细胞工厂培养原代地鼠肾细胞工艺的建立及优化  

Establishment and optimization of cell factory culture process for primary hamster kidney cells

作　　者：李军[1] 刘少祥 王玖强 金鹏[1] 侯良玉[1] 张莹[1] 时萍 林松 徐冬鑫 李俊明[1] Li Jun;Liu Shaoxiang;Wang Jiuqiang;Jin Peng;Hou Liangyu;Zhang Ying;Shi Ping;Lin Song;Xu Dongxin;Li Junming(No.1 Department of Viral Vaccine,Chengdu Institute of Biological Products Co.,Ltd.,Vaccine Engineering Research Center of Sichuan Province,Chengdu 610023,China;Chengdu Institute of Biological Products Co.,Ltd.,Vaccine Engineering Research Center of Sichuan Province,Chengdu 610023,China)

机构地区：[1]成都生物制品研究所有限责任公司病毒性疫苗一室、四川省疫苗工程技术研究中心,成都610023 [2]成都生物制品研究所有限责任公司、四川省疫苗工程技术研究中心,成都610023

出　　处：《国际生物制品学杂志》2022年第1期37-40,共4页International Journal of Biologicals

摘　　要：目的采用40层细胞工厂(40-layer cell factory,CF40)工艺培养原代地鼠肾(primary hamster kidney,PHK)细胞,优化乙脑减毒活疫苗的生产工艺。方法用不同浓度新生牛血清培养不同接种密度PHK细胞,比较各组PHK细胞数量,确定CF40工艺的PHK细胞接种密度和新生牛血清浓度。通过对比CF40和15 L转瓶采用不同新生牛血清的PHK细胞培养情况,比较两种工艺对新生牛血清的选择性。对4个厂家CF40培养的PHK细胞数量进行单因素方差分析,判定对CF40的选择性。结果CF40工艺的最优条件为PHK细胞接种密度6.1×10^(5)ml^(-1)、新生牛血清浓度6%。CF40工艺对新生牛血清的选择性较低,能比转瓶工艺更稳定地培养更多的PHK细胞。4个厂家CF40培养的PHK细胞数量之间的差异无统计学意义(F=0.23,P>0.05),对CF40选择性低。结论采用CF40可以制备出满足乙脑减毒活疫苗生产要求的PHK细胞。Objective To establish and culture primary hamster kidney cells(PHK cells)by using 40-layer cell factory(CF40)process,and optimize the production process of live attenuated Japanese encephalitis vaccine.Methods Cultured PHK cells of different inoculation densities with different concentrations of newborn calf serum.Compared the number of PHK cells in each group,and determined the PHK cell inoculation density and newborn calf serum concentration of the CF40 process.By comparing the PHK cell culture situations with different newborn calf sera in CF40 and 15 L rolling bottle,the selectivity of the two processes on newborn calf serum was compared.A one-way analysis of variance was performed on the number of PHK cells cultured in the cell factories of 4 manufacturers to determine the selectivity to the cell factories.Results The optimal inoculation density of PHK cell in the CF40 process was 6.1×10^(5)ml^(-1),and the optimal concentration of newborn calf serum was 6%.The CF40 process was less selective for newborn calf serum,and cultured more PHK cells more stably than the rolling bottle process.There was no statistically significant difference in the number of PHK cells cultured in the cell factories of the 4 manufacturers(F=0.23,P=0.873),and the selectivity to cell factories was low.Conclusion CF40 can be used to prepare PHK cells that meet the requirements for the production of live attenuated Japanese encephalitis vaccines.

关 键 词：细胞工厂 原代地鼠肾细胞 细胞培养 

分 类 号：R392[医药卫生—免疫学]