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目的比较轮状病毒(rotavirus,RV)在不同细胞上的培养效果,确定RV最适培养细胞株。方法采用原代牛肾细胞(PCKC)、MA104细胞、CV-1细胞、Vero细胞4种细胞培养RV,观察RV在不同细胞培养过程中细胞病变效应(cytopathic effect,CPE),并检测病毒收获液的病毒毒力及病毒RNA。结果 MA104和PCKC细胞较适宜RV的增殖培养,其次是CV-1细胞,而Vero细胞感染病毒后3 d才出现CPE,6 d才有约75%的细胞圆缩死亡,这时收获的病毒液滴度也不及前3种细胞的病毒收获液滴度,且收获液的病毒RNA电泳的条带较模糊。结论 MA104、PCKC、CV-1细胞较适宜RV的增殖培养,而应用于大规模生产疫苗的Vero细胞培养RV要想达到前3种细胞的效果,还需进一步完善感染和培养方法。
Objective To compare the culture effects of rotavirus (RV) on different cells and determine the optimum culture medium for RV. Methods The primary cultured bovine kidney cells (PCKC), MA104 cells, CV-1 cells and Vero cells were used to culture RV. The cytopathic effect (CPE) of RV in different cell cultures was observed and the virus harvest liquid Virus virulence and viral RNA. Results Compared with control group, the proliferation of PC12 cells and PC12 cells was more suitable for the proliferation of RV, followed by CV-1 cells. However, CPE occurred only 3 days after virus infection in Vero cells, and about 75% of the cells died in 6 days. At this time, The titers were also less than the titer of the virus harvested fluid of the first three cells, and the bands of the harvested liquid viral RNA electrophoresis were hazy. CONCLUSIONS: MA104, PCKC and CV-1 cells are more suitable for the proliferation of RV. However, the application of RV in large-scale production of Vero cells to achieve the effect of the first three kinds of cells requires further improvement of infection and culture methods.