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Human ECFCs: Cord Blood vs Peripehral Blood ECFCs

ABSTRACT: miRNA profiling was carried out using the miRCURY LNA™ microRNA Array (5th gen - hsa, mmu & rno). microRNA profiling of CB and PB-derived ECFCs from 3 independent donors from each cell source. To identify and understand the regulation of endothelial cell functions through comparing miRNA expression profiling of cord blood (CB) derived endothelial colony forming cells (ECFCs) and peripheral blood (PB) ECFCs

ORGANISM(S): Rattus norvegicus Mus musculus Homo sapiens

PROVIDER: GSE81574 | GEO | 2017/03/20

SECONDARY ACCESSION(S): PRJNA322029

REPOSITORIES: GEO

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Comparative analysis of TAL1 binding in TSA-treated versus non-treated Endothelial Colony Forming Cells (ECFCs)

Project description:Endothelial progenitors represent one of the most promising cell-based strategies for vascular repair of ischemic tissue damage, including limb ischemia, myocardial infarction and stroke. We have shown that the transcription factor TAL1 regulates a transcription program that drives the migration and adhesion of ECFCs. Furthermore, treatment of ECFCs with the HDAC inhibitor TSA increases the expression of TAL1-dependent genes and promotes the migration, chemotaxis and adhesion of ECFCs. Finally, ex vivo treatment with TSA also improves the vascular repair properties of ECFCs in vivo when these cells are transplanted in a mouse model of hindlimb ischemia. The goal of this experiment was to test whether TSA treatment of ECFCs affect TAL1 genomic binding. TAL1 ChIP-sequencing was performed from ECFCs that have been treated or not TSA. As negative controls, we performed Mock-ChIP-seq from the same samples using normal IgG instead of the TAL1 antibody. Overall, we find that there is no change in TAL1 genomic binding in ECFCs upon TSA treatment.

2014-05-06 | E-GEOD-53423 | biostudies-arrayexpress

shRNA mediated knock-down of Tal1 in human Endothelial Colony Forming Cells (ECFCs)

Project description:Endothelial colony-forming cells (ECFCs) have been reported as promising cells for regenerative medicine thanks to their angiorepair properties. Transcription factors are primary determinants of the functional capacity of the cells and TAL1 has been shown as a critical regulator of endothelial lineage in both development and adult life. However, only few (three) TAL1 targets have been identified so far in mouse and human endothelial cells. This microarray experiment, where TAL1 expression was knocked-down, was designed to identify TAL1-dependent genes in primary human endothelial stem/progenitor cells. ECFCs were isolated from three independent cord blood samples (n=3, biological replicates) and cultured in complete EGM-2 medium. The knockdown of TAL1 was induced by infection with lentiviruses expressing an anti-TAL1 shRNA. A scrambled shRNA was used as a negative control. Cells were then harvested for RNA extraction. DNA-free total RNA was isolated with RNeasy Mini Kit and hybridized to the Affymetrix Human Gene 1.0 ST gene expression microarray.

2014-05-06 | E-GEOD-44444 | biostudies-arrayexpress

Comparative analysis of TAL1 binding in TSA-treated versus non-treated Endothelial Colony Forming Cells (ECFCs)

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Human umbilical cord blood-derived endothelial colony forming cells under serum-free conditions

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2010-07-29 | E-GEOD-20504 | biostudies-arrayexpress

Transcriptome analysis of ECFCs treated with GSK-343 and Panobinostat

Project description:Human endothelial colony-forming cells (ECFCs) represent one of the most promising sources of adult stem cells for vascular regeneration after ischemia. Yet, the molecular mechanism underlying the unique properties of these cells to repair damaged blood vessels remains unclear, and new strategies to improve their regenerative function are needed. Here, we demonstrate that ex vivo treatment of ECFCs with a combination of epigenetic drugs (i.e. the histone H3K27 demethylase inhibitor GSK-343 and the histone deacetylase inhibitor panobinostat) improves the migration of these cells and their capacity to form capillary-like networks while also increasing their resistance to serum starvation-induced apoptosis. Furthermore, the kinetic of blood flow recovery is significantly increased upon transplantation of drug-treated ECFCs in a pre-clinical model of hind-limb ischemia. Gene expression profiling by RNA-sequencing reveals that these effects are mediated through the simultaneous activation of several major pro-angiogenic signaling pathways including VEGFR, CXCR4, WNT, NOTCH and SHH. At the molecular level, we provide evidence that the two drugs work through complementary mechanisms that entail the resolving of bivalently marked genes characterized by the presence of both negative (H3K27me3) and positive (H3K4me3 and H3-Ac) histone modifications to create a transcriptionally permissive chromatin environment. Thus, ex vivo treatment with epigenetic drugs increases the vascular repair properties of ECFCs through transient activation of major pro-angiogenic signaling pathways.

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Human umbilical cord blood-derived endothelial colony forming cells under serum-free conditions

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Functional Analysis and Gene Expression Profile of Umbilical Cord Blood Regulatory T Cells

Project description:Since the role of cord blood (CB) regulatory T cells (Tregs) for the suppression of the allogeneic T-cell response is under investigation, we analyzed and compared the functional properties and gene expression profile of Tregs expanded from CB units or from the peripheral blood (PB) of helathy donors. Tregs were purified from 23 CB units and from the PB of 13 donors and expanded for 6 days with anti-CD3, anti-CD28 and IL-2. Immunophenotypic analyses were performed, and suppressor activity of expanded Tregs was measured in mixed lymphocyte reaction (MLR) cultures. The IL-10 production capacity was tested and gene expression profile experiments were performed on 6 Tregs from PB and 4 from CB. CB and PB Tregs had similar immunophenotypic features. Tregs from CB presented a higher expansion capacity and genomic characterization showed in CB-derived Tregs a significant enrichments of genes involved in cell proliferation, chromatin modification and regulation of gene expression in CB-derived Tregs. All samples were positive for the Foxp3 gene and protein after expansion. CB and PB expanded Tregs exerted a comparable and potent suppressive function of MLR and presented a high in vitro IL-10 production capacity. Gene profile analysis also revealed for PB Tregs a significant enrichments of genes involved in the adaptive immune response.

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2018-08-08 | GSE118259 | GEO

shRNA mediated knock-down of Tal1 in human Endothelial Colony Forming Cells (ECFCs)

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