Hughes J H, Sheridan J F
Department of Medical Microbiology and Immunology, Ohio State University, Columbus.
J Virol Methods. 1988 Oct;22(1):75-80. doi: 10.1016/0166-0934(88)90089-4.
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.
与以每分钟2转（rpm）滚动培养或静止培养的情况相比，痘苗病毒载体在以每分钟96转滚动培养的细胞中产生的细胞病变效应（CPE）量显著增加。以96 rpm滚动培养的细胞产生的病灶（46.3±8.4）明显多于静止培养的细胞（16.3±6.5）或2 rpm滚动培养的细胞（27.6±2.8）。以96 rpm滚动培养的细胞中病灶的大小也明显大于以2 rpm滚动培养或静止培养的细胞中的病灶（F = 12.65，P<0.01）。低感染复数感染并以96 rpm滚动培养的细胞比静止培养的细胞提前4天出现最大CPE。与静止培养条件相比，以2 rpm滚动培养的细胞病毒产量几何平均数增加了7.0倍。然而，以96 rpm滚动培养的细胞病毒产量几何平均数增加了75倍。这些数据表明，接种痘苗病毒载体的细胞应进行滚动培养以最大化病毒复制。
