摘要
目的：以细胞工厂代替转瓶培养轮状病毒基因重配株Ls的可行性研究。方法采用细胞工厂与相应的转瓶培养工艺作对比，比较两种容器内细胞生长状态与病毒收获液滴度，并对细胞工厂培养条件进行了优化。结果以相同浓度接种细胞时，细胞工厂4 d长成单层，转瓶却需要7 d，经细胞仪计数后单位面积内细胞密度相当；以相同MOI接种病毒后，转瓶内的病毒于第7天病毒滴度达到峰值，细胞已完全脱落；细胞工厂于第3天病毒滴度达到峰值，并实现了3次收获。细胞工厂每次收获的病毒液滴度都稳定在一定范围，与转瓶相当。另外，细胞工厂培养条件优化结果表明，Vero细胞最佳接种浓度为3．0×104细胞/cm2，接种病毒的最适MOI为0．02～0．04。结论使用细胞工厂培养Ls株病毒不仅提高了效率，而且减少了培养空间，可替代转瓶规模化生产轮状病毒疫苗。
Objective To estimate the possibility of culturing the reassortant rotaviruses strain Ls on a cell factory instead of a conventional roller bottle .Methods The growths of cells and titers of harvested virus cultured by the two methods were compared.Furthermore, the condition for culture on the cell factory was optimized .Results The results indicate that the monolayer is formed on the surface of the cell factory within 4 d of cultivation , but on the surface of the roller bottle within 7 d, when the Vero cells are inoculated at the same cell concentration .We found that the cell density of two contain-ers is equal by using the cell counter .The titer of viruses in roller bottle reaches a peak value at 7th day, afterwards cells have completely exfoliated;Those cultured on the cell factory achieve the highest titer of viruses at the 3rd day, and could be harvested for 3 times.The titers of each virus harvest are stable within a certain range on the cell factory , which are equal to those of the roller culture.The optimal inoculation concentration and MOI are 3.0×104 cells/cm2 and 0.02-0. 04, respectively.Conclusion The efficiency of culturing the rotaviruses strain Ls on the cell factory is not only increased , but the culture space is decreased , which shows that the cell factory could be used in a large-scale production of rotavirus vaccine instead of the roller bottle .
出处
《微生物学免疫学进展》
2014年第1期25-30,共6页
Progress In Microbiology and Immunology
