant (median values (ng/ml): 11136.9 [IQR: 9294.1-
groups especially in PCOS patients than MF group. Cyclopamine significantly 15275.5] vs. 8305.6 [IQR: 4688.3-11238.9], p<0.05). In the group stimulated only
suppressed the expression of proteins. with FSH, we observed no differences in steroid levels for none of the analysed
Limitations, reasons for caution: Since human cells were used in study and ART outcomes, except for P levels, which were found to be predicted of successful
the cells of each patient do not exhibit the same characteristics, the lowest fertilization with an accuracy of 70.3±6.0%, P<0.005.
number of patient samples identified in the statistical power analysis were Limitations, reasons for caution: The low absolute number of embryos in
included in the study. the three quality categories and the paucity of women achieving pregnancy in
Wider implications of the findings: First time, the effect of FF-MAS via HH the two groups may have limited the statistical power regarding the role of FF
was evaluated on human GCs, which has been used in animal ART before. We have steroids in predicting embryo quality and pregnancy.
introduced a new approach to the literature on the molecular mechanisms of FF-MAS Wider implications of the findings: LH supplementation modifies the ovar-
and to be considered as a new approach in human IVF treatments of PCOS. ian steroidogenic activity better resembling the physiological steroid pattern
Trial registration number: not applicable likely improving ovarian response. Our study does not support a direct associ-
ation between FF steroid levels and ART outcomes, with the exception of
P-650 Steroid levels in single follicular fluid: relation to ovarian Progesterone, which predicts with fair sensitivity and specificity the attainment
stimulation protocol and destiny of the corresponding oocyte in of fertilization.
assisted reproduction Trial registration number: not applicable

