Key Laboratory of Education Ministry of China for Mammalian Reproductive Biology and Biotechnology, Inner Mongolia University, Hohhot, China.
In Vitro Cell Dev Biol Anim. 2011 Sep;47(8):593-600. doi: 10.1007/s11626-011-9446-z. Epub 2011 Aug 19.
Spermatogonial stem cells (SSCs) maintain gamete production in the testes throughout adult life by balancing self-renewal and differentiation. In vitro culture of SSCs is a crucial technique for gene manipulation of SSCs to generate transgenic animals, for transplantation of SSCs to restore male fertility for infertile man, and for generation of pluripotent stem cells from SSCs to differentiate into various cell lineages. Isolation of highly purified SSCs is an all-important component for development of these techniques. However, definitive markers for SSCs, which purify SSCs (100% enrichment), are unknown. SSCs of many species can colonize the mouse testis; thus, we reasoned that same molecules of SSCs are conserved between species. In mouse, undifferentiated spermatogonia express the surface marker E-cadherin. The hypothesis tested in this work was that E-cadherin (also known as CDH1) can be expressed by undifferentiated spermatogonia of rat testes. In this paper, cross-section immunohistochemistry and whole-mount immunohistochemistry of rat seminiferous tubules were conducted to show that E-cadherin-positive cells were small in number and there are single, paired, and aligned spermatogonia attached along the basement membrane. During in vitro culture period, the undifferentiated rat spermatogonial colonies co-expressed E-cadherin and glial-derived neurotrophic factor family receptor alpha-1 or E-cadherin and promyelocytic leukemia zinc finger. Data collected during the study demonstrate that E-cadherin is expressed by a small population of rat undifferentiated spermatogonia both in vivo and during in vitro culture period.
精原干细胞 (SSC) 通过自我更新和分化来维持成年期睾丸中的配子产生。SSC 的体外培养是对 SSC 进行基因操作以产生转基因动物、将 SSC 移植以恢复不育男性的生育能力以及从 SSC 生成多能干细胞以分化为各种细胞谱系的关键技术。高纯度 SSC 的分离是开发这些技术的重要组成部分。然而，用于纯化 SSC（100%富集）的 SSC 明确标志物尚不清楚。许多物种的 SSC 均可定植于小鼠睾丸；因此，我们推断 SSC 之间存在物种间保守的相同分子。在小鼠中，未分化的精原细胞表达表面标志物 E-钙黏蛋白。本研究检验的假设是 E-钙黏蛋白（也称为 CDH1）可以在大鼠睾丸的未分化精原细胞中表达。在本文中，通过对大鼠生精小管的切片免疫组织化学和全组织免疫组织化学进行研究，结果表明 E-钙黏蛋白阳性细胞数量较少，并且有单个、成对和沿基底膜排列的精原细胞附着。在体外培养期间，未分化的大鼠精原细胞集落共同表达 E-钙黏蛋白和神经胶质细胞衍生的神经营养因子家族受体 α-1 或 E-钙黏蛋白和早幼粒细胞白血病锌指。在研究过程中收集的数据表明，E-钙黏蛋白在体内和体外培养期间均由大鼠未分化的精原细胞中的一小部分表达。
