Microvesicles and exosomes: new players in metabolic and cardiovascular disease

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   Figure 1

   (A) Timeline (1956–2014) of the publications referring to extracellular vesicles (black line), microvesicles (blue line), and exosomes (red line). (B) Schematic representation of the mechanisms of formation of microvesicles, exosomes, and apoptotic bodies. Microvesicles (0.2–2.0 μm) originate via budding and shedding from the plasma membrane of cells and therefore may contain specific surface markers from the cell of origin. Exosomes (50–100 nm) on the other hand originate intracellularly through a sorting pathway involving intermediate organelles such as the early endosome and a late multivesicular body, which fuses with the plasma membrane to release exosomes via exocytosis. Apoptotic bodies (1–2 μm) originate via blebbing of the plasma membrane.
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   Figure 2

   The endothelial cells marker CD144 is absent from exosomes isolated from HUVEC endothelial cells (A), despite being detectable on the parent cells (B). HUVEC cells or HUVEC exosomes bound to 4 μm beads were labelled with anti-CD144 and fluorescent secondary antibody, before fluorescent detection using a BD AccuriC6 flow cytometer.
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   Figure 3

   Flow cytometry (FCM) allows direct analysis of microvesicles (MVs) and indirect (conjugated) analysis of exosomes. Nanoparticle tracking analysis (NTA) is the preferred technique for EV quantitation. Electron microscopy (EM) is the golden standard for EV visualization. (A) Direct flow cytometric analysis of MVs in plasma of rats fed chow or high fat diets (HFD; Heinrich et al. 2015) after staining for phosphatidyl serine exposure (Annexin V PE-Cy7.7) and CD106 (PE) to determine MV release from activated endothelial cells. Enumeration beads (red) and 1.1 μm sizing beads (green) were added as internal controls. (B) NTA of MVs from rats fed chow or HFD. (C) Indirect flow cytometric analysis of exosomes bound to aldehyde sulphate beads (4 μm) after staining for the tetraspannin CD63 and surface HSP70 (Vicencio et al. 2015). (D) NTA of human plasma exosomes isolated via ultraceintrifugation (black line) or using the Exo-spin (Cell Guidance Systems, Cambridge, United Kingdom) commercial kit (red line). (E) Electron micrograph of MVs and exosomes.

• © 2016 Society for Endocrinology