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C3H/10T1/2, Clone 8 (ATCC® CCL-226)

Organism: Mus musculus, mouse  /  Tissue: embryo  /  Disease: sarcoma

Permits and Restrictions

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Organism Mus musculus, mouse
Tissue embryo
Product Format frozen
Morphology fibroblast
Culture Properties adherent
Biosafety Level 1

Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.

Disease sarcoma
Age embryo
Strain C3H
Applications
This line is a suitable transfection host.
Storage Conditions liquid nitrogen vapor phase
Karyotype Mouse karyotype with a modal number of 80 chromosomes.
Images CCL-226 micrograph
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.
Clinical Data
The depositor recommends that the line be used between the 5th and 15th passages only.
Tumorigenic No
Effects
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.
Complete Growth 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).
Subculturing

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
Cryopreservation
Freeze medium: Complete growth medium 95%; DMSO, 5%
Storage temperature: liquid notrogen vapor temperature
Culture Conditions
Temperature: 37°C
Name of Depositor C Heidelberger
Deposited As Mus musculus
Passage History
The donor recommends that the line be used between the 5th and 15th passages only.
References

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

Reznikoff CA, et al. Establishment and characterization of a cloned line of C3H mouse embryo cells sensitive to postconfluence inhibition of division. Cancer Res. 33: 3231-3238, 1973. PubMed: 4357355

Jain MK, et al. Molecular cloning and characterization of SmLIM, a developmentally regulated LIM protein preferentially expressed in aortic smooth muscle cells. J. Biol. Chem. 271: 10194-10199, 1996. PubMed: 8626582

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation
Other Documentation
References

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

Reznikoff CA, et al. Establishment and characterization of a cloned line of C3H mouse embryo cells sensitive to postconfluence inhibition of division. Cancer Res. 33: 3231-3238, 1973. PubMed: 4357355

Jain MK, et al. Molecular cloning and characterization of SmLIM, a developmentally regulated LIM protein preferentially expressed in aortic smooth muscle cells. J. Biol. Chem. 271: 10194-10199, 1996. PubMed: 8626582