骨髓间充质干细胞移植治疗生精细胞损伤模型大鼠的实验研究

摘要

目的:探讨骨髓间充质干细胞(MMSCs)在睾丸内定向分化为生精细胞用于治疗实验性雄性不育模型大鼠的可行性.方法:复苏冻存的MMSCs-GFP并培养传代后经睾丸网移植到经白消安处理的生精细胞损伤模型大鼠睾丸曲细精管内,移植后8周采样,观察睾丸质量指数、组织学表现和供体细胞分布,运用免疫荧光检测生殖细胞表面特异标志c-kit的表达.结果:MMSCs-GFP移植组右侧受体睾丸质量指数与模型组大鼠及自体未细胞移植的左侧睾丸相比有差异(P＜0.01),与正常组大鼠睾丸质量指数相比无差异(P＞0.05)；光镜下显示曲精管内生精上皮明显恢复,荧光显微镜下观察MMSCs-GFP大部分迁移至曲精管基底部,免疫荧光结果显示移植组与自身未移植组相比,睾丸曲细精管内精原细胞表面特异标志基因c-kit表达有差异.结论:细胞冻存不影响MMSCs的增殖能力,MMSCs在体内生精微环境中能够定植并定向分化为生殖细胞.%Objective; To investigate whether the marrow mesenchymal stem cells ( MMSCs) could differentiate into spermatogenic cells in a rat model of male infertility. Methods: The frozen MMSCs - GFP were recovered and cultured, and then transplanted into the seminiferous tubule of the right testis of rat model of spermatogenic cells injure established with Busulfan (50mg/kg) , with the left testis untreated as self control. The testes were collected 8 weeks later. The testicle weight index, testicular histology and donor cell distribution were observed. Immunofluorescence was employed to test the germ cell - specific marker c - kit expression. Results: The testicle weight index of MMSCs - GFP transplanted testis was comparable to that of normal rat testes (P > 0.05) , but significantly different from that of self - control testis (P<0.01). No local or systemic in-flammartory reaction was observed, while the spermatogenic epithelium was regenerated in the the MMSCs - GFP transplanted testis. Fluorescence microscopy showed that most of MMSCs - GFP moved to the basilar zone of seminiferous tubule. The c -kit expression in the seminiferous tubule of the MMSCs - transplanted testis was significantly different from that of the self -control testis. Conclusion: Cryopreservation did not affect the proliferation ability of MMSCs. MMSCs transplanted into convoluted seminiferous tubule could differentiated into spermatogenic cells.