Abstract
A number of milk types and milk fractions were investigated as possible substitutes for serum in cell culture media. A filtrate of reconstituted nonfat dry milk showed promise. Culture fluids containing 5% of the nonfat dry milk filtrate were used to propagate primary and continuous cell cultures, and the cell growth from these cultures was compared with that of cells grown in a serum-containing medium. The nonfat dry milk filtrate-supplemented medium supported the growth of all epithelial cells tested, but two fibroblast-type cultures failed to replicate. Cells grown in the medium containing the milk filtrate grew slowly for 2 to 3 days and then propagated to confluency in 6 to 8 days. Viable cell counts of 9 days were comparable to those of serum-grown cells that had been propagated for 7 days. Cells grown in the milk filtrate could be split 1 to 4 when subcultures were prepared. Cell growth could be stimulated by refeeding on days 2 to 3 or by the addition of 30 microM 2-mercaptoethanol to the growth medium. Virus susceptibility and titer comparisons with poliovirus 1, coxsackievirus B2, echovirus 7, and herpes simplex virus indicated that approximately the same data were obtained when either the nonfat dry milk filtrate-treated or the serum-treated cells were studied. The nonfat dry milk filtrate is inexpensive, is easily prepared, and is a substitute for serum in epithelial cell culture media.
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Selected References
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- Barron A. L., Olshevsky C., Cohen M. M. Characteristics of the BGM line of cells from African green monkey kidney. Brief report. Arch Gesamte Virusforsch. 1970;32(4):389–392. doi: 10.1007/BF01250067. [DOI] [PubMed] [Google Scholar]
- Broome J. D., Jeng M. W. Growth stimulation of mouse leukemia cells by thiols and disulfides in vitro. J Natl Cancer Inst. 1972 Aug;49(2):579–581. [PubMed] [Google Scholar]
- Broome J. D., Jeng M. W. Promotion of replication in lymphoid cells by specific thiols and disulfides in vitro. Effects on mouse lymphoma cells in comparison with splenic lymphocytes. J Exp Med. 1973 Sep 1;138(3):574–592. doi: 10.1084/jem.138.3.574. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dahling D. R., Berg G., Berman D. BGM, a continuous cell line more sensitive than primary rhesus and African green kidney cells for the recovery of viruses from water. Health Lab Sci. 1974 Oct;11(4):275–282. [PubMed] [Google Scholar]
- GOCHENOUR A. M., BARON S. Viral sensitivity of cell cultures maintained with skim milk medium. Proc Soc Exp Biol Med. 1959 Oct;102:230–233. doi: 10.3181/00379727-102-25202. [DOI] [PubMed] [Google Scholar]
- Hasegawa K., Saito M., Taguchi F., Nagaki D. Storage-effects of chorio-allantoic fluid and skim milk on influenza virus infectivity. Kitasato Arch Exp Med. 1967 Dec;40(1):13–19. [PubMed] [Google Scholar]
- Hoffeld J. T., Oppenheim J. J. Enhancement of the primary antibody response by 2-mercaptoethanol is mediated by its action on glutathione in the serum. Eur J Immunol. 1980 May;10(5):391–395. doi: 10.1002/eji.1830100514. [DOI] [PubMed] [Google Scholar]
- Klagsbrun M. Bovine colostrum supports the serum-free proliferation of epithelial cells but not of fibroblasts in long-term culture. J Cell Biol. 1980 Mar;84(3):808–814. doi: 10.1083/jcb.84.3.808. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Larkin E. P., Tierney J. T., Sullivan R., Peeler J. T. Collaborative study of the glass wool filtration method for the recovery of virus inoculated into ground beef. J Assoc Off Anal Chem. 1975 May;58(3):576–578. [PubMed] [Google Scholar]
- Opitz H. G., Lemke H., Hewlett G. Activation of T-cells by a macrophage or 2-mercaptoethanol activated serum factor is essential for induction of a primary immune response to heterologous red cells in vitro. Immunol Rev. 1978;40:53–77. doi: 10.1111/j.1600-065x.1978.tb00401.x. [DOI] [PubMed] [Google Scholar]
- RABSON A. S., LEGALLAIS F. Y. Growth of human epidermoid carcinoma in tissue culture using nonfat milk medium. Arch Pathol. 1959 Sep;68:342–347. [PubMed] [Google Scholar]