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 equipment for developing mammalian fibroblastic telephone cultures and protecting tradition stipulations for epithelial, neuronal, and hematopoietic cells between others. consciousness is given to the range of tradition media and extracellular matrices had to continue the differential services of the classy cells. additionally describes tradition thoughts for either dwelling and stuck cells.

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Glass Bead Immobilized Beds (3,5). Apparatus-this type of culture system is easily fabricated in the laboratory (Fig. 5). A suitable Scale- Up of Animal Cells n A h 59 aiir I D Fig. 5. Glass-sphere immobilize&bed culture: (A) immobilized bed; (B) reservoir with (E) stirrer, pH and oxygen probes, air spar&g; (Cl pump; (D) inlet for inoculum/trypsin; water at 37OC. cylindrical or funnel-shaped glass container is packed with borosilicate glass spheres (minimum diameter 3 mm, optimum diameter 4 or 5 mm).

This will keep for 1 wk at 4°C. 2. B Cell Growth Factor (BCGF). 3. Clostridium perfringens neuraminidase. 4. EBV suspension, 5 x lo4 Transforming Units (TFU). 5. F(ab’), fragment of goat anti-human IgM (p-chain specific). 6. Ficoll-paque. 7. RPM1 growth medium contains 5,10, or 15% fetal calf serum, 2 mM Lglutamine, 100 pg/mL penicillin, and 100 mg/mL streptomycin. 8. Growth medium containing 10% FCS, and 2-Mercaptoethanol (2 x lo4 M). 9. Hanks’ balanced salt solution pH 4 (seeAppendix). 10. Helix pomatia Lectin-Sepharose 6MB.

The easiest means of perfusing is the spin filter (as described for suspension cells), but a much larger pore size can be used (60-100 pm). This allows much faster perfusion rates to be attained (1-2 vol/ h). Perfusion from a reservoir that is adequately gassed is an efficient means of oxygenating the culture. Spin filter systems are commercially available (LH Fermentation, New Brunswick). 4. Conclusion Microcarrier is undoubtably the most efficient scale-up method for anchorage-dependent cells currently available.

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