Damaging agitation intensities increase DNA synthesis rate and alter cell-cycle phase distributions of CHO cells
Article first published online: 19 FEB 2004
Copyright © 1992 John Wiley & Sons, Inc.
Biotechnology and Bioengineering
Volume 40, Issue 8, pages 978–990, 20 October 1992
How to Cite
Lakhotia, S., Bauer, K. D. and Papoutsakis, E. T. (1992), Damaging agitation intensities increase DNA synthesis rate and alter cell-cycle phase distributions of CHO cells. Biotechnol. Bioeng., 40: 978–990. doi: 10.1002/bit.260400814
- Issue published online: 19 FEB 2004
- Article first published online: 19 FEB 2004
- Manuscript Accepted: 11 JUN 1992
- Manuscript Received: 12 MAR 1992
- DNA synthesis rate;
- cell-cycle kinetics;
- flow cytometry;
- cell culture
The effects of fluid-mechanical force (agitation) on the cell cycle kinetics of Chinese hamster ovary (CHO) cells cultured in suspension in 2-L bioreactors has been examined. A two-color flow cytometry method was used to determine the fraction rate of DNA synthesis. With increased agitation intensity, cell viability decreased as a result of increased cell death. However, increased agitation induced the viable cells of the culture to a higher proliferative state relative to a control culture. The fraction of viable cells of the high-agitation culture (250 rpm) in S phase was higher (up to 45%) and in G1 phase was lower (up to 50%) compared with the viable cells of the control culture (80 rpm). The DNA synthesis rate per viable S-phase cell of the high-agitation culture was confirmed by recovery experiments, which were conducted to measure the apparent specific growth rate and the cell cycle kinetics of the high-agitation culture upon reduction in the agitation rate from 250 rpm back to 80 rpm. The apparent specific growth rate of the test culture, calculated for the first 12 h of the recovery period, was greater than the apparent specific growth rate of the control culture. Furthermore, the proliferative state of the viable cells of the test culture, which had become higher relative to the control culture during the high agitation period, gradually approached the level of the control culture during recovery. Results also show that the magnitude of the agitation intensity; the culture agitated at 250 rpm attained a greater proliferative state than a parallel culture agitated at 235 rpm. The 250-rpm culture had a higher fraction of S-phase and a lower fraction of G1-phase cells than the 235-rpm culture. The DNA sunthesis rate per viable S-phase cell of the 250-rpm culture was greater than of the 235-rpm culture. © 1992 John Wiley & Sons, Inc.