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Effects of defined medium, fetal bovine serum, and human serum on growth and chemosensitivities of human breast cancer cells in primary culture: Inference for in vitro assays

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Summary

We compared the effects of defined medium, fetal bovine serum (FBS) and human serum (HuS) on the growth and responses to chemotherapeutic agents of human breast cancer cells in primary culture. Normal and tumor tissues were dissociated to small aggregates and single cells and seeded onto collagen-gel-coated wells in defined medium or medium supplemented with 5% FBS or 5% HuS. In all cases examined, defined medium and medium containing HuS were superior to medium containing FBS in supporting growth of both normal and tumor cell cultures. However, cultures in defined medium showed an initial cell loss. Cells from the same tumor cultured in different media varied in their responses to chemotherapeutic agents. In light of these results, medium supplemented with HuS, which promoted attachment of these cells in culture and stimulated their growth, should be the most appropriate nutrient environment for determining the effects of therapeutic agents on cells as it most closely resembles the in vivo situation. Because there were also variations in growth rates and chemosensitivities of tumor cells cultured in different human serum samples, we suggest that optimal conditions in which to culture these cells include the serum of the patient whose tumor is removed. This serum may provide host factors that influence cell growth and interact with exogenous factors.

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References

  1. Angello, J. C.; Danielson, K. G.; Anderson, L. W., et al. Glycosaminoglycan synthesis by subpopulations of epithelial cells from a mammary adenocarcinoma. Cancer Res. 42:2207–2210; 1982.

    PubMed  CAS  Google Scholar 

  2. Barnes, D.; Sato, G. Methods for growth of cultured cells in serum-free medium. Anal. Biochem. 102:255–270; 1980.

    Article  PubMed  CAS  Google Scholar 

  3. Bertelsen, C. A.; Sondak, V. K.; Mann, B. D., et al. Chemosensitivity testing of human solid tumors. A review of 1582 assays with 258 clinical correlations. Cancer 53:1240–1245; 1984.

    Article  PubMed  CAS  Google Scholar 

  4. 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  PubMed  CAS  Google Scholar 

  5. 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 

  6. Emerman, J. T.; Worth, A. J. Phenotypic stability of mouse mammary tumor cells cultured on collagen gels. In Vitro 21:49–56; 1985.

    CAS  Google Scholar 

  7. Haeuptle, M.; Suard, Y. L. M.; Bogenmann, E., et al. Effect of cell shape change on the function and differentiation of rabbit mammary cells in culture. J. Cell Biol. 96:1425–1434; 1983.

    Article  PubMed  CAS  Google Scholar 

  8. Hammond, S. L.; Ham, R. G.; Stampfer, M. R. Serum-free growth of human mammary epithelial cells: Rapid clonal growth in defined medium and extended serial passage with pituitary extract. Proc. Natl. Acad. Sci. USA 81:5435–5439; 1984.

    Article  PubMed  CAS  Google Scholar 

  9. Heppner, G.; Miller, B.; Cooper, D. N., et al. Growth interactions between mammary tumor cells. In: Brenner, N. J.; McGrath, C. M.; Rich, M. A., eds. Cell biology of breast cancer. New York: Academic Press; 1981:161–172.

    Google Scholar 

  10. Honn, K. V.; Singley, J. A.; Chavin, W. Fetal bovine serum: a multivariable standard. Proc. Soc. Exp. Biol. Med. 149:344–347; 1975.

    PubMed  CAS  Google Scholar 

  11. Jones, C. A.; Tsukamoto, T.; O’Brien, P. C., et al. Soft agarose culture human tumour colony forming assay for drug sensitivity testing: [3H]Thymidine incorporation vs colony formation. Br. J. Cancer 52:303–310; 1985.

    PubMed  CAS  Google Scholar 

  12. Kirkland, W. L.; Yang, N. S.; Jorgensen, T., et al. Growth of normal and malignant human mammary epithelial cells in culture. JNCI 63:29–41; 1979.

