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Growth factor interactions between mouse mammary cell lines cocultured in collagen gels

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Summary

Three related mouse mammary cell lines were cultured in collagen gels and assayed for growth factor responsiveness and interaction via soluble factors. The CL-S1 cell line is nontumorigenic and grows poorly in collagen gel culture. The +SA and −SA cell lines exhibit different degrees of malignant behavior in vivo and have different growth properties in vitro. In collagen gel culture, +SA growth was stimulated by serum but not by epidermal growth factor (EGF), whereas both serum and EGF were required for optimal growth of −SA cells of early passage number as well as CL-S1 cells. −SA cells of later passage repeatedly exhibited a change so as to no longer require serum while retaining EGF responsiveness. [125I]EGF binding analyses indicated that CL-S1 cells bound EGF with less affinity than did −SA cells whereas +SA cells bound almost to ligand. When cell lines were maintained in separate collagen gels but shared the same culture medium, growth of +SA or −SA cells was slightly enhanced in the presence of CL-S1 cells and −SA cell growth was enhanced by the presence of +SA cells. Using the normal rat kidney fibroblast line NRK (clone 49F) as an indicator, serum-containing conditioned media from each cell line and from each pair of cell lines cultured in collagen gels were tested for transforming growth factor (TGF) activity. Both the −SA and CL-S1 lines tested positive for TGF-α production and possibly released a TGF-β activity. These results suggest mechanisms by which cell populations in and around tumors can modify one another’s growth characteristics.

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References

  1. Absher, M. Hemocytometer counting. In: Kruse, P. F., Jr.; Patterson, M. K., Jr., eds. Tissue culture methods and applications. New York: Academic Press; 1973:395–397.

    Google Scholar 

  2. Anderson, L. W.; Danielson, K. G.; Hosick, H. L. Epithelial cell line and subline established from premalignant mouse mammary tissue. In Vitro 15:841–843; 1979.

    Article  PubMed  CAS  Google Scholar 

  3. Danielson, K. G.; Anderson, L. W.; Hosick, H. L. Selection and characterization in culture of mammary tumor cells with distinctive growth properties in vivo. Cancer Res. 40:1812–1819; 1980.

    PubMed  CAS  Google Scholar 

  4. DeLarco, J. E.; Todaro, G. J. Growth factors from murine sarcoma virus-transformed cells. Proc. Natl. Acad. Sci. USA 75:4001–4005; 1978.

    Article  CAS  Google Scholar 

  5. Derynck, R. Transforming growth factor alpha. Cell 54:593–595; 1988.

    Article  PubMed  CAS  Google Scholar 

  6. Fidler, I. J.; Hart, I. R. Biological diversity in metastatic neoplasms: origins and implications. Science 217:998–1003; 1982.

    Article  PubMed  CAS  Google Scholar 

  7. Hallowes, R. C.; Bone, E. S.; Jones, W. A new dimension in the culture of human breast. In: Richards, R. J.; Rajan, K. T., eds. Tissue culture in medical research (II). Oxford and New York: Pergamon Press; 1980;213–220.

    Google Scholar 

  8. Heppner, G. H. Tumor heterogeneity. Cancer Res. 44:2259–2265; 1984.

    PubMed  CAS  Google Scholar 

  9. Imagawa, W.; Tomooka, Y.; Nandi, S. Serum-free growth of normal and tumor mouse mammary epithelial cells in primary culture. Proc. Natl. Acad. Sci. USA 79:4074–4077; 1982.

    Article  PubMed  CAS  Google Scholar 

  10. Jones, W.; Hosick, H. L. Collagen concentration as a significant variable for growth and morphology of mouse mammary parenchyma in collagen matrix culture. Cell Biol. Int. Rep. 10:277–286; 1986.

    Article  PubMed  CAS  Google Scholar 

  11. Kaplan, P. L.; Ozanne, B. Cellular responsiveness to growth factors correlates with a cell’s ability to express the transformed phenotype. Cell 33:931–938; 1983.

