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

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

CIP cites the sequence because the major access. Covers tools for growing mammalian fibroblastic mobilephone cultures and preserving tradition stipulations for epithelial, neuronal, and hematopoietic cells between others. consciousness is given to the variety of tradition media and extracellular matrices had to preserve the differential capabilities of the classy cells. additionally describes tradition ideas for either dwelling and stuck cells.

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Alternatively, freeze overnight in a polystyrene container in a -8OOCrefrigerator and then remove ampules to -196OC. 4. Virus Transformation Either: 1. Carefully thaw the ampule in a 37OCwater bath, and transfer contents to 10 mL of 10% FBS RPM1 1640 to remove DMSO. Centrifuge at 1OOg for 5 min, or 2. Centrifuge cells in RPM1 1640/heparin at 1508 for 4 min. Then: 3. 8) supernatant per 5 x lo6 cells. 4. 5h. Agitate the suspension at least once during the incubation period. 5. Centrifuge at 1OOgfor 5 min.

Approximately 60% of the bound cells are recovered. 6. , IL2 (seeNote 3). Kinchington 40 1. 2. 3. 4. 5. 6. 7. 8. 1. Maintenance of T-Lymphocytes Using IL2 Dilute the enriched T-cell suspension (with growth medium containing 10% FCS) to give a density of 5 x 1O5-1 O6cells /mL. Depending on the volume, place either 200 PL aliquots in a microtiter plate, 1 mL/ well in a 24-well plate, or 3-7 mL in a 25-cm* flask. Add PHA to give a final concentration of 2 pg/mL. Incubate at 37OC in a humidified 5% CO,/95% air incubator.

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. 212,213. , and Sjoquist, J, I. (1978) Separation of cells by affinity chromatography on SPA-Sepharose 6MB. 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.

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