Stem Cells

Clinical Applications

Dr T.V.Rao MD
Nerve Cell Cannot Regenerate ?
 In 1913 the great Spanish neuroscientist
Santiago Ramón y Cajal pronounced
“that in adult centres the nerve paths are
something fixed, ended, immutable.
Everything may die, nothing may be
regenerated”. For many years
neuroscientists believed not just that brain
damage was irreparable, but also that no
process to replace lost neurons existed in
our brain.
antiago Ramón y Cajal
(1852-1934)
Both beliefs turned out to be false as several
types of cells can regenerate
 Cell – A complex Organ
 Cell Theory: all living
things are composed of
one or more cells
 Cells fall into two basic
types prokaryotic and
eukaryotic.
 Prokaryotic cells are
smaller and lack much of
the internal
compartmentalizin
and complexity of
eukaryotic cells.
 History of Adult Stem Cell
Research
 Since the 1970’s, bone marrow
transplants have been used for
treatment of Immunodeficient
and leukemia.
 History of Human Embryonic
Stem Cell Research

Inner cell mass Isolate inner cell mass

(forms fetus)

Culture cells
Day 5-6
Blastocyst

In 1998, James Thomson (University of

Wisconsin-Madison) isolated cells from the inner
cell mass of the blastocyst, and developed the first
human embryonic stem cell line in culture.
 Stem Cell History
 1998 - Researchers first extract stem cells from human
embryos
 1999 - First Successful human transplant of insulin-making
cells from cadavers
 2001 - President Bush restricts federal funding for embryonic
stem-cell research
 2002 - Juvenile Diabetes Research Foundation International
creates $20 million fund-raising effort to support stem-cell
research
 2003?? - California ok stem cell research
 2004 - Harvard researchers grow stem cells from embryos
using private funding
 2004 - Ballot measure for $3 Billion bond for stem cells
History of Human Embryonic Stem Cell
Research
 In 1998, James Thomson (University of
Wisconsin-Madison) isolated cells from the
inner cell mass of the early embryo, and
developed the first human embryonic stem cell
lines,
 In 1998, John Gearhart (Johns
Hopkins University) derived human
embryonic germ cells from cells in
fetal gonadal tissue (primordial
germ cells).
 Pluripotent stem cell “lines” were
developed from both sources
 Stem Cell/Cloning Topics
 What are stem cells?
 History of stem
cell/cloning research
 Possible uses of the
technology
 Current
status/knowledge
 Questions and known
problems
 Legal considerations
 Politics
 Moral considerations
 What Are Stem Cells?
Stem cells are the raw
material from which all
of the body’s mature,
differentiated cells are
made. Stem cells give
rise to brain cells, nerve
cells, heart cells,
pancreatic cells, etc.
Stem Cell – Definition
 A cell that has the
ability to
continuously
divide and
differentiate
(develop) into
various other
kind(s) of
cells/tissues
 Stem Cell – are Dynamic
 Are undifferentiated Stem cell
“master” cell that do
not yet have a specific
function
 Can change to one or

Di
al

f fe
ew
several different cell

re
n
re

nt
lf-

iat
types (differentiate)

Se

e
under proper
conditions
 Can undergo unlimited Stem cell Specialized cell
cell division, self- (e.g., white blood cell)
renewal)
One Cell - Several lineages
 Embryogenesis and
Differentiation
 Specific regions of the embryo give rise to the
specific organ systems
 Ectoderm generates the outer layer of the

embryo and produces the surface layer

(epidermis) of the skin and forms the nerves
 Endoderm becomes the innermost layer of

the embryo and produces the digestive tube

and its associated organs (including the
lungs)
 Mesoderm becomes sandwiched between

the ectoderm and endoderm and generates

the blood, heart, kidney, gonads, bones, and
connective tissues.
Stages of Development
Ear l y Human
Development
An Overview of Early Development
modeled with Play-Dough
Fertilized egg
Totipotent: Can become any cell in
body or placenta

Totipotent Pluripotent: Can become any cell in

stem cells body
Fate Decision Multipotent: Can become any cell
within a specific germ layer or cell
Pluripotent
Blastocyst
lineage
stem cells Embryonic stem cells come from inner cell mass of blastocyst.
(3-5 days old)

Gastrulation (day 14) leads to

Primary Germ Cells
Fate Decision
Decision Endoderm (inner)digestive tract, resp. track
Mesoderm (middle)bones, blood cells, heart
Ectoderm (outer)skin, CNS
Implantation
Implantation Multipotent
 Pluripotent
Embryonal stem cell

Pluripotent
stem cells
Totipotent
stem cells Somatic Germ stem
stem cell cell

Somatic Cell
Primitive
Endoderm Ectoderm Mesoderm germ cell

Adult Multipotent
stem cell

Liver Skin Blood

Intestine Hair Muscle Gametes
Pancreas Nerves Bone
Cartilage
Courtesy of Dr F. Prosper
Cardoso.
Bone Marrow Stem Cells
How to Derive an Embryonic Stem Cell Line?

