Development of the optimal inoculation conditions for microcarrier cultures
Article first published online: 19 FEB 2004
Copyright © 1992 John Wiley & Sons, Inc.
Biotechnology and Bioengineering
Volume 39, Issue 3, pages 305–313, 5 February 1992
How to Cite
Forestell, S. P., Kalogerakis, N., Behie, L. A. and Gerson, D. F. (1992), Development of the optimal inoculation conditions for microcarrier cultures. Biotechnol. Bioeng., 39: 305–313. doi: 10.1002/bit.260390308
- Issue published online: 19 FEB 2004
- Article first published online: 19 FEB 2004
- Manuscript Accepted: 7 AUG 1991
- Manuscript Received: 19 MAR 1991
- inoculation protocol;
- anchorage-dependent cells;
- initial cell attachment
The environmental conditions under which anchorage-dependent mammalian cells are grown are not necessarily those under which a culture should be initiated. Cell attachment is a physical process, and those factors which affect forces involved in cell attachment differ from the biological factors which affect cell growth. We have conducted an extensive experimental study to define clearly the optimal environmental conditions for MRC-5 cell attachment onto microcarriers. These inoculation conditions are particularly important when the serial propagation of mammalian cells on microcarriers is considered as in a human vaccine production process. The conditions which were investigated are: initial serum content (% v/v), initial pH, inoculation level (cells/bead), agitation rate (rpm), and the concentration of microcarriers (g/L). The initial distribution of attached cells was found to have a significant affect on the overall efficiency of anchorage-dependent cell cultures, and was used to evaluate attachment efficiency. Based on the experimental results, we propose an optimized protocol for the inoculation of microcarrier cultures.