Skip to main content
SpringerLink
Log in

Morphogenetic behavior of simian virus 40-transformed human mammary epithelial stem cell lines on collagen gels

  • Regular Papers
  • Published:
In Vitro Cellular & Developmental Biology - Animal Aims and scope Submit manuscript

Summary

Transformation of primary cultures of human breast cells with simian virus 40 and clonal selection has yielded single-cell-cloned, epithelial cell lines, as well as myoepithelial-related cell lines. When grown on floating collagen gels, the epithelial cell lines give rise to branching rays of cells, thick fingerlike protrusions, saclike structures, and degenerating areas. The myoepithelial-related cell lines give rise only to the branching rays. Epidermal growth factor stimulates the production of the thick protrusions, whereas cholera toxin stimulates the production of the degenerating areas. Immunocytochemical staining of these cultures using reagents directed against the cell surface-extracellular matrix or the cellular cytoskeleton confirms the epithelial and myoepithelial nature of the cells, and demonstrates that the degenerating areas are undergoing squamous metaplasia. The fingerlike protrusions consist of cords of cells composed of inner, epithelial and outer, myoepithelial-related cells sometimes surrounding a central lumen reminiscent of ducts. The saclike structures resemble alveoli. Ultrastructural analysis confirms the identification of the basic cell types and also identifies indeterminate cells possessing features of both epithelial and myoepithelial cells. It is suggested that the epithelial cell lines represent human mammary stem cells that can undergo processes of morphogenesis and differentiation in vitro to form many of the three-dimensional structures found within the breast.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Bennett, D. C.; Peachey, L. A.; Durbin, H., et al. A possible mammary stem cell line. Cell 15:296–308; 1978.

    Article  Google Scholar 

  2. Bissell, M. J.; Hall, H. G. Form and function in the mammary gland: the role of the extracellular matrix. In: Neville, M. C.; Daniel, C. W., eds. Mammary gland: development, regulation, and function. New York: Plenum Press; 1987:97–146.

    Google Scholar 

  3. Burwen, S. J.; Pitelka, D. R. Secretory function of lactating mouse mammary epithelial cells cultured on collagen gels. Exp. Cell. Res. 126:249–262; 1980.

    Article  CAS  PubMed  Google Scholar 

  4. Chang, S. E.; Keen, J.; Lane, E. B., et al. Establishment and characterization of SV40-transformed human breast epithelial cell lines. Cancer Res. 42:2040–2053; 1983.

    Google Scholar 

  5. Clark, R.; Stampfer, M. R.; Milley, M. R., et al. Transformation of human mammary epithelial cells by oncogenic retroviruses. Cancer Res. 48:4689–4694; 1989.

    Google Scholar 

  6. Dairkee, S. H.; Blayney-Moore, C. M.; Smith, H. S., et al. Concurrent expression of basal and luminal epithelial markers in cultures of normal human breast analyzed using monoclonal antibodies. Differentiation 32:93–100; 1986.

    Article  CAS  PubMed  Google Scholar 

  7. Easty, G. C.; Easty, D. M.; Monaghan, P., et al. Preparation and identification of human epithelial cells in culture. Int. J. Cancer 26:577–584; 1980.

    Article  CAS  PubMed  Google Scholar 

  8. Edwards, P. A. W.; Brooks, I. M.; Bunnage, H. J., et al. Clonal analysis of expression of epithelial antigens in cultures of normal human breast. J. Cell. Sci. 80:91–101; 1986.

    CAS  PubMed  Google Scholar 

  9. Edwards, P. A. W.; Brooks, I. M.; Monaghan, P. Expression of epithelial antigen in primary cultures of normal breast analysed with monoclonal antibodies. Differentiation 25:247–258; 1984.

    Article  CAS  PubMed  Google Scholar 

  10. Emerman, J. T.; Burwen, S. J.; Pitelka, D. R. Substrate properties influencing ultrastructural differentiation of mammary epithelial cells in culture. Tissue & Cell 11:109–119; 1979.

    Article  CAS  Google Scholar 

  11. Emerman, J. T.; Enami, J.; Pitelka, D. R., et al. 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  CAS  PubMed  Google Scholar 

  12. Foster, C. S.; Smith, C. A.; Dinsdale, E. A., et al. Human mammary gland morphogenesisin vitro: the growth and differentiation of normal breast epithelium in collagen gel cultures defined by electron microscopy, monoclonal antibodies and autoradiography. Dev. Biol. 86:197–216; 1983.

    Article  Google Scholar 

  13. Hall, H. G.; Farson, D. A.; Bissell, M. J. Lumen formation by epithelial cell lines in response to collagen overlay: a morphogenetic model in culture. Proc. Natl. Acad. Sci. USA 79:4672–4676; 1982.

    Article  CAS  PubMed  Google Scholar 

  14. Hillman, E. A.; Valerio, M. G.; Halter, S. A., et al. Long-term explant culture of normal mammary epithelium. Cancer Res. 43:245–257; 1983.

    CAS  PubMed  Google Scholar 

  15. Jamieson, S.; Dunnington, D. J.; Ormerod, E. J., et al. Dedifferentiation of rat mammary myoepithelial-like cell lines after passagein vivo or cloningin vitro. JNCI 76:247–256; 1986.

    CAS  PubMed  Google Scholar 

  16. Ohtani, H.; Sasano, N. Myofibroblasts and myoepithelial cells in human breast carcinoma. Virchows Arch. A Pathol. Anat. 385:247–261; 1980.

