应用生物反应器培养Vero细胞制备EV71病毒初探

A microcarrier cell culture process for EV71 virus in Vero cells

目的应用生物反应器培养Vero细胞制备EV71病毒。方法以3 L生物反应器采用4 g/L、8 g/L Cytodex-1微载体培养比较Vero细胞比生长率,并以4 g/L微载体培养EV71病毒。结果 4 g/L微载体培养Vero细胞3～4 d微载体细胞密度达2.3×106/mL,按0.001的感染复数(MOI)接种EV71病毒,病毒收获液的滴度最高达7.90 lgPFU/mL,较静置培养平均高出0.92 lgPFU/mL。结论初步建立了3 L生物反应器微载体培养Vero细胞制备EV71病毒的工艺,为进一步放大生产规模奠定了基础。

Objective To prepare enterovirus 71(EV71) in Vero cell by using a bioreactor.Methods It is by using concentrations of microcarrier cytodex-1 for 4 g/L,8 g/L to cultivate Vero cell line respectively,so as to compare of growth rate,and producction of EV71 with a microcarrier concentration of 4 g/L.Results The EV71 infected at a MOI of 0.001 when cell density reached to 2.3×106/mL after virus growing on 4 g/L microcarriers for 3 to 4 days,and obtained a high-titer of 7.90 lgPFU/mL for EV71 production,which is about 0.92 lgPFU/mL in average higher than that of static culture.Conclusion It was initially established a microcarrier cell culture process for producing EV71 in 3L bioreactor,which provides a basis for a scale up production of EV71.