水牛精原干细胞体外诱导精子的研究

Mouse spermatogonial stem cells (SSCs) has recently made significant progress in research that the sperm can be induced in vitro by remodeling the SSCs niche microenvironment, however, the application of SSCs in big livestock need to carry out more research. In this research, the influence factors and mechanisms to be explored to establish buffalo's SSC and STO co-culture system, SSC and testis tissue (or testis cell) co-culture system. In order to facilitate the observation of changes in the differentiation process of SSC, GFP, as a visual custom tag, to be introduced into SSCs purified from the SSCs culture system in our lab. These SSCs with GFP to be investigated the feasibility of inducing into sperm in vitro in SSC and STO co-culture system, SSC and testis tissue (or testis cell) co-culture system. Finally, the function of induced Round sperm or sperm cells to be verified by the ICSI technique. The related technical systems derived from this study not only provide a visual operable method for exploring the mechanisms of homing to niche, self-renew and differentiation of SSCs, but also provide high-efficiency platform to maximize the use of male genetic resource and to breed new varieties of buffalo.

小鼠精原干细胞（spermatogonial stem cells，SSCs）研究最近取得了重大进展，通过对SSCs niche微环境重构，可以在体外诱导出精子，而大动物相关研究相对滞后，SSCs在家畜繁育中的应用尚需开展更多的研究。本项目拟通过探索影响水牛SSCs体外自我更新的影响因素及机制，建立水牛SSCs体外培养系统、水牛SSCs与睾丸组织及睾丸细胞共培养系统，通过向SSC导入GFP可视化标记，探索水牛SSCs在水牛睾丸组织共培养系统、水牛睾丸细胞共培养系统和水牛SSCs体外培养系统中化学诱导SSCs生成精子可行性，利用显微注射和胚胎移植技术对诱导的单倍体细胞或精子样细胞进行功能性验证。本项目拟建立的SSCs相关技术体系，不但能为大动物SSCs自我更新和分化等深层理论问题研究提供一个可视化的体外研究平台，同时有助于最大化利用水牛雄性遗传资源，加速水牛品种改良和新品种培育进程。

小鼠精原干细胞（spermatogonial stem cells，SSCs）研究最近取得了重大进展，通过对SSCs niche微环境重构，可以在体外诱导出精子，而大动物相关研究相对滞后，SSCs在家畜繁育中的应用尚需开展更多的研究。本项目通过探索影响水牛SSCs体外自我更新的影响因素及机制，优化了水牛SSCs体外富集、体外培养和体外诱导的方法，阐明了血清在水牛精原干细胞体外培养中并非必须要素，观察了Nano2基因在水牛精原细胞中的表达模式，Nano2在水牛精原细胞中的表达模式与其他动物既存在保守性，也存在差异性。同时我们以小鼠精原干细胞为模型探索精子生成的相关机制，证实了Zpac在精子生成过程的功能作用。这些研究成果为今后水牛精原干细胞的研究奠定了很好的基础，拓展了我们对哺乳动物精子关键机制的深入理解。水牛精原干细胞的研究有助于最大化利用水牛雄性遗传资源，加速水牛品种改良和新品种培育进程。