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Continuous growth of human tumor cell lines in serum-free media

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Summary

Five human tumor cell lines were studied for growth factor requirements and for replication in serum-free media. Of the five tumor lines HT-29 (colon carcinoma), TWI (melanoma), A-549 (lung carcinoma), Panc-1, (carcinoma of the pancreas) and EJ, (bladder carcinoma) only HT-29 and TWI grew in the serum-free medium (SFM). In a series of additional experiments, a combination of transferrin (5 μg/ml), insulin (5 μg/ml), triiodothyronine (2×10−10 M), epidermal growth factor (20 ng/ml), and selenium (5 ng/ml) was added to Chee’s essential medium (CEM) without serum (C-TITES medium). The C-TITES modification of CEM was found to allow optimal replication of HT-29 and TWI cells. Both HT-29 and TWI cells have replicated continuously in C-TITES medium for periods of more than 15 mo. These cells replicate with slightly lower doubling times than in CEM supplemented with 10% fetal bovine serum. Deletion of insulin or transferrin from the C-TITES medium resulted in cessation of cell growth of HT-29 and TWI. HT-29 assumed a somewhat rounded morphology, whereas TWI grew with the characteristic fibroblastic morphology in C-TITES medium. Cell line EJ did not grow in C-TITES medium. The other two cell lines, A-549 and Panc-1, grew in C-TITES medium but their growth rate was much slower than that in SSM. Availability of cell lines that can be propagated in serum-free, hormone-supplemented medium may aid in the study of the mechanisms by which hormones influence cell growth.

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References

  1. Antoniades, H. N.; Owen, A. J. Growth factors and regulation of cell growth. Ann. Rev. Med. 33:445–463; 1982.

    Article  PubMed  CAS  Google Scholar 

  2. Barnes, D.; Sato, G. Serum-free culture: a unifying approach. Cell 22:649–655; 1980.

    Article  PubMed  CAS  Google Scholar 

  3. 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 

  4. Barnes, D.; van der Bosch, J.; Masui, H.; Miyazaki, K., et al. The culture of human tumor cells in serum-free medium. Methods Enzymol. 79:368–391; 1981.

    Article  PubMed  CAS  Google Scholar 

  5. Barnes, D. W.; Sirbasku, D. A.; Sato, G. H., eds. Methods of serum-free culture of epithelial and fibroblastic cells. In: Cell culture methods for molecular and cell biology, vol. 3. New York: Alan R. Liss, Inc.; 1984.

    Google Scholar 

  6. Carney, D. N.; Bunn, P. A.; Gazdar, A. F., et al. Selective growth in serum-free hormone-supplemented medium of tumor cells obtained by biopsy from patients with small cell carcinoma of the lung. Proc. Natl. Acad. Sci. USA 78:3185–3189; 1981.

    Article  PubMed  CAS  Google Scholar 

  7. Cezard, J. P.; Forgue-Lafitte, M. E.; Chamblier, M. C., et al. Growth-promoting effect, biological activity and binding of insulin in human intestinal cancer cells in culture. Cancer Res. 41:1148–1153; 1981.

    PubMed  CAS  Google Scholar 

  8. Faulk, W. P.; Hsi, B. L.; Stevens, P. J. Transferrin and transferrin receptors in carcinoma of the breast. Lancet 2(8191), 390–392; 1980.

    PubMed  CAS  Google Scholar 

  9. Fogh, J.; Trempe, G. New human tumor cell lines. In: Fogh, J., ed. Human tumor cells in vitro. New York: Plenum Press; 1975:115–159.

    Google Scholar 

  10. Forgue-Lafitte, M. E.; Horvat, A.; Rosselin, G. E. Insulin binding by a cell line (HT-29) derived from human colonic cancer. Mol. Cell Endocrinol. 14:123–130; 1979.

    Article  PubMed  CAS  Google Scholar 

  11. Giard, D. J.; Aaronson, S. A.; Todaro, J. G., et al. In vitro cultivation of human tumors: Establishment of cell lines derived from a series of solid tumors. JNCI 51:1417–1423; 1973.

    PubMed  CAS  Google Scholar 

  12. Gospodarowicz, D.; Moran, J. S. Growth factors in mammalian cell culture. Ann. Rev. Biochem. 45:531–557; 1976.

    Article  PubMed  CAS  Google Scholar 

  13. Hayashi, I.; Sato, G. H. Replacement of serum by hormones permits growth of cells in defined medium. Nature 259:132–134; 1976.

    Article  PubMed  CAS  Google Scholar 

  14. Lieber, M.; Mazzetta, J.; Nelson-Rees, W., et al. Establishment of a continuous tumor-cell line (Panc-1) from a human carcinoma of the exocrine pancreas. Int. J. Cancer 15:741–747; 1975.

