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Establishment of mouse primitive endoderm stem cell lines [scRNA-Seq II]

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 Epi and TB, the currently available extraembryonic endoderm cells (XENC) do not fully recapitulate the developmental potential of PrE. Here we report derivation of primitive endoderm stem cells (PrESCs) in mice. PrESCs express both PrE and pluripotency marker genes like founder PrE. These cells are efficiently incorporated into PrE upon blastocyst injection, generate functionally competent PrE-derived tissues, and support fetal development of PrE-depleted blastocysts in chimeras. Establishment of PrESCs therefore represents a significant step-forward in elucidating the mechanisms for PrE specification and subsequent pre- and post-implantation development.

ORGANISM(S): Mus musculus

PROVIDER: GSE185461 | GEO | 2021/12/31

REPOSITORIES: GEO

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Primitive endoderm stem cell lines in mice

Project description:A blastocyst consists of three distinctive cell types: epiblast (EPI), trophoblast (TB), and primitive endoderm (PrE). Stem cell lines representing EPI and TB (embryonic stem (ES) cells and trophoblast stem (TS) cells) have been derived and they contribute to epiblast derivatives and trophoblast derivatives of stem cell-blastocyst chimeras, respectively. Although derived from PrE, extraembryonic endoderm (XEN) cells contribute to only a limited part of parietal endoderm (PE) but rarely to other PrE derivatives. Here we describe the establishment of primitive endoderm stem (PrES) cell lines in mice. PrES cells were derived and maintained in a serum-free media containing CHIR99021, FGF4, heparin, and PDGF-AA. RNA-seq analysis revealed that the transcriptome of PrES cells is globaly different from XEN cells: PrES cells express not only endoderm markers (Dab2, Gata4, Gata6, Sox17, etc.), but also pluripotent markers (Pou5f1, Cdh1, Nanog, Zfp42, etc.), and resembles in vivo founder PrE. PrES cells were rapidly and efficiently incorporated into PrE after blastocyst injection and efficiently contributed to all PrE derivatives, including PE, VE (visceral endoderm), and MZE (marginal zone endoderm) in chimeras. Importantly, PrES cells rescued embryonic lethality of PrE-depleted blastocysts by complementing all PrE derivatives. PrES cells thus not only contribute to understanding of the mechanisms of PrE specification but also provide a critical resource for artificial embryo reconstitution by stem cells alone.

2020-05-30 | GSE130957 | GEO

Establishment of primitive endoderm stem cell lines in mice [scRNA-Seq]

2021-12-31 | GSE159183 | GEO

Establishment of primitive endoderm stem cell lines in mice [bulk]

2021-12-31 | GSE159181 | GEO

Establishment of primitive endoderm stem cell lines in mice [coculture]

Project description:The mammalian blastocyst consists of three distinct cell types: epiblast, trophoblast, and primitive endoderm (PrE). Although stem cell lines that retain functional properties of epiblasts or trophoblasts have been established, establishment of stem cell lines that fully retain developmental potential of PrE has long been awaited. Here we report derivation of primitive endoderm stem (PrES) cells, which are capable to give rise functional PrE derived tissues and support fetal development of PrE-depleted blastocysts in chimeras, in mice. ES (#1M), TS cell (#1F), PrES (#6-5), and XEN (#6-5) cells were dissociated to single cells by incubation with 0.05% trypsin-EDTA at 37 °C for 3 min. The cells were resuspended in DMEM (Thermo Fisher Scientific) with 10% FBS, 1 × glutamine-penicillin-streptomycin, 0.1mM β-ME. The cells were replated alone or in combinations of each at a concentration of 20 cells per microwell (1.3×103 cells/dish) on 60mm EZSPHERE® SP dishes (Iwaki) and were placed at 37 °C and 5% CO2 for 1 day or 3 days.

2021-12-31 | GSE159182 | GEO

Stem cell lines of the early mouse embryo

Project description:Expression profiling of stem cell lines derived from the early embryo representing the trophoblast, primitive endoderm, early epiblast (inner cell mass E3.5) and late post-implantation epiblast (E5.5). Cells were grown without feeders and harvested. Comparisons were made to provide evidence of unique gene expression between the cell lines. Specifically, pre-implantation (ICM, epiblast and primitive endoderm, and trophoblast), as well as pre-implantation and post-implantation epiblasts.

2012-05-21 | E-GEOD-34799 | biostudies-arrayexpress

The immune checkpoint molecule, B7-H4/VTCN1, guides differentiation and suppresses proinflammatory responses and MHC class I expression in an embryonic stem cell-derived model of primitive human trophoblast III

Project description:VTCN1/B7-H4 is expressed in human embryonic stem cells (hESC) to an in vitro model of primitive trophoblast (TB) and first trimester but not term human placenta and that primitive trophoblast may be uniquely susceptible to certain pathogens.

2023-03-13 | GSE227096 | GEO

2023-03-13 | GSE227095 | GEO

2023-03-13 | GSE227094 | GEO

BMP4 signalling directs primitive endoderm-derived XEN cells to an extraembryonic visceral endoderm identity

Project description:The visceral endoderm (VE) is an epithelial tissue in the early postimplantation mouse embryo that encapsulates the pluripotent epiblast distally and the extraembryonic ectoderm proximally. In addition to facilitating nutrient exchange before the establishment of a circulation, the VE is critical for patterning the epiblast. Since VE is derived from the primitive endoderm (PrE) of the blastocyst, and PrE-derived eXtraembryonic ENdoderm (XEN) cells can be propagated in vitro, XEN cells should provide an important tool for identifying factors that direct VE differentiation. In this study, we demonstrated that BMP4 signalling induces the formation of a polarized epithelium in XEN cells. This morphological transition was reversible, and was associated with the acquisition of a molecular signature comparable to extraembryonic (ex) VE. Resembling exVE which will form the endoderm of the visceral yolk sac, BMP4-treated XEN cells regulated hematopoiesis by stimulating the expansion of primitive erythroid progenitors. We also observed that LIF exerted an antagonistic effect on BMP4-induced XEN cell differentiation, thereby impacting the extrinsic conditions used for the isolation and maintenance of XEN cells in an undifferentiated state. Taken together, our data suggest that XEN cells can be differentiated towards an exVE identity upon BMP4 stimulation, and therefore represent a valuable tool for investigating PrE lineage differentiation. Total RNA isolated in triplicate from XEN stem cell cultures that were untreated (samples 1-3) or treated with BMP4 growth factor (samples 4-6). Total RNA isolated in triplicate from XEN stem cells that were treated with BMP4 and were flow sorted as Afp::GFP-positive (samples 7-9) or Afp::GFP-negative (samples 10-12).

2011-09-13 | E-GEOD-32094 | biostudies-arrayexpress

Cell-to-cell gene expression variability followed by reinforcement progressively segregates lineages in the early mouse embryo

Project description:Mammalian embryonic development begins with the specification and segregation of the two extra-embryonic lineages, trophectoderm and primitive endoderm, from the pluripotent embryonic lineage, the epiblast. To establish a map of epiblast (EPI) versus primitive endoderm (PrE) lineage segregation which occurs within the inner cell mass (ICM), we comprehensively characterised the gene expression profiles of individual inner cells during blastocyst development. Lineage represents the two embryonic cell lineages: the epiblast (EPI), and the primitive endoderm (PE), which are segregated within the inner cell mass(ICM) during blastocyst development.

2013-11-26 | E-MTAB-1681 | biostudies-arrayexpress

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