Derivation of functional trophoblast stem cells from human primed pluripotent stem cells [bulk RNA-seq]

Trophoblast stem cells (TSC) have recently been derived from human embryos, and early first trimester placenta; however, aside from ethical challenges, the unknown disease potential of these cells limits their scientific utility. We have previously established a BMP4-based two-step protocol for differentiation of primed human pluripotent stem cells (hPSC) into functional trophoblast; however, those trophoblast could not be maintained in a self-renewing TSC-like state. Here, we use the first step from this protocol, followed by a switch to a newly-developed TSC media, to derive bona-fide TSC. We show that these cells resemble placenta- and naïve hPSC-derived TSC, based on their transcriptome, as well as in vitro and in vivo differentiation potential. We conclude that primed hPSC can be used to generate functional TSC, through a simple protocol which can be applied to a widely-available set of existing hPSC, including induced pluripotent stem cells derived from patients with known birth outcomes. We differentiated primed PSC into TE-like cells by treatment with BMP4/IWP2 for 4 days followed by adaptation in TSC media for at least 6 passages. Starting and final cells as well as various intermediate cell types were profiled by RNA-seq.