Development of the optimal inoculation conditions for microcarrier cultures

Authors

  • Sean P. Forestell,

    1. Pharmaceutical Production Research Facility (PPRF), 2500 University Drive N.W., University of Calgary, Calgary, Alberta T2N 1N4, Canada
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  • Nicolas Kalogerakis,

    1. Pharmaceutical Production Research Facility (PPRF), 2500 University Drive N.W., University of Calgary, Calgary, Alberta T2N 1N4, Canada
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  • L. A. Behie,

    Corresponding author
    1. Pharmaceutical Production Research Facility (PPRF), 2500 University Drive N.W., University of Calgary, Calgary, Alberta T2N 1N4, Canada
    • Pharmaceutical Production Research Facility (PPRF), 2500 University Drive N.W., University of Calgary, Calgary, Alberta T2N 1N4, Canada
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  • Donald F. Gerson

    1. Connaught Laboratories Ltd., 1755 Steeles Avenue West, Willowdale, Ontario M2R 3T4, Canada
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Abstract

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.

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