Transcription profiling of human umblical vein endothelial cells (HUVEC) undergoing tubulogenesis in a 3D fribrin matrix or cultured as monolayers
ABSTRACT: In order to determine genes that are differentially expressed during angiogenesis, Human Umbilical Vein Endothelial Cells (HUVEC) were cultured as 3D cultures undergoing tubulogenesis in a 3D fibrin matrix, or cultured as monolayers on top of a 3D fibrin matrix. RNA was then collected, reverse transcribed to cDNA and hybridized to glass slide oligo arrays containing 19k human genes. Differentially expressed genes in HUVECs undergoing tubulogenesis were then determined by comparing 2D to 3D culture samples.
Project description:Identification of the relationships of Kaposi sarcoma (KS), normal skin to various cell cultures. The effects of KS herpes virus, the infectious cause of KS, on infected endothelial cells are also investigated.
Project description:The aim of this study was to identify HDL and apoE-regulated genes in human placental endothelial cells (HPEC), which are exposed to fetal HDL. HPECs extracted from 5 human placentas were cultivated and treated for 16 h with 15ug/ml fetal HDL, 15ug/ml reconstituted HDL (rHDL), or endothelial basal medium (EBM) as vehicle control. Gene expression analysis from these 3 conditions (5 biological replicates) using 15 Applied Biosystems Human Whole Genome Survey V2.0 Microarrays was perfomed and significantly differentially expressed genes between two different groups (HDL vs control or rHDL vs control) were identified.
Project description:As susceptibility to many adult disorders originates in utero, we here hypothesized that fetal sex influences gene expression in placental cells and produces functional differences in human placentas. We found that fetal sex differentially affects gene expression in a cell-phenotype dependent manner among all four placental cell-phenotypes studied: cytotrophoblasts, syncytiotrophoblasts, arterial endothelial cells and venous endothelial cells. The markedly enriched pathways in males were identified to be signaling pathways for graft-versus-host disease as well as the immune and inflammatory systems, both supporting the hypothesis that there is reduced maternal-fetal compatibility for male fetuses. Our study is the first microarray study investigating sexual dimorphism in purified and characterized somatic cells from a single human tissue, the placenta, that underlines the importance of considering fetal sex as an independent variable in any work using human placenta. Arterial and venous endothelial cells were isolated from eight different placentas, four of each sex. A total of ten placentas were used for isolation of cytotrophoblasts and six for syncytiotrophoblasts, with equal numbers from each sex.
Project description:Collagen- and fibrin-based gels are extensively used to study cell behaviour. However, 2D-3D, collagen-fibrin, and in vivo-in vitro comparisons of gene expression, cell shape and mechanotransduction have not been reported. Here we compared chick tendon fibroblasts (CTFs) at three stages of embryonic development with CTFs cultured in collagen- or fibrin-based tissue engineered constructs (TECs).
