The influence of cell shape on the induction of functional differentiation in mouse mammary cells in vitro (original) (raw)

Summary

To define more clearly the in vitro conditions permissive for hormonal induction of functional differentiation, we cultured dissociated normal mammary cells from prelactating mice in or on a variety of substrates. Cultivation of an enriched epithelial cell population in association with living adult mammary stroma in the presence of lactogenic hormones resulted in both morphological and biochemical differentiation. This differentiation, however, was not enhanced over that seen when the cells were associated with killed stroma, provided that the killed stroma had a flexibility similar to that of the living stroma. Cells cultured in inflexible killed stroma usually did not differentiate. Cells cultured within the flexible environment of a collagen gel, but removed from the gas-medium interface, differentiated in a manner similar to those cultured in flexible stroma. Cells cultured on the surface of an attached collagen gel were squamous, and their basolateral surfaces were sequestered from the medium; they did not differentiate. Cells cultured on floating collagen gels were cuboidal-columnar, with basolateral surfaces exposed to the medium, and showed good functional differentiation. Cells cultured on inflexible floating collagen gels were extremely flattened and had exposed basolateral surfaces, and showed no evidence of functional differentiation. We infer that assumption of cuboidal to columnar shapes similar to those of mammary cells in vivo may be important to the induction of functional differentiation in vitro. The additional requirement of basolateral cell surface exposure also is important.

