特定转录因子诱导小鼠成纤维细胞直接重编程为精原干细胞样细胞

项目名称： 特定转录因子诱导小鼠成纤维细胞直接重编程为精原干细胞样细胞

项目编号： No.31301218

项目类型： 青年科学基金项目

立项/批准年度： 2014

项目学科： 生物科学

项目作者： 张仕强

作者单位： 西北农林科技大学

项目金额： 21万元

中文摘要： 精原干细胞（SSC）是雄性动物体内能将遗传信息传递给后代的一类数量稀少的成体干细胞。本课题拟尝试将体细胞直接重编程为精原干细胞样细胞（SSLC）。从15个SSC候选转录因子（Id4、Nanos2、Bcl6b、Etv5、Lhx1、Foxo1、Pou3f1、Pou5f1、Sox2、Klf4、c-Myc、Lin28、Utf1、Plzf和Taf4b）中筛选出特定组合，通过逆转录病毒载体pMXs将其导入雄性小鼠胎儿成纤维细胞和成体尾尖成纤维细胞，在添加胶质细胞神经营养因子（GDNF）的SSC培养条件下进行诱导，以期获得SSLC克隆。通过对表面标记、内源特征基因、转录表达谱、印记基因甲基化、核型、体外单倍体分化和体内睾丸移植分化等指标鉴定，筛选出具有典型SSC生物学特征和功能的SSLC克隆。本项目将从细胞重编程角度阐明SSLC命运决定的关键转录因子，为男性不育治疗和转基因动物生产提供新的途径。

中文关键词： 精原干细胞；iPS细胞；重编程；多能性；

英文摘要： Spermatogonial stem cells(SSC) are rare adult stem cells that can transfer genetic information to the offsprings in the body of male animals.This project aims to do direct reprogramming of somatic cells into spermatogonial stem cell-like cells (SSLC).First, a set of SSC transcription factors will be screened out for the direct reprogramming assay from 15 candidates that consist of Id4, Nanos2, Bcl6b, Etv5, Lhx1, Foxo1, Pou3f1, Pou5f1, Sox2, Klf4, c-Myc, Lin28, Utf1, Plzf and Taf4b; Second, the transcription factors will be introduced into mouse embryonic firoblasts or adult tail tip fibroblasts by retroviral vector pMXs; Third, the transduced fibroblasts will be maintained under SSC culture condition which contains glial cell line-derived neurotrophic factor(GDNF) to induce SSLC colonies; At last, the SSLC colonies with typical biological characteristics and fuctions of SSC will be screened out by the following criteria: expression of surface markers, expression of specific endogerous genes, transcriptome profile , methylation of imprinting genes, karyotype, haploid differentiation in vitro and differentiation in vivo via transplanting into testis. In conclusion, this project will clarify the pivotal transcription factors for determining SSLC fate from cellular reprogramming perspective; this project will also

英文关键词： spermatogonial stem cells；iPS cells；reprogramming；pluripotency；