生殖细胞核因子在精原干细胞体外培养分化中的表达

Expression of germ cell nuclear factor in vitro differentiation of spermatogonial stem cells

目的将分离纯化的小鼠精原干细胞(SSCs)体外培养并诱导分化,检测生殖细胞核因子(GCNF)在小鼠SSCs诱导分化前后的表达。方法用干细胞因子(SCF)诱导小鼠SSCs向精母细胞分化,通过间接免疫荧光染色和RT-PCR,检测GCNF在小鼠SSCs诱导分化前后的表达。结果小鼠SSCs向精母细胞诱导分化前后,在倒置相差显微镜下进行形态学观察,细胞形态未发生明显变化,仍然呈圆形或椭圆形,核较大;间接免疫荧光及RT-PCR结果均显示,原代培养的小鼠SSCs呈现GCNF阴性,但是向精母细胞诱导分化2d后,GCNF开始表达。结论 GCNF在体外培养小鼠SSCs向精母细胞分化的早期有表达,提示GCNF可能参与了SSCs的早期分化。

Objective To culture the mouse spermatogonium stem cells（SSCs） then separated and purified,to induce them differentiation in vitro, and to detect the germ cell nuclear factor （GCNF） expression of mouse SSCs before and after induction. Methods Stem cell factor（SCF） were used to induce spermatogonia to differentiate into the sperma tocytes. The immnofluoreseence staining and RT-PCR techniques were uesd in present studies in order to detect GCNF expression before and after differentiation of mouse SSCs. Results SSCs to spermatoeytes in mice before and after differentiation were observed by an inverted phase contrast microscope, the cells did not change significantly, they still had a round or oval nucleus that was relatively large; indirect immunofluorescenee and RT-PCR results were showed that primary cultured mouse SSCs were GCNF negative, but the 2 days later the spermatoeyte （after the induction of differentiation） GCNF began to express. Conclusion The mouse spermatogonium stem cells during the in vitro culture and differentiation process were able to express GCNF. The studies showed that GCNF might be involved in the early differentiation of spermatogonium stem cells.