Abstract
For industrial pharmaceutical protein production fast growing, high producing and robust cell lines are required. To select pH-shift permissive and faster growing sub-populations, the CHO DP-12 cell line was serially subcultured for more than four hundred days in shaker flasks. Initial adaptation to growth in suspension was carried out in chemically defined medium without hypoxanthine and thymidine (HT), while the final medium used for long term cultivation contains HT. Cell samples were cryopreserved at four different time points after 21, 95, 165 and 420 days. Cultivations of these four sub-populations (SP) in shaker flasks and bioreactors revealed considerable differences in specific growth rates and product formation as well as in the metabolism of glucose, lactate and several amino acids. For the elucidation of the intracelluar mechanism behind these alteration in growth characteristics and metabolism additional probes were analyzed using proteomic and metabolomic approaches [1].
Highlights
For industrial pharmaceutical protein production fast growing, high producing and robust cell lines are required
CHO DP-12 cells were cultivated in CD-ACF medium TC 42 (TeutoCell AG) and PowerCHO-2 (LONZA AG) for the first steps of suspension adaptation. 200 nM methotrexate was present at any time
The initial adaptation to growth in suspension of the CHO DP-12 cells led to considerable changes in average cell diameter and specific growth rate
Summary
For industrial pharmaceutical protein production fast growing, high producing and robust cell lines are required. To select pH-shift permissive and faster growing sub-populations, the CHO DP-12 cell line was serially subcultured for more than four hundred days in shaker flasks. Initial adaptation to growth in suspension was carried out in chemically defined medium without hypoxanthine and thymidine (HT), while the final medium used for long term cultivation contains HT. Cell samples were cryopreserved at four different time points after 21, 95, 165 and 420 days. Cultivations of these four sub-populations (SP) in shaker flasks and bioreactors revealed considerable differences in specific growth rates and product formation as well as in the metabolism of glucose, lactate and several amino acids. For the elucidation of the intracelluar mechanism behind these alteration in growth characteristics and metabolism additional probes were analyzed using proteomic and metabolomic approaches [1]
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