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.
Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:8 is recommended
Medium Renewal: 2 to 3 times per week
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.
Complete culture medium described above supplemented with 5% (v/v) DMSO. Cell culture tested DMSO is available as ATCC Catalog No. 4-X.
Evans CH, DiPaolo JA. Comparison of nude mice with the host species for evaluation of the tumorigenicity of guinea pig and hamster cells transformed in vitro by chemical carcinogens. Cancer Res. 36: 128-131, 1976. PubMed: 1247991
Evans CH, DiPaolo JA. Neoplastic transformation of guinea pig fetal cells in culture induced by chemical carcinogens. Cancer Res. 35: 1035-1044, 1975. PubMed: 1116142
Evans CH, et al. Colony inhibition mediated by nonimmune leukocytes in vitro and skin reactivity in vivo as indices of tumorigenicity of guinea pig cultures transformed by chemical carcinogens. Cancer Res. 35: 1045-1052, 1975. PubMed: 1116143
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.
