皮肤来源干细胞是一种取材广泛且易获得的成体干细胞。Tet1基因在生殖细胞的减数分裂调控、印迹基因擦除中起着重要作用。目前，尚无关于Tet1调控猪皮肤干细胞体外分化过程中减数分裂分子机制的研究报道。本项目拟借助RNA-Seq、microRNA芯片、RNAi、DNA甲基化和免疫共沉淀等技术，分析Tet1通过去甲基化途径对猪SDSCs减数分裂分化形成配子过程中的表观遗传修饰动态变化与基因/蛋白表达关系，深入研究Tet1基因通过表观修饰途径调控猪皮肤干细胞体外分化过程中进入减数分裂形成配子的作用机制。通过该研究，有望发现并系统描述Tet1通过表观修饰途径参与调控猪皮肤干细胞分化进程中减数分裂的调节机制。该项目在实践中可以成功构建无伦理学限制的皮肤干细胞体外诱导分化为配子的技术平台，在理论上能够探索干细胞来源的生殖细胞发育调节机制。

The differentiation of germ cells from stem cell became one of the hotspot in rencent years. Deriving adult stem cells, such as skin-derived stem cells (SDSCs), into germ cells will be an ideal way. Tet1 is expressed in embryonic stem cells and preimplantation embryos; it played an important role in PGC reprogramming and/or the maintenance of pluripotency of the ESC. Although several epigenetic regulators have been reported to be crucial for meiosis, the mechanisms by which Tet1 regulates the initiation of meiosis in the SDSCs through gene demethylation and how Tet1 expression contributes to PGCLCs meiosis remain poorly understood. In recent years, some articles analysis of the DNA methylation dynamics in reprogramming PGC indicates that Tet1 functions to wipe out remaining methylation, including imprinted genes, at the reprogramming stage. Stem cell derived from fetal porcine skin is a kind of stem cell that easy obtained. In the present study, applied with the technologies of RNA-Seq、microRNA、RNAi、DNA methylation and CHIP et al., investigated the influence of demethylation on dynamic changes of gene/protein expression during the period of pig SDSCs meiosis differentiate into gametes, then the role of Tet1 for meiotic initiation during primordial germ cells development should be determined. Briefly, we address function of Tet1 signal pathway for meiotic in SDSCs. We further focus on the relationship between Tet1 and gene methylation, and hope found the key upstream/downstream gene in porcine SDSCs differentiation to SLCs.

体外诱导干细胞分化为功能性配子是国际最前沿、最受关注的生物医学课题之一。目前干细胞“体外”分化配子最关键的问题就是如何引导有丝分裂向减数分裂转变，迄今尚无成功的报道。皮肤来源干细胞是一种取材广泛的成体干细胞。本课题以猪皮肤来源干细胞为实验材料，通过筛选不同成分，建立了由猪皮肤干细胞分化为生殖细胞样细胞的体外培养分化体系。在此基础上，我们检测了生殖细胞特异表达因子DAZL，VASA的表达，同时检测了表观遗传相关蛋白H3,5mc相关变化，推测猪SDSCs在向PGCLCs分化过程中存在甲基化状态变化情况，目前有关猪SDSCs向PGCLCs分化过程中相关基因甲基化情况国际上尚未见报道，我们为首次得到该研究结果。