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C3H/10T1/2, Clone 8

CCL-226

C3H/10T1/2, Clone 8 is a cell line exhibiting fibroblast morphology that was isolated from a line of C3H mouse embryo cells. This cell line was deposited by C Heidelberger.
Product category
Animal cells
Organism
Mus musculus, mouse
Classification
Eukaryota, Animalia, Metazoa, Chordata, Vertebrata, Tetrapod
Morphology
fibroblast
Tissue
Embryo
Disease
Sarcoma
Applications
3D cell culture
Product format
Frozen
Storage conditions
Vapor phase of liquid nitrogen

Documentation

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 (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.

Detailed product information

General

Specific applications
This line is a suitable transfection host.

Characteristics

Growth properties
Adherent
Derivation
C3H/10T1/2, Clone 8 was isolated by C. Reznikoff, D. Brankow and C. Heidelberger in 1972 from a line of C3H mouse embryo cells.
Age
embryo
Strain
C3H
Clinical data
The depositor recommends that the line be used between the 5th and 15th passages only.
Karyotype
Mouse karyotype with a modal number of 80 chromosomes.
Tumorigenic
No;
No, in immunosuppressed mice
Yes, in semisolid medium
Comments
The cells are very sensitive to post confluence inhibition of cell division, do not produce tumors in syngeneic mice, have no background of spontaneous transformation, nor do they contain overt endogenous transforming murine leukemia or sarcoma viruses.
The cells are contact sensitive.
There is no detectable background spontaneous transformation.
They are highly susceptible to transformation by chemical agents.
Tested and found negative for ectromelia virus (mousepox).
Note: the inoculation density, feeding and harvesting schedules must be followed rigidly if the line is to retain its essential characteristics.
The batch of serum used for growth and for transformation assays may affect both the morphology of this line and the results obtained.
Monolayers established and maintained for the standard transformation assay should be free of all foci after 6 weeks.
The depositor recommends that the line be used between the 5th and 15th passages only.

Handling information

Unpacking and storage instructions
  1. Check all containers for leakage or breakage.
  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 Eagle's Basal medium (Thermofisher Catalog No. 21010-046) supplemented with heat-inactivated fetal bovine serum to a final concentration of 10% and 2mM L-glutamine (ATCC 30-2214).
Temperature
37°C
Handling procedure

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.

  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 125 x g for 5 to 7 minutes.
  4. Resuspend cell pellet with the recommended complete medium (see the specific batch information for the culture recommended dilution ratio). 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 complete 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). pH (7.0 to 7.6).
  5. 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

Never allow the culture to become completely confluent.  Subcultivation of cultures must be performed before they reach confluence.  Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes. Corning® T-75 flasks (catalog #430641) are recommended for subculturing this product.

  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53 mM  EDTA solution to remove all traces of serum which contains trypsin inhibitor.
  3. Add 2.0 to 3.0 mL of Trypsin-EDTA solution 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.
  6. Incubate cultures at 37°C.
Subcultivation Ratio: Seed new flasks at 2000 viable cells/cm2.
Medium Renewal: Once between subcultures if necessary
Reagents for cryopreservation
Complete growth medium supplemented with 5% (v/v) DMSO (ATCC 4-X)

Quality control specifications

Mycoplasma contamination
Not detected

History

Deposited as
Mus musculus
Depositors
C Heidelberger

Legal disclaimers

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.
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.

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.

Frequently Asked Questions

References

Curated Citations

Reznikoff CA, et al. Quantitative and qualitative studies of chemical transformation of cloned C3H mouse embryo cells sensitive to postconfluence inhibition of cell division. Cancer Res. 33: 3239-3249, 1973. PubMed: 4796800

Terzaghi M, Little JB. Repair of potentially lethal radiation damage in mammalian cells is associated with enhancement of malignant transformation. Nature 253: 548-549, 1975. PubMed: 1167940

Mondal S, Heidelberger C. Transformation of C3H/10T1/2 CL8 mouse embryo fibroblasts by ultraviolet irradiation and a phorbol ester. Nature 260: 710-711, 1976. PubMed: 1264242

Smith GJ, et al. Clonal analysis of the expression of multiple transformation phenotypes and tumorigenicity by morphologically transformed 10T1/2 cells. Cancer Res. 53: 500-508, 1993. PubMed: 8425183

Rapp UR, et al. Endogenous oncornaviruses in chemically induced transformation. I. Transformation independent of virus production. Virology 65: 392-409, 1975. PubMed: 165619

View All Curated Citations for this Product

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