论文部分内容阅读
目的探讨人胚胎干细胞(hES)系H1生殖细胞分化相关基因的表达。方法免疫荧光、碱性磷酸酶检测和核型分析等方法联合鉴定H1的基本生物学特性,RT-PCR方法检测未分化H1的生殖细胞分化相关基因的基本表达模式,分别以人正常睾丸组织和人胚胎成纤维细胞株HFF-1作为阳性和阴性对照组织。结果H1保持正常核型并维持未分化状态。未分化H1表达减数分裂前生殖细胞标志基因STELLAR、DAZL、FRAGILIS、PUM1、PUM2和GDF3;不表达原始生殖细胞(PGCs)标志基因BLIMP-1、减数分裂特异标志基因SCP1、减数分裂启动标志基因STRA8以及生殖细胞分化后期标志基因VASA。结论未分化H1细胞表达生殖细胞减数分裂前标志基因,但不表达分化后期基因,BLIMP-1可能成为区分未分化hES细胞与PGCs的特异性标志基因。
Objective To investigate the expression of human germline stem cell (hES) lineage H1 germ cell differentiation related genes. Methods The basic biological characteristics of H1 were identified by immunofluorescence, alkaline phosphatase and karyotype analysis. The basic expression patterns of germ cell differentiation-related genes in undifferentiated H1 were detected by RT-PCR. The normal human testis Human embryonic fibroblast cell line HFF-1 was used as a positive and negative control tissue. Results H1 maintains normal karyotype and maintains an undifferentiated state. Undifferentiated H1 expression pre-meiotic markers of germ cell STELLAR, DAZL, FRAGILIS, PUM1, PUM2 and GDF3; does not express the original germ cell (PGCs) marker gene BLIMP-1, meiosis specific marker gene SCP1, meiosis start Marker gene STRA8 and germ cell differentiation late marker gene VASA. Conclusion Undifferentiated H1 cells express the pre-meiotic markers of germ cells, but do not express late-stage genes. BLIMP-1 may be a specific marker for differentiating undifferentiated hES cells from PGCs.