Project description:Studies of miRNA profiling in early and late endothelial progenitor cells treated or not by cardioprotective nucleoside adenosine. Early outgrowth endothelial progenitor cells were obtained by adhesion of peripheral blood mononuclear cells of healthy volunteers. Late endothelial progenitor cells were obtained by purification of CD34+ peripheral blood cells and were cultured and amplified in endothelial-specific medium containing growth factors. Both cell types were treated by adenosine (10micromol/L) for 6 hours. Total RNA was extracted using mirVana Kit and quantified by Nanodrop. RNA was labeled and hybridized using Agilent miRNA Complete Labeling and Hyb Kit. 3 to 4 arrays per sample were hybridized and scanned with the Genepix 4000B Scanner (Molecular Devices). Six independent experiments were performed.
Project description:Endothelial progenitor cells (EPCs) are a group of cells which can differentiate to mature endothelial cells in a certain culture condition, and were first isolated from adult human peripheral blood by Asahara et al. in 1997. EPCs have a critical role in the restoration of injured vessel endothelium and the neovascularization in the area of ischemia injury. Recently, the role of androgens in the proliferation, differentiation and adhesion of EPCs is more and more focused. Dihydrotestosterone (DHT) is a kind of unmetabolizable androgen. We used microarrays to detail the global programme of gene expression underlying cellularisation and identified distinct classes of regulated genes in DHT-treated EPCs.
Project description:The mechanisms underlying proangiogenic function of brain-derived neurotrophic factor (BDNF) are not fully understood. Current study was designed to explore the miRNA profile in human early endothelial progenitor cells (EPCs, also referred to as CFU-Hill cells) in response to BDNF treatment.
Project description:Gene expression profiling of immortalized human mesenchymal stem cells with hTERT/E6/E7 transfected MSCs. hTERT may change gene expression in MSCs. Goal was to determine the gene expressions of immortalized MSCs.
Project description:Background: In recent times a subset of bone marrow (BM) derived cells; endothelial progenitor cells (EPCs), have generated tremendous interest, as these cells are suggested to home to sites of neovascularization and neoendothelialization and differentiate into mature endothelial cells (ECs), a process referred to as postnatal vasculogenesis. EPCs are being considered as a potential regenerative tool for treating various pathophysiological disease states including cardiovascular disorder and as a possible target to restrict vessel growth in tumour pathology. However, conflicting results have been reported in the field due to lack of EPC specific biomarkers, and the identification, characterization, and the precise role of EPCs in vascular biology still remains a subject of great debate. Therefore, the objective of this study was to use a bioinformatics approach to identify putative novel EPC specific biomarkers. Methods: This study reports a detailed gene expression profile of human umbilical cord blood (UCB) derived non-adherent CD133 + cells at different time points during in vitro differentiation (day 4 and day 7) which were compared with differentiated donor matched human umbilical vein endothelial cells (HUVEC). Results: EPC gene expression was profiled at both days 4 and 7, using affymetrix human 1.0ST exon arrays. Affymetrix gene expression profiling revealed significant expression changes in genes associated with stem/progenitor cell properties such as adhesion, signalling, molecular transport, cell structure organisation and growth. Conclusions: This is the first study utilizing gene expression profiling to examine non adherent CD133+ progenitor cells. Alterations in the gene expression reported in this study may be involved in the cellular processes characteristic of EPC development and differentiation. Gene expression profiling was performed on EPCs cultured for D4 and D7, on Affymetrix human 1.0ST exon array chips. Gene Spring (version GX11) was used to perform the gene expression analysis on three experimental groups; (a) D4 EPCs versus HUVEC; (b) D7 EPCs versus HUVEC and (c) D4 versus D7 EPCs.
Project description:Transcriptional profiling of human mesenchymal stem cells comparing normoxic MSCs cells with hypoxic MSCs cells. Hypoxia may inhibit senescence of MSCs during expansion. Goal was to determine the effects of hypoxia on global MSCs gene expression.
Project description:Endothelial progenitor cells (EPCs) are a group of cells which can differentiate to mature endothelial cells in a certain culture condition, and were first isolated from adult human peripheral blood by Asahara et al. in 1997. EPCs have a critical role in the restoration of injured vessel endothelium and the neovascularization in the area of ischemia injury. Recently, the role of androgens in the proliferation, differentiation and adhesion of EPCs is more and more focused. Dihydrotestosterone (DHT) is a kind of unmetabolizable androgen. We used microarrays to detail the global programme of gene expression underlying cellularisation and identified distinct classes of regulated genes in DHT-treated EPCs. EPCs were isolated from mouse bone marrow. Male C57BL/6 mice (8 weeks age) mice were sacrificed by cervical dislocation. For microarray analysis, the cells were grown in culture medium supplemented with 10nmol/L DHT, which was changed on day 2. On day 7, cells was collected for RNA extraction and subsequent experiments.