By Simon P. Langdon
Professional researchers describe intimately their most efficient and updated equipment for transforming into melanoma cells within the laboratory. there are ways to symbolize and authenticate phone strains, to isolate and advance particular forms of melanoma cells, and to advance new cellphone line types. practical assays are supplied to guage clonogenicity, telephone proliferation, apoptosis, adhesion, migration, invasion, senescence, angiogenesis, and phone cycle parameters. different tools allow the amendment of melanoma cells for transfection, improvement of drug resistance, immortalization, and move in vivo; the co-culture of other mobilephone varieties; and the detection and therapy of infection.
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Extra info for Cancer Cell Culture. Methods and Protocols
2. Cell Growth Rates and Colony Forming Efficiencies Under regular growth conditions where the same sources of tissue culture reagents are in use, cell lines have relatively constant growth rates and saturation densities. Marked deviation from the regular values is an indication that something may be wrong. Characteristic of a cell line under defined conditions is the percentage of cells that are able to produce colonies, either in a matrix such as agar or on plastic. 3. Expression Profiles Certain proteins can be particularly informative in helping to characterize cells and their origins, and these can be measured by a wide variety of techniques such as Western blotting, FACS analysis, or immunocytochemistry.
1. Restriction Endonuclease Digestion 1. 2. 3. 4. 5. 6. 7. 8. 2-mL reaction tubes. Multiblock heater. Microcentrifuge. Disposable pipets. Restriction endonuclease Hinf I (high concentration 50 U/µL). 10X Hinf I restriction buffer. 5X gel loading buffer. Bidistilled water Verification of Cell Line Identity 47 9. Absolute isopropanol (–20°C). 10. 70% (v/v) ethanol (–20°C). 2. Agarose Gel Electrophoresis. 1. 500-mL Erlenmeyer flask. 2. 8 M Na-Acetate, 40 mM EDTA. 2 with glacial acetic acid). 3. Ultra Pure agarose.
Flandermeyer, R. , and Hawthorne P. K. (1974) Banded marker chromosomes as indicators of intraspecies cellular contamination. Science 184, 1093–1096. 6. Nelson-Rees, W. , and Flandermeyer, R. R. (1976) HeLa cultures defined. Science 191, 96–98. 7. Nelson-Rees, W. , Daniels, D. , and Flandermeyer, R. R. (1981) Cross-contamination of cells in culture. Science 212, 446–452. 8. 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.
Cancer Cell Culture. Methods and Protocols by Simon P. Langdon