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Feeder-Independent Derivation of Induced-Pluripotent Stem Cells From Peripheral Blood Endothelial Progenitor Cells

ABSTRACT: The objective of this study was to reprogram peripheral blood-derived late-endothelial progenitor cells (EPCs) to a pluripotent state under feeder-free and defined culture conditions. Late-EPCs were retrovirally-transduced with OCT4, SOX2, KLF4, c-MYC, and iPSC colonies were derived in feeder-free and defined media conditions. EPC-iPSCs expressed pluripotent markers, were capable of differentiating to cells from all three germ-layers, and retained a normal karyotype. Transcriptome analyses demonstrated that EPC-iPSCs exhibit a global gene expression profile similar to human embryonic stem cells (hESCs). We have generated iPSCs from late-EPCs under feeder-free conditions. Thus, peripheral blood-derived late-outgrowth EPCs represent an alternative cell source for generating iPSCs.

ORGANISM(S): Homo sapiens

PROVIDER: GSE42947 | GEO | 2012/12/16

SECONDARY ACCESSION(S): PRJNA183924

REPOSITORIES: GEO

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Feeder-Independent Derivation of Induced-Pluripotent Stem Cells From Peripheral Blood Endothelial Progenitor Cells

Project description:The objective of this study was to reprogram peripheral blood-derived late-endothelial progenitor cells (EPCs) to a pluripotent state under feeder-free and defined culture conditions. Late-EPCs were retrovirally-transduced with OCT4, SOX2, KLF4, c-MYC, and iPSC colonies were derived in feeder-free and defined media conditions. EPC-iPSCs expressed pluripotent markers, were capable of differentiating to cells from all three germ-layers, and retained a normal karyotype. Transcriptome analyses demonstrated that EPC-iPSCs exhibit a global gene expression profile similar to human embryonic stem cells (hESCs). We have generated iPSCs from late-EPCs under feeder-free conditions. Thus, peripheral blood-derived late-outgrowth EPCs represent an alternative cell source for generating iPSCs. Six samples were analyzed. The gene expression profile of four iPS clones were compared to the H9 human embryonic stem cell line and the parent endothelial progenitor cell line.

2012-12-16 | E-GEOD-42947 | biostudies-arrayexpress

Human iPSCs derived under feeder free conditions displays an unqiue cell cycle and DNA replication genotype

Project description:Induced pluripotent stem cells (iPSCs) have been generated from various somatic cells under feeder-layer conditions. These feeder-derived iPSCs generated in different labs exhibit greater variability than between different traditional embryo derived hESC lines. For that reason, it is important to develop a standard and defined system for deriving autologous patient stem cells. We have generated iPSCs under feeder-free conditions using Matrigel coated vessels in chemically defined medium, mTeSR1. These feeder-free derived iPSCs are in many ways similar to feeder-derived iPSCs and also to hESCs, with respect to their pluripotent gene expression (OCT4, NANOG, SOX2), protein expression (OCT4, NANOG, SSEA4, TRA160) and differentiation capabilities. We conducted a whole genomic transcript analysis using Affymetrix Human Gene 1.0 ST arrays to elucidate the important differences between traditional feeder-derived iPSCs and feeder-free derived iPSCs. We reveal that feeder-free iPSCs have over-represented terms belonging to DNA replication and cell cycle genes which are lacking in feeder-derived iPSCs. Feeder-free iPSCs are in many aspects more similar to hESCs including; apoptosis, chromatin modification enzymes and mitochondrial energy metabolism. We have also identified potential biomarkers for fully reprogrammed iPSCs (FRZB) and partially reprogrammed iPSCs (POTEG, MX2) based on their expression trends across all cell types. In conclusion, feeder-free derived iPSCs is transcriptomically more similar to hESCs than feeder derived iPSCs, in many biological functions.

2011-06-03 | GSE21655 | GEO

Human iPSCs derived under feeder free conditions displays an unqiue cell cycle and DNA replication genotype

2011-06-03 | E-GEOD-21655 | biostudies-arrayexpress

Bulk RNAseq anaysis of PSCs and differentiating cells under on-feeder and feeder-free cultures

Project description:Gene expression profiling of human iPSCs [201B7] under on-feeder, feeder-free [StemFit], and FGF inhibitor [100 nM PD173074] -treated cultures, and differentiating cells.

2022-03-09 | GSE195689 | GEO

Single cell RNA-seq analysis of feeder-free iPSC-derived gliogenic NS/PCs and neurogenic NS/PCs

Project description:We performed single cell RNA sequencing to analyze the transcriptional profile of gliogenic NS/PCs and neurogenic NS/PCs derived from the same parental feeder-free iPSCs.

