Abstract
With over 3000 cancer cell lines described in the literature and thousands in regular use, it has become essential to characterize and authenticate cell-line models. Characterization of the properties of cell lines is important for a number of reasons. First, the relationship of the cell line to the cells of origin should be established to confirm that the cell line is derived from and is representative of its tissue of origin. If the cell line is to have any value as a model it should reflect the properties of the cell type from which it was derived. For example, for a cell line established from a breast carcinoma it is helpful to show that the cell line has characteristics consistent with breast and epithelial origin. Although the genetic profile should remain constant, expression may change and features such as differentiation characteristics may be lost over time in culture. Similarly, as the culture develops, certain clones may emerge with selection and predominate. Particularly important is the need to check for purity and potential cross-contamination with other cell lines. The history of cell culture indicates that cross-contamination between cell lines is widely prevalent and continues to be an ongoing problem (1–3). During the 1970s and 1980s, multiple studies initiated by Stanley Gartler and Walter Nelson-Rees demonstrated that one in three cell lines were either contaminated or even totally replaced by other cell lines (4–8). The most frequent contaminant was the HeLa cervical carcinoma cell line which had been established in 1951 (9) and had been widely distributed to many research laboratories. As a result of its rapid growth rate, once mixed with other cell lines it would generally outgrow them. Unfortunately this problem has not disappeared, and contamination continues to be widespread (1,2,10). This area is covered in more detail in Chapter 30.
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References
O’Brien, S. J. (2000) Cell culture forensics. Proc. Natl. Acad. Sci. USA 98, 7656–7658.
Stacey, G. N., Masters, J. R. W., Hay, R. J., Drexler, H. G., Macleod, R. A. F., and Freshney, R. I. (2000) Cell contamination leads to inaccurate data: we must take action now. Nature 403, 356.
Macleod, R. A. F., Dirks, W. G., and Drexler, H. G. (2002) Persistent use of misidentified cell lines and its prevention. Genes Chromosomes Cancer 33, 103–105.
Gartler, S. M. (1968) Apparent HeLa contamination of human heterodiploid cell lines. Nature 217, 750–751.
Nelson-Rees, W. A., Flandermeyer, R. R., and Hawthorne P. K. (1974) Banded marker chromosomes as indicators of intraspecies cellular contamination. Science 184, 1093–1096.
Nelson-Rees, W. A., and Flandermeyer, R. R. (1976) HeLa cultures defined. Science 191, 96–98.
Nelson-Rees, W. A., Daniels, D. W., and Flandermeyer, R. R. (1981) Cross-contamination of cells in culture. Science 212, 446–452.
Nelson-Rees, W. A. and Flandermeyer, R. R. (1977). Inter-and intraspecies contamination of human breast tumor cell lines HBC and BrCa5 and other cell cultures. Science 195, 1343–1344.
Gey, G. O., Coffman, W. D., and Kubicek, M. T. (1952) Tissue culture studies of the proliferative capacity of cervical carcinoma and normal epithelium. Cancer Res. 12, 264–265.
MacLeod, R. A. F., Dirks, W. G., Matsuo, Y., Kaufmann, M., Milch, H., and Drexler, H. G. (1999) Widespread intraspecies cross-contamination of human tumor cell lines arising at source. Int. J. Cancer 83, 555–563.
UKCCCR (2000) UKCCCR guidelines for the use of cell lines in cancer research. Brit. J. Cancer 82, 1495–15091.
Furlong, M. T., Hough, C. D., Sherman-Baust, C. A., Pizer, E. S., and Morin, P. J. (1999) Evidence for the colonic origin of ovarian cancer cell line SW626. J. Natl. Cancer Inst. 91, 1327–1328.
Masters, J. R. (2002) HeLa cells 50 years on: the good, the bad and the ugly. Nat. Rev. Cancer 2, 315–319.
Gilbert, D. A., Reid, Y. A., Gail, M. H., Pee, D., White, C., Hay, R. J., et al. Application of DNA fingerprinting for cell-line individualization. Am. J. Hum. Genet. 47, 499–514.
Stacey, G., Bolton, B., Doyle, A., and Griffiths, B. (1992) DNA fingerprinting—a valuable new technique for the characterization of cell lines. Cytotechnol 9, 211–216.
Stacey, G. N., Bolton, B. J., and Doyle, A. (1991) The quality control of cell banks using DNA fingerprinting. EXS 58, 361–370.
Jeffreys, A. J., Wilson, V., and Thein, S. L. (1985) Hypervariable “minisatellite” regions in human DNA. Nature 314, 67–73.
Jeffreys, A. J., Wilson, V., and Thein, S. L. (1985) Individual-specific “fingerprints” of human DNA. Nature 314, 76–79.
