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Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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

ABSTRACT: Derivation and expansion of human umbilical cord blood-derived endothelial colony forming cells under serum-free conditions - a transcriptome analysis. Endothelial colony forming cells (ECFCs) were isolated from term umbilical cord blood units. ECFCs were expanded under standard, fetal bovine serum (FBS) containing endothelial medium, or transferred to chemically defined endothelial media without FBS. Microarray expression profiling was applied to compare the transcriptome profiles in FBS-containing versus FBS-free culture. Comparison of the expression patterns of ECFCs that were either cultured in FBS-containing medium or in serum-free medium (five replicates each).

ORGANISM(S): Homo sapiens

SUBMITTER: Agapios Sachinidis

PROVIDER: E-GEOD-20504 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Optimization of the culturing conditions of human umbilical cord blood-derived endothelial colony-forming cells under xeno-free conditions applying a transcriptomic approach.

Zeisberger Steffen M SM Zoller Stefan S Riegel Mariluce M Chen Shuhua S Krenning Guido G Harmsen Martin C MC Sachinidis Agapios A Zisch Andreas H AH

Genes to cells : devoted to molecular & cellular mechanisms 20100520 7

Establishment of fetal bovine serum (FBS)-free cell culture conditions is essential for transplantation therapies. Blood-derived endothelial colony-forming cells (ECFCs) are potential candidates for regenerative medicine applications. ECFCs were isolated from term umbilical cord blood units and characterized by flow cytometry, capillary formation and responsiveness to cytokines. ECFCs were expanded under standard, FBS-containing endothelial medium, or transferred to chemically defined endothelia ...[more]

PMID: 20497237

Similar Datasets

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

Project description:Derivation and expansion of human umbilical cord blood-derived endothelial colony forming cells under serum-free conditions - a transcriptome analysis. Endothelial colony forming cells (ECFCs) were isolated from term umbilical cord blood units. ECFCs were expanded under standard, fetal bovine serum (FBS) containing endothelial medium, or transferred to chemically defined endothelial media without FBS. Microarray expression profiling was applied to compare the transcriptome profiles in FBS-containing versus FBS-free culture.

2010-06-29 | GSE20504 | GEO

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

Genomic binding profile of TAL1 in human Endothelial Colony-Forming Cells

2014-05-06 | E-GEOD-44442 | biostudies-arrayexpress

Cardiovascular-related proteomic changes in ECFCs exposed to the serum of COVID-19 patients

Project description:Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection not only exerts a deep impact over the respiratory system, but it also affects the cardiovascular system, causing vascular damage and thromboembolic events in critical patients. The endothelial dysfunction taking place in response to COVID-19 represents one of the first steps in this cascade of events that might leads to long-term sequelae over the vascular function. However, despite the enormous efforts in the last two years, the molecular mechanisms involved in such processes remains poorly understood. Using a label free-quantitative proteomics approach, we profiled the proteome of endothelial colony forming cells (ECFCs), after its incubation with the serum from individuals infected with COVID-19, whether asymptomatic or critical patients. Specifically, ECFCs were incubated ex-vivo with the serum of either SARS-CoV-2 negative donors (PCR-/IgG-), SARS-CoV-2 positive asymptomatic donors, at different infective stages: PCR+/ IgG- and PCR-/IgG+, or hospitalized critical COVID-19 patients, followed by mass spectrometry analysis. Remarkably, in response to all infected serums, the differentially expressed protein were found to be correlated with alterations in viral infection, RNA metabolism or autophagy, among others. Moreover, the serum of critical patients promoted the highest number of protein alterations, many of which associated with cardiovascular related pathologies. Finally, machine learning algorithms confirmed the discriminating potential of some proteins differentially expressed between conditions.

2023-03-20 | PXD034620 | Pride

Distinct phenotype and proteomic signature of peripheral blood-endothelial colony-forming cells of subjects with varied cardiovascular disease risks

Project description:Endothelial progenitor cells (EPCs) are circulating endothelial precursors shown to incorporate into foci of neovascularisation. Herein, we describe phenotypic characteristics of an EPC sub-type called endothelial colony-forming cells (ECFCs). Peripheral blood-isolated ECFCs expressed endothelial and progenitor surface antigens and displayed cobblestone-patterned colonies with clonal proliferative and angiogenic capacities in vitro. ECFCs demonstrated endothelial cell-like shear stress responses including cell alignment and PECAM-1 expression. Proteomic comparison with an endothelial reference population (human umbilical vein endothelial cells) confirmed a similar proteomic profile. Hierarchical clustering revealed two distinct ECFC clusters with differences in cell growth, proliferation and angiogenesis capacities. The cluster with compromised functionality was also associated with elevated blood pressure and impaired lipid profile. Our findings described an endothelial-like phenotype of blood-derived ECFCs with distinctive proteomic signatures based on cellular and clinical characteristics. ECFCs may aid in identifying novel mechanisms associated with cardiovascular disease risk and new targets to enhance angiogenesis.

