In vitro analysis of proliferating epithelial cell populations from the mouse mammary gland: Fibroblast-free growth and serial passage (original) (raw)

Summary

Normal and neoplastic mouse mammary epithelial cells were cultured in nutrient medium containing D-valine substituted for L-valine. Fibroblast overgrowth was prevented and epithelial cell functions and morphology were retained in cultures maintained in, D-valine medium up to 2 months. A nonenzymatic technique was devised to dissociate epithelial cell monolayers. The combined use of this dissociation buffer and D-valine nutrient medium made it possible to passage serially normal and neoplastic mammary epithelial cells. Normal cells were derived from mammary glands of animals stimulated with exogenous hormones for various periods. The period of in vivo hormonal stimulation influenced the ability of normal mammary epithelial cells to attach and proliferate in primary and serially passaged cultures. A greater proportion of cells derived from glands following 2 to 4 weeks of hormonal stimulation were recovered after replating and showed higher labeling indices during serial passage than cells from unstimulated or 5- to 7-week stimulated groups.

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References

  1. Slemmer, G. 1974. Interactions of separate types of cells during normal and neoplastic mammary gland growth. J. Invest. Dermatol. 63: 27–47.
    Article Google Scholar
  2. Munford, R. E. 1974. Changes in the mammary glands of rats and mice during pregnancy, lactation and involution. J. Endocrinol. 28: 1–15.
    Article Google Scholar
  3. Nicoll, C. S., and H. A. Tucker. 1965. Estimates of parenchymal, stromal and lymph node DNA in mammary glands of C3H/Crgl/2 mice. Life Sci. 4: 993–1001.
    Article PubMed CAS Google Scholar
  4. Banerjee, M. R., J. E. Wagner, and D. L. Kinder. 1971. DNA synthesis in the absence of cell reproduction during functional differentiation of mouse mammary gland. Life Sci. 10: 867–877.
    Article CAS Google Scholar
  5. Traurig, H. H. 1967. Cell proliferation in the mammary gland during late pregnancy and lactation. Anat. Rec. 157: 489–504.
    Article Google Scholar
  6. Traurig, H. H., 1967. A radiographic study of cell proliferation in the mammary gland of the pregnant mouse. Anat. Rec. 159: 239–248.
    Article PubMed CAS Google Scholar
  7. Bresciani, F. 1965. Effect of ovarian hormones on duration of DNA synthesis of the C3H mouse mammary gland. Exp. Cell Res. 38: 13–32.
    Article PubMed CAS Google Scholar
  8. Bresciani, F. 1968. Topography of DNA synthesis in the mammary gland of the C3H mouse and its control by ovarian hormones: An autoradiographic study. Cell Tissue Kinet. 1: 51–63.
    CAS Google Scholar
  9. Traurig, H. H., and C. F. Morgan. 1964. The effects of ovarian and hypophysial hormones on mammary gland epithelial cell proliferation. Anat. Rec. 150: 423–434.
    Article PubMed CAS Google Scholar
  10. Stockdale, F. E., and Y. J. Topper. 1966. The role of DNA synthesis and mitosis in hormone dependent differentiation. Proc. Nat. Acad. Sci. U.S.A. 56: 1283–1289.
    Article CAS Google Scholar
  11. Oka, T., and Y. J. Topper. 1972. Is prolactin mitogenic for mammary epithelium? Proc. Nat. Acad. Sci. U.S.A. 69: 1693–1696.
    Article CAS Google Scholar
  12. Lasfargues, E. Y., and D. H. Moore. 1971. A method for the continuous cultivation of mammary epithelium. In Vitro 7: 21–25.
    PubMed CAS Google Scholar
  13. Owens, R. B., H. S. Smith, and A. J. Hackett. 1974. Epithelial cell cultures from normal glandular tissues of mice. J. Nat. Cancer Inst. 53: 262–269.
    Google Scholar
  14. Gilbert, S. F., and B. R. Migeon. 1975. D-valine as a selective agent for normal human and rodent epithelial cells in culture. Cell 5: 11–17.
    Article PubMed CAS Google Scholar
  15. Nandi, S., 1958. Endocrine control of mammary gland development and function in the C3H/HE Crgl mouse. J. Nat. Cancer Inst. 21: 1039–1063.
    PubMed CAS Google Scholar
  16. Fox, C. H., and K. K. Sanford. 1975. Chemical analysis of mammalian sera commonly used as supplements for tissue culture media. Procedure 94121. TCA Manual 1: 233–237.
    Article CAS Google Scholar
  17. Das, N. K., H. L. Hosick, and S. Nandi. 1974. Influence of seeding density on multicellular organization and nuclear events in cultures of normal and neoplastic mammary epithelium. J. Nat. Cancer Inst. 52: 849–861.
    PubMed CAS Google Scholar
  18. Daniel, C. W., and K. B. DeOme. 1965. Growth of mouse mammary glands_in vivo_ after monolayer culture. Science 149: 634–636.
    Article PubMed CAS Google Scholar

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Author notes

  1. Michael T. White
    Present address: Department of Medicine (Oncology Division), Stanford University Medical School, 94305, Stanford, California
  2. A. S. L. Hu
    Present address: Department of Biochemistry, University of Kentucky, 40506, Lexington, Kentucky

Authors and Affiliations

  1. Cencer Research Laboratory and Department of Zoology, University of California, 94720, Berkeley, California
    Michael T. White, A. S. L. Hu, Susan T. Hamamoto & S. Nandi

Authors

  1. Michael T. White
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  2. A. S. L. Hu
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  3. Susan T. Hamamoto
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  4. S. Nandi
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Additional information

This investigation was supported by Grant No. CA 05388 from the National Cancer Institute and by Cancer Research Funds of the University of California.

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White, M.T., Hu, A.S.L., Hamamoto, S.T. et al. In vitro analysis of proliferating epithelial cell populations from the mouse mammary gland: Fibroblast-free growth and serial passage.In Vitro 14, 271–281 (1978). https://doi.org/10.1007/BF02616036

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