Growth inhibition in animal cell culture
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Eight independent cell lines accumulated ammonia in culture to concentrations between 1.3 and 2.9 mM. The growth inhibition of such concentrations of ammonium chloride when added to culture medium was variable. The cell lines tested could be divided into 3 groups depending on their growth response to 2 mM added NH4CI. In the first group (293, HDF, Vero, and PQXB1/2) little (< 14%) or no growth inhibition occurred. In the second group (McCoy and MDCK) a reduction in final cell yield of 50-60% was observed. The third group (HeLa and BHK) was most sensitive to the effects of NH4CI with growth inhibition (>75%) compared to controls. The growth inhibitory effect of added lactate up to 20 mM was negligible (<10%) for 3 cell lines, although one cell line (PQXB1/2) showed greater sensitivity.
The interactive effects of ammonia and lactate were determined in a matrix experiment. At lactate (> 12 mM) and ammonia (1-4 mM), the growth inhibitory effects of the two components were synergistic. However, at low concentrations of lactate (< 12 mM) the toxic effect of ammonia was reduced. A proposed mechanism for the sparing effect of lactate on ammonia toxicity is discussed. This may have importance in developing strategies for the optimal growth of ammoniasensitive cell lines.
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- Growth inhibition in animal cell culture
Applied Biochemistry and Biotechnology
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