Abstract
Cytogenetic analysis forms an essential part of characterizing and identifying cell lines, in particular those established from tumors. In addition, karyotypic analysis can be used to distinguish individual subclones and to monitor stability. This chapter describes basic cytogenetic procedures suited to cells in continuous culture. The provision of unlimited material by cell lines encourages an heuristic approach to harvesting and hypotonic treatments to yield metaphase chromosome slide preparations of improved quality suitable for subsequent banding and fluorescence in situ hybridization (FISH) analysis. The experience of the writers with more than 500 different cell lines has shown that no single hypotonic harvesting protocol is adequate to consistently deliver satisfactory chromosome preparations. Thus, evidence-based protocols are described for hypotonic harvesting, rapid G-banding, and FISH analysis of cell cultures to allow troubleshooting and fine-tuning to suit the requirements of individual cell lines.
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References
Drexler, H. G. (2001) The Leukemia-Lymphoma Cell Line FactsBook, Academic, London, p. 2.
Heim, S. and Mitelman, F. (1995) Cancer Cytogenetics, 2nd ed., Wiley-Liss, New York, passim.
Reya, T., Morrison, S. J., Clarke, M. F., and Weissman, I. L. (2001) Stem cells, cancer, and cancer stem cells. Nature 414, 105–111.
Drexler, H. G., MacLeod, R. A. F., and Uphoff, C. C. (1999) Leukemia cell lines: in vitro models for the study of Philadelphia chromosome-positive leukemia. Leukemia Res. 23, 207–215.
Drexler, H. G., MacLeod, R. A. F., Borkhardt, A., and Janssen, J. W. G. (1995) Recurrent chromosomal translocations and fusion genes in leukemia-lymphoma cell lines. Leukemia 9, 480–500.
Marini, P., MacLeod, R. A. F., Treuner, C., et al. (1999) SiMa, a new neuroblas-toma cell line combining poor prognostic cytogenetic markers with high adrenergic differentiation. Cancer Genet. Cytogenet. 112, 161–164.
Drexler, H. G., Matsuo, Y., and MacLeod, R. A. F. (2000) Continuous hemato-poietic cell lines as model systems for leukemia-lymphoma research. Leukemia Res. 24, 881–911.
Tosi, S., Giudici, G., Rambaldi, A., et al. (1999) Characterization of the human myeloid leukemia-derived cell line GF-D8 by multiplex fluorescence in situ hybridization, subtelomeric probes, and comparative genomic hybridization. Genes Chromosomes Cancer 24, 213–221.
MacLeod, R. A. F., Spitzer, D., Bar-Am, I., et al. (2000) Karyotypic dissection of Hodgkin’s disease cell lines reveals ectopic subtelomeres and ribosomal DNA at sites of multiple jumping translocations and genomic amplification. Leukemia 14, 1803–1814.
Vanasse, G. J., Concannon, P., and Willerford, D. M. (1999) Regulated genomic instability and neoplasia in the lymphoid lineage. Blood 94, 3997–4010.
Kiippers, R. and Dalla-Favera, R. (2001) Mechanisms of chromosomal translocations in B cell lymphomas. Oncogene 20, 5580–5594.
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.
Drexler, H. G., Dirks, W. G., and MacLeod, R. A. F. (1999) False human hemato-poietic cell lines: cross-contaminations and misinterpretations. Leukemia 13, 1601–1607.
Nelson-Rees, W. A., Daniels, D. W., and Flandermeyer, R. R. (1981) Cross-contamination of cells in culture. Science 212, 446–452.
Markovic, O. and Markovic, N. (1998) Cell cross-contamination in cell cultures: the silent and neglected danger. In Vitro Cell Dev. Biol. Anim. 34, 1–8.
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.
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.
Caspersson, T., Zech, L., and Johansson, C. (1970) Differential binding of alky-lating fluorochromes in human chromosomes. Exp. Cell Res. 60, 315–319.
Sumner, A. T., Evans, H. J., and Buckland, R. A. (1971) New technique for distinguishing between human chromosomes. Nature New Biol. 232, 31–32.
Seabright, M. (1973) Improvement of trypsin method for banding chromosomes. Lancet 1, 1249–1250.
Rowley, J. D. (1973) A new consistent chromosomal abnormality in chronic myelogenous leukaemia identified by quinacrine fluorescence and Giemsa staining. Nature 243, 290–293.
Nowell, P. C. and Hungerford, D. A. (1960) A minute chromosome in human granulocytic leukemia. Science 132, 1497.
Cremer, T., Lichter, P., Borden, J., Ward, D. C., and Manuelidis, L. (1988) Detection of chromosome aberrations in metaphase and interphase tumor cells by in situ hybridization using chromosome-specific library probes. Hum. Genet. 80, 235–246.
Lichter, P., Cremer, T., Borden, J., Manuelidis, L., and Ward, D. C. (1988) Delineation of individual human chromosomes in metaphase and interphase cells by in situ suppression hybridization using recombinant DNA libraries. Hum. Genet. 80, 224–234.
Lichter, P. (1997) Multicolor FISHing: what’s the catch? Trends Genet. 12, 475–479.
Chang, H., Blondal, J. A., Benchimol, S., Minden, M. D., and Messner, H. A. (1995) p53 mutations, c-myc and bcl-2 rearrangements in human non-Hodgkin’s lymphoma cell lines. Leukemia Lymphoma 19, 165–171.
ISCN (1985) An International System for Human Cytogenetic Nomenclature: report of the Standing Committee on Human Cytogenetic Nomenclature. Birth Defects Orig. Artie. Ser. 21, 1–117.
Drexler, H. G., Dirks, W., MacLeod, R. A. F., Quentmeier, H., Steube, K. G., and Uphoff, C. C. (2001) DSMZ Catalogue of Human and Animal Cell Lines (8th ed.), DSMZ, Braunschweig, Germany.
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MacLeod, R.A.F., Drexler, H.G. (2005). Cytogenetic Analysis of Cell Lines. In: Helgason, C.D., Miller, C.L. (eds) Basic Cell Culture Protocols. Methods in Molecular Biology™, vol 290. Humana Press. https://doi.org/10.1385/1-59259-838-2:051
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DOI: https://doi.org/10.1385/1-59259-838-2:051
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