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Extra resources for Animal Cell Culture: A Practical Approach
G. 27 g oxygen (Table 1). This amount will support 108 cells for 450 h and is thus clearly adequate. The second factor is whether this oxygen can be made available to the cells. The transfer rate of oxygen from the gas phase into a liquid phase has been calculated at about 17 (jug/cm2/h (8). Again, this is well in excess of that required by cells in a 1 litre flask. However, if the surface is assumed to be 26 SCALING-UP OF ANIMAL CELL CULTURES Table 1. 21 = proportion of oxygen in air; 32 = molecular weight of oxygen; 22400 = gram molecular volume; x 10-4 = solubility of oxygen in water at 37°C when equilibrated with air.
This is exaggerated in monolayer cultures because as cells become more tightly packed together, the surface area available for nutrient uptake becomes smaller (4). Glucose is often another limiting factor as it is destructively utilized by cells and, rather than adding high concentrations at the beginning, it is more beneficial to supplement after two to three days. In order to maintain a culture, some additional feeding often has to be carried out either by complete, or partial, media changes or by perfusion.
Basic cell culture: a practical approach. IRL Press at Oxford University Press, Oxford. 3. Freshney, R. I. (2000). Culture of animal cells: a manual of basic technique, 4th edn. WileyLiss, New York. 4. Freshney, R. I. (1999). Freshney's culture of animal cells, a multimedia guide. Wiley-Iiss, New York. 5. Boyce, S. T. and Ham, R. G. (1983) J. Invest. Dermatol, 81 (Suppl 1), 33s. 6. Rooney, S. , Young, S. L, and Mendelson, C. R. (1994). , 8, 957. 7. McCormick, C. and Freshney, R. I. (2000). Br. J.