论文部分内容阅读
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 and differentiate isolated and purified mouse spermatogonial stem cells (SSCs) in vitro and to detect the expression of germ cell nuclear factor (GCNF) before and after differentiation of mouse SSCs. Methods SSCs were induced to differentiate into spermatocytes by using stem cell factor (SCF). The expression of GCNF was detected by indirect immunofluorescence staining and RT-PCR. Results The morphological changes of spermatogenic cells before and after differentiation were observed by inverted phase contrast microscope. The morphology of the cells did not change significantly. The morphology of the cells was still round or oval with larger nuclei. Indirect immunofluorescence and RT-PCR results The results showed that primary cultured mouse SSCs showed negative GCNF, but GCNF began to express after induced into spermatocytes for 2 days. CONCLUSIONS: The expression of GCNF in mouse SSCs in early stages of spermatocyte differentiation suggests that GCNF may be involved in the early differentiation of SSCs.