2022-02-15 | PXD020677 | Pride

Prospective surface marker-based isolation and expansion of fetal endothelial colony-forming cells from human term placenta.

Project description:The term placenta is a highly vascularized tissue and is usually discarded upon birth. Our objective was to isolate clinically relevant quantities of fetal endothelial colony-forming cells (ECFCs) from human term placenta and to compare them to the well-established donor-matched umbilical cord blood (UCB)-derived ECFCs. A sorting strategy was devised to enrich for CD45-CD34+CD31Lo cells prior to primary plating to obtain pure placental ECFCs (PL-ECFCs) upon culture. UCB-ECFCs were derived using a well-described assay. PL-ECFCs were fetal in origin and expressed the same cell surface markers as UCB-ECFCs. Most importantly, a single term placenta could yield as many ECFCs as 27 UCB donors. PL-ECFCs and UCB-ECFCs had similar in vitro and in vivo vessel forming capacities and restored mouse hind limb ischemia in similar proportions. Gene expression profiles were only minimally divergent between PL-ECFCs and UCB-ECFCs, probably reflecting a vascular source versus a circulating source. Finally, PL-ECFCs and UCB-ECFCs displayed similar hierarchies between high and low proliferative colonies. We report a robust strategy to isolate ECFCs from human term placentas based on their cell surface expression. This yielded much larger quantities of ECFCs than UCB, but the cells were comparable in immunophenotype, gene expression, and in vivo functional ability. We conclude that PL-ECFCs have significant bio-banking and clinical translatability potential. Data has been integrated into the Stemformatics web resource http://www.stemformatics.org/datasets/view/6306

2015-03-02 | E-MTAB-3277 | biostudies-arrayexpress

Questioning the purity of the media - Extracellular small non-coding RNA contaminants in fetal bovine serum and serum-free media

Project description:In the research field of extracellular vesicles (EVs), the use of EV-depleted fetal bovine serum (FBS) for in vitro studies is highly recommended to eliminate the confounding effects of media derived EVs. EV-depleted FBS may either be prepared by ultracentrifugation or bought commercially, nevertheless these depletion methods do not guarantee an RNA-free preparation. In this study we have addressed the RNA contamination issue in FBS, ultracentrifuged EV-depleted FBS, commercially available EV-depleted FBS, and also from our recently developed filtration based EV depleted FBS. Commercially available serum-free, xeno-free defined media were also screened for RNA contamination.

2019-04-30 | GSE120594 | GEO

Derivation of endothelial colony forming cells from human pluripotent stem cells

Project description:Human induced pluripotent stem (hiPS) cells and human embryonic stem (hES) cells differentiate into cells of the endothelial lineage, but derivation of cells with human umbilical cord blood endothelial colony forming cell (ECFC)-like properties has not been reported. Here we describe a novel serum- and stromal cell-free ECFC differentiation protocol for the derivation of clinically relevant numbers of ECFCs (> 108) from hiPS and hES cells. We identified NRP-1+CD31+ selected cells that displayed a stable endothelial phenotype exhibiting high clonal proliferative potential, extensive replicative capacity, formation of human vessels that inosculated with host vasculature upon transplantation, but lacking in teratoma formation in vivo. We also identified NRP-1-VEGF165-KDR-mediated activation of KDR as a critical mechanism for the emergence and derivation of ECFCs from hiPS and hES cells. This protocol advances the field by generating highly replicative but stable endothelial cells for use as a potential cell therapy for human clinical disorders. Transcriptome sequencing of undifferentiated day 0 hiPS cells, Day 3 differentiated hiPS-derived mesoderm proginator cells, Day 12 hiPS-derived NRP-1+CD31+ cells, Day 12 H9-hES-derived NRP-1+CD31+ cells and cord blood-derived Endothelial colony forming cells.

2014-07-18 | E-GEOD-49074 | biostudies-arrayexpress

Derivation of endothelial colony forming cells from human pluripotent stem cells

2014-07-18 | GSE49074 | GEO

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