Isolate inner cell mass

(destroys embryo)
ETHICS?
Inner cell mass

Culture cells
Day 5-6
Blastocyst

A stem cell line is

composed of a
population of cells
that can replicate
themselves for
long periods of
time in vitro
(out of the body) An embryonic
stem cell
clone
 The Science of Stem Cells
 Stem cells have the ability to continually
reproduce themselves while maintaining the
capacity to give rise to other more specialized
cells.
 Stem cells are found at all stages of development,
from embryonic stem (ES) cells that can
differentiate into all specialized cells found in the
human body, to adult stem cells capable of
regenerating their tissue of origin.
 Stem cells occur from the earliest stages of
development and provide the starting material
for every organ and tissues.
 Embryonic stem (ES) cells
 ES cells are found
at the blastocyst
stage, four to five
days after the
union of the sperm
and egg, before
the embryo
implants in the
uterus.
ES Cells are "pluripotent" - i.e. capable
of forming embryonic tissues
 Source of Stem cells
 Stem cells may be derived from
autologus, allogeneic or xenogenic
sources. Histocompatability is prerequisite
for transplantation of allogeneic stem
cells. Fatal tissue is the best current
tissue source for human neural stem
cells, however ethical issues are a major
concern.
Placenta a Source of Stem Cells
 Placental stem cells,
like umbilical cord
blood and bone
marrow stem cells,
can be used to cure
chronic blood-related
disorders such as
sickle cell disease,
Thalasemia, and
leukaemia.
 Placental Blood as a Source of
Hematopoietic Stem Cells for
Transplantation into Unrelated Recipients

 Report of preliminary
results of
transplantation using
partially HLA-
mismatched
placental blood from
unrelated donors.
. Joanne Kurtzberg,
M.D.et al
 Umbilical Cord Blood Stem Cell
Transplant
 Umbilical cord blood
stem cell transplants are
less prone to rejection
than either bone marrow
or peripheral blood stem
cells. This is probably
because the cells have
not yet developed the
features that can be
recognized and attacked
by the recipient's
immune system
 Kinds of Stem Cells
Stem cell type Description Examples

Totipotent Each cell can develop into a Cells from early (1-3
new individual days) embryos

Pluripotent Cells can form any (over 200) Some cells of

cell types blastocyst (5 to 14
days)

Multipotent Cells differentiated, but can Fetal tissue, cord

form a number of other tissues blood, and adult stem
cells
 What’s So Special About
Stem Cells?
• They have the potential to replace cell tissue
that has been damaged or destroyed by
severe illnesses.

They can replicate themselves over and over

for a very long time.

Understanding how stem cells develop into

healthy and diseased cells will assist the
search for cures.
 Two Kinds of Stem Cells

•Embryonic (also called “pluripotent”)

stem cells are capable of developing into
all the cell types of the body.

•Adult stem cells are less versatile and

more difficult to identify, isolate, and
purify.
Stages of Embryogenesis

Day 2
2-cell embryo Day 3-4
Day 1
Multi-cell embryo
Fertilized egg

Day 5-6
Day 11-14 Blastocyst
Tissue Differentiation
 Derivation and Use of
Embryonic Stem Cell Lines
Isolate inner cell mass
Outer cells (destroys embryo)
(forms placenta)
Inner cells
(forms fetus) Culture cells

Day 5-6
Blastocyst “Special sauce”
(largely unknown)

Liver
Heart
repaired
Kidney Heart muscle
Embryonic Stem Cells:
Researchers extract stem cells from a 5-7 days old blastocyst.
Stem cells can divide in culture to form more of their own kind,
thereby creating a stem cell line.

