Project description:Analysis of the effects of cell shape on human coronary artery endothelial cell transcription. The hypothesis is that defined alterations in endothelial cell shape uniquely affect the endothelial transcriptome. Human coronary artery endothelial cells were plated onto spatially defined micropatterns (Cytoo) forcing them to conform to Disc, Crossbow, H, Y, or L shapes. As a control, human coronary artery endothelial cells were plated on non-restrictive areas of the Cytoo growth plate. Cells were serum starved for 48 hours prior to mRNA collection. RNA was collected after 48 hours growth on the restricted or non-restricted growth patterns and subjected to whole genome microarray analysis.
Project description:Analysis of the effects of cell shape on human coronary artery endothelial cell transcription. The hypothesis is that defined alterations in endothelial cell shape uniquely affect the endothelial transcriptome. Human coronary artery endothelial cells were plated onto spatially defined micropatterns (Cytoo) forcing them to conform to Disc, Crossbow, H, Y, or L shapes. As a control, human coronary artery endothelial cells were plated on non-restrictive areas of the Cytoo growth plate. RNA was collected after 24 hours growth on the restricted or non-restricted growth patterns and subjected to whole genome microarray analysis.
Project description:Shear stress is known to regulate endothelial cell orientation along the direction of flow. We asked wither cellular patterning along, in the absence of shear could have similar biological effects as shear. We used DNA microarrays to examine the effect of cellular patterning on their transcriptome. Human microvascular endothelial cells were cultured in parallel micropatterned channels (30um wide channels, 30um apart) composed of polydimethylsiloxane, followed DNA Microarray analysis (Affymetrix 1.0 ST array)
Project description:Loss of contractility and acquisition of an epithelial phenotype of vascular smooth muscle cells (VSMCs) are key events in proliferative vascular pathologies such as atherosclerosis and post-angioplastic restenosis. There is no proper cell culture system allowing VSMC differentiation so that it is difficult to delineate the molecular mechanism responsible for proliferative vasculopathy. We investigated whether a micro-patterned substrate could restore the contractile phenotype of VSMCs in vitro. To induce and maintain the differentiated VSMC phenotype in vitro, we introduced a micro-patterned groove substrate to modulate the morphology and function of VSMCs.
Project description:Cell samples of undifferentiated human umbilical cord mesenchymal stem cells (1-3) and cells that have been cultured in smooth muscle differentiation medium for 6 hours (4-6) and 24 hours (7-9) were collected and subjected to miRNA array. Exploration of miRNA involved smooth muscle differentiation mechanism would offer potential therapeutic choices for improving performance of vascular grafts engineered with umbilical cord mesenchymal stem cells.
Project description:Angiogenesis is tightly regulated by both soluble growth factors and cellular interactions with the extracellular matrix (ECM). While cell adhesion via integrins has been shown to be required for growth factor signaling and downstream angiogenesis, the effects of quantitative changes in cell adhesion and spreading against the ECM remain less clear. Examining changes in global gene expression in limited versus high adhesion contexts in human umbilical vein endothelial cells, we demonstrated a VEGF-induced upregulation of genes associated with vascular invasion and remodeling when cell adhesion was restricted, whereas cells on highly adhesive surfaces upregulated genes associated with a proliferative response. HUVECs were cultured on micropatterned islands of fibronectin or allowed to spread fully for 2 hrs and then stimulated with 50ng/ml VEGF or no growth factor for 18 hrs in starve media. Sample RNA from three biological replicates was extracted and prepared for hybridization on Affymetrix microarrays.
Project description:Blood platelets destined for transfusion release panoply of molecules during preparation and storage. The leukoreduction process made the transfusion safer but did not completely abolish the adverse events. The rationale is to study the proteome profile of platelet components PPC (platelet pellets) involved in transfusion adverse events.
Project description:Blood platelets destined for transfusion release panoply of molecules during preparation and storage. The leukoreduction process made the transfusion safer but did not completely abolish the adverse events. The rationale is to study the proteome profile of platelet components SDA-PC (platelet pellets) involved in transfusion adverse events.
Project description:The liver is one of most important organs in our bodies. It performs many essential functions including metabolism, synthesis, secretion, detoxification, and storage. Hepatocytes are the principal cell type in the liver and are involved in multiple liver-specific functions. There have been several efforts to develop in vitro culture systems capable of maintaining hepatocyte-specific phenotype over long time periods. In hepatic tissue engineering, two commonly used culture systems are the collagen sandwich and monolayers of cells.  In this study, genome-wide gene expression profiles of primary hepatocytes were measured over an 8-day period for each cell culture system using Affymetrix GeneChips and analyzed via Gene Set Enrichment Analysis (GSEA), which is a powerful method to elicit biologically meaningful information from microarray data at the level of gene sets. Results indicate that the gene expression in hepatocytes in collagen sandwich cultures gradually diverges from that in monolayer cultures. Gene sets up-regulated in collagen sandwich cultures include those associated with liver metabolic and synthetic functions.  These functions are associated with lipid, amino acid, carbohydrate, and alcohol metabolism and bile acid synthesis. Nuclear receptors are up-regulated in collagen sandwiches 24 hours after seeding. Signals transmitted from these receptors may cause the up-regulation of other processes in subsequent days. Cytochrome-P450 monooxygenase expression was initially down-regulated but exhibited up-regulation after 72 hours. Our results provide a baseline for further explorations into the systems biology of engineered liver mimics as well as 2D and 3D co-cultures of primary hepatocytes and non-parenchymal cells. To better understand differences in quality of in vitro growth of rat hepatocytes between culture on a monolayer of collagen gel and sandwiched between two layers of gel, we measured gene expression in hepatocytes in these two culture conditions in triplicate for four time points: 1 day, 2 days, 3 days, and 8 days of culturing. Overall, we obtained Affymetrix microarray data for 24 samples, divided to 12 samples from monolayer and double layer cultures each, each of which are divided into four time points with 3 sample replicates.
Project description:Analysis of human alveolar epithelial cell signatures at gene expression level. The aim of the study was to identify candidate genes that are induced in human alveolar epithelial type II cells on collagen-coated dishes. Results provide important information about changes hAT2 cells undergo in the transformation to AT1-like cells. Total RNA obtained from human alveolar epithelial cells grown on collagen- or matrigel-coated dishes for 12, 24, 48 and 72 hr.