人用疫苗生产用人二倍体细胞株KMB17的鉴定  

Identification of human diploid cell KMB17 strain for production of vaccines for human use

作　　者：宋彩花 任芳芳[1] 刘杰[1] 段男 钱兴丽[1] 李娅娴 杨晓蕾[1] 洪超[1] SONG Cai-hua;REN Fang-fang;LIU Jie;DUAN Nan;QIAN Xing-li;LI Ya-xian;YANG Xiao-lei;HONG Chao(Institute of Medical Biology,Peking Union Medical College,Chinese Academy of Medical Science,Laboratory of Quality Control ＆ Yunnan Provincial Key Laboratory For Development of Vaccines against Major Infections Disease,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》2018年第10期1135-1141,共7页Chinese Journal of Biologicals

摘　　要：目的鉴定本所自主建立的人用疫苗生产用人二倍体细胞株KMB17的正确性。方法用染色体核型检查法和短串联重复序列(short tandem repeat,STR)基因分型法对本所疫苗生产用人二倍体细胞株KMB17进行鉴定。将疫苗生产用KMB17细胞经秋水仙素处理,积累中期相染色体后,低渗条件下释放出染色体,吉姆萨染料染色制片,镜检,观察染色体结构,精确计数100个细胞的染色体数目;选取人源细胞的20个STR位点对KMB17细胞进行DNA分型,并将获得的STR图谱与ATCC和DSMZ数据库进行比对分析。结果高倍镜下观察可见,样品细胞染色体组结构正常,无缺失和突变;精确计数100个细胞中,染色体数为46的细胞数有84个,无染色单体和染色体断裂细胞,无结构异常细胞,亚二倍体细胞14个,超二倍体细胞数2个。STR基因分型获得的特征性图谱清晰,未出现三单位基因,在ATCC和DSMZ数据库中均未找到与其细胞分型完全匹配的细胞株。结论本所自主建立的疫苗生产用人二倍体细胞株KMB17的细胞染色体结构和数目均正常,不存在污染和交叉污染,为正确细胞株。Objective To identify the human diploid cell KMB17 strain developed by the Institute of Medical Biology, Peking Union Medical College, Chinese Academy of Medical Science for production of vaccine for human use. Methods Chromosome karyotype and short tandem repeat （STR） genotyping were used. KMB17 cells for vaccine production were treated with colchieine to accumulate metaphase chromosomes and then release chromosomes at low permeability. The chromosomes were stained with Giemsa dye and observed for structure, of which the number in 100 cells were recorded precisely by microscopy. Twenty STR loci from human cells were subjected to DNA typing, and the obtained STR profiles were compared with those in ATCC and DSMZ data banks. Results The genome of sample cells showed normal structure under high power microscope, without deletion or mutation. Of the 100 cells counted, 84 with 46 chromosomes were observed. However, no chromatid, chromosome fragmentation or chromosome structural aberration were observed. Fourteen hypodiploid and two hyperdiploid cells were found. STR profiles were unambiguous and no triplicate peak was observed. No 100% matched cell lines were found in ATCC and DSMZ data banks. Conclusion The established human diploid cell KMB17 strain for vaccine production was identified as correct, of which the structure and number of chromosomes were normal, and no contamination or cross-contamination was found.

关 键 词：人二倍体细胞KMB17 染色体核型 短串联重复序列 

分 类 号：R392-33[医药卫生—免疫学]