C2C12

Product Sheet

CRL-1772™

Description
C2C12 is a myoblast cell line that is a subclone (produced by H. Blau, et al) of the
mouse myoblast cell line established by D. Yaffe and O. Saxel. The C2C12 cell line
differentiates rapidly, forming contractile myotubes and producing characteristic
muscle proteins. Treatment with bone morphogenic protein 2 (BMP-2) cause a shift
in the differentiation pathway from myoblastic to osteoblastic.
Organism: Mus musculus, mouse
Cell Type: myoblast
Tissue: Muscle
Morphology: myoblast
Growth properties: Adherent

Storage Conditions
Product format: Frozen
Storage conditions: Vapor phase of liquid nitrogen

Intended Use
This product is intended for laboratory research use only. It is not intended for any
animal or human therapeutic use, any human or animal consumption, or any
diagnostic use.

BSL 1
ATCC determines the biosafety level of a material based on our risk assessment as
guided by the current edition of Biosafety in Microbiological and Biomedical Laboratories

www.atcc.org
Page 1 of 7
C2C12
Product Sheet

CRL-1772

(BMBL), U.S. Department of Health and Human Services. It is your responsibility to

understand the hazards associated with the material per your organization’s policies
and procedures as well as any other applicable regulations as enforced by your local
or national agencies.

ATCC highly recommends that appropriate personal protective equipment is always

used when handling vials. For cultures that require storage in liquid nitrogen, it is
important to note that some vials may leak when submersed in liquid nitrogen and
will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid
nitrogen back to its gas phase may result in the vial exploding or blowing off its cap
with dangerous force creating flying debris. Unless necessary, ATCC recommends
that these cultures be stored in the vapor phase of liquid nitrogen rather than
submersed in liquid nitrogen.

Certificate of Analysis
For batch-specific test results, refer to the applicable certificate of analysis that can
be found at www.atcc.org.

Growth Conditions
Temperature: 37°C
Seeding density: 5.0 x 103 viable cells/cm2

Handling Procedures
Unpacking and storage instructions:
1. Check all containers for leakage or breakage.

www.atcc.org
Page 2 of 7
C2C12
Product Sheet

CRL-1772

2. Remove the frozen cells from the dry ice packaging and immediately place the
cells at a temperature below -130°C, preferably in liquid nitrogen vapor, until
ready for use.
Complete medium: The base medium for this cell line is ATCC-formulated Dulbecco's
Modified Eagle's Medium (ATCC 30-2002). To make the complete growth medium,
add the following components to the base medium: fetal bovine serum (ATCC 30-
2020) to a final concentration of 10%.
Handling Procedure:

Handling Procedure for Frozen Cells

To insure the highest level of viability, thaw the vial and initiate the culture as soon
as possible upon receipt. If upon arrival, continued storage of the frozen culture is
necessary, it should be stored in liquid nitrogen vapor phase and not at -70°C.
Storage at -70°C will result in loss of viability.

SAFETY PRECAUTION: ATCC highly recommends that protective gloves and clothing
always be used and a full face mask always be worn when handling frozen vials. It is
important to note that some vials leak when submersed in liquid nitrogen and will
slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen
back to its gas phase may result in the vessel exploding or blowing off its cap with
dangerous force creating flying debris.

1. Thaw the vial by gentle agitation in a 37°C water bath. To reduce the
possibility of contamination, keep the O-ring and cap out of the water.
Thawing should be rapid (approximately 2 minutes).
2. Remove the vial from the water bath as soon as the contents are thawed, and
decontaminate by dipping in or spraying with 70% ethanol. All of the
operations from this point on should be carried out under strict aseptic
conditions.
3. Transfer the vial contents to a centrifuge tube containing 9.0 ml complete
culture medium. and spin at approximately 150 to 400 x g for 8 to 12 minutes.
4. Discard the supernatant and resuspend the cell pellet in an appropriate
amount of fresh growth medium.
5. Transfer the cell suspension to an appropriate size vessel. It is important to
avoid excessive alkalinity of the medium during recovery of the cells. It is
suggested that, prior to the addition of the vial contents, the culture vessel
containing the growth medium be placed into the incubator for at least 15
minutes to allow the medium to reach its normal pH (7.0 to 7.6).

