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

ABSTRACT: 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.

ORGANISM(S): Homo sapiens

PROVIDER: GSE147839 | GEO | 2021/03/24

REPOSITORIES: GEO

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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 single cell RNA-seq

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Project description:We performed a microarray experiment to analyze the transcriptional profile of human fetal tissue derived neural stem/progenitor cells, human iPSCs, or human iPSC-derived neural stem/progenitor cells generated using xenogenic or xeno-free reagents. Gene expression patterns were compared among human fetal tissue derived neural stem/progenitor cells, human iPSCs, or human iPSC-derived neural stem/progenitor cells generated using xenogenic or xeno-free reagents. Samples with a title including "Xf" (201B7-lt-NES (Xf)-1, 201B7-lt-NES (Xf)-2, 1210B2-lt-NES (Xf)-1,1210B2-lt-NES (Xf)-2, 1231A3-lt-NES (Xf)-1, 1231A3-lt-NES (Xf)-2) correspond to human iPSC-derived neural stem/progenitor cells generated using xeno-free reagents, whereas samples with a title including "Con" (201B7-lt-NES (Con)-1, 201B7-lt-NES (Con)-2) correspond to those generated using xenogenic reagents.

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Genome-wide analysis of chromatin state in extended pluripotent stem cells, primed pluripotent stem cells, and naïve pluripotent stem cells

Project description:Among all known cultured stem cell types, pluripotent stem cells (PSCs) sit atop the landscape of developmental potency and are characterized by their unrestricted developmental potential, able to generate all cell types of an adult organism. However, PSCs show limited contribution to the extraembryonic (ExEm) tissues, in particular, those giving rise to the placenta in vivo. To date, it remains unknown whether stem cells with both embryonic and extraembryonic developmental potency can be captured and maintained in vitro. Here, we identify a new chemical cocktail that allows for the generation of stem cells with extended developmental potency from mouse and human, designated as extended pluripotent stem (EPS) cells, which is capable of chimerizing both embryonic and extraembryonic tissues. Importantly, a single mouse EPS (mEPS) cell shows widespread contribution to both embryonic and extraembryonic lineages in chimeric mouse conceptuses at late-gestation stages, and permits generation of high-grade germline competent chimeras as well as single EPS-derived viable mice by tetraploid complementation. Furthermore, human EPS (hEPS) cells contribute to embryonic and extraembryonic tissues in interspecies chimeric mouse conceptuses. Compared to known PSCs, EPS cells show unique gene modules that upregulate in embryonic cells from early preimplantation development. Further analysis shows that PARP1 inhibition is required for maintaining EPS potency. Our findings constitute a first step towards capturing pluripotent stem cells with extraembryonic developmental potentials in culture, and open new avenues for generating mammalian PSCs with robust chimeric competency for basic and translational research.

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

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