The research aims to induce these cells to generate healthy

tissue needed by patients.
How Many Human Embryonic Stem
Cell Lines are There?
 The actual number of human
embryonic stem cell lines is a matter
of some debate.
 To date, more than 100 human
embryonic stem cell lines have been
derived worldwide.
 However, most of those lines are not
adequately characterized yet.
 Only 22 cell lines are eligible for
federal funding in the USA.
 Autologous – Stem Cells
 Sources of the patient's
own stem cells (autologous)
are either the cells from
patient's own body or his or
her cord blood. For
autologous transplants
physicians now usually
collect stem cells from the
peripheral blood rather than
the marrow
 This procedure is easier,
unlike a bone marrow
harvest, it can take place
outside of an operating
room and the patient does
not have to be under
general anaesthesia.
Allogeneic – Stem Cells
 Sources of stem cells
from another donor
(allogeneic) are primarily
relatives (familial-
allogeneic) or completely
unrelated donors
(unrelated-allogeneic).
The stem cells in this
situation are extracted
from either the donor's
body or cord blood
 Xenogenic - Stem Cells
 In this stem cells from
different species are
transplanted, e.g. striatal
porcine fetal ventral
mesencephalic (FVM)
xenotransplants for
Parkinson's disease.
This has no major ethical
concerns and a large
amount of tissue is
available, however life
long immunosupression
and risk of rejection are
the major limitations
 How Does Cell Therapy Work?
 Stem cells can be
used to generate
healthy and
functioning
specialized cells,
which can then
replace diseased or
dysfunctional cells.
 It is similar to the
process of organ
transplantation only the
treatment consists of
transplanting cells
instead of organs.
How Does Cell Therapy Work?
 Bone marrow transplants are an example of
cell therapy in which the stem cells in a
donor's marrow are used to replace the blood
cells of the victims of leukemia.
 Cell therapy is also being used in experiments
to graft new skin cells to treat serious burn
victims, and to grow new corneas for the
sight-impaired.
 In all of these uses, the goal is for the
healthy cells to become integrated into the
body and begin to function like the
patient's own cells.
 What Diseases Can be
Cured by Stem Cell Therapies
 Any disease in
which there is
tissue degeneration
can be a potential
candidate for stem
cell therapies
 Major Progress in Several
Important Health problems
 Alzheimer’s disease
 Parkinson’s disease
 Spinal cord injury
 Heart disease
 Severe burns
 Diabetes
 Alzheimer’s disease and can
stem cells help?
 Stem cells could,
however, be genetically
modified so as to deliver
substances to the
Alzheimer brain, to stop
cells from dying and
stimulate the function of
existing cells. A recent
clinical trial (Phase I) has
shown this approach to
be of some benefit to
patients with Alzheimer’s
disease, by slowing
down the progression of
the disease.
 Drug Testing
Stem cells could allow scientists
to test new drugs using human
cell line which could speed up
new drug development.
Only drugs that were safe and
had beneficial effects in cell line
testing would graduate to whole
animal or human testing.
It would allow quicker and safer
development of new drugs.
 Major types of Stem Cells
 The two broad types of
mammalian stem cells
are: embryonic stem
cells that are isolated
from the inner cell mass
of blastocysts, and adult
stem cells that are
found in adult tissues. In
a developing embryo,
stem cells can
differentiate into all of the
specialized embryonic
tissues. I
 Stem cells act as Progenitor
cells
 In adult organisms,
stem cells and
progenitor cells act
as a repair system for
the body,
replenishing
specialized cells, but
also maintain the
normal turnover of
regenerative organs,
such as blood, skin
or intestinal tissues.
 History of Animal Cloning
 Since then, animals including mice (1998),
cows (1998), pigs (2000), cats (2001), and
rabbits (2002) were successfully cloned.

Cattle
MOUSE
PIG

CAT
RABBIT
 How Successful Was Animal
Cloning? Very low (~1-3%)
1 live birth out of 29 cloned
Dolly embryos
3%
(sheep) 31 live births out of 2468 cloned
Cloned mice embryos
1%
5 live births out of 335 cloned
Cloned pigs embryos
1%
3 live births out of 85 cloned
Cloned goats embryos
3%
30 live births out of 496 cloned
Cloned embryos
6%
cattle 1 live birth out of 87 cloned
Cloned cat embryos
1%
6 live births out of 371 of cloned
Cloned embryos
1%
rabbits
 First Success of Human Embryo
Cloning
On February 12, 2004, South
Korean scientists, Dr. Woo Suk
Hwang and Dr. Shin Young Moon of
Seoul National University, reported the
successful creation of 30 cloned
human embryos developed to the
blastocyst stage and then destroyed
by stem cell extraction, yielding one
embryonic stem cell line.
Source of Stem Cells for Medical
therapies
 Tens of thousands of
frozen embryos are
routinely destroyed
when couples finish
their treatment.
 These surplus embryos
can be used to produce
stem cells.
 Regenerative medical
research aims to
develop these cells into
new, healthy tissue to
heal severe illnesses.
Stem Cell Research Worldwide
Adult multipotent stem cells
Adult Stem Cells
Autologus – Stem Cells
 While most blood stem
cells reside in the bone
marrow, a small number
are present in the
bloodstream. These
multipotent peripheral
blood stem cells, or
PBSCs, can be used just
like bone marrow stem
cells to treat leukaemia,
other cancers and
various blood disorders
 Speculation

multipoten
t
Treatments becomes specific
 Applications of Stem Cells
 Cell Replacement Therapies
 Cells could be stimulated to develop into

specialized cells that represent renewable

sources of cells and tissue for transplantation.
 Cell replacement therapy could treat injuries

and various genetic and degenerative

conditions including muscular dystrophies,
retinal degeneration, Alzheimer disease,
Parkinson's disease, arthritis, diabetes, spinal
cord injuries, and blood disorders such as
hemophilia.
 Understanding Cell
Specialization

