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Chemically defined and growth factor-free system for highly efficient endoderm induction of human pluripotent stem cells [ATAC-seq]

ABSTRACT: Chemically defined and growth factor-free system for highly efficient endoderm induction of human pluripotent stem cells [ATAC-seq]

PROVIDER: PRJNA1147603 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR30227968_1.fastq.gz	Fastqsanger.gz
	SRR30227968_2.fastq.gz	Fastqsanger.gz
	SRR30227969_1.fastq.gz	Fastqsanger.gz
	SRR30227969_2.fastq.gz	Fastqsanger.gz
	SRR30227970_1.fastq.gz	Fastqsanger.gz

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Chemically defined and growth factor-free system for highly efficient endoderm induction of human pluripotent stem cells [RNA-seq]

Project description:Chemically defined and growth factor-free system for highly efficient endoderm induction of human pluripotent stem cells [RNA-seq]

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Chemically defined and growth factor-free system for highly efficient endoderm induction of human pluripotent stem cells [ATAC-seq]

Project description:Definitive endoderm (DE) derived from human pluripotent stem cells (hPSCs) holds great promise for cell-based therapies and drug discovery. However, current DE differentiation methods required undefined components and/or expensive recombinant proteins, limiting their scalable manufacture and clinical use. Homogeneous DE differentiation in defined and recombinant protein-free conditions remains a major challenge. Here, by systematic optimization and high-throughput screening, we report a fully synthetic, small molecule-based defined system that contains only four components (4C), enabling highly efficient and cost-effective DE specification of hPSCs in the absence of recombinant proteins. 4C-induced DE can differentiate into functional hepatocytes, lung epithelium, and pancreatic β cells in vitro and multiple DE-derivatives in vivo. Genomic accessibility analysis reveal that 4C reconfigures chromatin architecture to allow key DE transcription-factor binding and identify TEAD3 as a novel key DE regulator. This system may facilitate mass production of mature DE-derivatives for drug discovery, disease modelling, and cell therapy.

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Project description:Cardiomyocytes can be differentiated from human pluripotent stem cells (hPSCs) in defined conditions, but efficient and consistent cardiomyocyte differentiation often requires expensive reagents such as B27 supplement or recombinant albumin. Using a chemically defined albumin-free (E8 basal) medium, we identified heparin as a novel factor that significantly promotes cardiomyocyte differentiation efficiency, and developed an efficient method to differentiate hPSCs into cardiomyocytes. The treatment of heparin helped cardiomyocyte differentiation consistently reach at least 80% purity (up to 95%) from more than 10 different hPSC lines in chemically defined DMEM/F-12 based medium on either Matrigel or defined matrices like Vitronectin and Synthemax. One of heparinâs main functions was to act as a WNT modulator that helped promote robust and consistent cardiomyocyte production. Our study provides an efficient, reliable, and cost-effective method for cardiomyocyte derivation from hPSCs that can be used for potential large-scale drug screening, disease modeling, and future cellular therapies. 12 human pluripotent stem cells (hPSCs) at three different cardiac differentiation times (0 Days, 3 Days, 6 Days, 10 Days) under different culture conditions (+/- Heparin, +/- IWP2).

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