CCC-HPF-2细胞基因组DNA可作为STR分型标准物质

摘要

目的 检测细胞培养传代过程中的基因稳定性,筛选符合中国人遗传特性的DNA标准物质,建立DNA标准细胞库.方法 通过显微镜下观察、免疫组织化学、核型分析检测细胞传代过程中形态、分子表达及核型是否稳定；应用PCR方法检测细胞的外源微生物及种属来源；STR检测验证细胞在传代过程中STR表达谱的平衡性及稳定性.结果 CCC-HPF-2细胞呈成纤维细胞样；波形蛋白表达阳性,CK阴性；无外源微生物污染；细胞来源于人；所有15个STR基因座都得到扩增,呈现较好的扩增平衡性,各代细胞STR表达谱一致,表现了高度稳定性.结论 CCC-HPF-2细胞来源于人,30代内的该细胞DNA可作为STR分型标准物质.%Objective To establish and identify a cell line originated from Chinese as reference standards for STR profiling and to establish reference STR-cell bank. Methods The cell morphology of various passages were observed under microscope. The expression of vimentin and CK was detected immuno-cytochemically. The karyotype was analysed. Contamination of mycoplasma and the species origination of the cell line were checked by PCR. The stability and balance of STR profiling were analysed. Results The cell was fibroblast. The expression of vimentin was positive, CK is negative. No contamination of mycoplasma was found. All the 15 STR loci were well amplified and in good balance. STR profiling was completely identical among various passages. Conclusions The cell was originated from human. Under 30 passage DNA from these cells can be used as reference standards for STR profiling.