By Jeffrey W. Pollard, John M. Walker
Animal phone tradition, the newest quantity in Humana's hugely profitable equipment in Molecular Biology sequence, offers designated useful options for the tradition of a large spectrum of easy mobilephone mobile forms. Chapters supply hands-on equipment for growing mammalian fibroblastic mobilephone cultures and keeping tradition stipulations for epithelial, neuronal, and hematopoietic cells between others. cognizance is given to the variety of tradition media and extracellular matrices had to continue the differentiated services of the aesthetic cells. The book's unique energy lies in its descriptions of tradition strategies for either dwelling and glued cells. Chapters disguise innovations corresponding to: • cinematographic research • in situ mRNA hybridization • immunofluorescence • immunoelectron microscopy • somatic cellphone hybridization • DNA transformation • insect mobile tradition • production of hybridoma telephone strains • monoclonal antibody recommendations • new, really expert methodologies. an invaluable appendix lists the main widely used tradition media. finished in scope and assurance, and punctiliously up to date, Pollard and Walker's designated guide on ANIMAL mobile tradition is an essential source for either the amateur and the pro professional.
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Extra resources for Animal Cell Culture (Methods in Molecular Biology Vol 5)
H. (1986) Use of the virus to prepare human-derived monoclonal antibodies, in The Epstein-BarrVirus: RecentAdz~ances (Epstein, M. A. and Achong, B. ), William Heinemann Medical Books, London, p. 249. Adams, A. (1975) EBV Production, Concentration and Purification (Ablushi, D. , Aalesed, H. ), IARC, Lyon, France, p. 129. Miller, G. and Lipman, M. (1973) Release of infectious Epstein-Barr virus by transformed marmoset leucocytes. Nutl. Acud. USA 70,190. Chapter 6 Scale-Up of Suspension and Anchorage-Dependent Animal Cells J, Bryan Griffiths 1.
5. Check for proliferation after 7 d. 6. Remove medium and cells from wells, spin down, and gently resuspend transformed cells in fresh medium. Continue to culture in 24well plates. 7. These cells can be grown inlimiting dilution for B-cell characterization with monoclonal or polyclonal antibodies or functional assays. 8. Maintenance Using B-Cell Growth of B-CeZZs Factor Recently, B cells from peripheral blood from normal donors grown in the presence of B-cell growth factor (BCGF) and anti-IgM has led to the establishment of a long-term cultured human B cell line (5).
Notes 1. A major consideration is the source of EBV. 8 (4) as a source of virus, although in some cases cocultivation with the cell line QIMR-WIL has been employed. 8 cell lines distributed informally between research laboratories are mycoplasma contaminated. It is therefore of the utmost importance that the basic starting material for the technique, the virus source, is obtained from a reliable source, such as an established culture collection. 2. ) should be transferred from the syringe immediately into preservative-free heparin (final concentration, 10 U/ mL).