www.atcc.org
Page 3 of 7
C2C12
Product Sheet

CRL-1772

6. Incubate the culture at 37°C in a suitable incubator. A 5% CO2 in air

atmosphere is recommended if using the medium described on this product
sheet.
Subculturing procedure:
IMPORTANT - DO NOT ALLOW CULTURES TO BECOME CONFLUENT.
Cultures must not be allowed to become confluent as this will deplete the myoblastic
population in the culture.
Myotube formation is enhanced when the medium is supplemented with 10% horse
serum instead of fetal bovine serum.
1. Remove and discard culture medium.
2. Briefly rinse the cell layer with D-PBS (ATCC 30-2200) to remove all traces of
serum which contains trypsin inhibitor.
3. Add 2.0 to 3.0 mL of Trypsin-EDTA solution (ATCC 30-2101) to flask and observe
cells under an inverted microscope until cell layer is dispersed (usually within 5
to 15 minutes).
Note: To avoid clumping do not agitate the cells by hitting or shaking the flask
while waiting for the cells to detach. Cells that are difficult to detach may be
placed at 37°C to facilitate dispersal.
4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently
pipetting.
5. Add appropriate aliquots of the cell suspension to new culture vessels.
Inoculate at a cell concentration between 1.5 X 10 5 and 1.0 X 10 6 viable
cells/75 cm2. Corning T-75 flasks (catalog #430641) are recommended for
subculturing this product.
6. Incubate cultures at 37°C.
Medium Renewal: Every two to three days
Reagents for cryopreservation: Complete growth medium supplemented with 5%
(v/v) DMSO (ATCC 4-X)

Material Citation
If use of this material results in a scientific publication, please cite the material in the
following manner: C2C12 (ATCC CRL-1772)

References

www.atcc.org
Page 4 of 7
C2C12
Product Sheet

CRL-1772

References and other information relating to this material are available at

www.atcc.org.

Warranty
The product is provided 'AS IS' and the viability of ATCC® products is warranted for 30

days from the date of shipment, provided that the customer has stored and handled
the product according to the information included on the product information sheet,
website, and Certificate of Analysis. For living cultures, ATCC lists the media
formulation and reagents that have been found to be effective for the product.
While other unspecified media and reagents may also produce satisfactory results, a
change in the ATCC and/or depositor-recommended protocols may affect the
recovery, growth, and/or function of the product. If an alternative medium
formulation or reagent is used, the ATCC warranty for viability is no longer valid.
Except as expressly set forth herein, no other warranties of any kind are provided,
express or implied, including, but not limited to, any implied warranties of
merchantability, fitness for a particular purpose, manufacture according to cGMP
standards, typicality, safety, accuracy, and/or noninfringement.

Disclaimers

This product is intended for laboratory research use only. It is not intended for any
animal or human therapeutic use, any human or animal consumption, or any
diagnostic use. Any proposed commercial use is prohibited without a license from
ATCC.

While ATCC uses reasonable efforts to include accurate and up-to-date information
on this product sheet, ATCC makes no warranties or representations as to its
accuracy. Citations from scientific literature and patents are provided for
informational purposes only. ATCC does not warrant that such information has been
confirmed to be accurate or complete and the customer bears the sole responsibility
of confirming the accuracy and completeness of any such information.

www.atcc.org
Page 5 of 7
C2C12
Product Sheet

CRL-1772

This product is sent on the condition that the customer is responsible for and
assumes all risk and responsibility in connection with the receipt, handling, storage,
disposal, and use of the ATCC product including without limitation taking all
appropriate safety and handling precautions to minimize health or environmental
risk. As a condition of receiving the material, the customer agrees that any activity
undertaken with the ATCC product and any progeny or modifications will be
conducted in compliance with all applicable laws, regulations, and guidelines. This
product is provided 'AS IS' with no representations or warranties whatsoever except
as expressly set forth herein and in no event shall ATCC, its parents, subsidiaries,
directors, officers, agents, employees, assigns, successors, and affiliates be liable for
indirect, special, incidental, or consequential damages of any kind in connection with
or arising out of the customer's use of the product. While reasonable effort is made
to ensure authenticity and reliability of materials on deposit, ATCC is not liable for
damages arising from the misidentification or misrepresentation of such materials.

Please see the material transfer agreement (MTA) for further details regarding the
use of this product. The MTA is available at www.atcc.org.

Copyright and Trademark Information

© ATCC 2023. All rights reserved.
ATCC is a registered trademark of the American Type Culture Collection.

Revision
This information on this document was last updated on 2024-08-19

Contact Information
ATCC
10801 University Boulevard
Manassas, VA 20110-2209
USA

www.atcc.org
Page 6 of 7
C2C12
Product Sheet

CRL-1772

US telephone: 800-638-6597
Worldwide telephone: +1-703-365-2700
Email: tech@atcc.org or contact your local distributor

www.atcc.org
Page 7 of 7