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

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

PROVIDER: PRJNA660602 | ENA |

REPOSITORIES: ENA

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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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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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Robust production of human neural cells by establishing neuroepithelial-like stem cells from peripheral blood mononuclear cell-derived feeder-free iPSCs under xeno-free conditions

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