基于测序的人类白细胞抗原分型和PCR短串联重复序列技术在人胚胎干细胞鉴定中的应用

目的探讨基于测序的人类白细胞抗原分型(HLA-sequencing-based typing,HLA-SBT)和PCR短串联重复序列(short tandem repeat,STR)技术在人胚胎干细胞(human embryonic stem cell,hESC)应用前检测中的运用,建立人胚胎干细胞系的基因型档案。方法人胚胎干细胞系SYSU-1、SYSU-3,分别培养到20代、40代,应用PCR寡核苷酸特异测序探针(sequence specific olignucleotideprobe,SSO)技术检测两株细胞系的HLA-A、-B、-DR位点的低分辨分型,再利用HLA-SBT技术检测两株细胞系的HLA-A、-B、-DR位点的高分辨分型。应用PCR-STR技术检测两株细胞系的基因遗传标记。结果获得两株hESC细胞系的HLA高分辨分型和STR基因型。结论可以运用HLA-SBT和PCR-STR技术建立人胚胎干细胞应用前的基因型档案。

HLA-sequencing-based typing and short tandem repeat genetic locus detection of human embryonic stem cell lines

Objective To evaluate the application of HLA-sequencing-based typing(HLA-SBT)and short tandem repeat(STR)genetic locus detection in human embryonic stem cell(hESC)lines.Methods Two hESC lines were cultured and passaged routinely. PCR-sequence specific olignucleotide probe(SSO) was used to analyze the results of HLA antigens and alleles. HLA-SBT was used to reanalyze the results of HLA antigens and alleles. STR genetic locus of the cell lines was detected with PCR in two hESC lines.Results SBT was used to identify HLA-A,-B,-DRB1 alleles in two hESC lines and got high-resolution typing of HLA-A,-B and -DRB1 alleles. The genetype of STR in two hESC lines was different.Conclusion HLA sequencing-based typing and STR genetic locus detection may be used to evaluate the embryonic stem cell lines in genetics.