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ScRNA-seq of mouse EpiSCs (epiblast stem cells) differentiated to extraembryonic mesoderm in vitro, in a monolayer

ABSTRACT: scRNA-seq of mouse EpiSCs (epiblast stem cells) differentiated to extraembryonic mesoderm in vitro, in a monolayer

PROVIDER: PRJEB55467 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	ERR10093000.fastq.gz	Fastqsanger.gz
	ERR10093100.fastq.gz	Fastqsanger.gz
	ERR10093200.fastq.gz	Fastqsanger.gz
	ERR10093300.fastq.gz	Fastqsanger.gz
	ERR10093001.fastq.gz	Fastqsanger.gz

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scRNA-seq of mouse EpiSCs (epiblast stem cells) differentiated to extraembryonic mesoderm in vitro, in a monolayer

Project description:EpiSCs derived from epiblast of a gastrulating embryo differentiated towards extraembryonic mesoderm to test the efficiency of the differentiation protocol and identity and homogeneity of the resulting cell population.

2022-11-30 | E-MTAB-12107 | biostudies-arrayexpress

RNA-seq of wild-type and PGR (progesterone receptor) knockout pluripotent mouse EpiSCs (epiblast stem cells) and differentiated samples derived from these cell lines (primitive streak and mesoderm)

Project description:EpiSCs (epiblast stem cells) are murine pluripotent stem cells derived from the epiblast of a post-implantation embryo. These cells are primed to differentiate towards embryonic germ layers and are a useful model to study these early developmental events. PGR (progesterone receptor) is a nuclear receptor described mainly in the context of fertility and breast cancer. We noticed that it is also a TF that is expressed in primed EpiSCs, but not in naive mESCs and is further upregulated during mesoderm specification. We hypothesized that it is involved in differentiation to primitive streak and mesoderm progenitors. To test this hypothesis we generated EpiSCs with a genetic knockout of PGR using CRISPR-Cas9-mediated knock-in of an antibiotic resistance cassette. Differentiation of the wild-type and knockout cells and comparison of the different cell types allowed us to conclude that PGR is important for acquiring extraembryonic mesoderm fate and modulates cardiac differentiation, specifically it ensures that correct pools of FHF (first heart field) and SHF (second heart field) are being produced. Our study sheds light on previously unappreciated role of PGR in early embryonic development.

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RNA-seq of H9 cell line during differentiation from naïve extraembryonic endoderm to extraembryonic mesoderm.

Project description:Extraembryonic mesoderm (ExM) is one of the first cell types that emerges during embryogenesis and constitutes essential supportive tissues for the pregnancy. Primate ExM is known to form prior to gastrulation, unlike its murine counterpart which is derived from the primitive streak. Based on the embryonic morphology and the proximity of ExM to the extraembryonic endoderm (hypoblast), we hypothesised that ExM can be derived in vitro from the naïve extraembryonic endoderm (nEnd) cell line. We applied a mesoderm differentiation protocol, which has been reported to induce ExM from mouse epiblast stem cells, on human nEnd and analysed the transcriptome on day 0, 1, 2, 8 and 15.

2023-06-29 | E-MTAB-12517 | biostudies-arrayexpress

Developmental equivalence of epiblast stem cells (EpiSCs)

2013-09-15 | GSE46227 | GEO

Developmental equivalence of epiblast stem cells (EpiSCs)

Project description:Epiblast stem cells (EpiSCs) were derived from the epiblast or the ectoderm (epi/ect) of pre-gastrula stage to late-bud stage mouse embryos. To identify if the EpiSCs retain any original stage specific characteristics or which developmental stage of epi/ect they most closely related to, we performed microarray analysis to compare the gene expression profile of multiple EpiSC lines with that of epi/ect of 7 different stages. Eighteen EpiSC lines established from R1-129 embryos of different stages and 1 line (EpiSC9) imported were harvested in triplicate cultured separately for 3 days. The * marks the subline thawed at a different time and harvested but originated from the same embryo. EpiSC9_T is an RNA sample provided by Dr Tesar. ESCs, iPSCs and MEFs were prepared accordingly. For the epiblast samples, in order to avoid the averaging effect by pooling samples, we linearly amplified the RNA starting from a single epiblast or ectoderm.

