不同类型生物反应器培养Vero细胞制备柯萨奇病毒A10型的比较  

Comparison of Coxsackievirus A10 production in Vero cells cultured in different types of bioreactors

作　　者：万鑫 蔡潜 叶童 肖奥 戴旱雨 游磊 孟胜利 郭靖 王泽鋆 申硕 WAN Xin;CAI Qian;YE Tong;XIAO Ao;DAI Hanyu;YOU Lei;MENG Shengli;GUO Jing;WANG Zejun;SHEN Shuo(Viral Vaccine Research Laboratory,Wuhan Institute of Biological Products,Co.,Ltd.,Wuhan 430207,Hubei Province,China)

机构地区：[1]武汉生物制品研究所有限责任公司病毒性疫苗研究一室,国家联合疫苗工程技术研究中心湖北省疫苗技术创新中心,湖北武汉430207

出　　处：《中国生物制品学杂志》2024年第10期1153-1160,1166,共9页Chinese Journal of Biologicals

基　　金：国家“重大新药创制”科技重大专项(2015ZX09102021);湖北省技术创新重大专项(2017ACA078)。

摘　　要：目的比较摇床生物反应器、篮式生物反应器和NBS生物反应器培养Vero细胞制备柯萨奇病毒A10型(Coxsackievirus A10,CV-A10)的效果。方法分别采用2 L摇床生物反应器(预辐照片状载体袋)、15 L篮式生物反应器(片状载体)和40 L NBS生物反应器(Cytodex 1球状载体)培养Vero细胞,制备CV-A10,比较不同类型生物反应器细胞贴壁效率、营养代谢物质含量、细胞生长特性以及病毒滴度和抗原含量差异。将2 L摇床生物反应器放大至50 L培养Vero细胞和CV-A10,比较不同规模摇床生物反应器制备的病毒滴度和抗原含量。结果Vero细胞在片状载体上的贴壁效率远高于球状载体,不同类型生物反应器中葡萄糖和乳酸含量变化曲线保持一致;不同类型生物反应器均可用于Vero细胞的高密度培养,相比于使用球状载体培养的NBS生物反应器,使用片状载体培养的摇床和篮式生物反应器可获得更高的细胞密度;不同类型生物反应器均可用于CV-A10的高效制备,且制备的CV-A10抗原含量和病毒滴度无差异;50 L摇床生物反应器可成功放大培养Vero细胞和CV-A10,不同规模摇床生物反应器制备的病毒滴度和抗原含量无差异。结论摇床生物反应器、篮式生物反应器和NBS生物反应器均可用于Vero细胞的稳定培养和CV-A10的高效制备。Objective To compare the production of Coxsackievirus A10(CV-A10)in Vero cells cultured in WAVE bioreactor,Basket bioreactor and NBS bioreactor.Methods Vero cells were cultured in 2 L WAVE bioreactor,15 L Basket bioreactor and 40 L NBS bioreactor filled with pre-irradiated fibracel disk,fibracel disk and Cytodex 1 microcarriers,respectively,to prepare CV-A10.The adherence efficiency,nutrient metabolites and growth characteristics of Vero cells cultured in different types of bioreactors,as well as the virus titers and antigen yields of the prepared CV-A10,were compared.Then,the CVA10 production process in a 2 L WAVE bioreactor was scaled-up to a 50 L WAVE bioreactor,and the virus titers and antigen yields of CV-A10 prepared in different scale WAVE bioreactors were compared.Results The adherence efficiency of Vero cells on fibracel disk was much higher than that on microcarriers,and the content variation curves of glucose and lactate in different types of bioreactors were consistent.The WAVE bioreactor,Basket bioreactor and NBS bioreactor could all be used for high density culture of Vero cells,while the WAVE bioreactor and Basket bioreactor with fibracel disk could achieve much higher cell density than the NBS bioreactor with Cytodex 1 microcarriers.CV-A10 could be efficiently prepared in different types of bioreactors and there was no significant difference in the virus titers and antigen yields of the prepared CV-A10.Vero cells and CV-A10 were successfully scaled up in the 50 L WAVE bioreactor,and there was no significant difference in the virus titers and antigen yields of different scale WAVE bioreactors.Conclusion The WAVE bioreactor,Basket bioreactor and NBS bioreactor can all be used for the stable culture of Vero cells and the efficient preparation of CV-A10.

关 键 词：摇床生物反应器 篮式生物反应器 NBS生物反应器 片状载体 球状载体 Vero细胞 柯萨奇病毒A10型 

分 类 号：Q813.1[生物学—生物工程]