Transfection and Transformation of Human Thyroid Epithelial Cells

  • Nicholas Robert Lemoine
  • David Wynford-Thomas
Part of the Methods in Molecular Biology book series (MIMB, volume 5)


High efficiency gene transfer into mammalian fibroblasts is a routine procedure that can be performed by a variety of methods (1), but transfection of epithelial cells has been more difficult to achieve. Each of the standard techniques has associated problems; special equipment is required for electroporation (2) and also for direct microinjection (3). Protoplast fusion (4) is suitable only for cloned genes. Retroviral transduction of genes (5) is a method of high efficiency for animal cells, but again is applicable only to cloned genes of restricted size, and safety considerations are likely to restrict greatly the use of retroviral vectors suitable for human cells. Calcium phosphate coprecipitation has been used with success in several epithelial lines (6,7,8) and, more recently, strontium phosphate coprecipitation (9). These techniques are very simple to perform, require no special equipment, and can be applied to genomic DNA transfection.


Immunocytochemical Assay Mammalian Fibroblast High Efficiency Gene Transfer Transient Assay System Caesium Chloride 
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  1. 1.
    Pollard, J. W., Luqmani, Y., Bateson, A., and Chotai, K. (1984) DNA transformation of mammalian cells, in Methods in Molecular Biology, vol. 2 (Walker,J. M., ed.), Humana Press, NJ, pp. 321–332.Google Scholar
  2. 2.
    Tur-Kaspa, R., Teicher, L., Levine, B. J., Skoultchi, A. I., and Shafritz, D. A. (1986) Use of electroporation to introduce biologically active genes into primary rat hepatocytes. Mol. Cell Biol. 6, 716–718.PubMedGoogle Scholar
  3. 3.
    Garcia, I., Sordat, B., Rauccio-Farinon, E., Dunand, M., Kraehenbuhl, J.-P., and Diggelmann, H. (1986) Establishment of two rabbit mammary epithelial cell lines with distinct oncogenic potential and differentiated phenotype after microinjection of transforming genes. Mol. Cell Biol. 6, 1974–1982.PubMedGoogle Scholar
  4. 4.
    Yoakum, G. R, Lechner, J. F., Gabrielson, E. W., Korba, B. E., Malan-Shibley, L., Willey, J. C, Valerio, M. G., Shamsuddin, A. M., Trump, B. F., and Harris, C. C. (1985) Transformation of human bronchial epithial cells transfected by Harvey ras oncogene. Science 22, 1174–1179.CrossRefGoogle Scholar
  5. 5.
    Wolff, J. A., Yee J.-K., Skelly, H. F., Moores, J. C., Respess, J. G., Friedmann, T., and Leffert, H. (1987) Expression of retrovirally transduced genes in primary cultures of adult rat hepatocytes. Proc. Natl. Acad. Sci. USA 84, 3344–3348.PubMedCrossRefGoogle Scholar
  6. 6.
    Hynes, N. E., Jaggi, R., Kozma, S. C., Ball, R., Muellener, D., Wetherall, N. T., Davis, B. W., and Groner, B. (1985) New acceptor cell for transfected genomic DNA: oncogene transfer into a mouse mammary epithelial cell line. Mol. Cell Biol. 5, 268–272.PubMedGoogle Scholar
  7. 7.
    Storer, R. D., Stein, R. B., Sina, J. F., DeLuca, J. G., Allen, H. L., and Bradley, M. O. (1986) Malignant transformation of a preneoplastic hamster epidermal cell line by the EJ c-Ha-ras oncogene. Cancer Res. 46, 1458–1464.PubMedGoogle Scholar
  8. 8.
    Summerhayes, I. C., Malone, P., and Visvanathan, K. (1986) Altered growth properties and cell surface changes in ras transformed mouse bladder epithelium. Int. J. Cancer 37, 233–240.PubMedCrossRefGoogle Scholar
  9. 9.
    Brash, D. E., Reddel, R. R., Quanrud, M., Yang, K., Farrell, M. P., and Harris, C. C. (1987) Strontium phosphate transfection of human cells in primary culture: stable expression of the simian virus 40 large-T-antigen gene in primary human bronchial epithelial cells. Mot Cell Biol. 7, 2031–2034.Google Scholar
  10. 10.
    Lemoine, N. R. and Wynford-Thomas, D. (1987) The thyroid follicular cell as a recepient for DNA transfection. Br. J. Cancer 55, 342.CrossRefGoogle Scholar
  11. 11.
    Boffey, S. A. (1984) Plasmid DNA isolation by the cleared lysate method, in Methods in Molecular Biology, vol. 2 (Walker, J. M., ed.), Humana Press, pp. 177–183.Google Scholar
  12. 12.
    Kaiser, K. and Murray, N. E. (1985) The use of phage lambda replacement vectors in the construction of representative genomic DNA libraries, in DNA Cloning, vol. 1 (Glover, D. M., ed.), IRL, pp. 1–74.Google Scholar
  13. 13.
    Gluzman, Y., Sambrook, J. F., and Frisque, R. J. (1980) Expression of early genes of origin-defective mutants of SV40. Proc. Natl. Acad. Sci. USA 77, 3898–3902.PubMedCrossRefGoogle Scholar

Copyright information

© The Humana Press Inc. 1990

Authors and Affiliations

  • Nicholas Robert Lemoine
    • 1
  • David Wynford-Thomas
    • 1
  1. 1.University of Wales College of MedicineHeath ParkUK

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