Rat epiblast-derived pluripotent stem cells produce functional germ cells

In mammals, pluripotent cells transit through a continuum of distinct molecular and functional states en route to initiating lineage specification. Capturing pluripotent stem cells (PSCs) mirroring in vivo pluripotent states provides accessible in vitro models to study the pluripotency program and mechanisms underlying lineage restriction. Here, we develop optimal culture conditions to derive and propagate post-implantation epiblast-derived PSCs (EpiSCs) in rats, a valuable model for biomedical research. We show that rat EpiSCs can be reset toward the naïve pluripotent state with exogenous Klf4, albeit not with the other five candidate genes (Nanog, Klf2, Esrrb, Tfcp2l1, and Tbx3) effective in mice. Finally, we demonstrate that rat EpiSCs retain competency to produce authentic primordial germ cell-like cells that contribute to functional gametogenesis leading to the birth of viable offspring. Our findings in the rat model uncover conserved principles underpinning pluripotency and germline competency across species. Overall design: RNA-seq of rat embryonic stem cells (ESCs), ESC-like cells (ESCLCs), epiblast stem cells (EpiSCs), epiblast-like cells (EpiLCs), and primordial germ cell like cells (PGCLCs).