GENETIC ANALYSIS OF GERM CELL FORMATION

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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.