Serum-free culture conditions for the growth of normal rat mammary epithelial cells in primary culture |
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Authors: | Ethier Stephen P |
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Institution: | (1) Department of Chemical Carcinogenesis, The Michigan Cancer Foundation, 110 E. Warren Avenue, 48201 Detroit, MI |
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Abstract: | Summary A monolayer culture system has recently been developed for the extended growth and serial passage of normal rat mammary epithelial
(RME) cells. In this system the cells undergo greater than 20 population doublings when grown on type I collagen-coated tissue
culture dishes in Ham's F12 medium supplemented with insulin, hydrocortisone, epidermal growth factor, prolactin, progesterone,
cholera toxin, and 5% fetal bovine serum (FBS). The purpose of the present studies was to define additional growth factors
that would allow equivalent RME cell proliferation in serum-free medium. Ethanolamine (EA) was effective at reducing the FBS
requirements for RME cell proliferation and at its optimum concentration did so by greater than 20-fold. Even with optimum
levels of EA there was essentially no cell proliferation in the absence of FBS. However, addition of bovine serum albumin
(BSA) to the hormone, growth factor, and EA-supplemented medium resulted in substantial proliferation in the absence of serum,
and the further addition of transferrin (T) potentiated this effect. Thus, in this culture system, replacement of FBS with
EA, BSA, and T resulted in RME cell proliferation in primary culture which was equivalent to that obtained in the 5% FBS-containing
medium.
This work was supported by grant RR-05529 from the Division of Research Resources, National Institutes of Health, Bethesda,
MD, and by Public Health Service grant CA40064-01 from the National Cancer Institute, Bethesda, MD. |
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Keywords: | mammary epithelial serum-free medium |
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