牛雄性生殖系干细胞的体外诱导分化

Differentiation of bovine male germ-line stem cells in vitro

雄性生殖系干细胞(Male germ-line stem cells,mGSCs)是一群具有高度自我更新能力和分化潜能的细胞,是雄性成体内唯一可复制的二倍体永生细胞。转基因技术与雄性生殖系干细胞异体及异种移植技术相结合,将会为克隆动物、转基因动物生产及一些人类遗传性疾病的基因治疗提供新的机遇与途径。本试验采用组合酶消化和选择贴壁法,对5月龄、6月龄牛胎儿及新生牛雄性生殖系干细胞体外培养及分化进行了研究。试验结果显示,睾丸支持细胞对雄性生殖系干细胞体外增殖、分化所必需的,同时对数期睾丸支持细胞对雄性生殖系干细胞贴壁、增殖与分化效果明显;共培养16 d后,牛雄性生殖系干细胞分化为长形精子细胞,试验建立了牛雄性生殖系干细胞体外诱导培养分化体系。

Male germ-line stem cells （mGSCs） have the capability of self-renewal and latent capability of differentiation, mGSCs is the unique diploid immortal cell which can transfer genetic information to filial generation. The combination of transgenic technology and mGSCs heterotransplanting will supply new opportunities and paths to cloning animal, transgenic animal and gene therapy of some human hereditary disease. We studied the isolation and cultivation of mGSCs that were isolated and purified from 5-6 month old bovine fetal testis, new born bovine testis by adopting mixed enzymes digestion and different attaching velocities methods. The results showed that sertoli cells were indispensable to mGSC＇s proliferation and differentiation in vitro. The sertoli cells in logarithmic phase have a significant effect on mGSC＇s attaching, proliferation and differentiation. Co-culture with Sertoli cells, mGSCs differentiated to long sperm after 16 days. A preliminary system for mGSC＇s inducing differentiation was established.