CAJ | 학술논문

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。
목적：대류감역묘생산용신형세포기질MDCK세포진행전면지질량공제검측，병대검측항목급결과진행회총화분석。방법근거《중국약전》삼부，대대검MDCK세포진행세포감별、내、외원인자화치류성검측，동시근거국제신요구대MDCK세포적렬해물급세포DNA적치암성진행검측。결과대검MDCK세포위첩벽、상피양세포형태，견세포래원，평균염색체수위80±1；미발현세균、진균、지원체오염，내、외원병독적오염검측결과현시，대검세포중미발현유치세포병변、산생혈흡부、혈응혹치동물사망적비특이성병독，역전록병독、우원성병독화견특이성병독검측균위음성；취107개활세포접충매지라서，관찰12주후，접충부위미발현결절，대체해부현시주요장기역미출현결절，병리검사현시여대조조무명현차이，세포미현시구유치류성。이등량세포적렬해물화세포DNA접충라서，균미치라서산생결절，병리검사결과여대조조무명현차이，제시세포미현시치암성。결론대검MDCK세포구유생산류감역묘적잠재가능성。
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 .