人用疫苗生产用工作细胞库Vero细胞的鉴别  

Authentication of working cell bank for production of vaccines for human use

作　　者：钱兴丽[1] 宋彩花 任芳芳[1] 赵勇 段男 李娅娴 俞建昆[1] 洪超[1] 杨晓蕾[1] 

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质量检定室云南省重大传染病疫苗研发重点实验室,云南昆明650118

出　　处：《中国生物制品学杂志》2017年第10期1022-1027,共6页Chinese Journal of Biologicals

摘　　要：目的采用两种不同的方法进行人用疫苗生产用工作细胞库Vero细胞的鉴别,为提高人用疫苗生产用细胞的准确性及疫苗的安全性提供保障。方法采用短串联重复序列(short tandem repeat,STR)-PCR基因分型法对本所疫苗生产用Vero细胞的8个STR位点(D8S1106、D1S518、D6S1017、D17S1304、D4S2408、D5S1467、D19S245和DYS389)进行测定,并与文献报道的结果进行比对分析;采用染色体核型检查法,将Vero细胞经Giemsa染料染色,于显微镜下精确计数100个细胞的染色体后,统计染色体数为58或60条的细胞所占比例。结果 STR基因分型得到的特征性图谱与文献报道的结果完全相同,未出现三单位基因,且STR的重复数完全相同;高倍镜下精确计数100个细胞染色体数为58或60条的细胞所占比例为79%。结论本所疫苗生产用工作细胞库Vero细胞为正确细胞株,不存在污染或交叉污染的情况,为本所人用疫苗的安全性和准确性提供了保障。Objective To authenticate the working cell bank for production of vaccines for human use by two methods,and provide a guarantee for improving the accuracy of cell lines for production of vaccines for human use and the safety of vaccines. Methods Short tandem repeat（STR） markers were used to sequence the eight loci, i. e. D8S1106, D1S518,D6S1017, D17S1304, D4S2408, D5S1467, D19S245 and DYS389, in Vero cell lines, and the profiles were compared with those reported in documents. The prepared Vero cell lines for vaccine production were treated with stained with Giemsa staining, and the chromosomes of 100 cells were counted precisely under microscope, based on which the proportion of cells with 58 or 60 chromosomes was calculated. Results The profiles of the Vero cell lines are identical to those reported in documents. No three unit genes appeared, and the numbers of repeats were identical. The proportion of cells with 58 or 60 chromosomes was 79%. Conclusion The Vero cell lines from the working cell bank for vaccine production were correct cell lines without contamination or cross-contamination, which provided a guarantee for the safety and accuracy of produced vaccines for human use.

关 键 词：非洲绿猴肾细胞 工作细胞库 细胞鉴别 STR图谱 染色体核型分析 

分 类 号：R392-33[医药卫生—免疫学]