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Capturing human trophoblast development with naive pluripotent stem cells in vitro

ABSTRACT: Capturing human trophoblast development with naive pluripotent stem cells in vitro

PROVIDER: PRJNA705591 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR11050200.fastq.gz	Fastqsanger.gz
	SRR11050169.fastq.gz	Fastqsanger.gz
	SRR11050170.fastq.gz	Fastqsanger.gz
	SRR11050171.fastq.gz	Fastqsanger.gz
	SRR11050172.fastq.gz	Fastqsanger.gz

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Project description:Recent reports suggested human embryonic development is not always consistent with mouse. However, experimental approach to in vivo human embryos is extremely limited. Therefore, it is important to develop an appropriate in vitro cell culture system to analyse human pre-implantation development. In this report, we use human naive pluripotent stem cells (PSCs), which share features with pre-implantation epiblasts and show in vitro lineage specification of human trophoblast lineages. We successfully differentiated naive PSCs into trophectoderm, cytotrophoblast, syncytiotrophoblast, and extravillous trophoblast. Cytotrophoblasts could be cultured for long term as stem cells.

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Polycomb Repressive Complex 2 Restricts Human Naive Pluripotent Stem Cell to Trophoblast Fate Induction

Project description:Human naive pluripotent stem cells have unrestricted lineage potential. Underpinning this property, naive cells are thought to lack chromatin-based lineage barriers. However, this assumption has not been tested. Here, we apply multi-omics to comprehensively define the chromatin-associated proteome, histone post-translational modifications and transcriptome of human naive and primed pluripotent stem cells. Integrating the chromatin-bound proteome and histone modification data sets reveals differences in the relative abundance and activities of distinct chromatin modules, identifying a strong enrichment of Polycomb Repressive Complex 2 (PRC2)-associated H3K27me3 in naive pluripotent stem cell chromatin. Single-cell approaches and human blastoid models reveal that PRC2 activity acts as a chromatin barrier restricting the differentiation of naive cells towards the trophoblast lineage, and inhibiting PRC2 promotes trophoblast fate induction and cavity formation. Our results establish that human naive pluripotent stem cells are not epigenetically unrestricted, but instead possess chromatin mechanisms that oppose the induction of alternative cell fates.

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Project description:We report global transcriptional alteration and transcriptional trajectories from naive human pluripotent stem cells-derived trphectoderm to cytotrophoblast. By graph-based cell identities of total samples on the Seurat platform, we identified 9 main clusters, and further revealed the pseudotime differentiation during trophectoderm-cytotrophoblast transition.

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Project description:In human embryos, naive pluripotent cells of the inner cell mass (ICM) generate epiblast, primitive endoderm and trophectoderm (TE) lineage, whence trophoblast cells derive. In vitro, naive pluripotent stem cells (PSCs) retain this potential and efficiently generate trophoblast stem cells (TSCs), while conventional PSCs form TSCs at low efficiency. Transient histone deacetylase and MEK inhibitions with LIF stimulation is used to chemically reset conventional to naive PSCs. Here we report that chemical resetting induces expression of both naive and TSC markers and of placental imprinted genes. A modified chemical resetting protocol allows for the fast and efficient conversion of conventional PSCs into TSCs, entailing shutdown of pluripotency genes and full activation of the trophoblast master regulators, without induction of amnion markers. Chemical resetting generates a plastic intermediate state, characterised by co-expression of naive and TSC markers, after which cells steer towards one of the two fates in response to the signalling environment. The efficiency and rapidity of our system will be useful to study cell fate transitions, and to generate models of placental disorders.

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Project description:Human naive pluripotent stem cells have unrestricted lineage potential. Underpinning this property, naive cells are thought to lack chromatin-based lineage barriers. However, this assumption has not been tested. Here, we define the chromatin-associated proteome, histone post-translational modifications and transcriptome of human naive and primed pluripotent stem cells. Our integrated analysis reveals differences in the relative abundance and activities of distinct chromatin modules. We identify a strong enrichment of Polycomb Repressive Complex 2 (PRC2)-associated H3K27me3 in naive pluripotent stem cell chromatin, and H3K27me3 enrichment at promoters of lineage-determining genes, including trophoblast regulators. PRC2 activity acts as a chromatin barrier restricting the differentiation of naive cells towards the trophoblast lineage, while inhibition of PRC2 promotes trophoblast fate induction and cavity formation in human blastoids. Together, our results establish that human naive pluripotent stem cells are not epigenetically unrestricted, but instead possess chromatin mechanisms that oppose the induction of alternative cell fates.

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