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.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Hu W-S. and Peshwa MV (1991) Animal cell bioreactor — Recent advances and challenges to scale-up. Can. J. Chem. Eng. 69: 409–420.
Hu W-S and Piret JM (1992) Mammalian cell culture processes. Curr. Opinion Biotechnol. 3: 110–114.
Tokashiki M, Arai T, Hamamoto K and Ishimaru K (1990) High density culture of hybridoma cells using a perfusion culture vessel with an external centrifuge. Cytotechnology 3: 239–244.
Kitano K, Shintani Y, Ichimori Y, Tsukamoto K, Sasai S and Kida M (1986) Production of human monoclonal antibodies by heterohy-bridomas. Appl. Microbiol. Biotechnol. 24: 282–286.
Seamans TC and Hu W-S (1990) Kinetics of growth and antibody production by a hybridoma cell line in a perfusion culture. J. Ferm. Bioeng. 70: 241–245.
Peshwa MV, Kyung Y-S, McClure DB and Hu W-S, (1993) Cultivation of mammalian cells as aggregates in bioreactors—effect of calcium concentration on spatial distribution of viability. Biotechnol. Bioeng. 41: 179–187.
Nikolai TJ, Peshwa MV, Goetghebeur S and Hu W-S, (1991) Improved microscopic observation of mammalian cells on microcarriers by fluorescent staining. Cytotechnology 5: 141–146.
Fleischaker RJ Jr and Sinskey AJ (1981) Oxygen demand and supply in cell culture. Eur. J. Appl. Microbiol. Biotechnol. 12: 193–197.
Hu W-S and Oberg MG (1990) Monitoring and control of animal cell bioreactors: Biochemical engineering considerations. In: Lubineicky AS (ed.) Large Scale Mammalian Cell Culture Technology (pp. 451–481) Marcel Dekker, Inc., New York, NY.
Konstantinov KB, Pambayun R, Matanguihan R, Yoshida T, Perusich C and Hu W-S (1992) On-line monitoring of Hybridoma cell growth using a laser turbidity sensor. Biotechnol. Bioeng. 40: 1337–1342.
Konstantinov KB and Yoshida T (1992) Knowledge-based control of fermentation process. Biotechnol. Bioeng. 39: 479–486.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Kyung, YS., Peshwa, M.V., Gryte, D.M. et al. High density culture of mammalian cells with dynamic perfusion based on on-line oxygen uptake rate measurements. Cytotechnology 14, 183–190 (1994). https://doi.org/10.1007/BF00749615
Issue Date:
DOI: https://doi.org/10.1007/BF00749615