生物反应器大规模培养脊髓灰质炎病毒

Large-scale Culture of Poliomyelitis Virus in Bioreactor

目的应用生物反应器大规模培养脊髓灰质炎病毒。方法用7L、75L和550L生物反应器逐级放大培养Vero细胞,每天取样进行细胞计数。在550L生物反应器中培养脊髓灰质炎病毒,观察细胞病变,并检测病毒感染性滴度。结果一级细胞培养(灌流培养法)、二级细胞培养(循环培养法)和三级细胞培养(批培养法)的平均细胞密度分别为2.79×106、2.38×106和1.04×106个/ml,一级培养的细胞倍增数最高;病毒培养体积达350L,病毒收获液滴度大于7.625lgCCID50∕ml。结论通过三级放大工艺,可大规模培养脊髓灰质炎病毒。

Objective To culture poliomyelitis virus in a large scale in bioreactor. Methods Vero cells were cultured in 7 L bioreactor, which was scaled up to 75 and 550 L bioreaetors, and counted daily. Poliomyelitis virus was inoculated into Vero cells cultured in 550 L bioreactor, then the CPE was observed and the infectious titer was determined. Results The densities of Veto cells cultured in 7 L （perfused culture）, 75 L （circulating culture） and 550 L （batch culture） bioreactors were 2. 79 × 10^6, 2. 38×10^6 and 1.04×10^6cells/ml respectively. The multiplication number of Vero cells in 7 L bioreactor was higher than those in 75 and 550 L bioreactors. In 550 L bioreactor, the volume of virus for culture was 350 L, and the titer of harvested virus liquid was more than 7. 625 lgCCID50/ml. Conclusion The scaled procedure was suitable for the large-scale culture of poliomyelitis virus.