诱导多能干细胞体外向生精细胞自发分化潜能的研究  

Spontaneous differentiation potency of induced pluripotent stem cells into male germ cells in vitro

作　　者：李朋[1,2,3] 胡洪亮[1,2,3] 张玲玲[4] 丁慧[1,2,3] 张伟[1,2,3] 田汝辉 刘平[1,2,3] 史占平[1,2,3] 郭熙志[4] 李铮[1,2,3] 黄翼然[1,2,3] 

机构地区：[1]上海交通大学医学院附属仁济医院泌尿外科 [2]上海市男科学研究所精子发育与遗传学实验室 [3]上海市人类精子库,上海200001 [4]上海交通大学Bio-X中心,上海200240

出　　处：《中华男科学杂志》2011年第1期8-12,共5页National Journal of Andrology

基　　金：国家重点基础研究发展计划(973计划;2010CB945200);上海市教委重点课题(10ZZ70);上海交通大学医学院自然基金课题(2008XJ017)~~

摘　　要：目的:探讨诱导多能干细胞(iPS)体外培养自发分化过程中生精细胞相关基因的表达,评估iPS体外向生精细胞自发分化的潜能。方法:经类胚体(EB)形成,体外诱导iPS向生精细胞分化,实时定量PCR和PCR检测生精细胞相关基因的表达。结果:实时定量PCR和PCR结果显示iPS经EB形成诱导分化后生精细胞不同时期的相关基因均有不同程度表达。iPS体外培养自发分化后生精细胞相关基因出现4种时间表达特征:Oct4基因表达量呈波浪状上升;Dppa3和Stra8基因表达量随诱导时间延长而下降;Dazl基因表达量呈波浪状下降;减数分裂前期基因Tex14、Msy2,减数分裂期基因Scp1、Scp3以及单倍体基因Akap3随着诱导时间延长先表达增加,而后表达下降。结论:iPS在经EB自发分化过程中表达生精细胞不同时期的相关基因,并且表达雄性配子单倍体基因,具有向雄性配子的分化潜能。Objective：To explore the expression profile of male germ cell-associated genes during the spontaneous differentiation of induced pluripotent stem cells（iPS） and assess the potency of their spontaneous differentiation into male germ cells in vitro.Methods：Embryoid body（EB） formation was used to promote the spontaneous differentiation of iPS into male germ cells,and the expressions of germ cell-associated genes were detected by real-time PCR and PCR.Results：Real-time PCR and PCR revealed different expression levels of relevant genes at different times of iPS spontaneous differentiation into male germ cells.Each of the 9 genes analyzed exhibited one of the four temporal expression patterns：wavelike increase of Oct4,progressive decrease of Dppa3 and Stra8,wavelike decrease of Dazl,and decrease following initial increase of Tex14,Msy2,Scp1,Scp3 and Akap3.Conclusion：Induced pluripotent stem cells express male germ cell-associated genes and male haploid genes during their spontaneous differentiation through EB formation,and have the potency of differentiating into male gametes.

关 键 词：诱导多能干细胞 类胚体 生精细胞 基因表达 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]