Department of Dairy and Animal Science, Center for Reproductive Biology and Health, The Pennsylvania State University, University Park, Pennsylvania 16802, USA.
Reproduction. 2010 May;139(5):893-903. doi: 10.1530/REP-09-0513. Epub 2010 Feb 12.
The undifferentiated spermatogonial population consists of stem and progenitor germ cells which function to provide the foundation for spermatogenesis. The stem cell component, termed spermatogonial stem cells (SSCs), is capable of self-renewal and differentiation. These unique attributes have made them a target for novel technologies to enhance reproductive function in males. With bulls, culture and transplantation of SSCs have the potential to enhance efficiency of cattle production and provide a novel avenue to generate transgenic animals. Isolation of SSCs is an essential component for the development of these techniques. In rodents and non-human primates, undifferentiated spermatogonia and SSCs express the surface marker THY1. The hypothesis tested in this study was that THY1 is a conserved marker of the undifferentiated spermatogonial population in bulls. Flow cytometric analyses showed that the THY1+ cell fraction comprises a rare sub-population in testes of pre-pubertal bulls. Immunocytochemical analyses of the isolated THY1+ fraction for expression of VASA showed that this cell population is comprised mostly of germ cells. Additionally, expression of the undifferentiated spermatogonial specific transcription factor promyelocytic leukemia zinc finger (PLZF, ZBTB16) protein was found to be enriched in the isolated THY1+ testis cell fraction. Lastly, xenogeneic transplantation of bull testis cells into seminiferous tubules of immunodeficient mice resulted in greater than sixfold more colonies from isolated THY1+ cells compared to the unselected total testis cell population indicating SSC enrichment. Collectively, these results demonstrate that THY1 is a marker of undifferentiated spermatogonia in testes of pre-pubertal bulls, and isolation of THY1+ cells results in their enrichment from the total testis cell population.
未分化精原细胞群体由干细胞和祖细胞组成，其功能是为精子发生提供基础。干细胞成分，称为精原干细胞（SSC），能够自我更新和分化。这些独特的特性使它们成为新技术的目标，以增强雄性的生殖功能。对于公牛，SSC 的培养和移植有可能提高牛的生产效率，并为产生转基因动物提供新途径。SSC 的分离是开发这些技术的重要组成部分。在啮齿动物和非人类灵长类动物中，未分化的精原细胞和 SSC 表达表面标记物 THY1。本研究测试的假设是，THY1 是公牛未分化精原细胞群体的保守标记物。流式细胞分析表明，THY1+细胞群在未成熟公牛的睾丸中构成了一个罕见的亚群。对分离的 THY1+细胞群进行 VASA 表达的免疫细胞化学分析表明，该细胞群主要由生殖细胞组成。此外，在分离的 THY1+睾丸细胞群中发现未分化精原细胞特异性转录因子早幼粒细胞白血病锌指（PLZF，ZBTB16）蛋白的表达丰富。最后，将公牛睾丸细胞异种移植到免疫缺陷小鼠的生精小管中，与未选择的总睾丸细胞群相比，从分离的 THY1+细胞中产生的集落数量增加了六倍以上，表明 SSC 富集。总之，这些结果表明，THY1 是未成熟公牛睾丸中未分化精原细胞的标记物，分离 THY1+细胞可从总睾丸细胞群中富集它们。
