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Global transcriptional analysis of xeno-free and feeder-cultured human extended pluripotent stem cells by single cell RNA-seq

ABSTRACT: Global transcriptional analysis of xeno-free and feeder-cultured human extended pluripotent stem cells by single cell RNA-seq

PROVIDER: PRJNA659528 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR12522006_1.fastq.gz	Fastqsanger.gz
	SRR12522006_2.fastq.gz	Fastqsanger.gz
	SRR12522007_1.fastq.gz	Fastqsanger.gz
	SRR12522007_2.fastq.gz	Fastqsanger.gz
	SRR12522008_1.fastq.gz	Fastqsanger.gz

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Project description:Developing an effective xeno-free and defined system for human keratinocyte culture has been one of the fundamental measures in current cellular therapy. For keratinocytes, in particular, such system will not only fulfill clinical safety and quality criteria, it will allow for the management of less severe burns and other skin injuries such as chronic wounds. To date, the expansion of autologous keratinocytes in the clinical settings still relies on 3T3 feeder co-culture system. Here we report a completely xeno-free and defined culture system by using human recombinant laminins as feeder replacement. We have thoroughly characterized the cells cultured in this new system both in vitro and in vivo. We found that laminin-511/-521, and the unanticipated laminin-411/-421, but not -332, -111, -121, -211, or LN-221 support adult human skin keratinocytes and could maintain their self-renewal properties comparable to the 3T3 system. We believe our new culture method should facilitate broader use of cultured epithelial cell products for today’s regenerative medicine.

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Project description:This dataset features transcriptomic profiles of feeder-free extended pluripotent stem cells (ffEPSCs) and their parentalderived from human embryonic stem cells (ESCs). ffEPSCs were generated using a Matrigel-based feeder-free system combined with a chemical cocktail to convert conventional ESCs into ffEPSCs. Generated using Smart-seq2 and sequenced on Illumina HiSeq 2000, the dataset explores transcriptional profiles and heterogeneity of ESCs and ffEPSCs, providing insights into early human development and stem cell biology.

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