| Comments |
The cells are responsive to ACTH and other steroidogenic agents. SBAC is a normal mammalian endocrine cell type that proliferates in culture, with or without serum, and maintains regulation of its differentiated functions. Fibroblast growth factor (FGF) stimulates cell division. The cells have a finite life span in culture (60 to 70 population doublings). Recent tests for bovine viral diarrhea virus (BVDV) indicate that the line is producing both detectable BVDV antigens and infectious BVDV virions (J. Virol Methods 48:211-221, 1994). |
| 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.
- Remove and discard culture medium.
- Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53mM EDTA solution to remove all traces of serum which contains trypsin inhibitor.
- 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.
- Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
- Add appropriate aliquots of the cell suspension to new culture vessels.
- Incubate cultures at 37°C.
Subcultivation Ratio: 1:3 to 1:6
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. |
| References |
Hornsby PJ, Gill GN. Hormonal control of adrenocortical cell proliferation. Desensitization to ACTH and interaction between ACTH and fibroblast growth factor in bovine adrenocortical cell cultures. J. Clin. Invest. 60: 342-352, 1977. PubMed: 194925
Simonian MH, et al. Clonal growth and culture life span of bovine adrenocortical cells in a serum-free medium. In Vitro Cell. Dev. Biol. 23: 247-252, 1987. PubMed: 3032889
Bolin SR, et al. Survey of cell lines in the American Type Culture Collection for bovine viral diarrhea virus. J. Virol. Methods 48: 211-221, 1994. PubMed: 7989438
Simonian MH, et al. Growth and function of cultured bovine adrenocortical cells in a serum-free defined medium. Endocrinology 111: 919-927, 1982. PubMed: 6213403
Gospodarowicz D, et al. Control of bovine adrenal cortical cell proliferation by fibroblast growth factor. Lack of effect of epidermal growth factor. Endocrinology 100: 1080-1089, 1977. PubMed: 189990
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.
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