Evaluation of a microcarrier process for large-scale cultivation of attenuated hepatitis A |
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Authors: | B H Junker F Wu S Wang J Waterbury G Hunt J Hennessey J Aunins J Lewis M Silberklang B C Buckland |
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Institution: | (1) Merck Research Laboratories, Rahway, New Jersey, USA;(2) Department of Chemical Engineering, University of Minnesota, Minneapolis, Minnesota, USA |
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Abstract: | Microcarrier culture was investigated for the propagation of attenuated hepatitis A vaccine in the anchorage-dependent human
fibroblast cell line, MRC-5. Cells were cultivated at 37°C for one to two weeks, while virus accumulation was performed at
32°C over 21 to 28 days. The major development focus for the microcarrier process was the difference between the cell and
virus growth phases. Virus antigen yields, growth kinetics, and cell layer/bead morphology were each examined and compared
for both the microcarrier and stationary T-flask cultures. Overall, cell densities of 4–5×106 cells/ml at 5–10 g/l beads were readily attained and could be maintained in the absence of infection at either 37°C or 32°C.
Upon virus inoculation, however, substantial cell density decreases were observed as well as 2.5 to 10-fold lower per cell
and per unit surface area antigen yields as compared to stationary cultures. The advantages as well as the problems presented
by the microcarrier approach will be discussed. |
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Keywords: | microcarrier hepatitis A virus propagation cell culture MRC-5 cells |
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