ABSTRACT: Using oligonucleotide microarray analysis, we identified 56 genes that were transcriptionally up-regulated and 69 that were suppressed upon exposure of endothelial cells to C. albicans. Among the regulated genes those attributed to the categories ‘chemotaxis’, ‘signaling’, and ‘transcription and translation’ were remarkably overrepresented. Keywords: endothelial stress response Overall design: We performed 4 independent experiments comparing the expression profiles of untreated HUVEC monolayer with those of candida-infected HUVEC monolayer.
INSTRUMENT(S): [HG-U133A] Affymetrix Human Genome U133A Array
Project description:Using oligonucleotide microarray analysis, we identified 56 genes that were transcriptionally up-regulated and 69 that were suppressed upon exposure of endothelial cells to C. albicans. Among the regulated genes those attributed to the categories ‘chemotaxis’, ‘signaling’, and ‘transcription and translation’ were remarkably overrepresented. Experiment Overall Design: We performed 4 independent experiments comparing the expression profiles of untreated HUVEC monolayer with those of candida-infected HUVEC monolayer.
Project description:To delineate specific patterns of signaling networks activated by H5N1 we used a comparative systems biology approach analyzing gene expression in endothelial cells infected with three different human and avian influenza strains of high and low pathogenicity. Overall design: HUVECs were infected with either PR8, FPV or H5N1 virus. We used wildtype HUVEC or HUVEC transfected with a dominant negative mutant of IKK2 (block of the NF-kB signaling pathway). In case of FPV infections we also blocked p38 MAP kinase with an SB-inhibitor.
Project description:We expressed a constitutively active mutant of MEK5 (MEK5D) in human primary endothelial cells (EC) to study the transcriptional and functional responses to Erk5 activation under static conditions. Overall design: HUVEC were infected with either empty vector or constitutively active MEK5D and RNA was processed for microarray analysis 40 h post infection.
Project description:Analysis of umbilical vein endothelial cells (HUVEC) treated with Egr-3 siRNA under the VEGF treatment for 0,1, and 4 h. Egr-3, a member of early growth response family, is immediately and dramatically induced by VEGF in HUVEC, which regulates expression of many genes related to endothelial activation. Overall design: Total 21 samples were derived from triplicate arrays of VEGF-treated si-Control-transfected cells and duplicate arrays of each of the VEGF-treated si-Egr-3-transfected cells.
Project description:To profile changes in gene expression in human endothelial cells in response to VEGF-A165 and phenotypic changes during vascular network formation in vitro 16 samples of human umbilical vein endothelial cells were analysized, four distinct biological samples were used in each condition. The four conditions included: VEGF treatment in Matrigel culture, Mock treatment in Matrigel culture, VEGF treatment in 2D monolayer, and mock treatment in 2D monolayer.
Project description:Angiogenesis, the formation of new capillaries by sprouting from preexisting vessels, is mainly induced by VEGF-A. To identify genes which are induced by VEGF-A in endothelial cells and genes which are differently expressed or VEGF-A induced in HUVEC and CEP, HUVEC and CEP were starved and induced by VEGF-A165 for 30, 60 and 150min. RNA of induced and uninduced cells was isolated and subjected to microarray analysis using Affymetrix microarray. Overall design: Human umbilical vein endothelial cells and circulating endothelial progenitor cells were grown in complete EGM-2 medium in dense culture for four days without change of medium and before the stimulation in EGM-2 medium without growth factors over night. The cells were stimulated with 100ng/ml VEGF-A165 for 30, 60 and 150 min. CD34 positive cells were isolated by MACS from cord blood. RNA was isolated with Trizol according to the manufacturer's instructions.
Project description:Transcriptional profiling of Human Umbilical Vein Endothelial Cells (HUVEC) comparing untreated control cells with IL-1-treated cells with or without pre-treatment with DHA. Three condition experiment: DHA treated HUVEC cells (25 µmol/L DHA for 48 hours) vs control; IL-1 treated HUVEC cells (5ng/ml for 3 hours) vs control; HUVEC treated with 25 µmol/L DHA for 48 hours and then stimulated with 5 ng/mL IL-1β for 3 hours. For each condition, 3 replicates
Project description:We expressed a constitutively active mutant of MEK5 (MEK5D) in human primary endothelial cells (EC) to study the transcriptional and functional responses to Erk5 activation under static conditions. HUVEC were infected with either empty vector or constitutively active MEK5D and RNA was processed for microarray analysis 40 h post infection.
Project description:Fundamental research and drug development for personalized medicine necessitates cell cultures from defined genetic backgrounds. However, providing sufficient numbers of authentic cells from individuals poses a challenge. Here, we present a new strategy for rapid cell expansion that overcomes current limitations. Using a small gene library, we expanded primary cells from different tissues, donors and species. Cell type specific regimens that allow the reproducible creation of cell lines were identified. In depth characterization of a series of endothelial and hepatocytic cell lines confirmed phenotypic stability and functionality. Applying this technology enables rapid, efficient and reliable production of unlimited numbers of personalized cells. As such, these cell systems support mechanistic studies, epidemiological research and tailored drug development. In these experiments primary HUVEC were compared to immortalized HUVEC with respect to their global gene expression pattern. Overall design: The global gene expression profile of primary and immortalized HUVEC was analyzed. In addition primary human fibroblasts (4 samples; from three different donors) as control cells were included in this experiment. The immortalized HUVEC lines are derived from the same donor as the primary cells and were treated with the following gene combinations (MYC, ID1, ID2 - e-hUVEC-2); (MYC, ID1, FOS - e-hUVEC-8); (MYC, ID2, FOS - e-hUVEC-9). Each HUVEC sample (primary or immortalized) was run in duplicate.
Project description:Human umbilical vein endothelial cells (HUVEC) were isolated from umbilical cords from four different donors, as previously described (Cooke et al., 1993). Cells were maintained in Medium 199 (M199, Invitrogen) containing 20% fetal calf serum, 28g/ml gentamycin, 2.5g/ml amphotericin B, 1ng/ml epidermal growth factor and 1g/ml hydrocortisone (all from Sigma) for 48 hours prior to processing. HUVEC cultures isolated using this method were 96-98% pure, determined by positive staining by flow cytometry of CD105, CD31 and vWF and the expression of elevated levels of intracellular adhesion molecule (ICAM-1) and E-selectin following stimulation with the inflammatory cytokine interleukin-1. Total HUVEC RNA was isolated using the RNeasy mini kit with QIAshredder (Qiagen) according to the manufacturers instructions. RNA integrity number was >8.0 for all samples.