不同代次牛肾原代细胞培养轮状病毒的比较研究

口服轮状病毒活疫苗(LLR株)生产用细胞基质为新生小牛肾原代细胞。原始的初代细胞(P0)产量小,一对牛肾平均生产7瓶细胞。将原始的初代细胞传代可使细胞产量显著增加,传代后(P2代)细胞产量可由7瓶增加为96~112瓶,细胞核型检查传至P5代的细胞染色体数目与初代细胞一致。细胞培养物均一性提高。P0代与P2代细胞病毒培养物滴度分别在6.2±1.5和6.5±0.5lgCCID50/ml,使用P2代细胞培养病毒,产量增加10~15倍。提高了疫苗生产的可控性和质量,生产规模显著放大,经济效益明显。

Comparison of producing the live attenuated rotavirus vaccine with the primary and subcultured calf kidney cells

The live attenuated rotavirus vaccine(LLR strain)was prepared using primary calf kidney cell.The resource of calf kidney,however,restricted the size of the vaccine production(7 bottles of cell can be gotten from a pair of kidney).To overcome this chokepoint,the cell subculture processing technology was developed,which improved fairly the cell production and equality.The results of the karyotype analysis showed that the chromosomes of subcultured cell are consistent with that of primary cell.The virus were propagated on secondary cell instead of primary cell,which enlarged the virus yields about 10~15 folds.The condition of virus titers are better than those of prepared from primary cell,the mean of virus titer is 6.5±0.5 and 6.2±1.5 lgCCID50/ml,respectively.Generally,the cell subculture technology improves the control of process,decreases production cycle time,increases virus yield,and more cost-effective.