Gene expression profile of endothelial cells maturated into brain-like endothelial cells
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ABSTRACT: To understand the molecular mechanisms during the maturation of cord blood-derived endothelial cells into blood brain barrier capillary endothelial cells (BCECs), we have employed whole genome microarray expression profiling to identify genes responsible for the maturation process. Hematopoietic stem cells were isolated from cord-blood samples and differentiated into endothelial cells. The endothelial cells were further maturated into BCECs by co-culturing with blood-brain barrier (BBB) specific cells (pericytes) for 3 days and 6 days. The gene expression in human hematopoietic stem cell-derived endothelial cells was measured at 3 and 6 days after co-culture with pericytes. Three independent experiments were performed at each time (3 or 6 days). The RNA obtained from different experiments were pooled together for each group before microarray studies.
Project description:Analysis of neutrophil proteomic alterations induced by migration towards inflamed joints in juvenile idiopathic arthritis (JIA). In this experiment neutrophil proteomes were investigated after migration towards JIA synovial fluid in an in vitro model of a synovial membrane, compared to neutrophils incubated in synovial fluid without migration. The migration model consisted of transwell inserts with human knee synoviocytes on the undersides and HMEC endothelial cells on the insides, placed in wells containing medium with 10 % JIA synovial fluid.
Project description:Goal of the study was to identify (using proteomics) the consequences of the lipocine 19,20-DHDP on the proteins enriched in the junctions between endothelial cells and pericytes.
Project description:Using RNA sequencing to map differentially expressed genes in the 3D model of the blood-brain barrier ( composed of human brain endothelial cells, human astrocytes, and human pericytes) challenged with WNV.
Project description:Using RNA sequencing to map differentially expressed genes in the 3D model of the blood-brain barrier ( composed of human brain endothelial cells, human astrocytes, and human pericytes) challenged with TBEV.
Project description:Using RNA sequencing to map differentially expressed genes in the 3D model of the blood-brain barrier ( composed of human brain endothelial cells, human astrocytes, and human pericytes) challenged with Borrelia garini .
Project description:Abnormal tumor vessels promote metastasis and impair chemotherapy. Hence, tumor vessel normalization (TVN) by targeting endothelial cells (ECs) is emerging as anti-cancer treatment. Here, we show that tumor ECs (TECs) have a hyper-glycolytic metabolism, shunting glycolytic intermediates to nucleotide synthesis. EC haplo-deficiency or blockade of the glycolytic activator PFKFB3 did not affect tumor growth, but reduced cancer cell intra- and extravasation and metastasis by normalizing tumor vessels, which improved vessel maturation and perfusion. Mechanistically, PFKFB3 inhibition tightened the vascular barrier by reducing VE-cadherin endocytosis in ECs and rendering glycolytic pericytes more quiescent; it also lowered the expression of cancer cell adhesion molecules in ECs. Additionally, PFKFB3-blockade treatment improved chemotherapy. Considering TEC metabolism for anti-cancer treatment might thus merit further attention.
Project description:This study aims to investigate differentially expressed proteins in tumor pericytes derived from colorectal cancer patients with or without liver metastasis. Tumor pericytes were isolated from tumor of colorectal cancer patients with or without liver metastasis. Then, tumor pericytes were cultured and subjected to proteomic analysis. TCAF2 was significantly increased in tumor pericytes from liver metastasis patients.
Project description:The objective of this array was to determine the global gene expression profile of human placental pericytes for comparison with other publicly available arrays of pericytes and mesenchymal stromal cells isolated from various human tissues. Pericytes are critical cellular components of the microvasculature that play a major role in vascular development and pathologies, yet their study has been hindered by lack of a standardized method for their isolation and growth. Here we report a method for culturing human pericytes from a readily available tissue source, placenta, and provide a thorough characterization of resultant cell populations. We developed an optimized protocol for obtaining pericytes by outgrowth from microvessel fragments recovered after enzymatic digestion of human placental tissue. We characterized outgrowth populations by immunostaining, by gene expression analysis, and by functional evaluation of cells implanted in vivo. Our approach yields human pericytes that may be serially expanded in culture and that uniformly express the cellular markers NG2, CD90, CD146, α-SMA, and PDGFR-β, but lack markers of smooth muscle cells, endothelial cells, and leukocytes. When co-implanted with human endothelial cells into C.B-17 SCID/bg mice, human pericytes invest and stabilize developing human endothelial cell-lined microvessels. We conclude that our method for culturing pericytes from human placenta results in the expansion of functional pericytes that may be used to study a variety of questions related to vascular biology. Total RNA from three different pericyte isolations at subculture 1 was collected and examined for relative gene expression.
Project description:To understand the the effect of poly(lactic-co-glycolic acid) nanoparticles (NPs) encapsulating a fluorine contrast agent on human umbilical vein endothelial cells (HUVECs), we have employed whole genome microarray expression profiling. The NPs used in this study were prepared in our lab. HUVECs were ordered from Lonza (Cat# C2517A). HUVECs were incubated with NPs for 4 hours. After 4 h, the cells were washed couple of times to remove the NPs and gene expression profiles were detected after 20 h (total time: 24 h) and 7 d in the absence of NPs. HUVECs after 24 h without nanoparticles were used as controls. The gene expression in HUVECs was measured at 24 h and 7 d after incubation with nanoparticles. Three independent experiments were performed at each time.
Project description:The interplay of cerebromicrovascular endothelial cells, pericytes and perivascular macrophages is central to the recruitment of neutrophils across the blood-brain barrier into the brain, and therefore to the CNS inflammatory response. This experiment examined the endothelial response to direct stimulation in the presence or absence of pericytes to explore the effect of co-culture on the endothelial inflammatory response. It also assessed the endothelial response to signalling by monocyte-derived macrophages, modelling perivascular macrophages, the only canonical innate immune cell found within the perivascular space.