小鼠雌性生殖干细胞体外分化及其分子机制

Mouse female germline stem cells (FGSCs) were isolated and cultured as well as applied in generation of transgenic mice. How FGSCs are differentiated in vitro and how they contributed to the follicle pool is an important issue for the female reproductive biology. According to the past, embryonic stem cells and spermatogonial stem cell (SSC) differentiation method and technology platform in vitro as well as our previous results and work, to elucidate the molecular mechanisms of FGSC in vitro differentiation, this project will be conducted from both in vitro differentiation and simulated ovarian micro-environment differentiation of FGSCs. Using gene expression of high flux detection, transcriptome analysis and chromosome immunoprecipitation sequencing (CHIP-seq), the regulation of the FGSC differentiation of the signal pathway and epigenetic networks will be determined. The finding of this project will provide the theoretical and experimental basis for oogenesis abnormalities related to disease diagnosis and treatment of infertility, spontaneous abortion and birth defects as well as the improvement of assisted reproductive technology.

小鼠雌性生殖干细胞(FGSC)的分离培养及其在转基因小鼠应用领域已经展开，然而FGSC在体内外是如何分化的，对卵泡池的贡献如何是当前雌性生殖生物学面临的重要课题。根据以往胚胎干细胞和精原干细胞(SSC)体外分化的方法和技术平台以及项目组前期研究基础，为了阐明FGSC体外分化的分子机制，本项目将从体外直接分化和模拟卵巢微环境分化两方面着手，通过基因表达高通量检测，转录组分析技术和染色体免疫共沉淀测序技术(CHIP-seq)，确定调控FGSC分化的信号通路及表观遗传网络。为卵子发生异常相关疾病的诊治，不孕不育、自发流产和出生缺陷以及辅助生殖技术的完善提供理论和实验依据。

小鼠雌性生殖干细胞(FGSC)的分离培养及其在转基因小鼠应用领域已经展开，然而 FGSC 在体内外是如何分化的，对卵泡池的贡献如何是当前雌性生殖生物学面临的重要课题。本项目经过3年的努力, 完成了预期目标, 取得下列成果: 1)建立CD-1小鼠雌性生殖干细胞系以及体外分化系统并获得GV期卵母细胞同时揭示重要分化因子; 2) 定位出生后大鼠雌性生殖干细胞并建立大鼠雌性生殖干细胞系以及体内外分化系统同时筛选出重要分化因子; 3) 筛选和阐明PCGF6对生殖细胞分化起重要调控作用及其机制; 4) 构建了生殖干细胞发育的重要调控网络。发表SCI论文4篇, 培养研究生6名, 获上海市自然科学一等奖, 上海市优秀博士论文1人次。为卵子发生异常相关疾病的诊治提供理论基础和技术平台。