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microRNA expression profiles in far-infrared treated human endothelial precursor cells

Project description:High glucose impairs the angiogenic activities of late endothelial precursor cells (EPC). We found that far infrared (FIR) treatment restored partially the activity of late EPC. However, the mechanisms are unclear. We applied microRNA expression microarrays to assess the microRNA expression profiles of high glucose-treated late EPC with or without FIR treatment. Late EPCs isolated from peripheral blood were cultured in high glucose condition for 48 hours, and then treated with or without FIR radiation for 30 minutes. Total RNA were extracted and evaluated with the Agilent Human miRNA Oligo Microarray R14 V2 chips.

2014-04-03 | E-GEOD-37043 | biostudies-arrayexpress

mRNA expression profiles in far-infrared treated human endothelial precursor cells

Project description:High glucose impairs the angiogenic activities of late endothelial precursor cells (EPC). We found that far infrared (FIR) treatment restored partially the activity of late EPC. However, the mechanisms are unclear. We performed gene expression microarray analysis to assess the expression profiles of high glucose-treated late EPC with or without FIR treatment. Late EPCs isolated from peripheral blood were cultured in high glucose condition for 48 hours, and then treated with or without FIR radiation for 30 minutes. Total RNA were extracted and evaluated with the Affymetrix GeneChip Human U133 plus 2.0 arrays.

2014-04-03 | E-GEOD-37044 | biostudies-arrayexpress

Single-cell RNAseq anaysis of 4-month-old brain organoids from feeder-free iPSCs with PD173074 treatment

Project description:Single-cell gene expression profiling of 4-month-old brain organoids from feeder-free iPSCs with PD173074 treatment

2022-03-09 | GSE195690 | GEO

Proteome Profiling of PBMC-originated, iPSC-derived Lentoid Bodies at differentiation day 35

Project description:Differentiation of pluripotent stem cells into lentoid bodies is important for the understanding of the lens development and investigating the processes critical for lens morphogenesis. This Study was initiated to investigate a comprehensive proteome profiling of the peripheral blood mononuclear cell (PBMC)-originated, induced pluripotent stem cell (iPSC)-derived lentoid bodies through mass spectrometry-based protein sequencing. Briefly, a small aliquot of blood sample was ascertained to collect PBMCs that were reprogrammed to iPSCs using the Sendai-virus delivery system. The PBMC-originated, iPSCs were differentiated into lentoid bodies employing the “fried egg” method using feeder-free conditions. The quantitative real-time PCR (qRT-PCR) confirmed the expression of lens-associated markers, which exhibited at least an order magnitude increased expression in lentoid bodies at differentiation day 35. Subsequently, the total cellular protein was extracted from lentoid bodies at day 35, digested with trypsin, fractionated into 96 fractions and subjected to an mass spectrometry-based label-free quantitative proteomics. mass spectrometry-based proteome profiling revealed 9,717 proteins in iPSC-derived lentoid bodies at differentiation day 35. In here, we report a comprehensive proteome of PBMC-originated, iPSC-derived lentoid bodies at day 35, which will help in better understanding processes critical for the development of the ocular lens.

2022-02-28 | PXD009633 | Pride

Proteome Profiling of PBMC-originated, iPSC-derived Lentoid Bodies at differentiation day 25

Project description:Differentiation of pluripotent cells to generate lentoid bodies is important for the understanding of the lens development and investigating the processes critical for lens morphogenesis. This Study was initiated to investigate a comprehensive proteome profiling of the peripheral blood mononuclear cell (PBMC)-originated, induced pluripotent stem cell (iPSC)-derived lentoid bodies through mass spectrometry-based protein sequencing. Briefly, a small aliquot of blood sample was ascertained to collect PBMCs that were reprogrammed to iPSCs using the Sendai-virus delivery system. The PBMC-originated, iPSCs were differentiated into lentoid bodies employing the “fried egg” method using feeder-free conditions. The quantitative real-time PCR (qRT-PCR) confirmed the expression of lens-associated markers, which exhibited at least an order magnitude increased expression in lentoid bodies at differentiation day 25. Subsequently, the total cellular protein was extracted from lentoid bodies at day 25, digested with trypsin, fractionated into 24 fractions and subjected to an mass spectrometry-based label-free quantitative proteomics. mass spectrometry-based proteome profiling revealed 9,473 proteins in iPSC-derived lentoid bodies at differentiation day 25. In here, we report a comprehensive proteome of PBMC-originated, iPSC-derived lentoid bodies at day 25, which will help in better understanding processes critical for the development of the ocular lens.

2022-03-01 | PXD011143 | Pride

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