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

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

PROVIDER: PRJNA622253 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR11459602_1.fastq.gz	Fastqsanger.gz
	SRR11459602_2.fastq.gz	Fastqsanger.gz
	SRR11459603_1.fastq.gz	Fastqsanger.gz
	SRR11459603_2.fastq.gz	Fastqsanger.gz
	SRR11459604_1.fastq.gz	Fastqsanger.gz

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Chromatin accessibility of xeno-free and feeder-cultured human extended pluripotent stem cells by ATAC-seq

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Chromatin accessibility of xeno-free and feeder-cultured human extended pluripotent stem cells by ATAC-seq

Project description:Extended pluripotent stem (EPS) cells have shown great applicative potentials in generating synthetic embryos, directed differentiation and disease modeling. However, the lack of a xenofree culture condition has significantly limited their wide applications. We developed a chemically defined and xeno-free culture condition for culturing and deriving human EPS cells, which can be long-term stably propagated in vitro, as well as preserve their embryonic and extraembryonic developmental potentials.

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

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

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

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Xeno-free and Chemically Defined Human System for Culturing Human Epidermal Keratinocytes

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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Single-Cell RNA Sequencing of Human Embryonic Stem Cells and Feeder-free Extended Pluripotent Stem Cells

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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Project description:In this study, we sought to examine whether an extracellular matrix (ECM)-based xeno-free culture system that we recently established could be used together with a microRNA-enhanced mRNA reprogramming method for the generation of clinically safe iPS cells. The notable features of this method are (1) the use of a xeno-free/feeder-free culture system for the generation and expansion of iPS cells rather than the conventional labor-intensive culture systems using human feeder cells or human feeder-conditioned medium and (2) the enhancement of mRNA-mediated reprogramming via the delivery of microRNAs. Strikingly, we observed the early appearance of iPS cell colonies (~11 days), substantial reprogramming efficiency (~0.2-0.3%), and a high percentage of ESC-like colonies among the total colonies (~87.5%), indicating enhanced kinetics and reprogramming efficiency. Therefore, the combined method established in this study provides a valuable platform for the generation and expansion of clinically safe (i.e., integration- and xeno-free) iPS cells, facilitating immune-matched cell therapy in the near future.

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Project description:Here, we examined the effect of LDN-193189, an inhibitor of the BMP signaling pathway, on pig embryonic stem cells (ESCs) in the absence of feeder cells to develop a feeder-free culture system. Feeder cells play an important role in supporting the pluripotency of pig ESCs by blocking trophoblast and mesodermal differentiation through inhibition of the BMP pathway. Transcriptome analysis revealed that LDN-193189 treatment instead of coculturing with feeder cells could help maintain the pluripotency and homogenous cell population of pig ESCs by stimulating the secretion of chemokines and suppressing differentiation. We confirmed that LDN-193189-treated pig ESCs cultured without feeder cells maintained pluripotency for an extended period and were capable of differentiating in vivo and in vitro. The pig ESCs represented distinct characteristics of pig epiblasts as an in vivo counterpart at a transcriptomic level. In summary, we developed feeder-free culture systems for pig ESCs, and our findings will improve the understanding of pluripotency gene networks and comparative embryogenesis.

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