    Article  PubMed  CAS  Google Scholar 

  12. Laemmli, U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685; 1973.

    Article  Google Scholar 

  13. Macpherson, I. Soft agar techniques. In: Kruse, P. F., Jr.; Patterson, M. K., eds. Tissue culture methods and applications. New York: Academic Press; 1973:276–280.

    Google Scholar 

  14. Massague, J. The TGF-beta family of growth and differentiation factors. Cell 49:437–438; 1987.

    Article  PubMed  CAS  Google Scholar 

  15. McClure, D. B. Anchorage-independent colony formation of SV40 transformed BALB/c-3T3 cells in serum-free medium: role of cell- and serum-derived factors. Cell 32:999–1006; 1983.

    Article  PubMed  CAS  Google Scholar 

  16. Miller, B. E.; Miller, F. R.; Heppner, G. H. Assessing tumor drug sensitivity by a new in vitro assay which preserves tumor heterogeneity and subpopulation interactions. J. Cell. Physiol. (Suppl.) 3:105–116; 1984.

    Article  CAS  Google Scholar 

  17. Miller, B. E.; Miller, F. R.; Leith, J., et al. Growth interaction in vivo between tumor subpopulations derived from a single mouse mammary tumor. Cancer Res 40:3977–3981; 1980.

    PubMed  CAS  Google Scholar 

  18. Moses, H. L.; Tucker, R. F.; Leof, E. B., et al. Type beta transforming growth factor is a growth stimulator and a growth inhibitor. In: Feramisco, J.; Ozanne, B.; Stiles, C.; eds. Cancer cells 3: growth factors and transformation. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory; 1985:65–71.

    Google Scholar 

  19. Nowell, P. C. The clonal evolution of tumor cell populations. Science 194:23–28; 1976.

    Article  PubMed  CAS  Google Scholar 

  20. Peehl, D. M.; Stanbridge, E. J. Anchorage-iddependent growth of normal human fibroblasts. Proc. Natl. Acad. Sci. USA 78:3053–3057; 1981.

    Article  PubMed  CAS  Google Scholar 

  21. Pircher, R.; Lawrence, D. A.; Jullien, P. Latent beta-transforming growth factor in nontransformed and Kirsten sarcoma virus-transformed normal rat kidney cells, clone 49F. Cancer Res. 44:5538–5543; 1984.

    PubMed  CAS  Google Scholar 

  22. Roberts, A. B.; Anzano, M. A.; Lamb, L. C., et al. Isolation from murine sarcoma cells of novel transforming growth factors potentiated by EGF. Nature 295:417–419; 1982.

    Article  PubMed  CAS  Google Scholar 

  23. Scatchard G. The attractions of proteins for small molecules and ions. Ann. NY Acad. Sci. 51:660–672; 1949.

    Article  CAS  Google Scholar 

  24. Schnebli, H. P.; Burger, M.; Strasser, F. F., et al. Marked instability of character of cloned epithelial cells. Exp. Cell Biol. 45:24–33; 1977.

    PubMed  CAS  Google Scholar 

  25. Schreiber, A. B.; Winkler, M. E.; Derynck, R. Transforming growth factor-alpha: a more potent angiogenic mediator than epidermal growth factor. Science 232:1250–1253; 1986.

    Article  PubMed  CAS  Google Scholar 

  26. Wray, W.; Boulikas, T.; Wray, V. P., et al. Silver staining of proteins in polyacrylamide gels. Anal. Biochem. 118:197–203; 1981.

    Article  PubMed  CAS  Google Scholar 

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The work was supported by a grant from the American Institute for Cancer Research, by American Cancer Society Institutional grant IN-119, by funds from the Poncin Trust (Seattle-First National Bank), and by grants CA-39611 and CA46885 from the National Institutes of Health, Bethesda, MD.

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Hamner, S., Jones, W., Starkey, J.R. et al. Growth factor interactions between mouse mammary cell lines cocultured in collagen gels. In Vitro Cell Dev Biol 25, 1107–1113 (1989). https://doi.org/10.1007/BF02621261

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