P-651 AMP-activated protein kinase (AMPK) controls dormancy of 1

Aarhus University, Department of Biomedicine, Aarhus C, Denmark ;
the primordial follicle pool in murine ovarian tissue 2
Aarhus University, Department of Biomedicine and Department of Clinical
J. Nielsen1, M. Amoushahi1, E.H. Ernst2, K. Lykke-Hartmann1,3,4 Medicine, Aarhus C, Denmark ;
3
1 Aarhus University Hospital, Department of Clinical Genetics, Aarhus, Denmark
2
Herning Region Hospital, Department of Obstetrics and Gynaecology, Herning,
Study question: We sought to improve and more precisely determine the
Denmark ;
3
follicle distribution of primordial, primary and secondary follicles in ovaries from
Aarhus University Hospital, Department of Clinical Genetics, Aarhus C, Denmark ;
4
juvenile mice by automated counting.
Aarhus University, Department of Clinical Medicine, Aarhus C, Denmark
Summary answer: Fluorescent labeling of oocytes and quantification of fol-
licles by automatic analysis software generates comparable data with conven-
Study question: Can inhibition of AMPK stimulate growth of primordial fol-
tional methods and ensures objective results.
licles in ovarian tissue?
What is known already: Knowledge of the number of ovarian follicles is of
Summary answer: AMPK signaling is an essential signaling pathway governing
interest to investigators studying mammalian reproductive biology. One of the most
dormancy of primordial follicles and inhibition of AMPK enhances the number
widely used approaches for estimating the number of follicles in different develop-
of activated follicles.
mental stages is based on histological sampling of the total ovarian mass followed
What is known already: Molecular signaling pathways including PI3K/AKT,
by manual classification and counting using a microscope. This procedure is time-con-
AKT/mTOR, and HIPPO contribute to regulate the primordial-to-primary follicle
suming and highly operator dependent. Improving the accuracy of the follicle clas-
transition in the mammalian follicle development. It is, however, widely acknowl-
sification and quantification will reduce time and produce more reliable results.
edged that we are only beginning to elucidate the molecular mechanisms main-
Study design, size, duration: In this pilot study, whole ovaries isolated from
taining dormancy and activation of primordial follicles. Recently, our team
juvenile mice (7-8-days old) were used for quantification of the number of fol-
performed transcriptomic profiles of human primordial and primary follicles
licles in three different developmental stages; primordial, primary and secondary
providing molecular clues towards which alterations in gene expression that
follicles. Ovaries were kept in organ culture for four days in basic medium in a
dictate dormancy and activation of primordial and primary follicles. One prom-
well-insert culture system.
ising gene candidate identified was AMPK; a kinase responsible for restoring
Participants/materials, setting, methods: Ovaries were fixed, processed,
cellular energy homeostasis.
and embedded in paraffin. Samples were cut in 5-μm sections and stained with
Study design, size, duration: Whole murine ovaries were excised from
hematoxylin and eosin (H&E) or immunofluorescence (IF) staining. The number
female juvenile mice (C57BLxCBA F1) and randomly divided into two groups
of follicles at three developmental stages was either counted manually using an
of equal size and grown in a primary three-dimensional culture system in culture
inverted research microscope or automatically using high-content image analysis
medium supplemented with an inhibitor of AMPK (IHB) or without (NT). After
software (MetaXpress). The follicle distribution of every 5th section of each
five to seven days of culture, ovaries were collected.
ovary was counted. Germ-cell marker MVH was used to visualize oocytes for IF.
Participants/materials, setting, methods: In vitro-cultured ovaries were
Main results and the role of chance: IF staining results showed a strong
collected for histological analysis to address the balance between follicular acti-
MVH signal in oocyte cytoplasm of follicles in all three developmental stages
vation and quiescence after drug-exposure. Additionally, we addressed important
present in the cultured juvenile mouse ovaries. The average distribution of fol-
parameters that can evaluate the follicle quality after drug-exposure. Towards
licles classified and counted manually was 83% (SD 3%), 13% (SD 2%) and 4%
this, we performed assays e.g. TUNEL, BAX-to-BCL-2 ratio, cytochrome c oxi-
(SD 1%) among primordial, primary, secondary follicles, respectively. The pre-
dase activation, and ROS generation. Furthermore, follicles from the drug-ex-
liminary customized analysis module in MetaXpress was programmed to recog-
posed and NT ovaries were mechanically isolated and cultured in an alginate
nize all MVH-positive follicles with a certain intensity and with a visible nucleus.
hydrogel, culminating in meiotic competent oocytes.
Only about 5% of the visible follicles were not recognized by the analysis module.
Main results and the role of chance: We found that in vitro culture with an
Since stage-specific IF markers were still not identified, the preliminary results
AMPK inhibitor for five days significantly increased the number of activated follicles
of the analysis module were based on the number of follicles rather than clas-
(NT: 18.97%±1.017 vs. IHB: 39.73±1.28%, p<0.0001; n=17/15). As the level of
sification of the follicles. By setting a diameter range for the different follicular
activated AMPK were significantly decreased (p<0.05, n=18), and the activation
of PI3K-AKT signaling was unaffected (p<0.05; n=18) by the drug-exposure, our stages, the analysis software discovered 10% (SD 3%) more follicles in the pri-
mordial stage compared to manual counting. This method is a way of eliminating
results indicated that inhibition of AMPK can trigger follicular growth synergistically
workload and subjectivity in microscopic image analysis, however, human assess-
with PI3K-AKT signaling. Furthermore, we assessed the inhibitor’s impact on
ment of the health of the ovary is always important.
oocyte quality parameters. We showed that inhibitor-exposure increased the
Limitations, reasons for caution: The main limitations of this study are the
survival rate by conducting a TUNEL analysis (number of apoptotic cells NT:7.48%
lack of stage-specific markers for the different pre-antral developmental stages.
vs. IHB:3.30, p<0.05, n=5-6) and the BAX-to-BCL-2 ratio were also significantly
Additionally, this method will not provide the total number of follicles of the
decreased after drug-exposure (p<0.01, n=12). Moreover, we showed that the
whole ovary, but the method can be used as an estimation of the effect of an
activity of cytochrome c oxidase (p>0.05, n=12) and ROS generation (p>0.05,
experimental intervention.
n=6) were not significantly affected by the drug-exposure. In line with these obser-
Wider implications of the findings: Automated image analysis provides an
vations, we elucidated that follicles isolated from drug-exposed ovaries had the
important advantage by significantly reducing manual counting variability. IF meth-
ability to release meiotic competent oocytes in response to an ovulating dose of
ods also efficiently enhance the visibility of primordial follicles that are difficult
human chorionic gonadotropin, with the same survival rate as the control-exposed
to see and ensures accurate recording of the number of follicles. Focus on finding
ovaries (NT: 76.47% vs. IHB: 73.33%; n=34/30).
stage-specific markers would further improve the classification of follicles.
Limitations, reasons for caution: The study is for now limited to murine
tissue and further studies are needed to assess if the murine findings are trans-
latable into the human reproductive system. Moreover, the inhibitor has previ-
ously been reported to promote promiscuous AMPK-independent affairs, P-653 Human oocytes and granulosa cells from primordial and
therefore the results must be interpreted with potential off-targets in mind. primary follicles revealed expression patterns of genes encoding
Wider implications of the findings: This study furthers our understand on Toll-like receptors (TLRs)
cellular signaling which dictate the balance between follicular activation and dor- G. Saritas1, E.H. Ernst2, M. Amoushahi1, K. Lykke-Hartmann1,3,4,
mancy, and understanding the details of ovarian biomechanics may have sub- T. Wenzel Kragstrup1, A.S. Sørensen1
stantial clinical implications. 1
Herning Region Hospital, Department of Obstetrics and Gynaecology, Herning,
Denmark ;
P-652 Automated quantification of the distribution of pre-antral 3
Aarhus University Hospital, Department of Clinical Genetics, Aarhus N,
ovarian follicles to facilitate accurate and unbiased counting Denmark ;
E. Bruun1, M. Amoushahi1, K. Lykke-Hartmann2,3 4
Aarhus University, Department of Clinical Medicine, Aarhus C, Denmark