    Article  CAS  Google Scholar 

  17. Ormerod, E. J.; Rudland, P. S. Mammary gland morphogenesisin vitro: formation of branched tubules in collagen gels by a cloned rat mammary cell line. Dev. Biol. 91:360–375; 1982.

    Article  CAS  PubMed  Google Scholar 

  18. Ormerod, E. J.; Rudland, P. S. Isolation and characterization of cloned epithelial cell lines from normal rat mammary glands. In Vitro Cell. Dev. Biol. 21:143–155; 1985.

    Article  CAS  PubMed  Google Scholar 

  19. Ormerod, E. J.; Rudland, P. S. Mammary gland morphogenesis in vitro: extracellular requirements for the formation of tubules in collagen gels by a cloned rat mammary epithelial cell line. In Vitro Cell. Dev. Biol. 24:17–27; 1988.

    Article  CAS  PubMed  Google Scholar 

  20. Ozzello, L. Ultrastructure of the human mammary gland. Pathol. Annu. 6:1–58; 1971.

    CAS  PubMed  Google Scholar 

  21. Peterson, O. W.; van Deurs, B. Growth factor control of myoepithelial cell differentiation in cultures of human mammary gland. Differentiation 39:197–215; 1988.

    Article  Google Scholar 

  22. Rudland, P. S.; Barraclough, R. Differentiation of simian virus 40-transformed human mammary epithelial stem cell lines to myoepithelial-like cells is associated with increased expression of viral large T antigen. J. Cell. Physiol. 142:657–665; 1990.

    Article  CAS  PubMed  Google Scholar 

  23. Rudland, P. S.; Hughes, C. M. Immunocytochemical identification of cell types in human mammary gland: variations in cellular markers are dependent on glandular topography and differentiation. J. Histochem. Cytochem. 37:1087–1100; 1989.

    CAS  PubMed  Google Scholar 

  24. Rudland, P. S.; Hughes, C. M.; Ferns, S. A., et al. Characterization of human mammary cell types in primary culture: immunofluorescent and immunocytochemical indicators of cellular heterogeneity. In Vitro Cell. Dev. Biol. 25:23–36; 1989.

    Article  CAS  PubMed  Google Scholar 

  25. Rudland, P. S.; Ollerhead, G.; Barraclough, R. Isolation of simian virus 40-transformed human mammary epithelial stem cell lines that can differentiate to myoepithelial-like cells in culture andin vivo. Dev. Biol. 136:167–180; 1989.

    Article  CAS  PubMed  Google Scholar 

  26. Russo, J.; Reina, D.; Frederick, J., et al. Expression of phenotypic changes by human breast epithelial cells treated with carcinogensin vitro. Cancer Res. 48:2837–2857; 1988.

    CAS  PubMed  Google Scholar 

  27. Russo, J.; Tay, L. K.; Russo, I. H. Differentiation of the mammary gland and susceptibility to carcinogenesis. Breast Cancer Res. Treat. 2:5–73; 1982.

    Article  CAS  PubMed  Google Scholar 

  28. Schaeffer, F. V.; Custer, R. P.; Sorof, S. Induction of abnormal development and differentiation in cultured mammary glands by cyclic adenine nucleotide and prostaglandins. Nature 286:807–810; 1980.

    Article  Google Scholar 

  29. Shannon, J. M.; Pitelka, D. R. Influence of cell shape on the production of functional differentiation in mouse mammary cells in vitro. In Vitro 17:1016–1028; 1981.

    Article  CAS  PubMed  Google Scholar 

  30. Shirasuna, K.; Watatani, K.; Sugiyama, M., et al. Isolation and characterization of different clones including myoepithelial-like variants from cloned neoplastic epithelial cell line of human salivary gland origin. Cancer Res. 46:1418–1426; 1986.

    CAS  PubMed  Google Scholar 

  31. Stampfer, M.; Hallowes, R. C.; Hackett, A. J. Growth of normal human mammary cells in culture. In Vitro 16:415–425; 1980.

    Article  CAS  PubMed  Google Scholar 

  32. Vorherr, H. Development of the female breast. In: The breast: morphology, physiology, and lactation. New York: Academic Press; 1974:1–18.

    Google Scholar 

  33. Yang, J.; Guzman, R.; Richards, J., et al. Primary culture of human mammary epithelial cells embedded in collagen gels. JNCI 65:337–343; 1980.

    CAS  PubMed  Google Scholar 

  34. Yang, N. S.; Kube, D.; Park, C., et al. Growth of human mammary epithelial cells on collagen gel surfaces. Cancer Res. 41:4093–4100; 1981.

    CAS  PubMed  Google Scholar 

  35. Yang, J.; Richards, J.; Guzman, R., et al. Sustained growth in primary culture of normal mammary epithelial cells embedded in collagen gel. Proc. Natl. Acad. Sci. USA 77:2088–2092; 1980.

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Cancer and Polio Research Fund Laboratories, Department of Biochemistry, University of Liverpool, P.O. Box 147, L69 3BX, Liverpool, United Kingdom

    Philip S. Rudland, Gillian E. Ollerhead & Angela M. Platt-Higgins

Authors
  1. Philip S. Rudland
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Gillian E. Ollerhead
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Angela M. Platt-Higgins
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

This work was supported by the North West Cancer Research Fund and the Cancer and Polio Research Fund.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Rudland, P.S., Ollerhead, G.E. & Platt-Higgins, A.M. Morphogenetic behavior of simian virus 40-transformed human mammary epithelial stem cell lines on collagen gels. In Vitro Cell Dev Biol - Animal 27, 103–112 (1991). https://doi.org/10.1007/BF02630995

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02630995

Key words

Search

Navigation