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Rapid and robust directed differentiation of mouse epiblast stem cells into definitive endoderm and forebrain organoids I

ABSTRACT: Rapid and robust directed differentiation of mouse epiblast stem cells into definitive endoderm and forebrain organoids I

PROVIDER: PRJNA784930 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR17074100_1.fastq.gz	Fastqsanger.gz
	SRR17074100_2.fastq.gz	Fastqsanger.gz
	SRR17074101_1.fastq.gz	Fastqsanger.gz
	SRR17074101_2.fastq.gz	Fastqsanger.gz
	SRR17074102_1.fastq.gz	Fastqsanger.gz

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Rapid and robust directed differentiation of mouse epiblast stem cells into definitive endoderm and forebrain organoids II

Project description:Rapid and robust directed differentiation of mouse epiblast stem cells into definitive endoderm and forebrain organoids II

| PRJNA784929 | ENA

Rapid and robust directed differentiation of mouse epiblast stem cells into definitive endoderm and forebrain organoids II

Project description:Mouse is the pre-eminent model system for studying genetic regulation of embryonic development. Directed differentiation of pluripotent stem cells (PSCs) has emerged as a powerful complement to in vivo studies of development, allowing for examination of developmental mechanisms at the molecular and cellular level. However, the limited availability of high-quality protocols for directed differentiation of mouse PSCs into defined lineages and organoids has prevented the widespread application of mouse PSC directed differentiation models. Here, we examine the potential of mouse epiblast stem cells (EpiSCs) cultured in media containing Wnt pathway inhibitors (primed ground state conditions) as a starting point for directed differentiation. First, we systematically optimized conditions to generate definitive endoderm that are significantly faster and more efficient than current state-of-the-art protocols for mouse PSCs. Second, we developed a robust protocol for generation of forebrain-patterned organoids. These new models can complement and empower in vivo studies in the mouse and can inform human PSC-based studies of development, which cannot readily be compared to analogous stages in vivo.

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Rapid and robust directed differentiation of mouse epiblast stem cells into definitive endoderm and forebrain organoids I

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Rapid and robust directed differentiation of mouse epiblast stem cells into definitive endoderm and forebrain organoids.

Project description:Directed differentiation of pluripotent stem cells (PSCs) is a powerful model system for deconstructing embryonic development. Although mice are the most advanced mammalian model system for genetic studies of embryonic development, state-of-the-art protocols for directed differentiation of mouse PSCs into defined lineages require additional steps and generates target cell types with lower purity than analogous protocols for human PSCs, limiting their application as models for mechanistic studies of development. Here, we examine the potential of mouse epiblast stem cells cultured in media containing Wnt pathway inhibitors as a starting point for directed differentiation. As a proof of concept, we focused our efforts on two specific cell/tissue types that have proven difficult to generate efficiently and reproducibly from mouse embryonic stem cells: definitive endoderm and neural organoids. We present new protocols for rapid generation of nearly pure definitive endoderm and forebrain-patterned neural organoids that model the development of prethalamic and hippocampal neurons. These differentiation models present new possibilities for combining mouse genetic tools with in vitro differentiation to characterize molecular and cellular mechanisms of embryonic development.

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Saracatinib Promotes Definitive Endoderm Differentiation by Inhibition of FAK-YAP Signaling Axis

Project description:Optimizing the efficiency of definitive endoderm differentiation is significant for the generation of diverse organ-like structures. In this study, we utilized saracatinib to enhance definitive endoderm differentiation in pluripotent stem cells. We found saracatinib significantly improved the definitive endoderm differentiation at low concentrations. To investigate the impact of 0.5 μM saracatinib on definitive endoderm differentiation of ESC H1 cells, we conducted RNA-seq analysis with differentiated cells with or without 0.5 μM saracatinib treatment.

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Project description:Understanding the molecular mechanisms controlling early cell fate decisions in mammals is a major objective towards the development of robust methods for the differentiation of human pluripotent stem cells into clinically relevant cell types. Here, we used human embryonic stem cells (hESCs) to study specification of definitive endoderm in vitro. Using a combination of whole genome expression and ChIP-seq analyses, we established a hierarchy of transcription factors regulating endoderm specification. Importantly, pluripotency factors, namely NANOG, OCT4 and SOX2 have an essential function in this network by actively directing differentiation. Indeed, these transcription factors control the expression of EOMES, which marks the onset of endoderm specification. In turn, EOMES interacts with SMAD2/3 to initiate the transcriptional network governing endoderm formation. Together, these results provide for the first time a comprehensive molecular model connecting the transition from pluripotency to endoderm specification during mammalian development.

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A distinct microRNA signature for definitive endoderm derived from human embryonic stem cells

Project description:This SuperSeries is composed of the following subset Series: GSE16678: MicroRNA expression data from differentiation of human Cyt49 ESCs into definitive endoderm in feeder-free conditions GSE16681: mRNA expression data from differentiation of human ESCs into definitive endoderm, Cyt49 on matrigel GSE16687: MicroRNA expression data from differentiation of human Cyt49 ESCs into definitive endoderm on MEF feeder layers GSE16689: MicroRNA expression data from differentiation of human H9 ESCs into definitive endoderm on MEF feeder layers Refer to individual Series

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Pluripotency Factors Regulate Definitive Endoderm Specification through Eomesodermin

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Project description:Eomes related gene regulation in Definitive Endoderm

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