Study question: Does the early hormone-independent, primordial-to-primary Study question: What is the relationship between selenium level and prema-
follicle development involve inflammatory-like responses mediated through TLRs? ture ovarian insufficiency?
Summary answer: The hormone-independent transition from primordial to Summary answer: Patients with abnormal low level of selenium have low
primary follicles in human folliculogenesis revealed oocyte- and granulosa ovarian reserve.
cell-specific expression patterns and biological functions of TLR3 and TLR4. What is known already: Selenium is a co-factor of the antioxidant enzymes
What is known already: Later stages of follicle development and ovulation, responsible for prevention of ROS production. Glutathione peroxidases (GPxs)
encompassing the maturation of secondary to antral/preovulatory follicles, are the main selenoprotein enzymes associated with reproductive and pregnancy
involves inflammatory-like responses similar to the innate immune-related cell processes.The importance of serum selenium assessment in infertile women
surveillance system including: inflammation, rupture (or ovulatory) event and with POI lies in the fact that antioxidants, including selenoprotein enzymes such
reparation. Cumulus oocyte complex, which appears in antral follicles, secretes as GPxs, are highly sensitive to changes in selenium status and decrease signifi-
luteinizing hormone and amphiregulin inducing expression of immune-related cantly with selenium deficiency. The present study was therefore conducted to
receptors as TLRs. Murine granulosa and cumulus cells exposed to these hor- assess serum selenium level and GPx activity (main objectives) and nutrition
mones revealed gene expression levels of various immune components, e.g. status in terms of selenium intake (secondary objective) in infertile women with
TLR2, TLR4 and TLR9. The impact of immune system on the primordial-to-pri- POI compared to healthy fertile women.
mary follicle transition remains unknown which could clarify molecular mecha- Study design, size, duration: Study design: Case-control study.
nisms protecting these early follicle stages. size: Case group consisted of 29 infertile women with idiopathic POI, amen-
Study design, size, duration: Expression data of TLR transcripts was orrhea and FSH >40 mIU/ml and three occult OI with oligomenorrhea and
extracted using bioinformatic tools from previous global RNA transcriptome AMH <0.3 ng/ml. Control group cosisted of 31 healthy fertile women matched
studies in human Laser Capture Microdissection (LCM)-isolated oocytes and with the case group in terms of age and BMI.
granulosa cells from primordial (n= 539) and primary (n= 261) follicles. Ovarian Duration: from August 2018 to Feb 2019.
tissues were donated by three women undergoing ovarian removal surgery for Participants/materials, setting, methods: Case group consisted of 29
cryopreservation before gonadotoxic treatment of non-gynecological cancer. infertile women with premature ovarian insufficiency (POI), amenorrhea and
As a proof of concept, intrafollicular localization and functionality of selected FSH >40 mIU/ml.
TLRs investigated by immunohistochemical and interleukin 6 (IL-6)-specific ELISA Infertility referral centers and tertiary hospital.
analyses. methods: The serum selenium level was measured by an atomic absorption
Participants/materials, setting, methods: qPCR was performed on LCM- spectrophotometer, and plasma GPx activity was measured by a Glutathione
isolated oocytes and granulosa cells from two independent patient samples to Peroxidase Activity Assay Kit after about 12 hours of fasting.
confirm TLR transcriptome data. Intrafollicular localization of TLR3 and TLR4 Main results and the role of chance: There was a significant reduction in
were studied in two patient samples using immunohistochemistry with anti-TLR3 the serum selenium level in the case group compared to the control group
and -TLR4 antibodies. For ELISA analysis, supernatants were extracted from (Adjusted Mean Difference (AMD) = -15.1 µg/ml, 95% CI: -24.8 to -5.3). The
juvenile murine ovaries in vitro cultured without or either with polyinosinic:poly- plasma GPx activity was lower in the case group compared to the control group,
cytidylic acid (poly I:C), or with lipopolysaccharides (LPS) (known as ligands for although not significantly (AMD = -67.0 U/ml, 95%CI: -194.5 to 60.3).
TLR3 and TLR4, respectively). Limitations, reasons for caution: present study limitations were selenium
Main results and the role of chance: Among other TLR transcripts, TLR3, measurement fee.
TLR4 and TLR5 were differentially and stage-specific consistently expressed genes Wider implications of the findings: This study showed a significant decline
(SSCEG) in both/either oocytes and/or granulosa cells from primordial and pri- in selenium status in infertile women with POI. It could weaken the selenium-de-
mary follicles. Inconsistent TLR5 expression across the three patient samples was pendent antioxidant defense against reactive oxygen species and may lead to POI.
the cause to exclude from this study. The qPCR analysis on TLR3 transcript aligned Trial registration number: IRCTID: IRCT20160410027311N6
