流感疫苗生产用新型细胞基质 MDCK细胞的质量控制研究  

Study on quality control of MDCK cells used for the production of influenza vaccine

作　　者：吴雪伶[1] 冯建平[1] 樊金萍[1] 李秀华[1] 付瑞[1] 孟淑芳[1] 

机构地区：[1]中国食品药品检定研究院卫生部生物技术产品检定方法及其标准化重点实验室,北京100050

出　　处：《中华微生物学和免疫学杂志》2013年第12期943-949,共7页Chinese Journal of Microbiology and Immunology

基　　金：国家科技支撑计划（2008BAl54800-4）;国家“十二五”863计划课题（2012AA02A402）;国家科技基础条件平台-国家实验细胞资源共享平台

摘　　要：目的：对流感疫苗生产用新型细胞基质MDCK细胞进行全面地质量控制检测，并对检测项目及结果进行汇总和分析。方法根据《中国药典》三部，对待检MDCK细胞进行细胞鉴别、内、外源因子和致瘤性检测，同时根据国际新要求对MDCK细胞的裂解物及细胞DNA的致癌性进行检测。结果待检MDCK细胞为贴壁、上皮样细胞形态，犬细胞来源，平均染色体数为80±1；未发现细菌、真菌、支原体污染，内、外源病毒的污染检测结果显示，待检细胞中未发现有致细胞病变、产生血吸附、血凝或致动物死亡的非特异性病毒，逆转录病毒、牛源性病毒和犬特异性病毒检测均为阴性；取107个活细胞接种每只裸鼠，观察12周后，接种部位未发现结节，大体解剖显示主要脏器亦未出现结节，病理检查显示与对照组无明显差异，细胞未显示具有致瘤性。以等量细胞的裂解物和细胞DNA接种裸鼠，均未致裸鼠产生结节，病理检查结果与对照组无明显差异，提示细胞未显示致癌性。结论待检MDCK细胞具有生产流感疫苗的潜在可能性。Objective To conduct extensive quality control tests on Madin-Darby Canine Kidney （ MDCK） cells used for the production of influenza vaccine .Methods Tests for characteristics , extraneous agents, endogenous agents and tumorigenicity were performed on MDCK cells according to Chinese Pharma -copeia Book III .Cell lysate and DNA of MDCK cells were tested for oncogenicity in the light of new interna -tional requirements .Results The MDCK cells extracted from canis were adherent cells with an epithelial morphology, whose average number of chromosome was 80±1.No bacteria, fungi and mycoplasma contami-nation were detected . The detection for extraneous and endogenous virus showed that there was no nonspecific virus causing cytopathic effect , hemadsorption , hemagglutination or animal death .Tests for re-verse transcriptase , bovine viruses and canine viruses were all negative .Each nude mouse was injected with 107 viable cells to observe their tumorigenicity .Twelve weeks after cell injection , no node was found at the injection site and in large organs by gross anatomy .There was no significant difference between test group and negative control group .The test for tumorigenicity of viable cells was negative .Cell lysate and cellular DNA collected from equivalent amount of cells were respectively injected into nude mice , and no node forma-tion was found.There was no significant difference between the test cells and negative controls in pathology indicating that the tested MDCK cells were non-oncogenic .Conclusion It showed the possibility of using MDCK cells for the production of influenza vaccine .

关 键 词：MDCK 细胞鉴别 内外源病毒 致瘤性和致癌性 

分 类 号：R392.1[医药卫生—免疫学]