低血清培养Vero细胞和流感病毒条件的优化  

Optimization of Condition for Culture of Influenza Virus in Vero Cells in Low Serum Medium

作　　者：刘铮[1] 孙明波[1] 高菁霞[1] 马磊[1,1] 林小瑞[1] 燕景恩[1] 张勇[1] 吴忠香[1] 刘雅灵[1] 姚忠萍[1] 廖国阳[1] 李卫东[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生物制品五室,昆明650118

出　　处：《中国生物制品学杂志》2009年第6期593-595,共3页Chinese Journal of Biologicals

基　　金：国家863科技计划项目(2006AA02Z409;2008GA007);云南省自然科学基金资助项目(2004C0720M;2006YX29);云南省技术创新人才项目(2003PY07)

摘　　要：目的优化低血清培养Vero细胞和流感病毒的条件,为开发Vero细胞流感疫苗奠定基础。方法分别在DMEM/F12和SLM-199培养基中,添加不同浓度的血清,培养Vero细胞,观察细胞连续传代的生长状态;接种流感病毒后,检测病毒培养液不同pH值、不同浓度TPCK-胰酶和病毒不同接种量对病毒血凝效价的影响。结果用SLM-199在1.0%血清浓度下培养的Vero细胞接种流感病毒血凝效价较高,病毒培养液pH值为7.2,TPCK-胰酶含量为1.0μg/ml,接种病毒MOI为1.0时,72h收获的病毒血凝效价最高。结论已筛选出低血清培养Vero细胞和流感病毒的适宜条件。Objective To optimize the condition for culture of influenza virus in Vero cells in low serum medium and lay a foundation of developing Vero cells-based influenza vaccine. Methods The newborn calf sera at various concentrations were added into DMEM/F12 and SLM-199 media separately, in which Vero cells were cultured and observed for growth. Influenza virus was inoculated into the cultured Vero cells at various MOIs and incubated at various pH values and TPCK-trypsin concentrations, then determined for HI titers. Results The influenza virus incubated in Veto cells cultured in SLM-199 medium containing 1. 0% newborn calf serum reached a high HI titer. The optimal MOI, pH value and TPCK-trypsin concentration for culture of influenza virus in Vero cells were 1. 0, 7. 2 and 1. 0μg/ml respectively. Conclusion The condition for culture of influenza virus in Vero cells in low serum medium was optimized.

关 键 词：流感病毒 VERO细胞 低血清培养基 

分 类 号：Q2[生物学—细胞生物学] Q2-33