ABSTRACT: Transcription profiling of human umbilical cord blood derived Outgrowth Endothelial Cells (OECs) at early and late passages. Outgrowth endothelial cells were isolated from the mononuclear fraction of umbilical cord blood and cultured on collagen coated flasks. Once OEC colonies emerged they were expanded in culture to study cell growth kinetics. Total RNA was extracted from OECs at an early passage and OECs at a late passage which were becoming senescent . The purpose of this experiment was to were compared early and late passage OECs to determine how senescence affects the gene expression profile of OECs.
Project description:The Loeys-Dietz syndrome (LDS) is an inherited connective tissue disorder caused by mutations in the transforming growth factor β (TGF-β) receptors TGFBR1 or TGFBR2. Most patients with LDS develop severe aortic aneurysms resulting in early need of surgical intervention. We investigated circulating outgrowth endothelial cells (OEC) from the peripheral blood of LDS to gain further insight into the pathophysiology of the disorder. We performed gene expression profiling using microarray analysis followed by quantitative PCR for verification of gene expression. OECs isolated from age- and sex-matched healthy donors served as reference control. 3 OEC clones from different LDS patients were compared to their respective age- and sex-matched OEC clone isolated from healthy donors.
Project description:Gene expression profiles during the differentiation of EPCs into OECs were analyzed. Experiment Overall Design: RNA was isolated from Human cord blood derived- EPC and differentiated OEC from EPC. HG-U133A 2.0 microarray (54675 human genes; Affymetrix Santa Clara, CA, USA) was performed.
Project description:We analyzed the changes in the spinal cord transcriptome after a spinal cord contusion injury and MSC or OEC transplantation. The cells were injected immediately or 7 days after the injury. The mRNA of the spinal cord injured segment was extracted and analyzed by microarray at 2 and 7 days after cell grafting. 52 total samples were analyzed in 13 different groups. Each group include 4 samples and each one were analyzed as a biological replica. The intact animals were used as control of injury. The vehicle (VHC) groups were used as control of transplantation procedure. The MSC or OEC graft were injected at the day of injury (acute graft) or seven days after injury (delayed graft). The samples from engrafted animals were obtained at 2 or 7 days after cell transplantation. To determine the effects of MSC or OEC transplantation, the expression value of each engrafted sample were compared with correspondent VHC group.
Project description:Human umbilical vein endothelial cells (HUVECs) formed capillary structures when co-cultured with normal human dermal fibroblasts (NHDFs). HUVEC competence and NHDF supportiveness of cord formation were found to be highly cell-passage dependent with the early passage cells forming more angiogenic cord structures. We thus profiled gene expression in NHDFs with different passages to understand the molecular mechanisms underlying the in vitro angiogenesis control. Keywords: Time course
Project description:Analysis of DNA methylation and gene expression changes during regulated endothelial progenitor cells (EPCs) to outgrowth endothelial cells (OECs). Results provide information of DNA methylation and gene expression pattern during cord-blood derived EPCs differentiation. Taken together, we discovered specific set of genes regulated by hyper- and hypo-methylation during differentiation.
Project description:Analysis of DNA methylation and gene expression changes during regulated endothelial progenitor cells (EPCs) to outgrowth endothelial cells (OECs). Results provide information of DNA methylation and gene expression pattern during cord-blood derived EPCs differentiation. Taken together, we discovered specific set of genes regulated by hyper- and hypo-methylation during differentiation.
Project description:Human umbilical vein endothelial cells (HUVECs) formed capillary structures when co-cultured with normal human dermal fibroblasts (NHDFs). HUVEC competence and NHDF supportiveness of cord formation were found to be highly cell-passage dependent with the early passage cells forming more angiogenic cord structures. We thus profiled gene expression in NHDFs with different passages to understand the molecular mechanisms underlying the in vitro angiogenesis control. Experiment Overall Design: NHDFs cultured to various passages were assayed for their gene expression using the GeneChip technology.
Project description:Genome -wide analysis of EPC subtypes obtained from both umbilical cord and peripheral blood. This experiment sought to compare the expression levels of eEPCs and OECs against monocytes and human dermal microvascular cells (HDMECs) in order to establish if these cells are linked to a monocytic or endothelial lineage. This is very important as the term EPC is currently used to describe cell populations which are quite dissimilar in terms of biological properties.
Project description:To search for genes regulated by microRNA-100 in endothelial cells, we transfected human umbilical cord endothelial cells (HUVECs) with miR-100 precursor oligonucelotides. Human umbilical vein endothelial cells (HUVECs) were isolated from donated umbilical cords, pooled from two donors and cultivated up to passage 5. For transfection with pre-miR microRNA precursor molecules cells were cultured to 70% confluence and transfected with 8nM pre-miR-100 or an irrelevant control oligonucleotide (both from Ambion) using Lipofectamin RNAiMax (Invitrogen) according to the manufacturers instructions. Total RNA was isolated 48h after transfection.
Project description:To search for genes regulated by microRNA-100 in endothelial cells, we transfected human umbilical cord endothelial cells (HUVECs) with miR-100 precursor oligonucelotides. Human umbilical vein endothelial cells (HUVECs) were isolated from donated umbilical cords, pooled from two donors and cultivated up to passage 5. For transfection with pre-miR microRNA precursor molecules cells were cultured to 70% confluence and transfected with 8nM pre-miR-100 or an irrelevant control oligonucleotide (both from Ambion) using Lipofectamin RNAiMax (Invitrogen) according to the manufacturers instructions. Total RNA was isolated 48h after transfection.