 Studying human pluripotent stem cells can lead to
the identification of factors responsible for
differentiation of stem cells into specialized cell
types.
− these factors may ultimately be used to drive tissue
regeneration and repair if administered therapeutically.
 This work will provide basic knowledge on cell
determination and differentiation, human
development, genomic imprinting and somatic cell
aging.
 Development and Testing of
Drugs

 Researchers could
study the
beneficial and
toxic effects of
new medications
on human
pluripotent stem
cells that have
been developed to
mimic the disease
processes.
Can Sex Make difference in Stem
cell Therapy ?
 Are there sex-specific
differences in the biology

of stem cells? (short-long
term
 • How do sex-specific
differences play out in
terms of self-renewal and
differentiation? (mid-long
 XX
term)  vs.

XY
• Is there existing
evidence that the sex of
stem cells affects 
success of the
transplant?
Stem cells – Blindness
 In clinical trials at
Moorfields Eye
Hospital in London,
surgeons restored
eye sight for six
patients who lost
their sight after
chemical accidents
and genetic
diseases. The
patients went under
successful stem-cell
transplant.
 Limbal stem Cell therapy
 The treatment is known
as limbal stem cell
therapy, and the patients
who received the
treatment suffered from
chemical burn or genetic
disease know as aniridia
a By replacing the limbal
stem cells, the cornea
begins to clear up as the
cells are replaced with
the healthy transparent
layer again.
 Current possible uses
 Research in stem cells has opened up
new horizons in the area of treatment of
disorders such as stroke, epilepsy, neuro-
degeneration and trauma. Current
research is aimed at finding the
appropriate source of stem cells for a
given indication, ways of expanding and
perpetuating these cells in culture, best
route of administration of these cells and
methods to overcome rejection
 Possible Uses of Stem Cell
Technology
 Replaceable tissues/
organs
 Repair of defective
cell types
 Delivery of genetic
therapies
 Delivery
chemotherapeutic
agents
 Future –Making cells and
replacing the diseased cells ?
 Obstacles of Stem Cell Research
How to find the right type of stem cells?
How to put the stem cells into the right
place?
Will the stem cells perform the desired
function in the body?
Differentiation protocols for many cell
types have not been developed.
Embryonic Stem Cells are Unstable
and Mutate in Culture
 Like ordinary cells,
stem cells
accumulate
significant numbers
of mutations over
time, including
several that could
cause them to
become tumors.
 Ethical debate
 Harvesting ES cells
destroys the blast
cyst
 “This is murder”
 ES cell research
requires human cells
 Could create a
commercial market
for human cells
 “This devalues life”
Service
Reproduced by permission of Dave Catrow and Copley News
Destroying life to cure some one
– Ethical ?
 If stem cells have such potential to relieve
suffering, why are so many people so
upset about their use? The reason is that
the most powerful type of stem cell �
embryonic stem (ES) cells � can only be
obtained from human embryos. Many
people think that it's wrong to create and
destroy human embryos to treat disease
 Religious Debate over Harvesting
Embryonic Stem Cells
 The pro-life group generally
believes that:
 Personhood happens at, or
shortly after, conception.
 Thus, they consider the
removal of stem cells from an
embryo -- a procedure which
kills the stem cells -- to be a
form of murder of a human
being.
 They argue that no potential
health benefits to even
hundreds of millions of people Day 5-6
can justify the murder of other Blastocyst
humans.
 Religious Debate over Harvesting
Embryonic Stem Cells
 The pro-choice group generally
believes that:
 Personhood is attained much later in
pregnancy, perhaps when the fetal
brain develops consciousness during
the third trimester.
 Thus, extracting stem cells from an
five or ten-day old pre-embryo is
not murder.
 Killing a pre-embryo, which is only
a potential human being, is justified
if it has the potential to cure
diseases and extend the lives of
people.

Day 5-6
Blastocyst
 Why we should support
Can help several disabled
 Human embryonic
stem cell (HESC)
research offers
great promise of
cures for
otherwise
incurable
conditions: spinal
cord injuries, ALS,
Alzheimer’s,
Parkinson’s, etc.
 Shall be Clone Humans ?
 Arguments for and
against human cloning
research. Should we ban
human cloning? Why
investors are moving
away from human
cloning and why human
cloning now looks a last-
century way to fight
disease. Why some
people want to clone
themselves or even to
clone the dead.
Research on Stem Cells is progressing
in spite of several restrictions
Created for awareness to
Medical and Paramedical
Medical Students in
Developing World
Dr.T.V.Rao MD
Email
doctortvrao@gmail.com