Generation of spermatogonia from human and non-human primate pluripotent stem cells

Failures in germline development drive male infertility, but the lack of model systems that recapitulate human spermatogenesis hampers therapeutic development. Here, we develop a system to differentiate human-induced pluripotent stem cells (iPSCs) into primordial germ cell-like cells that self-organize with mouse fetal testicular cells into seminiferous tubule-like structures within a xenogeneic reconstituted testis (xrTestis). Subsequent transplantation of xrTestes into immunodeficient mice results in efficient generation of male germ cells up to meiotic onset, including spermatogonia with evidence of prepachytene PIWI-interacting RNA (piRNA) biogenesis, as well as differentiating spermatogonia and rare preleptotene spermatocytes exhibiting transcriptomic and phenotypic similarities to their in vivo counterparts. As future clinical applications will require testing in non-human primates, we apply a similar strategy to differentiate rhesus iPSCs through fetal germ cell stages into spermatogonia and differentiating spermatogonia. Together, these platforms provide a foundation for studying primate germ cell lineages and represent a step toward in vitro gametogenesis.