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Dynamics of single cell property distributions in Chinese hamster ovary cell cultures monitored and controlled with automated flow cytometry |
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| Authors: | Kacmar James Srienc Friedrich |
| Institution: | aDepartment of Chemical Engineering and Materials Science, University of Minnesota, 151 Amundson Hall, 421 Washington Avenue S.E., Minneapolis, MN 55455-0312, USA bBioTechnology Institute, University of Minnesota, 240 Gortner Laboratory, 1479 Gortner Avenue, St. Paul, MN 55108, USA |
| Abstract: | Two important variables that are often not measured online in Chinese hamster ovary (CHO) cell cultures are cell number concentration and culture viability. We have developed an automated flow cytometry system that measured the cell number concentration, single cell viability based on propidium iodide (PI) exclusion, and single cell light scattering from bioreactor samples every 30 min. The bioreactor was monitored during batch growth, and then the cell number concentration was controlled at a set point during cytostat operation. NH4Cl was added during steady state operation in cytostat mode to monitor the transient cell population response to adverse growth conditions. The automated measurements correlated well to cell concentration and viability determined manually using a hemacytometer. The described system provides a method to study mammalian cell culture physiology and dynamics in great detail. It presents a new method for the monitoring and control of animal cell culture. |
| Keywords: | Automated flow cytometry Bioreactor control Bioreactor monitoring Cytostat Mammalian cell culture Single cell heterogeneity |
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