2013-09-15 | E-GEOD-46227 | biostudies-arrayexpress

Expression profiling by high throughput sequencing (RNA sequencing)

Project description:RNAseq of CLDN6 sorted Epiblast Stem Cells (EpiSCs), WT vs Pbx1-KO lines in mouse embryonic stem cells (ES (2i/LIF and serum/LIF) and EpiSCs, as well as across differentiation (EpiSCs differentiated towards Posterior Primitive Streak (PPS) and Extraembryonic Mesoderm (ExM). Also EpiSCs treated with PD03 (MEK inhibitor) and 6h of WNT stimulation (CHIR).

2024-11-29 | E-MTAB-13618 | biostudies-arrayexpress

Modeling human extraembryonic mesoderm cells using naive pluripotent stem cells [scRNA_seq_timecourse_naiveTSC]

Project description:time course scRNAseq of naïve to trophoblast stem cells and extraembryonic mesoderm conversion and D70 sorted extraembryonic mesoderm. Samples were collected at day 0, 1, 2, 4, 8, 13 and 18.

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Project description:The epiblast is the first cell type that forms apical-basal polarity de novo as the mouse embryo implants into the maternal uterus, while the extraembryonic neighbours of the epiblast - trophectoderm and primitive endoderm - retain their pre-established polarity beyond implantation [1]; however, it is still unclear how the epiblast establishes apical-basal polarity de novo. Here, we focused on Rap1 signaling pathway, which is activated during the transition of the epiblast from the naïve to primed state of pluripotency during implantation [2]. Through the preestablished in vitro three-dimensional culture system [3], genetic knockouts and proximity-biotinylation analyses, we found that Rap1 integrates multiple signals that contribute to de novo formation of apical-basal polarity. Importantly, formation of apical-basal polarity in the epiblast is essential for its correct patterning and proper communication with the extraembryonic lineages. Altogether, these results not only dissect molecular details of de novo apical-basal polarity formation, but also have broader implications for epithelial polarity and development.

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scRNA-seq for mEpiSCs (epiblast stem cells) differentiated to posterior primitive streak (PPS) in vitro, in a monolayer

Project description:EpiSCs established from murine epiblast were differentiated to posterior primitive streak (PPS) and analyzed using scRNA-seq to investigate identity and homogeneity of the resulting population.

2022-11-30 | E-MTAB-12106 | biostudies-arrayexpress

Wnt signalling sustains an EpiSCs subpopulation similar to primitive streak with increased mesendodermal potency

Project description:During gastrulation, epiblast cells are pluripotent and their fate is thought to be constrained principally by their position. Cell fate is progressively restricted by localised signalling cues from areas including the primitive streak (PS). However, it is unknown whether this restriction accompanies, at the single cell level, a reduction in potency. Investigation of these early transition events in vitro is possible via the use of Epiblast Stem Cells (EpiSCs), self-renewing pluripotent cell lines equivalent to the postimplantation epiblast. Strikingly, EpiSCs express various early lineage-specific markers in self-renewing conditions. However, it is unknown whether cells that express these markers are pluripotent, spontaneously differentiated, or biased towards specific lineages. Here we show that EpiSC are inherently heterogeneous and contain two major and mutually exclusive subpopulations with PS and neural characteristics respectively. Using differentiation assays and embryo grafting we demonstrate that PS-like EpiSCs are biased towards mesoderm and endoderm differentiation but they still retain their pluripotent character. The acquisition of a PS character by undifferentiated EpiSC is mediated by paracrine Wnt signalling. Elevation of Wnt activity promotes further restriction into PS-associated cell fates which occurs via the generation of distinct clonal mesendodermal and neuromesodermal precursors. Collectively, our data suggest that primed pluripotency encompasses a range of reversible lineage-biased states reflecting the birth of pioneer lineage precursors from a pool of uncommitted EpiSCs similar to the earliest cell fate restriction events taking place in the gastrula-stage epiblast.

2014-03-04 | GSE48476 | GEO

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