GENETIC ANALYSIS OF GERM CELL FORMATION

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得主要资金，因此可以在其他CRISP条目中表示。所列机构为中心，不一定是研究者所在机构。生殖细胞可以从人胚胎干（ES）细胞中分化出来。因此，我们假设人ES细胞系统可用于特异性地探测人生殖细胞形成和分化的遗传学。此外，我们假设，下调关键基因，映射到一个区域的Y染色体，这是通常删除不育男性，AZFc区，将废除的能力，形成和/或分化的人类男性生殖细胞在体外和体内的移植系统。为了探索这一假设，我们提出：1）表征NIH批准的人ES细胞系在体外和体内对生殖细胞谱系的贡献的能力。为此，我们将使用ES细胞系H1、H14和HSF 1（NIH代码：WA 01、WA 14和UC 01）。2)沉默映射到AZFc缺失区间的Y染色体基因，并在体外评估生殖细胞形成。3)沉默Y染色体基因，映射到AZFc缺失区间，并评估生殖细胞的能力，殖民非人灵长类动物生精小管，随后分化。人类胚胎干细胞来源的雄性生殖细胞在非人灵长类动物生精小管中定植的能力将是ONPRC研究的重点。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Germ cells can be specified and differentiated from human embryonic stem (ES) cells. Thus, we hypothesize that the human ES cell system can be used to specifically probe the genetics of human germ cell formation and differentiation. Moreover, we hypothesize that the downregulation of key genes that map to a region of the Y chromosome that is commonly deleted in infertile men, the AZFc region, will abolish the ability to form and/or differentiate human male germ cells in vitro and in vivo in a transplant system. To explore this hypothesis, we propose to: 1) Characterize the ability of NIH-approved human ES cell lines to contribute to the germ cell lineage in vitro and in vivo. For this purpose, we will use ES cell lines H1, H14 and HSF1 (NIH codes: WA01, WA14 and UC01). 2) Silence the Y chromosome genes that map to the AZFc deletion interval and assess germ cell formation in vitro. 3) Silence the Y chromosome genes that map to the AZFc deletion interval and assess the ability of germ cells to colonize nonhuman primate spermatogenic tubules and subsequently differentiate. The ability of human ES derived male germ cells to colonize nonhuman primate spermatogenic tubules will be the focus of research conducted at ONPRC.