Murine hybridomas were cultivated in tissue culture flasks. Dissolved oxygen tensions in the gas and liquid phases during cell growth were measured non-invasively by an optical oxygen sensor. Readings were made with caps both cracked open and completely closed. During cell growth, gas phase oxygen concentrations remained near atmospheric levels, while the oxygen tension at the bottom of the flasks eventually reached zero. These results suggest that the widespread practice of cracking open tissue culture flask caps during cell growth with a view to supplying adequate oxygen to cells is ineffective and unnecessary. The mass transfer characteristics of the tissue culture flask indicate the dominant resistance to oxygen mass transfer to the cells was the liquid media. The mass transfer rates through the liquid layer under standard laboratory conditions were found to be greater than those predicted by diffusion alone, suggesting microscale mixing. Volumetric and specific oxygen consumption rates were calculated from the sensor data, and were comparable to published values. A recently developed single fiber optic oxygen sensor is described. This new sensor will provide oxygen concentrations at various levels in the tissue culture flasks, allowing more accurate modeling of oxygen diffusion.
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