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Appl Environ Microbiol. 1975 September; 30(3): 433-438
Copyright © 1975 American Society for Microbiology. All Rights Reserved.

Adaptation of BHK-21 Cells to Growth in Shaker Culture and Subsequent Challenge by Japanese Encephalitis Virus

The Hormel Institute, University of Minnesota, Austin, Minnesota 55912

ABSTRACT

Baby hamster kidney (BHK-21) cells were adapted to grow in shaker culture using Waymouth medium 752/1 containing 20 mM N-2-hydroxyethyl-piperazine-N'-2'-ethanesulfonic acid buffer and supplemented with 2.5% (vol/vol) calf serum, 0.002% (wt/vol) sodium oleate, and 0.2% fatty acid-free bovine serum albumin (WO_2.5). Infectivity of Japanese encephalitis virus grown in the cells adapted to WO_2.5 approached 2 x 10⁸ plaque-forming units per ml. The culture volume of infected cells was reduced fivefold 12 h after infection. This step resulted in a 10-fold increase in infectivity over that obtained from infected cultures not subjected to volume reduction.