Project description:We hypothesized that generating spinal motor neurons (sMNs) from human induced pluripotent stem cell (hiPSC)-derived neural aggregates (NAs) using a chemically-defined differentiation protocol would be more effective inside of 3D fibrin hydrogels compared to 2D poly-L-ornithine(PLO)/laminin-coated tissue culture plastic surfaces. We performed targeted RNA-Seq using next generation sequencing to determine the substrate-specific differences in gene expression that regulate cell phenotype. Cells cultured on both substrates expressed sMN genes CHAT and MNX1, though persistent WNT signaling contributed to a higher expression of genes associated with interneurons in NAs cultured in 3D fibrin scaffolds. Cells in fibrin also expressed lower levels of astrocyte progenitor genes and higher levels of the neuronal-specific gene TUBB3, suggesting a purer population of neurons compared to 2D cultures. STATEMENT OF SIGNIFICANCE: Fibrin scaffolds can support the neuronal differentiation of pluripotent stem cells. This study provides insight into how fibrin hydrogels affect neuronal induction by analyzing of the signaling pathways activated during the differentiation process. These insights can then be used to tailor the properties of these hydrogels to optimize the generation of sMNs for regenerative medicine applications. Overall design: 45 samples and 6 sample groups. Minimum of 6 replicates per sample group. Sample groups can be differentiated by day and substrate
Project description:Unrestricted Somatic Stem Cells (USSC) are a cell population derived from umbilical cord blood. They have been shown to have greater plasticity than mesenchymal stem cells, and are being investigated as a therapeutic option for many disease states, including cardiovascular disease.<br><br>In this study, we aimed to evaluate the efficacy of USSC on treating acute myocardial infarction (MI). We compared the effectiveness of USSC to the more widely studied bone marrow derived mesenchymal stem cell. Finally, we investigated whether pre-conditioning USSC prior to administration enhances their effectiveness. We determined effectiveness primarily using cardiac ultrasound, and supported these findings with histological analyses.<br><br>We observed that pre-conditioned (guided) USSC had the most significant beneficial effect on cardiac function. (Pre-conditioned cells were grown in serum-free F12 media was supplemented with 50ng/ml Basic Fibroblast Growth Factor (bFGF), 20ng/ml Hepatocyte Growth Factor (HGF) and 20ng/ml Bone Morphogenetic Protein 2 (BMP2)). We performed microarray analysis of guided USSC and unmodified USSC to compare the gene expression profile of both cell populations, in order to evaluate whether any specific genes, or gene groups, were involved in mediating this beneficial effect.<br>
Project description:From two donors of human umbilical vein endothelial cells, in vitro cell lines were established. Both cell lines were grown in vitro until irreversible growth arrest was observed (replicative senescence). Total RNA from young (replicating) cells as well as senescent cells was harvested and used for hybridization of microRNA chips (MRC) from TU Graz based on Sanger miRBase 9.2
Project description:Fenofibrate is a synthetic ligand for the nuclear receptor peroxisome proliferator-activated receptor (PPAR) alpha, but there are reports that fenofibrate affects endothelial cells in PPARa-independent manner. In order to identify PPARa-dependently and PPARa-independently regulated transcripts we generated microarray data from human endothelial cells treated with fenofibrate with and without siRNA-mediated knock-down of PPARa. In this study, we generated microarray data from human umbilical vein endothelial cells (HUVECs) treated with fenofibrate. Time 0 indicates the start point of observation immediately prior to exposure to the 25umol fenofibrate. There are five time points (2, 4, 6, 8, and 18hours) (n=3 at each time point). The control is untreatment HUVEC (n=4).
Project description:Tumour cell extravasation is a key event for metastatic colonisation1. However, little is known about the signalling pathways that govern tumour cell adhesion to the endothelium and subsequent transmigration through the blood vessel wall. Here, we combine Stable Isotope Labelling by Amino acids in Cell culture (SILAC) and global phosphoproteomic analysis to identify cell-specific signalling pathways regulated between interacting tumour and endothelial cells. We identified over 2,000 unique phosphorylation sites, where 77 were specifically regulated in the tumour cells and 43 in the endothelial cells.
Project description:Acidic tissue microenvironment is commonly found in a variety of pathophysiological conditions. GPR4 is a proton-sensing G protein-coupled receptor that is fully activated by acidic extracellular pH but has lesser activity at the physiological pH 7.4 and minimal activity at more alkaline pH. To determine the effects of GPR4 activation by acidosis on vascular endothelial cells, we examined the global gene expression of the acidosis response in primary human umbilical vein endothelial cells (HUVEC) with varying level of GPR4. HUVEC with endogenous or overexpressed GPR4 level (designated as HUVEC/Vector & HUVEC/GPR4 cells). Two treatment conditions: pH 6.4 vs. pH 8.4 for 5 h. Biological replicates: 4 HUVEC/Vector replicates (pH6.4 vs pH 8.4), and 4 HUVEC/GPR4 replicates (pH6.4 vs pH 8.4).