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Transcriptomically-guided mesendoderm induction of human pluripotent stem cells using a systematically defined culture scheme

ABSTRACT: Transcriptomically-guided mesendoderm induction of human pluripotent stem cells using a systematically defined culture scheme

PROVIDER: PRJNA531831 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR8874340.fastq.gz	Fastqsanger.gz
	SRR8874350.fastq.gz	Fastqsanger.gz
	SRR8874360.fastq.gz	Fastqsanger.gz
	SRR8874341.fastq.gz	Fastqsanger.gz
	SRR8874351.fastq.gz	Fastqsanger.gz

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Transcriptomically-guided mesendoderm induction of human pluripotent stem cells using a systematically defined culture scheme

Project description:Human pluripotent stem cells (hPSCs) are an essential cell source in tissue engineering, studies of development, and disease modeling. Efficient and broadly amenable protocols for rapid lineage induction of hPSCs are of great interest in the stem cell biology field. We describe a simple yet robust method for differentiation of hPSCs into mesendoderm in defined conditions utilizing single-cell seeding and BMP4 and Activin A (BA) treatment. Gene sets and gene ontology terms related to mesoderm and endoderm differentiation were enriched after 48 hours of BA treatment. After 14 days of differentiation, BA-treated cells expressed genes indicative of mesoderm and endoderm-derived tissue types, including lung, kidney and adipose tissue. BA treatment was readily incorporated into existing protocols for chondrogenic and endothelial progenitor cell differentiation. Teratomas formed from BA-treated cells contained higher ratios of mesoderm and endoderm to ectoderm tissue compared to PSC-derived teratomas. Collectively, these data demonstrate that single-cell seeding with BA treatment is a powerful method for induction of mesendoderm that can be integrated into protocols for mesoderm and endoderm differentiation.

2019-04-11 | GSE129570 | GEO

Transcriptomically Guided Mesendoderm Induction of Human Pluripotent Stem Cells Using a Systematically Defined Culture Scheme.

Project description:Human pluripotent stem cells (hPSCs) are an essential cell source in tissue engineering, studies of development, and disease modeling. Efficient, broadly amenable protocols for rapid lineage induction of hPSCs are of great interest in the stem cell biology field. We describe a simple, robust method for differentiation of hPSCs into mesendoderm in defined conditions utilizing single-cell seeding (SCS) and BMP4 and Activin A (BA) treatment. BA treatment was readily incorporated into existing protocols for chondrogenic and endothelial progenitor cell differentiation, while fine-tuning of BA conditions facilitated definitive endoderm commitment. After prolonged differentiation in vitro or in vivo, BA pretreatment resulted in higher mesoderm and endoderm levels at the expense of ectoderm formation. These data demonstrate that SCS with BA treatment is a powerful method for induction of mesendoderm that can be adapted for use in mesoderm and endoderm differentiation.

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Mesendoderm induction

Project description:ES cells are differentiated into mesenderm cell linage by Activin A treatment. During this differentaition, gene expression profile of ES cells, day 4 mesendoderm cell, day 5 mesendoderm cells and mesoderm cells are examined.

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Transcriptomically-guided pharmacological experiments in neocortical and hippocampal NPY-positive GABAergic interneurons

Project description:Cortical GABAergic interneurons have been shown to fulfil important roles by inhibiting excitatory principal neurons. Recent transcriptomic studies have confirmed seminal discoveries that used anatomical and electrophysiological methods highlighting the existence of multiple different classes of GABAergic interneurons. However, individual studies have rarely addressed regional specific differences in gene expression in a given subclass of neuron. Using single-cell Patch-RNAseq, we characterised neuropeptide Y (NPY)-positive GABAergic interneurons in superficial layers of the primary auditory cortex and in distal layers of area CA3. We found that more than 300 genes are differentially expressed in NPY-positive neurons between these two brain regions. For example, the AMPA receptor auxiliary subunit Shisa9/CKAMP44 and the 5-HT2a receptor are significantly higher expressed in auditory NPY-positive neurons. These findings guided us to perform pharmacological experiments that revealed a role for 5-HT2a receptors in auditory NPY-positive neurons. Specifically, although the application of 5-HT led to a depolarisation of both auditory and CA3 NPY-positive neurons, the 5-HT2a receptor antagonist ketanserin only reversed membrane potential changes in auditory NPY-positive neurons. Our study demonstrates the potential of single-cell transcriptomic studies in guiding directed pharmacological experiments.

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Project description:FSA RM Scheme 2023/2024 - AFBI Seq

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