Growth of Chikungunya Virus in Baby Hamster Kidney Cell (BHK-21-Clone 13) Suspension Cultures
FORT DETRICK FREDERICK MD
Pagination or Media Count:
The report describes the methods used to obtain high titers of chikungunya virus with suspension culture of BHK-21-clone 13 cells. The cells were grown at 37 C to a cell concentration of one to two million per ml. After maximum cell growth, the cells were inoculated with chikungunya virus at a multiplicity of 1 to 2 50 suckling mouse intracerebral lethal doses SMICLD50 per cell in the spent Eagles minimum essential medium for suspension cultures MEMS, or the cell cultures were centrifuged at 200 X g and resuspended in either fresh MEMS or medium 199 prior to inoculation. The medium used had no effect on virus titer. The inoculated cultures were incubated at 34 C until the cell viability dropped to 30, which usually occurred 28 to 30 hr postinoculation. After these procedures, chikungunya virus titers of log10 10.3 to 11.8 SMICLD50 per ml were obtained.