轮状病毒LH9株在Vero细胞中的培养条件优化研究  

Culture conditions optimization of rotavirus LH9 on Vero cells

作　　者：包红[1] 寇桂英[1] 王名强[1] 韩平[1] 余黎[1] 周旭[1] 

机构地区：[1]兰州生物制品研究所,兰州730046

出　　处：《微生物学免疫学进展》2011年第1期15-19,共5页Progress In Microbiology and Immunology

摘　　要：为优化轮状病毒株在Vero细胞上的培养条件,将轮状病毒基因重配株LH9按0.1MOI分别接种于不同规格的细胞培养瓶(100ml、2000ml、3L、15L)。病毒接种采用吸附与未吸附两种方式、病毒收获采取低温冻融后离心与直接离心两种方法,观察分析对病毒滴度的影响。实验中,用CASY细胞计数仪分析活细胞率,病毒接种后逐日观察细胞病变(CPE)并取样,采用细胞半数感染量测定病毒滴度。结果表明,使用不同规格细胞培养瓶经吸附法培养接种、低温破碎法收获的LH9株病毒滴度高,其中以15L立瓶培养滴度最高(6.0～7.0 logCCID50/ml)。In order to optimize the culture conditions of rotavirus strains on Vero cells,the strain LH9 was inoculated in cell culture bottles with different volume(100ml,2000ml,3L,15L)by 0.1 MOI.The two inoculation methods(adsorption or non-adsorpttion) and two harvesting methods(direct centrifugalizing and centrifugalizing after freeze-thaw) were adopted to observe the effects of these methods on the virus titers.The viability of the cell was analyzed using the CASY Cell Counter,the CPE of Vero cells after inoculation was observed day by day and the virus titers were determined by CCID50 test.The results showed that the virus titers are higher when using adsorption method and freeze-thaw technique.The virus titer is the highest(6.0 - 7.0 logCCID50/ml).in the case of 15L cell culture bottles.

关 键 词：轮状病毒 基因重配株 VERO细胞 高滴度 

分 类 号：R373.2[医药卫生—病原生物学]