不同材质细胞培养容器培养甲型肝炎病毒的比较

Comparison of culture of hepatitis A virus in vessels of various materials

目的比较中性玻璃转瓶和聚苯乙烯细胞工厂两种材质的培养容器培养人二倍体细胞和甲型肝炎病毒(hepatitis A virus,HAV)的差异。方法采用相同的培养条件,在中性玻璃转瓶和聚苯乙烯细胞工厂中分别培养人胚肺二倍体细胞KMB17,并接种HAV H2株,倒置光学显微镜下观察细胞的形态变化;细胞接种病毒后第0、7、14、18、22、26、30天时取样,ELISA法测定病毒的感染性滴度,分析病毒的增殖动力学。结果两种材质的容器培养的KMB17细胞的生长状态和形态无明显差异;HAV H2株增殖动力学相似;单位体积病毒收获液中的病毒产量无显著差异;聚苯乙烯细胞工厂培养的HAV的单位培养容积产量是中性玻璃转瓶的7.28倍。结论聚苯乙烯细胞工厂和中性玻璃转瓶培养的人二倍体细胞和HAV均能良好生长,细胞工厂可替代转瓶成为新的疫苗规模化生产的细胞培养技术。

Objective To comp are the difference in culture of hepatitis A virus（HAV） in neutral glass spinner and polystyrene cell factory. Methods Human embryonic lung diploid cells（KMB17）were simultaneously cultured in neutral glass spinner and polystyrene cell factory respectively and inoculated with HAV H2 strain,then observed for morphological change under inverted microscope. Samples were taken from the cells on days 0,7,14,18,22,26 and 30 after inoculation and determined for virus titer by ELISA,based on which the kinetics of virus proliferation was analyzed. Results There was no difference in growth states and morphologies of cells cultured in neutral glass spinner and polystyrene cell factory,while the proliferation kinetics were similar. No significant difference was observed between the virus yields per unit volume of the harvests from vessels of two materials. However,the area of KMB17 cells cultured in polystyrene cell factory was 7. 28 times of that in neutral glass spinner. Conclusion Both human diploid cells and HAV grew well in neutral glass spinner and polystyrene cell factory,indicating that cell factory might be used as a novel cell culture technique for large-scale production of vaccine instead of spinner.