Abstract
A high-cell-density perfusion culture process, using a novel centrifuge, was developed. The centrifuge has spiral multiple settling zones to separate cells from culture medium. Because of the multiple zones, the separation area can be efficiently increased without enlarging the diameter of the centrifuge. The centrifuge used in this study had a separation capacity of 2600 ml culture medium min−1 at 100g of the centrifugal force. A new cell separation and withdrawal method was also developed. The cells separated in the centrifuge can be withdrawn easily from the centrifuge with no cell clogging by feeding a liquid carrier such as a perfluorocarbon into the centrifuge and pushing the cells out with the liquid carrier. By this culture process, monoclonal antibodies were produced with mouse-human hybridoma X87X at a cell density of about 8 × 106 cells ml−1 for 25 days. This centrifuge culture shows promise as a large-scale perfusion culture process.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Apelman S (1992) Separation of animal cells in continuous cell culture systems. In: Murakami H, Shirahata S, Tachibana H (eds) Animal cell technology: basic and applied aspects. Kluwer, Dordrecht, pp 149–154
Feder J, Tolbert WR (1983) The large-scale cultivation of mammalian cells. Scientific American 248:24–31
Hamamoto K, Ishimaru K, Tokashiki M (1989) Perfusion culture of hybridoma cells using a centrifuge to separate cells from culture mixture. J Ferment Bioeng 67:190–194
Himmelfarb P, Thayer PS, Martin HE (1969) Spin filter culture: the propagation of mammalian cells in suspension. Science 164:555–557
Jäger V (1992) High density perfusion culture of animal cells using a novel continuous flow centrifuge. In: Murakami H, Shirahata S, Tachibana H (eds) Animal cell technology: basic and applied aspects, Kluwer, Dordrecht, pp 209–216
Murakami H, Shimomura T, Nakamura T, Ohashi H, Shinohara K, Omura H (1984) Development of a basal medium for serum-free cultivation of hybridoma cells in high density. J Agric Chem Soc Jpn 58:575–583
Tokashiki M, Arai T (1989) High-density culture of mouse-human hybridoma cells using a perfusion culture apparatus with multisettling zones to separate cells from the culture medium. Cytotechnology 2:5–8
Tokashiki M, Arai T, Hamamoto K, Ishimaru K (1990) High density culture of hybridoma cells using a perfusion culture vessel with an external centrifuge. Cytotechnology 3:239–244
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Takamatsu, H., Hamamoto, K., Ishimura, K. et al. Large-scale perfusion culture process for suspended mammalian cells that uses a centrifuge with multiple settling zones. Appl Microbiol Biotechnol 45, 454–457 (1996). https://doi.org/10.1007/BF00578455
Received:
Revised:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00578455