By Simon P. Langdon (auth.), Simon P. Langdon (eds.)
The starting to be of melanoma cells within the laboratory has been a strong device for learning and realizing the biology of melanoma and the results of gear on melanoma cells. In melanoma cellphone tradition: tools and Protocols, specialist researchers describe intimately their best and up to date equipment for transforming into melanoma cells within the laboratory. compatible for amateur and skilled researchers alike, those simply reproducible recommendations resolve a various variety of experimental difficulties. there are ways to represent and authenticate cellphone traces, to isolate and boost particular kinds of melanoma cells, and to improve new telephone line types. useful assays are supplied to guage clonogenicity, mobilephone proliferation, apoptosis, adhesion, migration, invasion, senescence, angiogenesis, and mobile cycle parameters. different tools allow the amendment of melanoma cells for transfection, improvement of drug resistance, immortalization, and move in vivo; the coculture of other cellphone varieties; and the detection and remedy of illness. each one totally proven protocol is defined in step by step aspect by way of a longtime specialist within the box and features a historical past creation explaining the primary in the back of the procedure, gear and reagent lists, and pointers on troubleshooting and fending off recognized pitfalls.
complete and hugely sensible, melanoma phone tradition: tools and Protocols offers either uncomplicated scientists and scientific researchers with a gold-standard number of complicated recommendations for culturing melanoma cells successfully of their laboratories.
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Extra info for Cancer Cell Culture: Methods and Protocols
A modification of this process is to soak the initial fragments overnight at 4°C in the trypsin solution, allowing effective tissue penetration, and a single 30-min treatment at 37°C will be more effective. The cell suspension obtained is then centrifuged and cell culture medium (containing serum) added. 7. The other enzyme widely used for this step is collagenase as collagen is one of the major extracellular matrix proteins present in many stroma. Collagenase is added to culture medium (containing serum) at 200–1000 U/mL and can be left for 2–5 d.
Hough, C. , Sherman-Baust, C. , Pizer, E. , and Morin, P. J. (1999) Evidence for the colonic origin of ovarian cancer cell line SW626. J. Natl. Cancer Inst. 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.
Cell cultures should also be tested for the presence of mycoplasma. 2. Initial Establishment from Cell Suspensions If the clinical tissue is a fluid such as an ascites, aspirate, or effusion the following method can be used. 1. Collect freshly obtained clinical fluids, for example, ovarian ascites, from the patient and transfer to a sterile environment. For an ovarian ascites, the volume is typically of the order of 1 L. 2. Centrifuge the fluids for 20 min at 3000g and 4°C to produce a cell pellet.