转瓶培养与生物反应器微载体培养乙脑病毒的比较  

Comparisons of the cell cultivation between using microcarrier in a 15L bioreactor and roller bottle for the preparation of JEV vaccine

作　　者：李薇[1] 孙振鹏[1] 孙燕[1] 白萍[1] 王玉琳[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》2008年第4期11-15,共5页Progress In Microbiology and Immunology

摘　　要：分别用15L转瓶与15L生物反应器微载体（2．5g／L CytodexⅢ）系统培养Vero细胞并接种乙型脑炎病毒（简称乙脑病毒）。转瓶培养Vero细胞7～8d，细胞数最高能达到8×10^8；当单层细胞长至3．0—4．5×10^8时接种乙脑病毒，病毒滴度能达到6．5～6．98Ig PFU／ml，并能够连续收获4～5次；采用微载体系统培养Vero细胞，细胞密度最高能达到170×10^8；当单层细胞长至60～70×10^8时接种乙脑病毒，病毒滴度能达到7～7．51gPFU／ml，并能够连续收获13～15次。两种方式培养的乙脑病毒收获液分别经灭活、浓缩、柱层析纯化后制备Vero细胞乙脑纯化疫苗，各项检定指标均符合《中国药典》的相关要求。Japanese encephalitis virus（ JEV） were separately cultivated in Vero cells with 15 L roller bottle and 15 L bioreactor system （2. Sg/L microcarrier. Cytodex Ⅲ） . A higher cells density was recovered from cultures using 15Lbioreactor （170×10^8 cells） ,compared with using the roller bottle mode（8 ×10^8cells）. JEV were innoculated into vero cells when the cell yields to 60-70 ×10^8 in the bioreactor and 3- 4.5 ×10^8 in roller bottle, respectively. After 3- 4 days the virus titres reached to 7-7.51gPFU/ml and the harvest times were 13-15 for the bioreactor while the virus titres were 6.5-6.98 1g PFU/ ml and the harvest times were 4-5 for the roller bottle. The JEV vaccine （ final bulk ） were prepared separately using the harvested virus by inactivation, concentration and purification and the results detected could comply with the requirements of Chinese pharmacopoeia.

关 键 词：转瓶 生物反应器 乙脑病毒 Veto细胞 

分 类 号：R373.31[医药卫生—病原生物学]