TCMK-1 (ATCC® CCL-139)

Organism: Mus musculus, mouse  /  Cell Type: SV40 transformed  /  Tissue: kidney  /  Disease: normal

Organism Mus musculus, mouse
Tissue kidney
Cell Type SV40 transformed
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 2 [Cells contain polyomavirus DNA sequences]
Disease normal
Age weanling
Strain C3H/Mai
Storage Conditions liquid nitrogen vapor phase
Karyotype modal number = 78; range = 64 to 157.
All cells showed 2-6 acrocentric chromosomes with secondary constrictions and 1-4 biarmed chromosomes per cells. Sixty percent of the cells had one or more minute chromosomes.
Derivation The cells are SV40 transformed and are positive for SV40 T-antigen.
Antigen Expression
H-2k
Tumorigenic No
Effects
No, in immunosuppressed mice
Yes, in semisolid medium
Virus Susceptibility Vesicular stomatitis, Glasgow (Indiana)
Human poliovirus 1
Virus Resistance
poliovirus 1
Comments

About 2% of the cells are positive for SV40 viral antigen, but virus is not recoverable.

Tested and found negative for ectromelia virus (mousepox).

Complete Growth Medium The base medium for this cell line is ATCC-formulated Eagle's Minimum Essential Medium, Catalog No. 30-2003. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.
Subculturing Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
  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: A subcultivation ratio of 1:2 to 1:8 is recommended
Medium Renewal: Every 2 to 3 days

Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994.

Cryopreservation
Freeze medium: Complete growth medium 95%; DMSO, 5%
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Temperature: 37°C
Name of Depositor PH Black, W Rowe
Year of Origin 1963
References

Black PH, Rowe WP. SV-40 induced proliferation of tissue culture cells of rabbit, mouse, and porcine origin. Proc. Soc. Exp. Biol. Med. 114: 721-727, 1963. PubMed: 14120333

Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.

Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988.

Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.

Biosafety in Microbiological and Biomedical Laboratories, 5th ed. HHS. U.S. Department of Health and Human Services, Centers for Disease Control and Prevention. Washington DC: U.S. Government Printing Office; 2007. The entire text is available online at http://www.cdc.gov/OD/ohs/biosfty/bmbl5/bmbl5toc.htm

Basic Documentation
References

Black PH, Rowe WP. SV-40 induced proliferation of tissue culture cells of rabbit, mouse, and porcine origin. Proc. Soc. Exp. Biol. Med. 114: 721-727, 1963. PubMed: 14120333

Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.

Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988.

Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.

Biosafety in Microbiological and Biomedical Laboratories, 5th ed. HHS. U.S. Department of Health and Human Services, Centers for Disease Control and Prevention. Washington DC: U.S. Government Printing Office; 2007. The entire text is available online at http://www.cdc.gov/OD/ohs/biosfty/bmbl5/bmbl5toc.htm