Masters, J. R., Thomson, J. A., Daly-Burns, B. et al., (2001) Short tandem repeat profiling provides an international reference standard for human cell lines. Proc. Natl. Acad. Sci. USA 98, 8012–8017.
Rowley, J. D. (1973) A new consistent chromosomal abnormality in chronic myelogenous leukaemia identified by quinacrine fluorescence and Giemsa staining. Nature 243, 290–293.
Seabright, M. (1973) Improvement of trypsin method for banding chromosomes. Lancet 1, 1249–1250.
Caspersson, T., Zech, L., and Johansson, C. (1970) Differential binding of alkylating fluorochromes in human chromosomes. Exp. Cell Res. 60, 315–319.
Pardue, M. L. and Gall, J. G. (1970) Chromosome localization of mouse satellite DNA. Science 168, 1356–1358.
Dutrillaux, B., Finaz, C., de Grouchy, J. and Lejeune, J. (1972) Comparison of banding patterns of human chromosomes obtained with heating, fluorescence, and proteolytic digestion. Cytogenetics 11, 113–116.
Lichter, P. (1997) Multicolor FISHing: what’s the catch. Trends Genet 13, 475–479.
Speicher, M. R., Ballard, S. G., and Ward, D. C. (1996) Karyotyping human chromosomes by combinatorial multifluor FISH. Nature Genet. 12, 368–375.
Schrock, E., du Manoir, S., Veldman, T. et al. (1996) Multicolor spectral karyotyping of human chromosomes. Science 273, 494–497.
Henegariu, O., Heerema, N. A., Bray-Ward, P. et al. (1999) Colour-changing Karyotyping: an alternative to M-FISH/SKY. Nat. Genet. 23, 263–264.
Tanke, H. J., Wiegant, J., van Gijlswijk, R. P. M. et al. (1999) New strategy for multicolour fluorescence in situ hybridisation: COBRA: combined binary ratio labeling. Eur. J. Hum. Genet. 7, 2–11.
O’Brien, S. J., Kleiner, G., Olson, R., and Shannon, J. E. (1977) Enzyme polymorphisms as genetic signatures in human cell cultures. Science 195, 1345–1348.
O’Brien, S. J., Shannon, J. E., and Gail, M. H. (1980) A molecular approach to the identification and individualization of human and animal cells in culture: isozyme and allozyme genetic signatures. In Vitro 16, 119–135.
Nims, R. W., Shoemaker, A. P., Bauernschub, M. A., Rec, L. J., and Harbell, J. W. (1998) Sensitivity of isoenzyme analysis for the detection of interspecies cell line cross-32. contamination. In Vitro Cell. Dev. Biol. Anim. 34, 35–39.
Steube, K. G., Grunicke, D., and Drexler, H. G. (1995) Isoenzyme analysis as a rapid method for the examination of the species identity of cell cultures. In Vitro Cell. Dev. Biol. Anim. 31, 115–119.
Lin, M. A., Latt, S. A., and Davidson, R. L. (1974) Identification of human and mouse chromosomes in human-mouse hybrids by centromere fluorescence. Exp. Cell Res. 87, 429–433.
Friend, K. K., Dorman, B. P., Kucherlapati, R. S., and Ruddle, F. H. (1976) Detection of interspecies translocations in mouse-human hybrids by alkaline Giemsa staining. Exp. Cell Res. 99, 31–36.
Stulberg, C. S. (1973) Extrinsic cell contamination of tissue culture. In: Fogh J (ed). Contamination in Cell Cultures, Academic Press, New York, pp. 1–23.
Thacker, J. (1993) Fingerprinting of mammalian cell lines with a single PCR primer. Biotechniques 16, 252–253.
Nelson, D. L., Ledbetter, S. A., Corbo, L. et al. (1989) Alu polymerase chain reaction: A method for rapid isolation of human-specific sequences from complex DNA sources. Proc. Natl. Acad. Sci. USA 86, 6686–6690.
Puts J. J., Vooijs G. P., Huysmans A. et al. (1986) Cytoskeletal proteins as tissue-specific markers in cytopathology. Exp. Cell Biol. 54, 73–79.
Quentmeier H., Osborn M., Reinhardt J., et al. (2001) Immunocytochemical analysis of cell lines derived from solid tumors. J. Histochem. Cytochem. 49, 1369–1378
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Langdon, S.P. (2004). Characterization and Authentication of Cancer Cell Lines. In: Langdon, S.P. (eds) Cancer Cell Culture. Methods in Molecular Medicine™, vol 88. Humana Press. https://doi.org/10.1385/1-59259-406-9:33
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DOI: https://doi.org/10.1385/1-59259-406-9:33
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