目的  采用40层细胞工厂（CF40）替代15 L转瓶工艺制备乙型脑炎（乙脑）减毒活疫苗。方法  分别采用CF40和15 L转瓶工艺制备乙脑减毒活疫苗，通过显微镜观察和细胞计数来考察细胞质量。病毒收获时观察细胞病变情况，并对2种工艺制备的原液、半成品以及成品进行病毒滴度检测对比，成品37 ℃放置7 d考察成品热稳定性。结果  两种工艺相比，细胞工厂内单位面积内细胞数量差异存在统计学意义（P＜0.05》），生长一致性和单只地鼠制备的细胞数量均优于15 L转瓶；CF40和15 L转瓶制备的单一病毒收获液平均滴度分别为7.59 lgPFU/ml和7.56 lgPFU/ml，没有显著差异(P＞0.05)，但CF40单一病毒收获液收率一致性更好；两种工艺制备的原液、半成品、成品滴度以及7 d后37 ℃热稳定性均符合企业注册标准规定。结论  使用CF40制备乙脑减毒活疫苗是可行的，并且在产品收率和一致性方面优于15 L转瓶工艺。

Objective  To prepare l live attenuated Japanese encephalitis vaccine（JEV）by 40-layer cell factory（CF40） instead of 15 L roller bottle. Method  JEV was prepared by CF40 and 15 L roller bottle separately. Cell quality was checked by microscope and cell counting. Growth of cells was observed when virus was harvested. Virus titers of the bulk, final bulk and final product prepared by two processes were compared. The thermal stability of the final product was investigated after being placed at 37 ℃ for 7 days. Results  Among two processes, there was significant difference in the number of cells per unit area in the cell factory (P＜0.05) . The growth consistency and the number of cells prepared by a single hamster were better than that of 15 L roller bottle. The average titers of CF40 and 15L roller bottle single virus harvested solution were 7.59 lgPFU/ml and 7.56 lgPFU/ml, respectively, with no significant difference (P ＞0.05 ) ,but the yield of CF40 single virus harvested solution was more consistent. The titers of the bulk, final bulk and final product and the heat stability at 37 ℃ after 7 days prepared by the two processes all meet the provisions of enterprise registration standards.  Conclusion  It is feasible to prepare live attenuated Japanese encephalitis vaccine by CF40, and its yield and consistency of product are better than those of 15 L roller bottle.