Comparison of gene expression between Human umbilical vein endothelial cells and Human liver sinusoidal endothelial cells
ABSTRACT: To identify leukocyte adhesion receptors which differentially regulate recruitment in human liver sinusoidal endothelial cells compared to a protoptypic venular endothelium Gene expression was measured in four groups Group 1: cultured human liver sinusoidal endothelial cells (HSEC) Group 2: cultured human umbilical vein endothelial cells (HUVEC) Group3: Interferon gamma and tumour necrosisfactor alpha treated HSEC and Group 4: Interferon gamma and tumour necrosisfactor alpha treated HUVEC. Two replicates were used for each group.
The recruitment of lymphocytes via the hepatic sinusoidal channels and positioning within liver tissue is a critical event in the development and persistence of chronic inflammatory liver diseases. The hepatic sinusoid is a unique vascular bed lined by hepatic sinusoidal endothelial cells (HSECs), a functionally and phenotypically distinct subpopulation of endothelial cells. Using flow-based adhesion assays to study the migration of lymphocytes across primary human HSECs, we found that lymphocyt ...[more]
Project description:mRNA expression after Ezh2 knock down was analyzed to identify genes regulated by Ezh2. Human umbilical vein endothelial cells (HUVEC) were transfected with 25 nmol/L of control small interfering RNA (siRNA) (Silencer Select Negative Control Ambion, Austin, TX) or siRNA directed against Ezh2 (s4918; Ambion) using Oligofectamine (Invitrogen). Total RNA was harvested 72 hours after transfection.
Project description:To test our hypothesis that human embryonic stem cell-derived endothelial cells(hESC-ECs) represent their in vivo counterparts, we performed transcriptional profiling experiments in which we compared the gene expression profiles of d6 CD34+KDR+ cells, d12-14 hESC-ECs and human umbilical vein endothelial cells(HUVECs). Significantly upregulated genes were identified using significance analysis of microarrays (SAM), by comparing each of these populations to undifferentiated hESCs. Differentially regulated genes from each of these populations were examined using overlap analysis;to identify which genes are uniquely expressed in each cell type and which are expressed in common, followed by functional annotation clustering of the conserved core group of genes.
Project description:To further investigate the mechanism how the decline in Notch signaling induces premature senescence in endothelial cells, we performed microarray analysis and identified Id1 nd DUSP1 as the downstream molecules of Notch pathway. In quantitative PCR and western blot analyses, the expression level of Id1 and DUSP1 increased in Notch1 over-expressing endothelial cells and decreased in knockdown similar to the result of microarray. The gene expression of human unbilical endothelial vein cells (HUVEC) infected with retroviral vectors encoding Jagged1, Jagged1-shRNA, or Notch1-shRNA. HUVEC infected with empty vector was used as a control. In each genotypes, three independent lines at passage 8 were performed.
Project description:Effect of TNF-alpha on microRNAs levels in Human Umbilical Endothelial Cells (HUVECs). HUVEC that were treated or not for 2 or 24 hours with TNF (10 ng/ml). Duplicate samples (1 or 2) of two different isolations of HUVEC (A or B)
Project description:Whole transcriptome comparisons of proliferating pure cultures of neonatal dermal microvacsular endothelial cells to infantile hemangioma endothelial cells. The total RNA was obtained from human dermal microvascular endothelial cells and infantile hemangioma endothelial cells. Illumina microarrays were performed to determine the whole genome expression differences between the cell lines.
Project description:To compare the gene expression profiles of unpassaged, proliferating HUVEC and human iris, retinal and choroidal microvascular endothelial cells. Gene expression profiling revealed significant differences between HUVEC and ocular microvascular endothelial cells suggesting that HUVE cells may not be a suitable surrogate when studying pathophysiological mechanisms of ocular disorders. There were significant differences in the gene expression of important cell signalling pathways in human retinal and choroidal ECs. These differences may be important in the mechanisms and treatment of choroidal and retinal neovascularisation. 12 arrays are included. Endothelial cells were derived from 4 tissues: iris, retina, choroid and human umbilical vein. RNA extracts from cells were hybridised to Affymetrix HGU133plus2 arrays in triplicate.
Project description:We report the transcriptome human primary hepatocytes and liver sinusoidal endothelial cells. Hepatocytes were obtained from commercial sources. LSECs were isolated based on the coexpression of Tie2 and CD32b, te strategy of purification controlled by RNA-Seq. Comparison of the expression of the Tie-2, CD32b, SELP, FVIII, VWF, Alb, Fg, F7genes
Project description:Class IIa histone deacetylases (HDACs) are signal-responsive regulators of gene expression involved in vascular homeostasis. To investigate the differential role of class IIa HDACs for the regulation of angiogenesis, we used siRNA to specifically suppress the individual HDAC isoenzymes. Among the HDAC isoforms tested, silencing of HDAC5 exhibited a unique pro-angiogenic effect evidenced by increased endothelial cell migration, sprouting and tube formation. Consistently, overexpression of HDAC5 decreased sprout formation, indicating that HDAC5 is a negative regulator of angiogenesis. The anti-angiogenic activity of HDAC5 was independent of MEF2 binding and its deacetylase activity, but required a nuclear localization indicating that HDAC5 might affect the transcriptional regulation of gene expression. To identify putative HDAC5 targets, we performed microarray expression analysis. Silencing of HDAC5 increased the expression of fibroblast growth factor 2 (FGF2) and angiogenic guidance factors including Slit2. Antagonization of FGF2 or Slit2 reduced sprout induction in response to HDAC5 siRNA. ChIP assays demonstrate that HDAC5 binds to the promoter of FGF2 and Slit2. In summary, HDAC5 represses angiogenic genes, like FGF2 and Slit2, which causally contribute to capillary-like sprouting of endothelial cells. The de-repression of angiogenic genes by HDAC5 inactivation may provide a useful therapeutic target for induction of angiogenesis. Experiment Overall Design: 6 samples: 3x siSCRAMBLED transfected HUVEC (control) + 3x siHDAC5 transfected HUVEC 24h after transfection