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STR图谱分析方法扩大用于人源细胞的鉴别研究

STR profiling on authentication of human cell lines in cell banks

摘要目的 采用短串联重复序列(STR)图谱分析的方法,建立本所细胞库中所有人源细胞的鉴别图谱,并进行细胞交叉污染及细胞误判现状分析的研究.方法 采用16个位点的STR图谱分析方法,检测本所细胞库中所有收集保存的及其他单位来源的共计61株人源细胞,并将STR图谱检测结果与目前国际细胞库公布的数据进行比对,分析细胞的正确性及交叉污染情况.对未检出信号的细胞株,采用同工酶法对细胞的种属进行鉴别.结果 被检测的61株细胞中,41株细胞呈现特征性STR图谱,不存在与其他细胞的交叉污染现象,其中36株细胞与ATCC或JCRB保存的同一名称细胞在相应的9个STR位点的数据完全一致,5株细胞均仅在vWA位点与ATCC数据不完全相同,可判定为正确细胞;7株细胞尚无国际可比对数据,但STR图谱特异,可认定为新建细胞.11株细胞(占18.0%)被鉴定为错误的细胞,其中舌癌细胞Tca8113及肝癌细胞HHCC(现更名为FHCC98)的STR图谱数据分别与HeLa及HeLa S3细胞的数据完全一致;2株HuT-102的数据与ATCC的数据完全不同;4株细胞未检测到信号,同工酶结果显示均为鼠源细胞;同时还发现2株细胞为交叉污染细胞.结论 细胞误判及细胞交叉污染现象较为严重,正确鉴别细胞,特别是对国内自建细胞重新鉴定,对保证科学研究的可信性及可重复性极为重要.

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abstractsObjective To study the profiling and authentication of human cell lines in cell bank of our department using short tandem repeat (STR) loci and to analyze the situation of cell contamination and misjudgment. Methods Sixty-one human cells including cells collected and preserved by our cell bank and cells from the other departments were detected by the 16 STR loci-method. To analyze the cross contamina-tion between human cell lines, the results were aligned with profiles published by international cell banks. Isoenzyme detection was employed to authenticate the cell species when the STR signal can not be detected. Results Among the 61 cells, specific profiles were produced by 41 ceils and there was no cross contamina-tion. Thirty-six cells had the completely same STR profiles in 9 STR loci with the same cells preserved by ATCC or JCRB, while 5 cells have different profiling just in the vWA loci. The cells referred above can be recognized as correct cells; Eleven cells (18.0%) were the false cells. Among them, cancer cells of tongue named Tca8113 and cancer cell of liver named HHCC(changed to FHCC98 now) had the same profile with HeLa and HeLa S3 respectively; Two ceils both named HUT-102 have the completely different profiles with ATCC; The signal of 4 cells was not be detected, and all of them were determined as hamster cell lines by u-sing isoenzyme detection. Also 2 cells were identified to be mixed cells. Conclusion The phenomenon of cell misjudgment and cross contamination between cells is serious. Authentication of cell lines correctly, es-pecially for the re-authenticatian of domestic self-established cells, is very important for the guarantee of the reliability and reproducibility for scientific researches.

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