Access this article

Log in via an institution

Subscribe and save

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Banerjee, M. R. Responses of mammary cells to hormones. Intl. Rev. Cytol. 47: 1–97; 1976.
    Article CAS Google Scholar
  2. Saacke, R. C.; Heald, C. W. Cytological aspects of milk formation and secretion. Larson, B. L.; Smith, V. R. eds. Lactation—A comprehensive treatise, Vol. 2. New York: Academic Press; 1974: 147–189.
    Google Scholar
  3. Forsyth, I. A. Organ culture techniques and the study of hormone effects on the mammary gland. J. Dairy Res. 3: 419–444; 1971.
    Google Scholar
  4. Lasfargues, E. Y. Cultivation and behavior_in vitro_ of the normal mammary epithelium of the adult mouse. Anat. Rec. 127: 117–130; 1959.
    Article Google Scholar
  5. Lasfargues, E. Y. Cultivation and behavior_in vitro_ of the normal mammary epithelium of the adult mouse. II. Observations on the secretory activity. Exp. Cell Res. 13: 553–562; 1957.
    Article PubMed CAS Google Scholar
  6. Feldman, M. K.De novo synthesis of casein and proteins in cell cultures of mouse mammary epithelial cells. Am. Zool. 11: 677; 1971.
    Google Scholar
  7. Ceriani, R. L. Hormone induction of specific protein synthesis in midpregnant mouse mammary cell culture. J. Exp. Zool. 196: 1–12; 1976.
    Article PubMed CAS Google Scholar
  8. Emerman, J. T.; Pitelka, D. R. Maintenance and induction of morphological differentiation in dissociated mammary epithelium on floating collagen membranes. In Vitro 13: 316–328; 1977.
    Article PubMed CAS Google Scholar
  9. Emerman, J. T.; Enami, J.; Pitelka, D. R.; Nandi, S. Hormonal effects on intracellular and secreted casein in cultures of mouse mammary epithelial cells on floating collagen membranes. Proc. Natl. Acad. Sci. USA 74: 4466–4470; 1977.
    Article PubMed CAS Google Scholar
  10. Michalopoulos, G.; Pitot, H. C. Primary cultures of parenchymal liver cells on collagen membranes. Exp. Cell Res. 94: 70–78; 1975.
    Article PubMed CAS Google Scholar
  11. Emerman, J. T.; Burwen, S. J.; Pitelka, D. R. Substrate properties influencing ultrastructural differentiation of mammary epithelial cells in culture. Tissue and Cell 11: 109–119; 1979.
    Article PubMed CAS Google Scholar
  12. Folkman, J.; Moscona, A. A. Role of cell shape in growth control. Nature 273: 345–349; 1978.
    Article PubMed CAS Google Scholar
  13. Gospodarowicz, D.; Greenburg, G.; Birdwell, C. R. Determination of cellular shape by the extracellular matrix and its correlation with the control of cellular growth. Cancer Res. 38: 4155–4171; 1978.
    PubMed CAS Google Scholar
  14. Lasfargues, E. Y.; Moore, D. H. A method for the continuous cultivation of mammary epithelium. In Vitro 7: 21–25; 1971
    PubMed CAS Google Scholar
  15. DeOme, K. B.; Faulkin, L. J., Jr.; Bern, H. A.; Blair, P. B. Development of mammary tumors from hyperplastic alveolar nodules transplanted into gland-free mammary fat pads of female C3H mice. Cancer Res. 19: 515–520; 1959.
    PubMed CAS Google Scholar
  16. Richardson, K. C.; Jarett, L.; Finke, E. H. Embedding in epoxy resins for ultrathin sectioning in electron microscopy. Stain Technol. 35: 313–323; 1960.
    PubMed CAS Google Scholar
  17. Cardiff, R. D. Quantitation of mouse mammary tumor virus (MTV) virions by radioimmunoassay. J. Immunol. 111: 1722–1729; 1973.
    PubMed CAS Google Scholar
  18. Enami, J.; Nandi, S. A sensitive radioimmunoassay for a component of mouse casein. J. Immunol. Methods 18: 235–244; 1977.
    Article PubMed CAS Google Scholar
  19. Hinegardner, R. T. An improved fluorometric assay for DNA. Anal. Biochem. 39: 197–201; 1971.
    Article PubMed CAS Google Scholar
  20. Stockdale, F. E.; Topper, Y. J. The role of DNA synthesis and mitosis in hormone-dependent differentiation. Proc. Natl. Acad. Sci. USA 56: 1283–1289; 1966.
    Article PubMed CAS Google Scholar
  21. Pickett, P. B.; Pitelka, D. R.; Hamamoto, S. T.; Misfeldt, D. S. Occluding junctions and cell behavior in primary cultures of normal and neoplastic mammary gland cells. J. Cell Biol. 66: 316–332; 1975.
    Article PubMed CAS Google Scholar
  22. Emerman, J. T. Morphological and behavioral characteristics of normal and neoplastic mouse mammary epithelial cells in culture. Berkeley, CA: Univ. California; 1977. Thesis.
    Google Scholar
  23. Enami, J. Hormonal control of milk, protein synthesis in mouse mammary epithelial cells in vitro. Berkeley, CA: Univ. California; 1977. Thesis.
    Google Scholar
  24. Cereijido, M.; Ehrenfeld, J.; Mesa, I.; Martinez-Palomo, A. Structural and functional membrane polarity in cultured monolayers of MDCK cells. J. Membrane Biol. 52: 147–159; 1980.
    Article CAS Google Scholar
  25. Misfeldt, D. S.; Hamamoto, S. T.; Pitelka, D. R. Transepithelial transport in cell culture. Proc. Natl. Acad. Sci. USA 73: 1212–1216; 1976.
    Article PubMed CAS Google Scholar

Download references

Author information

Author notes

  1. John M. Shannon
    Present address: Department of Anatomy, University of Colorado Health Sciences Center, 4200 East Ninth Ave., 80262, Denver, Colorado

Authors and Affiliations

  1. Department of Zoology, University of California, 94720, Berkeley, California
    John M. Shannon & Dorothy R. Pitelka
  2. Cancer Research Laboratory, University of California, 230 Warren Hall, 94720, Berkeley, California
    John M. Shannon & Dorothy R. Pitelka

Authors

  1. John M. Shannon
  2. Dorothy R. Pitelka

Additional information

This work was supported by U.S. Public Health Service Grants CA-05045 and CA-09041 from the National Cancer Institute, Bethesda, MD.

Rights and permissions

About this article

Cite this article

Shannon, J.M., Pitelka, D.R. The influence of cell shape on the induction of functional differentiation in mouse mammary cells in vitro.In Vitro 17, 1016–1028 (1981). https://doi.org/10.1007/BF02618428

Download citation

Key words