    Article  PubMed  CAS  Google Scholar 

  15. Lin, C-W.; Lin, J. C.; Prout, G. R. Establishment and characterization of four human bladder tumor cell lines with different degrees of malignancy. Proc. Am. Assoc. Cancer Res. 25:31; 1984.

    Google Scholar 

  16. Mather, J. P.; Sato, G. H. The use of hormone-supplemented serum-free media in primary culture. Exp. Cell Res. 124:215–221; 1979.

    Article  PubMed  CAS  Google Scholar 

  17. Messing, E. M.; Fahey, J. L.; DeKernion, J. B., et al. Serum-free medium for the in vitro growth of normal and malignant urinary bladder epithelial cells. Cancer Res. 42:2392–2397; 1982.

    PubMed  CAS  Google Scholar 

  18. Murakami, H.; Masui, H. Hormonal control of human colon carcinoma cell growth in serum-free medium. Proc. Natl. Acad. Sci. USA 77:3464–3468; 1980.

    Article  PubMed  CAS  Google Scholar 

  19. Pichon, J.; Marchis-Mouren, G.; Marvaldi, J. Growth of a human colonic adenocarcinoma cell line in a serum-free medium: Effect of the vasointestinal peptide (VIP). 31st Falk symposium. In: Colonic carcinogenesis. Lancaster, England: MT Press; 1982:297–307.

    Google Scholar 

  20. Reid, L.; Sato, G. Replacement of serum in cell culture by hormones. In: Rickenberg, H. V., ed. Biochemistry and mode of action of hormones II. Baltimore, MD: University Park Press; 1978:219–251.

    Google Scholar 

  21. Simms, E.; Gazdar, A. F.; Abrams, P. G., et al. Growth of human small cell (oat cell) carcinoma of the lung in serum-free growth factor supplemented medium. Cancer Res. 40:4356–4363; 1980.

    PubMed  CAS  Google Scholar 

  22. Sutherland, R.; Delia, D.; Schneider, C., et al. Ubiquitous cell-surface glycoproteins on tumor cells in proliferation associated receptor for transferrin. Proc. Natl. Acad. Sci. USA 78:4515–4519; 1981.

    Article  PubMed  CAS  Google Scholar 

  23. Trowbridge, I. S.; Omary, M. B. Human cell-surface glycoprotein related to cell proliferation is the receptor for transferrin. Proc. Natl. Acad. Sci. USA 78:3039–3043; 1981.

    Article  PubMed  CAS  Google Scholar 

  24. Turner, M. W.; Hulme, B., eds. The plasma proteins; an introduction. London: Pitman; 1971.

    Google Scholar 

  25. Uittenbogaat, C. H.; Cantor, Y.; Fahey, J. L. Growth of human malignant lymphoid cell lines in serum-free medium. In Vitro 19:67–72; 1983.

    Article  Google Scholar 

  26. van der Bosch, J.; Masui, H.; Sato, G. Growth characteristics of primary tissue cultures from heterotransplanted human colorectal carcinomas in serum-free medium. Cancer Res. 41:611–618; 1981.

    PubMed  Google Scholar 

  27. Yang, J.; Guzman, R.; Richard, J., et al. Growth factors and cyclic nucleotide-induced proliferation of normal and malignant mammary epithelial cells in primary culture. Endocrinology 107:35–41; 1980.

    Article  PubMed  CAS  Google Scholar 

  28. Young, D. A.; Prout, G. R.; Lin, C-W. Production and characterization of a mouse monoclonal antibody to a human bladder tumor cell surface antigen. Proc. Am. Assoc. Cancer Res. 25:255; 1984.

    Google Scholar 

  29. Zirvi, K. A.; van der Bosch, J.; Masui, H., et al. Activity of cancer chemotherapeutic agents against human colorectal carcinomas grown as primary tissue culture. (Submited).

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Authors and Affiliations

  1. Surgical Service, VA Medical Center, 07109, East Orange, New Jersey

    Karimullah A. Zirvi & George J. Hill

  2. Scott Oncology Laboratories, 02823, West Warwick, Rhode Island

    Darwin O. Chee

  3. Department of Surgery, Room G595, Division of Surgical Oncology, UMDNJ-New Jersey Medical School, 100 Bergen Street, 07103, Newark, NJ

    Karimullah A. Zirvi & George J. Hill

Authors
  1. Karimullah A. Zirvi
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  2. Darwin O. Chee
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  3. George J. Hill
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Additional information

This work was supported by Veterans Administration Research Awards to two of the authors (Karimullah A. Zirvi and George J. Hill) and grant no. CA-37138 from the National Cancer Institute.

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Zirvi, K.A., Chee, D.O. & Hill, G.J. Continuous growth of human tumor cell lines in serum-free media. In Vitro Cell Dev Biol 22, 369–374 (1986). https://doi.org/10.1007/BF02623524

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  • DOI: https://doi.org/10.1007/BF02623524

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