Vero细胞培养流感病毒的低血清培养基的筛选  

Screening of low serum medium for culture of influenza virus in Vero cells

作　　者：耿兴良 戴宗祥[1] 段盼盼[1] 郭琦[1] 宋绍辉[1] 寸怡娜[1] 姜述德[1] 廖国阳[1] 

机构地区：[1]中国医学科学院北京协和医学院医学生物学研究所云南省重大传染病疫苗研发重点实验室,云南昆明650118

出　　处：《中国生物制品学杂志》2014年第5期687-690,共4页Chinese Journal of Biologicals

基　　金：国家国际合作项目"细胞流行性感冒病毒减毒活疫苗合作研究"(2011DFR30420)

摘　　要：目的筛选Vero细胞生长的最适低血清培养基,用于流感病毒的细胞培养。方法分别以MEM、M199、DMEM/F12、DMEM/F12与MEM混合物(1∶1)为基础培养基,添加5%、2%、1%的小牛血清传代培养Vero细胞,通过细胞形态观察、细胞计数及MTT法检测细胞的增殖活力筛选最适基础培养基;将流感病毒接种低血清适应的Vero细胞,检测病毒收获液的血凝效价及残留BSA含量,电子显微镜观察病毒形态。结果以DMEM/F12与MEM混合物(1∶1)为基础培养基,添加2%小牛血清培养的Vero细胞长势良好,细胞数量较多,增殖活力最强。低血清适应的Vero细胞培养的流感病毒血凝效价最高可达1∶512,平均值为1∶384;病毒液中的残留BSA含量约为正常血清培养条件下的1/18;病毒形态完整。结论成功筛选出Vero细胞培养流感病毒的低血清培养基,为更具生物安全性的流感疫苗的研发奠定了基础。Objective To screen the low serum medium for culture of Vero cells and use for culture of influenza virus. Methods MEM,M199,DMEM / F12 and mixture of DMEM / F12 and MEM（1 ∶ 1）were used as basal media,in which 5%,2% and 1% calf sera were added separately,and Vero cells were cultured. The cultured cells were observed for morphology,counted and determined for proliferative activity,based on which the optimal basal medium was screened. Influenza virus was inoculated into the Vero cells adapted in low serum medium,and the HA titer and residual BSA content in harvest were determined,while the virus morphology was observed by electron microscopy. Results The mixture of DMEM / F12 and MEM（1 ∶ 1） was used as a basal medium and added with 2% calf serum,in which Vero cells grew well in a large quantity and showed high proliferative activity. The HA titer of influenza virus cultured in low serum-adapted Vero cells reached 1 ∶ 512 at most,with a mean of 1 ∶ 384. The residual BSA content in virus liquid was about 1 / 18 of that cultured in normal serum medium. The virus particles in low serum medium were intact,of which the morphology showed no significant difference with those in normal serum medium. Conclusion Low serum medium for culture of influenza virus in Vero cells was screened,which laid a foundation of development of influenza vaccine with higher biosafety.

关 键 词：VERO细胞 流感病毒 血清 培养基 生物安全性 

分 类 号：R373.13[医药卫生—病原生物学] R392-33[医药卫生—基础医学]