High density culture of mammalian cells with dynamic perfusion based on on-line oxygen uptake rate measurements

• Published in: Cytotechnology (Dordrecht) Vol. 14; no. 3; p. 183
• Main Authors: Kyung, Y S, Peshwa, M V, Gryte, D M, Hu, W S
• Format: Journal Article
• Language: English
• Published: United States 1994
• Subjects: Cell Line - metabolism; Cell Survival - physiology; Humans; Online Systems; Oxygen - metabolism; Perfusion
• ISSN: 0920-9069
• DOI: 10.1007/BF00749615

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 10(8) 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.