Skip to main content
Log in

Assessment of chemical carcinogen-induced transforming activity using BALB/c-3T3 cells

  • Published:
Journal of tissue culture methods

Summary

The BALB/c-3T3 cell transformation assay evaluates the morphologic transforming potential of test chemicals using cultured mammalian cells as targets. The assay is semiquantitative and employs a contact-inhibited clone of cells derived from the original BALB/c-3T3 mouse cell line. Transforming activity, or a focus, is recognized as a dense layer of morphologically altered cells superimposed on a monolayer of the normal cell phenotype. The induction of transforming activity in a standard assay has been reported to correlate well with the carcinogenicity of many test chemicals in rodent bioassay; however, the standard assay does not detect carcinogens of all chemical classes. This report describes an operational protocol for the standard BALB/c-3T3 cell transformation assay. In addition, it provides assay acceptance criteria and a discussion of a method to evaluate transforming activity data.

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

Access this article

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. Bailey, N. T. J. Statistical methods in biology. New York: John Wiley & Sons, Inc.; 1959:50.

    Google Scholar 

  2. Bertram, J. S. Effects of serum concentration on the expression of carcinogen-induced transformation in the C3H10T 1/2 CL8 cell line. Cancer Res. 37:514–523; 1977.

    PubMed  Google Scholar 

  3. Bertram, J. S. Reduction in the formation of carcinogen-induced transformed foci by pencillin G sodium in the C3H10T 1/2 CL8 line. Cancer Lett. 7:289–298; 1979.

    PubMed  Google Scholar 

  4. Brusick, D. Genotoxic effects in cultured mammalian cells produced by low pH treatment conditions and increased ion concentrations. Environ. Mutagen. Vol. 8, 879–886; 1986.

    PubMed  Google Scholar 

  5. DiPaolo, J. A.; Donovan, P. J.; Nelson, R. L. Quantitative studies of in vitro transformation by chemical carcinogens. JNCI 42:867–878; 1969.

    PubMed  Google Scholar 

  6. Dunkel, V. C.; Pienta, R. J.; Sivak, A., et al. Comparative neoplastic transformation responses of BALB/c-3T3 cells, Syrian hamster embryo cells and Rauscher murine virus-infected Fischer's 344 rat embryo cells to chemical carcinogens. JNCI 67:1303–1315; 1981.

    PubMed  Google Scholar 

  7. Fisher, R. A. Statistical methods for research workers, 13th edition. Edinburgh and New York: Hafer Publishing Co.; 1932:99.

    Google Scholar 

  8. Galloway, S. M.; Bean, C. L.; Armstrong, M. A., et al. False positive in vitro chromosome aberration tests with non-mutagens at high concentrations and osmolalites. Environ. Mutagen Soc. Abst. FC-4; 1985.

  9. Heidelberger, C.; Freeman, A. E.; Pienta, R. J., et al. Cell transformation by chemical agents—a review and analysis of the literature. Mutat. Res. 114:283–385; 1983.

    PubMed  Google Scholar 

  10. Kakunaga, T. A quantitative system for assay of malignant transformation by chemical carcinogens using a clone derived from BALB/c-3T3. Int. J. Cancer 12:463–473; 1973.

    PubMed  Google Scholar 

  11. Reznikoff, C. A.; Bertram, J. S.; Brankow, D. W., et al. Quantitative and qualitative studies to chemical transformation of cloned C3H mouse embryo cells sensitive to post confluence inhibition of cell division. Cancer Res. 33:3239–3249; 1973.

    PubMed  Google Scholar 

  12. Rubin, H.; Chu, B. M. Solute concentration effects on the expression of cellular heterogeneity of anchorage-independent growth among spontaneously transformed BALB/c-3T3 cells. In Vitro 20:585–596; 1984.

    PubMed  Google Scholar 

  13. Rundell, J. O.; Guntakatta, M.; Matthews, E. J. Criterion development for the application of BALB/c-3T3 cells to routine testing for chemical carcinogenic potential. In: Waters, Sandhu, Lewtas, et al., eds. Short-term bioassays in the analysis of complex environmental mixtures III. New York: Plenum Publishing Co.; 1983:302–327.

    Google Scholar 

  14. Rundell, J. O. In vitro transformation assays using mouse embryo cell lines: BALB/c-3T3 cells. In: Douglas, J. F., ed. Carcinogenesis and mutagenesis testing. Clifton, New Jersey: Humana Press; 1984:279–285.

    Google Scholar 

  15. Whorton, E. B.; Ward, J. B.; Morris, D. L. Experimental design and statistical analysis considerations for in vitro mammalian cell transformation assays with BALB/c-3T3 cells. Environ. Mutagen. 4:595–603; 1982.

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Matthews, E.J. Assessment of chemical carcinogen-induced transforming activity using BALB/c-3T3 cells. Journal of Tissue Culture Methods 10, 157–164 (1986). https://doi.org/10.1007/BF01405078

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01405078

Key words

Navigation