不同细胞工厂制备F基因型腮腺炎减毒活疫苗(人二倍体细胞)的比较  

Comparison of live attenuated mumps vaccine(human diploid cells)of genotype F prepared in different cell factories

作　　者：杨振清 刘喜苹 文韬 李涛 艾晗 段黎恒 张玉平 王文慧 王晶晶[1] YANG Zhen-qing;LIU Xi-ping;WEN Tao;LI Tao;AI Han;DUAN Li-heng;ZHANG Yu-ping;WANG Wen-hui;WANG Jing-jing(Institute of Medical Biology,Chinese Academy of Medical Sciences&Peking Union Medical College,Kunming 650118,Yunnan Province,China)

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,云南昆明650118

出　　处：《中国生物制品学杂志》2022年第8期918-921,927,共5页Chinese Journal of Biologicals

基　　金：云南省科技人才和平台计划(2019HB073).

摘　　要：目的比较两种不同厂牌聚苯乙烯细胞工厂制备F基因型腮腺炎减毒活疫苗(人二倍体细胞)的差异。方法采用不同浓度血清(10%、8%、5%),在两组不同厂牌细胞工厂(A组:无锡耐思生物科技有限公司,B组:美国康宁公司)中分别培养人胚肺二倍体细胞KMB17,观察细胞形态变化并计数;在细胞长成致密单层后,接种SP-A株F基因型腮腺炎病毒,37℃培养7~9 d收获病毒液,经过滤后,获得疫苗原液。将病毒收获液、疫苗原液样品于4℃放置14 d、37℃放置7 d后检测病毒滴度,并按《中国药典》三部(2020版)要求检测相关安全性指标。结果两种不同厂牌细胞工厂培养的KMB17细胞生长状态和形态无明显差异,在细胞培养基的血清浓度为5%时,可获得较高滴度的病毒收获液;在该血清浓度下,两种不同厂牌细胞工厂培养的病毒收获液和疫苗原液感染性滴度均值为6.50 LgCCID50/mL,4℃放置14 d和37℃放置7 d后,A组与B组细胞工厂病毒滴度下降范围差异均无统计学意义(P>0.05);两种不同厂牌细胞工厂制备的病毒收获液和疫苗原液的无菌、分枝杆菌、支原体和细菌内毒素检查指标均符合《中国药典》三部(2020版)要求。结论两种不同厂牌细胞工厂均可用于制备SP-A株F基因型腮腺炎减毒活疫苗,本研究为F基因型腮腺炎减毒活疫苗的规模化生产提供了实验依据。Objective To compare the differences of live attenuated mumps vaccine(human diploid cells)of genotype F in polystyrene cell factories with different brands.Methods Human diploid cells KMB17 were cultured at various serum concentrations(10%,8%and 5%)in cell factories from Wuxi NEST Biotechnology Co.,Ltd.(group A)and Corning Incorporated(group B)separately,observed for the morphological changes and counted.The cells were inoculated with mumps virus SP-A strain of genotype F after formation of a dense monolayer and cultured at 37℃for 7~9 d.The virus liquid was harvested and filtrated to obtain the bulk.The virus harvest and bulk were stored at 4℃for 14 d and at 37℃for 7 d,determined for titer and for the indexes of safety according to the requirements in Chinese Pharmacopoeia(VolumeⅢ,2020 edition).Results No obvious differences were observed in the growths and morphologies of KMB17 cells cultured in the two cell factories.A higher virus titer was obtained when the serum concentration in cell culture medium was 5%.At this serum concentration,the mean virus titer of virus harvest and bulk prepared in two cell factories was 6.50 LgCCID50/mL.After storage at 4℃for 14 d and at 37℃for 7 d,the decrease of virus titers in groups A and B showed no significant difference(P>0.05).The results of sterility test as well as examinations of mycobacterium,mycoplasma and bacterial endotoxin of virus harvest and bulk prepared in two cell factories met the requirements in Chinese Pharmacopoeia(VolumeⅢ,2020 edition).Conclusion Both the cell factories may be used for preparation of live attenuated mumps vaccine of genotype F,which provides an experimental data for large-scale production of the vaccine.

关 键 词：细胞工厂 F基因型腮腺炎减毒活疫苗 人二倍体细胞 病毒滴度 

分 类 号：R373.1[医药卫生—病原生物学]