with the transcriptome expression levels indicating highly upregulation only in
granulosa cells from both follicle stages. Immunohistochemistry indicated a halo- P-655 Bilirubin in follicular fluid: a biochemical signature of female
shaped TLR3 staining in the interspace between surrounding granulosa cells and infertility
the oocyte in primordial and primary follicles, respectively. Primary follicles showed
R. Pallisco1, G. Bilotta2, I. Listorti2, P. Bilotta3, G. Lazzarino4, B.
a more intense TLR3 staining correlating with the significant mRNA levels from Tavazzi5, T. Capozzolo3, A. Guarino3, G. Lazzarino6, R. Mangione7
the transcriptome data. TLR4 staining indicated presence in both the oocyte and 1
Alma Res fertility center, Laboratory of Embriology, roma, Italy ;
granulosa cells from primordial and primary follicles, however a weaker staining in 2
Alma Res Fertility Center, Laboratory of andrology and embriology, Rome, Italy ;
primordial follicle than observed for TLR3 in primordial follicle. ELISA revealed 3
Alma Res Fertility Center, Obstetrics and Gynecology, Rome, Italy ;
higher IL-6 production in ovarian supernatants treated with LPS than the level in 4
University of Catania, Department of Biomedical and Biotechnological Sciences-
untreated and poly I:C-treated supernatants. These TLRs are clearly present in
Division of Medical Biochemistry, Catania, Italy ;
the earliest hormone-independent follicle stages in human ovary which may have 5
Catholic University of Rome and Fondazione Policlinico Gemelli, Institute of
a potential defensive role (especially in murine ovaries).
Biochemistry and Clinical Biochemistry, Rome, Italy ;
Limitations, reasons for caution: This is a descriptive study of TLR tran- 6
Saint Camillus International University of Health Sciences, Biochemistry, Roma, Italy ;
scripts in human ovary supported with their biological functions in murine ovary. 7
Catholic University, Institute of Biochemistry and Clinical Biochemistry, Rome, Italy
The number of human samples was limited which eliminates the considerations
of the natural biological variance in the samples according to the experimen-
Study question: To assess whether bilirubin in follicular fluid (FF) of fertile
tal design.
females differs from infertile. To correlate FF bilirubin concentration with in vitro
Wider implications of the findings: This study has indicated, for the first
fertilization (IVF) outcomes.
time, presence and functional role of pattern recognition receptors as TLR3 and
Summary answer: Despite normal plasma levels, FF bilirubin concentrations
TLR4 in human oocytes and granulosa cells from the hormone-independent
in infertile females are higher than in fertile females. FF bilirubin concentrations
primordial and primary follicle stages.
negatively correlate with main IVF measures.
What is known already: Successful in vitro fertilization (IVF) are unexplainably
low (about 32%), not only in females with full-blown pathologies, but also in
P-654 Serum selenium level in women with idiopathic premature those known as idiopathic. Studies indicated that the composition of FF of
ovarian insufficiency: A case-control study infertile female may show altered levels of proteins, lipids, hormones respect to
B. Sadeghzadeh1, M. Bazrafkan2, P. Shahabi3 values found in fertile females. Such changes were associated to endometriosis,
1
Tabriz university of Medical sciences, midwifery Department, Tabriz, Iran ; polycystic ovarian syndrome (PCOs), diminished ovarian reserve (DOR). To
2
Reproductive Biotechnology Research center- Avicenna Research Institute, date, no studies have been dedicated either to determine potential differences
Reproductive Biology, Tehran, Iran ; in the FF concentration of bilirubin in fertile and infertile females or to evaluate
3
Tabriz University of Medical Sciences, Physiology, Tabriz, Iran possible correlations with main IVF outcome measures.

Abstracts of the 36th Annual Meeting of the

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Abstracts

36th Virtual Annual Meeting of the

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Published for the

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Executive Committee (2019 – 2021) Committee of National Nebojsa Radunovic (Serbia)

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Monday, 06 July 2020

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Tuesday 07 July 2020

Wednesday 08 July 2020

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ESHRE 2020 / Oral presentations

O-001 Harnessing the power of data: Application of data science

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O-035 Ovarian function recovery in newly diagnosed advanced

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INVITED SESSION INVITED SESSION

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O-051 Endometrial evaluation for all?

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Study question: Do smooth endoplasmic reticulum clusters (sERCs) influence

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O-069 Human ovarian ageing can be caused by Reactive Oxygen

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O-083 Incidence of mosaic embryos in translocation carriers:

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