Establishment of mouse stem cells that can recapitulate the developmental potential of primitive endoderm

Abstract

The mammalian blastocyst consists of three distinct cell types: epiblast, trophoblast (TB), and primitive endoderm (PrE). Although embryonic stem cells (ESCs) and trophoblast stem cells (TSCs) retain the functional properties of epiblast and TB, respectively, stem cells that fully recapitulate the developmental potential of PrE have not been established. Here, we report derivation of primitive endoderm stem cells (PrESCs) in mice. PrESCs recapitulate properties of embryonic day 4.5 founder PrE, are efficiently incorporated into PrE upon blastocyst injection, generate functionally competent PrE-derived tissues, and support fetal development of PrE-depleted blastocysts in chimeras. Furthermore, PrESCs can establish interactions with ESCs and TSCs and generate descendants with yolk sac–like structures in utero. Establishment of PrESCs will enable the elucidation of the mechanisms for PrE specification and subsequent pre- and postimplantation development.

A chemically defined system supports two distinct types of stem cell from a single blastocyst and their self‐assembly to generate blastoid

“…To further investigate the molecular features of ACL cultured cells, We compared ACL‐ESCs and ACL‐XEN cells with ESCs, TSCs, 67 XEN cells, 67 E4.5‐Epi 66 and E4.5_PrE. UHC analysis demonstrated that ACL‐ESCs were close to ESCs (Figure S3A ), in line with the IF staining and in vivo chimera assays results.…”

“… 65 HTSeq (0.6.1) was used for reads counting, and the RefSeq gene expression level was estimated using the reads per kilobase transcriptome per million reads (RPKM) method. In vivo data for mouse embryos E4.5 Epi (E‐MTAB‐2958), 66 E4.5 PrE, 66 ESCs (GSE119985), 60 TSCs 67 and XEN cells (GSE159181) 67 were downloaded and processed identically. DEGs in different samples were determined using the edgeR package, with a fold change ≥2 and p ≤ 0.5.…”

“…To further investigate the molecular features of ACL cultured cells, We compared ACL-ESCs and ACL-XEN cells with ESCs, TSCs, 67 XEN cells, 67 E4.5-Epi 66 trophectoderm-related makers (Cdx2, Gata2, Tead4, etc.) were highly expressed in TSCs, however, endoderm-related makers (Gata4, Sox17, Foxa2, etc.)…”

Cell fate decision by a morphogen-transcription factor-chromatin modifier axis

“…2a) and bulk RNA-seq (Supplementary Fig. 2b) show that genes such as Sox17, Gata4/6 are highly enriched in PrECLCs 16,17 . To see if these genes play any potential role in the bifurcating decision between pluripotent and PrE fates, we add each factor to JGES and show that Gata4 stands out as a critical inhibitor for pluripotent reprogramming (Fig.…”

Chromatin as a sensor of metabolic changes during early development

“…This deep knowledge of the signaling milieu allowed researchers to recapitulate many stages of mouse and human embryonic development in vitro . By isolating cells from the mouse blastocyst, naïve pluripotent stem cells (mESC), trophoblast stem cells (mTSC) and extraembryonic primitive endoderm stem cells (nEnd) mimicking the ICM, the TE and the PrE respectively, can be obtained ( Tanaka et al, 1998 ; Ying et al, 2008 ; Anderson et al, 2017 ; Ohinata et al, 2022 ). Culture of naïve mESCs in vitro is achieved by the addition of GSK3β and MAPK/ERK inhibitors (2i) and leukemia inhibitory factor (LIF) in the so-called serum-free 2i/L media ( Ying et al, 2008 ).…”