The protein composition of these vesicles differs, depending on the stimulus used and might indicate differential roles during cardiac preconditioning.
In addition, we discovered that CD63 also has potential to discriminate between EVs derived from normal or from stressed endothelial cells.
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Human umbilical vein endothelial cells (HUVEC) were incubated with concentrations of glucose (between 6 and 50 mM) for 24 hours, followed by collection of medium to measure MV numbers.
Moreover, it has become clear that the cargo of EVs is tightly linked to the stage of a disease.
Since endothelial cells are activated in a range of inflammatory pathologies from atherogenesis to metastatic cancer, it is plausible that EVs may act as a signal for monocyte mobilization and recruitment.
We have previously demonstrated the selective extracellular export of specific regulatory miRNAs into extracellular vesicles (EVs) which may contribute to this response.
The aim of this study was to design a simple and easy protocol for THP depletion using polyvinylidenedifluoride (PVDF) membrane filters for urinary EVs for proteomic analysis based on 2 dimensional electrophoresis (2DE).
Canine valve interstitial cells (VICs) from diseased heart valves have a greater prevalence of cells with a myofibroblastic phenotype, growth retardation and lower viability in vitro.
In dogs with myxomatous mitral valvular disease (MMVD), the histological changes and functional consequences are virtually identical to those in humans.
HP mice had reduced expressions of markers of muscle differentiation indicating that palm oil had a deep impact on muscle homeostasis in addition to IR.
Our results support the idea that labelled and unlabelled biomarker based approaches for EV profiling can be used to study stress related responses in cells of patients with different risk profiles.
Highly aggregated THP molecules were trapped in the PVDF membrane upon filtration, leading to better resolution of urinary EV proteins in 2DE analysis.
Additionally, 74 subjects were hospitalized repetitively due to worsening CHF and also 16 subjects were readmitted in the hospital due to other cardiovascular reasons.
Among the 89 proteins identified in the present proteome analysis, 35 proteins have not been published or listed in the major database of urinary EVs, indication that these 35 proteins were newly identified in this study.
Adipocytes have been shown to release extracellular vesicles (EVs) in vitro that can promote angiogenesis in vivo.
MSC CM improve both cell growth and viability for fibroblastic and myofibroblatic VICs and have potential therapeutic benefits for the treatment of MMVD.
Recent publications indicate that exosomes might be involved in hypoxic preconditioning, but mechanistic data are still missing.
Lately, the crosstalk between different cardiac cell types (cardiomyocytes, fibroblasts, endothelial cells) has come into focus of current research.
These conditions are both important risk factors for the development of atherosclerosis, which is a chronic inflammatory disease of the large arteries, involving activation of the vascular endothelium, vascular inflammation and oxidative stress.
Future studies need to validate these reference ranges in other healthy individuals for comparison to patient populations or asymptomatic individuals at risk for cardiovascular, thrombotic or other diseases.
This result suggested that SkM might transfer specific signals through the exosomal route to key metabolic tissues.
The aim of this study was to determine if IR induces microvesicle (MV) release from human macrophages and if these can directly cause endothelial dysfunction in vitro.
Transmission electron microscopy (TEM) and dynamic light scattering were used to study the size distribution and the morphology of isolated EVs from different cell lines and from primary cultures of endothelial cells derived from patients with different risks for cardiovascular disease.
EVs tested using THP1 macrophages in a Dunn chamber, and their ability to stimulate a phenotypic shift towards an M1 phenotype using qPCR.
This result was confirmed in vitro as increasing the amount of palmitate in the medium of C2C12 induced an increase in exosome release.
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Methods: Twenty morbidly obese individuals were studied before and 3 months after weight loss due to gastric bypass.
In addition, it remains to be investigated, whether other preconditioning stimuli, like volatile anaesthetics also trigger the release of vesicles and if these exosomes differ in their protein composition and play a role in preconditioning.
We observed that stress applied to cells is resulting in changes in the number and biomarker profile of secreted EVs.
For this reason, we developed an enzyme linked immune sorbent assay (ELISA) that requires less starting material, allows more simultaneous measurements and could ultimately be applicable for clinical use.
Fluorescence in the lower chamber was measured over 2 hours, and the number of cells migrated quantified by comparison to a standard curve.
EV size and concentration were measured using nanoparticle tracking analysis, and changes in fatty acid composition were analyzed using gas chromatography with flame ionization detection.
This hypoxia is a strong stimulus for angiogenesis to supply the expanding tissue bed with new blood vessels.
Supernatants were collected at different time points after preconditioning, and exosomes were isolated via differential centrifugation with a final ultracentrifugation step.
Introduction: Healthcare systems are facing increasing costs for the diagnosis and treatment of patients with cardiovascular disease.
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