Derivation of pluripotent epiblast stem cells from mammalian embryos

Abstract

Although the first mouse embryonic stem (ES) cell lines were derived 25 years ago using feeder-layer-based blastocyst cultures, subsequent efforts to extend the approach to other mammals, including both laboratory and domestic species, have been relatively unsuccessful. The most notable exceptions were the derivation of non-human primate ES cell lines followed shortly thereafter by their derivation of human ES cells. Despite the apparent common origin and the similar pluripotency of mouse and human embryonic stem cells, recent studies have revealed that they use different signalling pathways to maintain their pluripotent status. Mouse ES cells depend on leukaemia inhibitory factor and bone morphogenetic protein, whereas their human counterparts rely on activin (INHBA)/nodal (NODAL) and fibroblast growth factor (FGF). Here we show that pluripotent stem cells can be derived from the late epiblast layer of post-implantation mouse and rat embryos using chemically defined, activin-containing culture medium that is sufficient for long-term maintenance of human embryonic stem cells. Our results demonstrate that activin/Nodal signalling has an evolutionarily conserved role in the derivation and the maintenance of pluripotency in these novel stem cells. Epiblast stem cells provide a valuable experimental system for determining whether distinctions between mouse and human embryonic stem cells reflect species differences or diverse temporal origins.

Distinct Developmental Ground States of Epiblast Stem Cell Lines Determine Different Pluripotency Features

“…S1B). Lines E3, T9, and C1a1 have previously been characterized using this assay [2][3][4]. Taken together, these data demonstrate that these six cell lines were pluripotent EpiSC lines.…”

“…For example, mESCs undergo self-renewal in the presence of leukemia inhibitory factor (LIF) and inhibitors of fibroblast growth factor (FGF) signaling [1]. In contrast, EpiSCs require FGF and Activin signaling to sustain self-renewal [2][3][4]. Intriguingly, human embryonic stem cells (hESCs), although derived from preimplantation embryos, appear to share more characteristics with EpiSCs than with mESCs.…”

Esrrb extinction triggers dismantling of naïve pluripotency and marks commitment to differentiation

“…Indeed, ESCs cultured in LIF/FCS have been proposed to contain a mixture of pluripotent cells that have progressed to different extents toward a primed state (Hackett & Surani, 2014). Such a primed state can also be captured in vitro as EpiSC, by explantation and culture of post‐implantation epiblast cells (Brons et al , 2007; Tesar et al , 2007). In contrast, ESCs cultured in LIF supplemented with inhibitors of both MEK and GSK3b (two inhibitors or 2i; Ying et al , 2008) may represent the opposite end of the range of pluripotent states that can be maintained in vitro (Hackett & Surani, 2014).…”

Concise Review: Induced Pluripotent Stem Cells Versus Embryonic Stem Cells: Close Enough or Yet Too Far Apart?

“…Epiblast stem cells present the next stage in development and therefore have a more limited developmental potential. They show poor success in generating chimeras and can manifest expression of early lineage commitment markers [20,21]. Thus, the similarity of mouse EpiESC to human ESC/iPSC, together with the limited pluripotency tests available in human lines, raise questions on whether those cells are capable of producing whole embryos and about their general level of pluripotency.…”