By Jeffrey W. Pollard, John M. Walker
Animal mobilephone tradition, the most recent quantity in Humana's hugely profitable equipment in Molecular Biology sequence, presents unique functional innovations for the tradition of a vast spectrum of simple cellphone cellphone kinds. Chapters provide hands-on equipment for growing mammalian fibroblastic mobile cultures and holding tradition stipulations for epithelial, neuronal, and hematopoietic cells between others. awareness is given to the variety of tradition media and extracellular matrices had to hold the differentiated features of the classy cells. The book's precise energy lies in its descriptions of tradition suggestions for either residing and stuck cells. Chapters conceal innovations corresponding to: • cinematographic research • in situ mRNA hybridization • immunofluorescence • immunoelectron microscopy • somatic mobile hybridization • DNA transformation • insect phone tradition • production of hybridoma mobile strains • monoclonal antibody strategies • new, really expert methodologies. an invaluable appendix lists the main standard tradition media. accomplished in scope and insurance, and carefully up to date, Pollard and Walker's targeted instruction manual on ANIMAL phone tradition is an critical source for either the amateur and the pro professional.
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Additional info for Animal Cell Culture (Methods in Molecular Biology Vol 5)
3. IL2 (T-cell growth factor) also shows variation and ideally should be assayed on established cell lines: increased growth is monitored by increased [3H] thymidine uptake. References I. 2. 3. 4. 5. , Milton Keynes, MK9 3HP, England). , Dillner, M. , and Perlmann, I’. (1976) The interaction of nonmitogenic and mitogenic lectins with T-lymphocytes: association of cellular receptor sites. J. Immunol. 5,45-55. Hudson, L. and Hay, F. C. (1980) Practical Immunology (Blackwell Scientific Publications), pp.
J. Immunol. Methods 21,133-141. , Rossio, J. , Ruscetti, F. , and Oppenheim, J. J. (1986) Establishment of a human B cell line that proliferates in response to B cell growth factor. J. Immunol. Methods 90,111-123. Chapter 5 Establishment of Lymphoblastoid Cell Lines A. Doyle 1. Introduction The ability to establish long-term B lymphocyte cultures from patients carrying particular genetic characteristics or with the ability to secrete specific antibodies (I) is an extremely valuable technique. However, there are several basic principles to follow in the approach to this technology.
4. 8 cell culture supernatant. 43 44 Lymphoblastoid Cell Lines 5. 5% PHA Wellcome. 6. Ficoll-hypaque or lymphroprep. 1. Production 3. Methods of Epstein-Barr Virus Stock (3) 1. 8 cell line (Note 1) is cultured routinely in 5% FBS RPM1 1640. Routine passage consists of diluting the cells between 1 in 2 and 1 in 5 at weekly intervals. A culture in the logarithmic growth phase would usually yield lo6 cells/ml at 37°C. 2. 2 x 106/mL in 2% FBS RPM1 1640. 3. Incubate at 33°C for 2 w without any medium additions or changes.