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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.

2021-03-24 | GSE157237 | GEO

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

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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

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.

2016-08-22 | E-GEOD-76900 | biostudies-arrayexpress

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

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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Extracellular matrix-dependent generation of integration- and xeno-free iPS cells using a modified mRNA transfection method

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

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

| PRJNA660602 | ENA

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

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

| PRJNA622253 | ENA

Generation of Blastocyst-like Structures from Extended Pluripotent Stem Cells

Project description:A single mouse blastomere from until 8-cell embryo can generate an entire blastocyst. Whether blastomere-like extended pluripotent stem cells (EPS cells) retain a similar generative capacity remains unknown. Here, we established a 3D differentiation system that enabled the generation of blastocyst-like structures from EPS cells (EPS-blastoids) through lineage segregation and self-organization. EPS-blastoids resembled blastocysts in morphology and cell lineage allocation. EPS-blastoids formation recapitulated key morphogenetic events during preimplantation and early postimplantation development in vitro. Upon transfer, some EPS-blastoids underwent implantation, induced decidualization, and generated live, albeit disorganized, tissues in utero. Single cell and bulk RNA-sequencing analysis revealed that EPS-blastoids contained all three blastocyst cell lineages and shared transcriptional similarity with natural blastocysts. We also provide proof-of-concept that EPS-blastoids can be generated from somatic cells via cellular reprograming. EPS-blastoids provide a unique platform for studying early embryogenesis, and pave the way to generate viable synthetic embryos using cultured cells.

2019-08-03 | GSE135289 | GEO

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