Enhanced production of poxvirus vectors by high speed rolling

The amount of CPE produced by vaccinia vectors in cultures rolled at 96 revolutions per minute (rpm) was significantly enhanced when compared to cultures rolled at 2 rpm or held stationary. Cultures rolled at 96 rpm had significantly more foci (46.3 +/- 8.4) than cultures held stationary (16.3 +/- 6.5) or rolled at 2 rpm (27.6 +/- 2.8). The size of foci in cultures rolled at 96 rpm was also significantly larger than foci in cultures rolled at 2 rpm or held stationary (F = 12.65, P less than 0.01). Cultures infected with a low multiplicity of infection and rolled at 96 rpm developed maximum CPE 4 days earlier than stationary cultures. When compared to stationary conditions, cultures rolled at 2 rpm had a 7.0-fold geometric mean increase in viral yield. However, cultures rolled at 96 rpm had a 75-fold geometric mean increase in viral yield. These data suggest that cultures inoculated with vaccinia vectors should be rolled to maximize viral replication.