Summary
The generation of cell strains and established cell lines from human papillomavirus (HPV)-infected cervical biopsies and ano-genital warts is best achieved by the application of conventional protocols for keratinocyte cell culture. The optimal protocol that permits growth at clonal density and serial cultivation involves the use of an inactivated 3T3 fibroblast feeder layer and medium supplemented with hydrocortisone and fetal bovine serum. Modifications in terms of additives and serum concentration are required for optimal culture of neoplastic and malignant HPV-containing keratinocytes.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Stanley, M. A. (2001) Pathobiology of human papillomaviruses. In Viruses, Cell Transformation and Cancer (Grand, R. A., ed.), Elsevier, Amsterdam: pp. 129–144.
De Villiers, E. M. (1994) Human pathogenic papillomavirus types: an update. Curr. Top. Microbiol. Immunol. 186, 1–12.
zur Hausen, H. (1999) Immortalization of human cells and their malignant conversion by high risk human papillomavirus genotypes. Semin. Cancer Biol. 9, 405–411.
Rheinwald, J. G. and Green, H. (1977) Epidermal growth factor and the multiplication of cultured human keratinocytes. Nature 265, 421–424.
Green, H. (1978) Cyclic AMP in relation to the epidermal cell: a new view. Cell 15, 801–811.
Green, H., Kehinde, O., and Thomas, J. (1979) Growth of cultured human epidermal cells into multiple epithelia suitable for grafting. Proc. Natl. Acad. Sci. USA 76, 5665–5668.
Taichmann, L., Reilly, S., and Garant, P. R. (1979) In vitro cultivation of human oral keratinocytes. Arch. Oral Biol. 24, 258–268.
Stanley, M. A. and Parkinson, E. K. (1979) Growth requirements of human cervical epithelial cells in culture. Int. J. Cancer 24, 407–414.
Rheinwald, J. G. and Green, H. (1975) Serial cultivation of strains of human epidermal keratinocytes: the formation of keratinizing colonies from single cells. Cell 6, 331–344.
Wilbanks, G. D. and Fink, C. G. (1976) Tissue and organ culture of cervical epithelium. In The Cervix (Jordan, J. A. and Singer, A., eds.), W.B. Saunders & Co., London, pp. 429–441.
Stanley, M. A. and Dahlenburg, K. (1984) Factors controlling the growth of human cervical epithelium in culture. In Vitro 20, 144–151.
Peehl, D. M. and Ham, R. G. (1980) Growth and differentiation of human keratinocytes without a feeder layer or conditioned medium. In Vitro 16, 516–525.
Tsao, M. C., Walthall, B. J., and Ham, R. G. (1982) Clonal growth of normal human epidermal keratinocytes in a defined medium. J. Cell Physiol. 110, 219–229.
Boyce, S. T. and Ham, R. G. (1986) Normal human epidermal keratinocytes. In In Vitro Models for Cancer Research (Webber, M. M. and Sekely, L. I., eds.), CRC Press, Boca Raton, FL, pp. 211–232.
Fusenig, N. E., Amer, S. M., Boukamp, P., and Worst, P. K. (1978) Characteristics of chemically transformed mouse epidermal cells in vitro and in vivo. Bull. Cancer 65, 271–279.
Lillie, J. H., MacCallum, D. K., and Jepsen, A. (1980) Fine structure of subcultivated stratified squamous epithelium grown on collagen rafts. Exp. Cell Res. 125, 153–165.
Kopan, R., Traska, G., and Fuchs, E. (1987) Retinoids as important regulators of terminal differentiation: examining keratin expression in individual epidermal cells at various stages of keratinization. J. Cell Biol. 105, 427–439.
Asselineau, D., Bernard, B. A., Bailly, C., Darmon, M., and Prunieras, M. (1986) Human epidermis reconstructed by culture: is it “normal”? J. Invest. Dermatol. 86, 181–186.
Meyers, C. and Laimins, L. A. (1994) In vitro systems for the study and propagation of human papillomaviruses. Curr. Top. Microbiol. Immunol. 186, 199–215.
Stanley, M. A., Browne, H. M., Appleby, M., and Minson, A. C. (1989) Properties of a non tumorigenic human cervical keratinocyte cell line. Int. J. Cancer 43, 672–676.
Sterling, J., Stanley, M., Gatward, G., and Minson, T. (1990) Production of human papillomavirus type 16 virions in a keratinocyte cell line. J. Virol. 64, 6305–6307.
Alazawi, W., Pett, M., Arch, B., et al. (2002) Changes in cervical keratinocyte gene expression associated with integration of human papillomavirus 16. Cancer Res. 62, 6959–6965.
Bedell, M. A., Hudson, J. B., Golub, T. R., et al. (1991) Amplification of human papillomavirus genomes in vitro is dependent on epithelial differentiation. J. Virol. 65, 2254–2260.
Meyers, C., Frattini, M. G., Hudson, J. B., and Laimins, L. A. (1992) Biosynthesis of human papillomavirus from a continuous cell line upon epithelial differentiation. Science 257, 971–973.
Hietanen, S., Auvinen, E., Grenman, S., et al. (1992) Isolation of two keratinocyte cell lines derived from HPV positive dysplastic vaginal lesions. Int. J. Cancer 52, 391–398.
Peitsaro, P., Hietanen, S., Johansson, B., Lakkala, T., and Syrjanen, S. (2002) Single copy heterozygote integration of HPV 33 in chromosomal band 5p14 is found in an epithelial cell clone with selective growth advantage. Carcinogenesis 23, 1057–1064.
Grassmann, K., Rapp, B., Maschek, H., Petry, K. U., and Iftner, T. (1996) Identification of a differentiation inducible promoter in the E7 open reading frame of human papillomavirus type 16 (HPV-16) in raft cultures of a new cell line containing high copy numbers of episomal HPV-16 DNA. J. Virol. 70, 2339–2349.
Kelland, L. R., Burgess, L., and Steel, G. G. (1987) Characterization of four new cell lines derived from human squamous carcinomas of the uterine cervix. Cancer Res. 47, 4947–4952.
Pal, A. K., Pratap, M., and Mitra, A. B. (1992) Cervix tumour cell line established from tumour after long term passages as heterotransplant in nude mice. Indian J. Exp. Biol. 30, 655–656.
Waggoner, S. E. and Wang, X. (1994) Effect of nicotine on proliferation of normal, malignant, and human papillomavirus transformed human cervical cells. Gynecol. Oncol. 55, 91–95.
Ho, T. H., Chew, E. C., Tam, J. S., et al. (1993) Biological characteristics of a newly established human cervical carcinoma cell line. Anticancer Res. 13, 967–971.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2005 Humana Press Inc.
About this protocol
Cite this protocol
Stanley, M.A. (2005). Establishing HPV-Containing Keratinocyte Cell Lines From Tissue Biopsies. In: Davy, C., Doorbar, J. (eds) Human Papillomaviruses. Methods in Molecular Medicine, vol 119. Humana Press. https://doi.org/10.1385/1-59259-982-6:129
Download citation
DOI: https://doi.org/10.1385/1-59259-982-6:129
Publisher Name: Humana Press
Print ISBN: 978-1-58829-373-2
Online ISBN: 978-1-59259-982-0
eBook Packages: Springer Protocols