H1N1疫苗株在MDCK和MDCK-G1细胞中的培养条件比较

Comparative analysis of conditions for culturing influenza virus H1N1 vaccine strain in MDCK and MDCK-G1 cell lines

目的比较H1N1流感病毒疫苗株在MDCK和MDCK-G1细胞中的最佳培养条件、产毒量、病毒滴度和细胞代谢情况。方法通过棋盘法摸索两种细胞的最佳种毒条件,然后比较2株细胞在最佳条件种毒后的血凝滴度、半数组织感染剂量(TCID50)、葡萄糖和乳酸的代谢情况。结果MDCK-G1细胞以感染复数(MOI)=0.001、1μg/ml TPCK胰酶接种H1N1流感病毒后,72 h时血凝滴度达到峰值(1∶512),病毒滴度为107.4TCID50/ml;MDCK细胞以MOI=0.0001、1μg/ml TPCK胰酶接种H1N1流感病毒后,72 h时血凝滴度达到峰值(1∶256),病毒滴度为106.6TCID50/ml。结论MDCK-G1比MDCK细胞更适合流感病毒的增殖。本研究为细胞基质流感疫苗的研究提供了参考数据。

Objective To compare the optimal conditions,virus yield,viral titer and cell metabolism between culturing influenza virus H1N1 vaccine strain in MDCK and MDCK-G1 cells.Methods The optimal culture conditions were investigated using chessboard method.The hemagglutination titer,half of the tissue infection dose(TCID50)and the metabolism of glucose and lactic acid were monitored and compared between the two cell lines.Results After MDCK-G1 cells were inoculated with H1N1 at the multiplicity of infection(MOI)of 0.001 with the presence of 1μg/ml of trypsin,the hemagglutination titer reached the peak of 1∶512 at 72 h and the viral titer was 107.4TCID50/ml.In the MDCK cell line group,the hemagglutination titer reached the peak of 1∶256 at 72 h and the viral titer was 106.6TCID50/ml when using H1N1 at MOI=0.0001 and 1μg/ml of trypsin.Conclusions MDCK-G1 cells were more suitable than MDCK cells for the proliferation of influenza virus.This study provided reference data for further research on cell-derived influenza vaccine.