    PubMed  CAS  Google Scholar 

  13. Kraehenbuhl, J. P. Dispersed mammary gland epithelial cells: isolation and separation procedures. J. Cell Biol. 72:390–405; 1977.

    Article  PubMed  CAS  Google Scholar 

  14. Labarca, C.; Paigen, K. A simple, rapid, and sensitive DNA assay procedure. Anal. Biochem. 102:344–352; 1980.

    Article  PubMed  CAS  Google Scholar 

  15. Lawler, E. M.; Miller, F. R.; Heppner, G. H. Significance of three-dimensional growth patterns of mammary tissues in collagen gels. In Vitro 19:600–610; 1983.

    Article  PubMed  CAS  Google Scholar 

  16. Matsushima, Y.; Kanzawa, F.; Hoshi, A., et al. Time-schedule dependency of the inhibiting activity of various anticancer drugs in the clonogenic assay. Cancer Chemother. Pharmacol. 14:104–107; 1985.

    Article  PubMed  CAS  Google Scholar 

  17. McGrath, C. M. Augmentation of the response of normal mammary epithelial cells to estradiol by mammary stroma. Cancer Res. 43:1355–1360; 1983.

    PubMed  CAS  Google Scholar 

  18. Miller, F. R.; Medina, D.; Heppner, G. H. Preferential growth of mammary tumors in intact mammary fatpads. Cancer Res. 41:3863–3867; 1981.

    PubMed  CAS  Google Scholar 

  19. Morgan, D.; Freshney, R. I.; Darling, J. L., et al. Assay of anticancer drugs in tissue culture: cell cultures of biopsies from human astrocytoma. Br. J. Cancer 47:205–214; 1983.

    PubMed  CAS  Google Scholar 

  20. Page, M. J.; Field, J. K.; Everett, N. P., et al. Serum regulation of the estrogen responsiveness of the human breast cancer cell line MCF-7. Cancer Res. 43:1244–1250; 1983.

    PubMed  CAS  Google Scholar 

  21. Pigott, D. A.; Grimaldi, M. A.; Dell’aquila, M. L., et al. Growth inhibitors in plasma derived human serum. In Vitro 18:617–625; 1982.

    PubMed  CAS  Google Scholar 

  22. Richards, J.; Pasco, D.; Yang, J., et al. Comparison of the growth of normal and neoplastic mouse mammary cells on plastic, on collagen gels and in collagen gels. Exp. Cell Res. 146:1–14; 1983.

    Article  PubMed  CAS  Google Scholar 

  23. Roobol, C.; Sips, H. C. M.; Theunissen, J., et al. In vitro assessment of cytotoxic agents in murine cancers: comparison between antiproliferative and antimetabolic assays. JNCI 72:661–666; 1984.

    PubMed  CAS  Google Scholar 

  24. Sato, G. The role of serum in culture. In: Litwak, G., ed. Biochemical action of hormones, vol. 3. New York: Academic Press; 1975:391–396.

    Google Scholar 

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

    PubMed  CAS  Google Scholar 

  26. Yang, J.; Larson, L.; Flynn, D., et al. Serum-free primary culture of human normal mammary epithelial cells in collagen gel matrix. Cell Biol. Int. Rep. 6:969–975; 1982.

    Article  PubMed  CAS  Google Scholar 

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This work was supported by a grant from the National Cancer Institute of Canada and funds contributed by Mr. B. T. Wharton in memory of his wife, Nadia. J. T. Emerman is a research scholar of the National Cancer Institute of Canada.

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Emerman, J.T., Fiedler, E.E., Tolcher, A.W. et al. Effects of defined medium, fetal bovine serum, and human serum on growth and chemosensitivities of human breast cancer cells in primary culture: Inference for in vitro assays. In Vitro Cell Dev Biol 23, 134–140 (1987). https://doi.org/10.1007/BF02623594

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  • DOI: https://doi.org/10.1007/BF02623594

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