基于鸡ESCs分化为雄性生殖细胞调控关键基因和信号通路筛选诱导剂的研究

项目名称： 基于鸡ESCs分化为雄性生殖细胞调控关键基因和信号通路筛选诱导剂的研究

项目编号： No.31272429

项目类型： 面上项目

立项/批准年度： 2013

项目学科： 农业科学

项目作者： 李碧春

作者单位： 扬州大学

项目金额： 80万元

中文摘要： 精原干细胞的可塑性使其能替代ESCs用于治疗和遗传修饰，而不涉及伦理和免疫排斥问题，故引起生物学界极大关注。然怎样能大量获得该类细胞？科学家们为此进行了干细胞定向诱导分化研究，虽取得了突破性进展，但因诱导方案迥异，生殖细胞分化机制及关键基因与诱导剂互作调控细胞分化的机理仍不清楚，导致诱导重复性差和效率低，体外难以获得功能齐全的生殖细胞，满足不了研究和实践需要。本项目旨在利用鸡基因表达谱芯片和免疫组化分析技术，筛选参与雄性生殖细胞分化的关键基因和信号通路，并探究其分子作用机理；在此基础上依据关键基因和信号通路关键分子的启动子及其转录因子，利用荧光素酶报告基因系统和哺乳动物细胞单杂交技术建立细胞筛选模型，对诱导剂进行筛选；比较候选诱导剂不同添加方式和不同添加组合的作用效果，建立适宜的诱导体系，为弄清生殖细胞分化机制提供理论依据，为提高细胞诱导效率及医学临床药物筛选建立细胞模型提供实践技术支撑。

中文关键词： 雄性生殖干细胞；高通量测序；差异表达基因；信号通路；诱导剂

英文摘要： Spermatogonial stem cells could be used to replace embryonic stem cells for disease therapy and genetic modification since it has the merits of pluripotency and do not involve in ethical and immune rejection problem, therefore, it caused great concern in the biological community. Although the scientists has performed the research about the stem differentiation for solving the problem how does abtain a large number of spermatogonial stem cells in vitro and achieved breakthrough, it is still unclear about the regulation mechanism of cells induced differentiation: how does the inducer interact with the key regulation gene, how does the germ cell differentiate into the specific cell types and what kind of inducing strategy was the most optimal. All these problem resulted in the inefficient induction, therefore, it is very hard to obtain fully function germ cells in vitro and could not meet the requirements of research and practice. The purpose of this project is to screen the key genes and signaling pathways involved in the germ cells differentiation by gene expression spectrum and immunohistochemical, verify its role of molecular regulation mechanism. Base on its molecular mechanism, the critical transcription factor and promoter related to this pathway was used to establish the cells screening model by Luciferase

英文关键词： Male germ stem cells；High throughput sequencing；Differentially expressed genes；signaling pathway；Inducer