By Simon P. Langdon
Specialist researchers describe intimately their most efficient and up to date equipment for turning out to be melanoma cells within the laboratory. there are ways to symbolize and authenticate telephone traces, to isolate and improve particular forms of melanoma cells, and to advance new mobile line types. useful assays are supplied to judge clonogenicity, phone proliferation, apoptosis, adhesion, migration, invasion, senescence, angiogenesis, and mobile cycle parameters. different equipment enable the amendment of melanoma cells for transfection, improvement of drug resistance, immortalization, and move in vivo; the co-culture of other telephone forms; and the detection and therapy of illness.
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Additional info for Cancer Cell Culture: Methods and Protocols (Methods in Molecular Medicine)
2. Confirmation of cell line identity between different passages of an intensively used cell line, e. , the human embryonic kidney cell line 293 (HEK 293; see Note 3). 3. Evaluation of “purity” of a given cell line and detection of the percentage of cell mixture in an early stage of contamination. 4. Characterization of somatic hybrid cell lines involving human cells. 5. Mapping of loss of heterozygosity of chromosomal regions, for example, to detect tumor suppressor genes. The following subheadings present the technique of multiplexed PCR amplification fragment length polymorphism (AmpFLP) of six prominent and highly polymorphic minisatellite VNTR loci (see Table 1) and one additional locus for sex determination using the detection of the SRY gene on the Y chromosome.
Many cell lines will have reattached almost completely within 24 h but others take longer. As the cell density is relatively low after subculturing it is important to ensure that the media is well gassed; otherwise, it will become alkaline as the pH increases. When growing in suspension live cells are typically bright when viewed under phase contrast. ). Most cell lines will readily form single-cell suspension by repeated agitation with a pastette. If this is not sufficient then a 10-mL syringe with an attached needle should be used.
91, 1327–1328. 13. Masters, J. R. (2002) HeLa cells 50 years on: the good, the bad and the ugly. Nat. Rev. Cancer 2, 315–319. 14. Gilbert, D. , Reid, Y. , Gail, M. , Hay, R. , et al. Application of DNA fingerprinting for cell-line individualization. Am. J. Hum. Genet. 47, 499–514. 15. , and Griffiths, B. (1992) DNA fingerprinting—a valuable new technique for the characterization of cell lines. Cytotechnol 9, 211–216. 16. Stacey, G. , Bolton, B. , and Doyle, A. (1991) The quality control of cell banks using DNA fingerprinting.
Cancer Cell Culture: Methods and Protocols (Methods in Molecular Medicine) by Simon P. Langdon