The Transcriptional and Functional Properties of Mouse Epiblast Stem Cells Resemble the Anterior Primitive Streak

Abstract

Mouse epiblast stem cells (EpiSCs) can be derived from a wide range of developmental stages. To characterize and compare EpiSCs with different origins, we derived a series of EpiSC lines from pregastrula stage to late-bud-stage mouse embryos. We found that the transcriptomes of these cells are hierarchically distinct from those of the embryonic stem cells, induced pluripotent stem cells (iPSCs), and epiblast/ectoderm. The EpiSCs display globally similar gene expression profiles irrespective of the original developmental stage of the source tissue. They are developmentally similar to the ectoderm of the late-gastrula-stage embryo and behave like anterior primitive streak cells when differentiated in vitro and in vivo. The EpiSC lines that we derived can also be categorized based on a correlation between gene expression signature and predisposition to differentiate into particular germ-layer derivatives. Our findings therefore highlight distinct identifying characteristics of EpiSCs and provide a foundation for further examination of EpiSC properties and potential.

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

“…This analysis was restricted to the top 500 differentially expressed gene identified at successive developmental stages sorting by Hotelling T² score, as described in Kojima et al (2014). …”

“…The distribution of expression levels of the two sets of differentially expressed genes is shown for Esrrb Hi (H), Esrrb Med (M), and Esrrb Neg (Neg) E‐GFPd1 ESCs and embryo‐derived (late bud stage) TNG E‐2a‐tdT EpiSCs (Epi). Gene expression distributions in cavity (CAV), prestreak (PS), late midstreak (LMS), late streak (LS), no bud (OB), early bud (EB), or late bud (LB) stage embryos from Kojima et al (2014) are shown for comparison. Boxes span the inter‐quartile range (IQR) from the first to the third quartile.…”

“…This supports the notion that ESCs that have lost Esrrb expression have initiated the transition from naïve to primed pluripotency. Comparison of the gene expression profile of sorted Esrrb‐GFP populations and EpiSCs with the transcriptome of epiblast cells from different stages of post‐implantation development (Kojima et al , 2014) revealed that Esrrb‐GFP negative cells are closer transcriptionally to in vivo epiblast cells prior to gastrulation. In contrast, EpiSCs more closely resemble epiblast cells from later gastrulating embryos (Fig 3H).…”

“…A coordinated downregulation of naïve pluripotency markers is reminiscent of the process occurring during the transition from pre‐ to post‐implantation development (Boroviak et al , 2014, 2015; Kojima et al , 2014). The period of loss of NANOG expression occurring at peri‐implantation (Chambers et al , 2003; Acampora et al , 2013) may enable cells of the epiblast to downregulate a number of pivotal naïve pluripotency determinants, including ESRRB (Adachi et al , 2013).…”

Cdx2Efficiently Induces Trophoblast Stem-Like Cells in Naïve, but Not Primed, Pluripotent Stem Cells

“…Thus, unlike 129-derived ESC/iPSCs, 129-derived EpiSCs are not competent to respond to overexpressed Cdx2 by activating TSC gene expression, pointing to differences in developmental potential. Consistent with this proposal, EpiSCs are thought to be primed to differentiate into germ layer derivatives and express markers of the late epiblast, including Sox17, Gata6, Foxa2, and T [5,6,13,44,45]. We therefore hypothesized that the degree of lineage priming could differ between ESC/iPSCs that do not efficiently produce TSC-like cells and ESC/iPSCs that do efficiently produce TSC-like cells.…”