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Loss of E-cadherin promotes extraembryonic fates accompanying multilineage differentiation

ABSTRACT: Loss of E-cadherin promotes extraembryonic fates accompanying multilineage differentiation

PROVIDER: PRJNA676122 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR13036156_1.fastq.gz	Fastqsanger.gz
	SRR13036156_2.fastq.gz	Fastqsanger.gz
	SRR13036157_1.fastq.gz	Fastqsanger.gz
	SRR13036157_2.fastq.gz	Fastqsanger.gz
	SRR13036158_1.fastq.gz	Fastqsanger.gz

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Loss of E-cadherin promotes extraembryonic fates accompanying multilineage differentiation

Project description:Embryonic development involves a range of complex morphogenic events as the embryo generates various cell types that go onto form functional tissues. This process is heavily reliant on symmetry breaking events that control the emergence of separate lineages that then spatial organize and physically arranging themselves within the embryo often via regulation of cell-cell adhesions. This work examines how changing adhesions in 3D aggregates of human pluripotent stem cells (hPSCs) encapsulated in hydrogel mixed with extracellular matrix allows for emergent population and morphological behaviors reminiscent of early embryonic behaviors and morphologies. In this context, knockdown of the cell adhesion molecule CDH1 generated aggregates with protrusion morphologies that did not arise in aggregates cultured without encapsulation. Genomic analysis revealed that the combination of CDH1 knockdown and encapsulation promoted emergence of extra-embryonic lineages. Further the transcriptomes of encapsulated CDH1(-) hPSCs were comparable to previously published single cell RNA-sequencing data of human embryos. Overall, these results establish a hPSC culture system that resembles early embryonic fate decisions and morphologies. Additionally, these studies highlight a potential relationship between changing adhesions, microenvironment, and acquisition of specific lineage fates, progressing our understanding of early human embryonic biology.

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Protein Kinase C Establishes the Signaling Boundary Between Mesodermal and Extraembryonic Lineages in Human Embryonic Stem Cell Differentiation

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OSKM induce extraembryonic endoderm stem (iXEN) cells in parallel to iPS cells

Project description:While the reprogramming factors OCT4, SOX2, KLF4, and MYC (OSKM) can reactivate the pluripotency network in terminally differentiated cells, they also regulate expression of non-pluripotency genes in other contexts, such as the mouse primitive endoderm. The primitive endoderm is an extraembryonic lineage established alongside the pluripotent epiblast in the blastocyst, and is the progenitor pool for extraembryonic endoderm stem (XEN) cells. Several studies have shown that endodermal genes are upregulated in fibroblasts undergoing reprogramming, although whether endodermal genes promote or inhibit acquisition of pluripotency is unclear. We show that, in fibroblasts undergoing conventional reprogramming, OSKM-induced expression of endodermal genes leads to formation of induced XEN (iXEN) cells, which possess key properties of blastocyst-derived XEN cells, including morphology, transcription profile, self-renewal, and multipotency. Our data show that iXEN cells arise in parallel to iPS cells, indicating that OSKM are sufficient to drive cells to two distinct fates during reprogramming. Sequence-based mRNA transcriptional profiling of three different cell lines (MEF, XEN, iXEN) with multiple biological replicates, under two different growth medium conditions (ESC medium, XEN medium) for XEN and iXEN cells.

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