Formative pluripotent stem cells show features of epiblast cells poised for gastrulation

The pluripotency of mammalian early and late epiblast could be recapitulated by naïve embryonic stem cells (ESCs) and primed epiblast stem cells (EpiSCs), respectively. However, these two states of pluripotency may not be sufficient to reflect the full complexity and developmental potency of the epiblast during mammalian early development. Here we report the establishment of self-renewing formative pluripotent stem cells (fPSCs) which manifest features of epiblast cells poised for gastrulation. fPSCs can be established from different mouse ESCs, pre-/early-gastrula epiblasts and induced PSCs. Similar to pre-/early-gastrula epiblasts, fPSCs show the transcriptomic features of formative pluripotency, which are distinct from naïve ESCs and primed EpiSCs. fPSCs show the unique epigenetic states of E6.5 epiblast, including the super-bivalency of a large set of developmental genes. Just like epiblast cells immediately before gastrulation, fPSCs can efficiently differentiate into three germ layers and primordial germ cells (PGCs) in vitro. Thus, fPSCs highlight the feasibility of using PSCs to explore the development of mammalian epiblast. We established formative pluripotent stem cells (fPSCs) with long-term self-renewal capacities from mESCs, and mouse pre-gastrula epiblast cells, through mimicking the in vivo environment of postimplantation developing embryos combined with suppression of the Wnt/β-catenin pathway. Then we performed transcriptome and epigenome analyses to investigate whether fPSCs represent a stable intermediate state between naïve ESCs and primed EpiSCs.