Animal Cell Culture by Walker J.M. (ed.), Pollard J.W. (ed.)

By Walker J.M. (ed.), Pollard J.W. (ed.)

CIP cites the sequence because the major access. Covers tools for developing mammalian fibroblastic mobilephone cultures and preserving tradition stipulations for epithelial, neuronal, and hematopoietic cells between others. awareness is given to the range of tradition media and extracellular matrices had to retain the differential services of the aesthetic cells. additionally describes tradition options for either residing and glued cells.

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Phytohemagglutinin (PHA) (1 mg/mL stock). 20. Protein A-Sepharose 6MB( Pharmacia). 21. 1%. 3. 1. Separation from Blood of hmphocytes Using Ficoll Commercial products such as Ficoll-paque mixtures are available to separate human mononuclear cell populations contaminated with RBC and granulocytes. 1. tg/mL preservative free heparin. Primary T and B Lymphocytes 35 2. Dilute blood 1:2 in serum-free medium. 3. Layer the diluted blood carefully onto the Ficoll-paque using a Pasteur pipet to produce a clean interface between the two layers.

Above 10 L, glass vessels become inconvenient to handle and the progression to in situ stainless steel vessels should be considered. , perfusion, media changes, and so on) increases. For this level of sophistication, a fermentation system needs to be used. The main differences are that (a) stirring is by a direct-drive mechanism with a motor, and (b) the vessel has a complex top that allows the inclusion of a range of sensors, probes, feed supply lines, and sampling devices for contamination-free monitoring and control.

Allow the cells to settle out at 4OC. 5. Centrifuge the supernatant at low speed (1508 for 5 min) in order to clarify. 6. 2 ym filter. 7. Aliquot the filtered supernatant in 2 mL sterile plastic ampules, and store either short-term (1 mo) at -20°C, medium-term (6mo) at -7O”C, or preferably in a gaseous phase liquid nitrogen container at -13OOC. 8. A sample of each batch should be examined for microbiological purity (bacteria, fungi,mycoplasma) and also tested for transforming ability. 2. EB-Virus Transformation: Blood Sample Preparation 1.

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