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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

ABSTRACT: 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.

ORGANISM(S): Homo sapiens

PROVIDER: GSE76900 | GEO | 2016/08/22

SECONDARY ACCESSION(S): PRJNA308904

REPOSITORIES: GEO

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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

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

2021-03-24 | GSE147839 | GEO

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

2021-03-24 | GSE156920 | GEO

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

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.

2016-04-21 | GSE68035 | GEO

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

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A novel xeno-free and feeder-cell-free system for human pluripotent stem cell culture.

Project description:While human induced pluripotent stem cells (hiPSCs) have promising applications in regenerative medicine, most of the hiPSC lines available today are not suitable for clinical applications due to contamination with nonhuman materials, such as sialic acid, and potential pathogens from animal-product-containing cell culture systems. Although several xeno-free cell culture systems have been established recently, their use of human fibroblasts as feeders reduces the clinical potential of hiPSCs due to batch-to-batch variation in the feeders and time-consuming preparation processes. In this study, we have developed a xeno-free and feeder-cell-free human embryonic stem cell (hESC)/hiPSC culture system using human plasma and human placenta extracts. The system maintains the self-renewing capacity and pluripotency of hESCs for more than 40 passages. Human iPSCs were also derived from human dermal fibroblasts using this culture system by overexpressing three transcription factors-Oct4, Sox2 and Nanog. The culture system developed here is inexpensive and suitable for large scale production.

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

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

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Single-cell RNA-seq of human cardiovascular progenitor cells generated with a chemically defined, xeno-free laminin based differentiation protocol

Project description:Regeneration of injured human heart muscle is limited and an unmet clinical need. There are no methods for the reproducible generation of clinical quality stem-cell-derived cardiovascular progenitors (CVPs). We identified laminin-221 (LN-221) as the most likely expressed cardiac laminin. We produced it as human recombinant protein, and showed that LN-221 promotes differentiation of pluripotent hESCs towards cardiomyocyte lineage and downregulates pluripotency and teratoma associated genes. We developed a chemically defined, xeno-free laminin-based differentiation protocol to generate CVPs. Single-cell RNA sequencing of CVPs derived from hESC lines supported reproducibility and identified three main progenitor subpopulations. These CVPs were transplanted into myocardial infarction mice, where heart function was measured by echocardiogram and human heart muscle bundle formation was identified histologically. This method may provide clinical quality cells for use in regenerative cardiology.

2019-03-20 | GSE127955 | GEO

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