High density culture of mammalian cells with dynamic perfusion based on on-line oxygen uptake rate measurements |
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Authors: | Yun-Seung Kyung Madhusudan V Peshwa David M Gryte Wei-Shou Hu |
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Institution: | (1) Department of Chemical Engineering & Materials Science, University of Minnesota, 55455-0132 Minneapolis, MN, USA |
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Abstract: | In a continuous culture with cell retention the perfusion rate must be adjusted dynamically to meet the cellular demand. An automated mechanism of adjusting the perfusion rate based on real-time measurement of the metabolic load of the bioreactor is important in achieving a high cell concentration and maintaining high viability. We employed oxygen uptake rate (OUR) measurement as an on-line metabolic indicator of the physiological state of the cells in the bioreactor and adjusted the perfusion rate accordingly. Using an internal hollow fiber microfiltration system for total cell retention, a cell concentration of almost 108 cells/mL was achieved. Although some aggregates were formed during the cultivation, the viability remained high as examined with confocal microscopy after fluorescent vital staining. The results demonstrate that on-line OUR measurement facilitates automated dynamic perfusion and allows a high cell concentration to be achieved. |
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Keywords: | Cell culture high cell density oxygen consumption bioreactor |
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