title: Isolation and Characterization of Microvesicles from Peripheral Blood.
abstract: The release of extracellular vesicles (EVs) including small endosomal-derived exosomes (Exos, diameter < 100 nm) and large plasma membrane-derived microvesicles (MVs, diameter > 100 nm) is a fundamental cellular process that occurs in all living cells. These vesicles transport proteins, lipids and nucleic acids specific for their cell of origin and in vitro studies have highlighted their importance as mediators of intercellular communication. EVs have been successfully isolated from various body fluids and especially EVs in blood have been identified as promising biomarkers for cancer or infectious diseases. In order to allow the study of MV subpopulations in blood, we present a protocol for the standardized isolation and characterization of MVs from peripheral blood samples. MVs are pelleted from EDTA-anticoagulated plasma samples by differential centrifugation and typically possess a diameter of 100 - 600 nm. Due to their larger size, they can easily be studied by flow cytometry, a technique that is routinely used in clinical diagnostics and available in most laboratories. Several examples for quality control assays of the isolated MVs will be given and markers that can be used for the discrimination of different MV subpopulations in blood will be presented.
authors.affiliation: Department of Hematology/Medical Oncology, University Medical Center Göttingen; kerstin.menck@inserm.fr.
authors.forename: Kerstin
authors.initials: K
authors.lastname: Menck
authors.affiliation: Department of Hematology/Medical Oncology, University Medical Center Göttingen.
authors.forename: Annalen
authors.initials: A
authors.lastname: Bleckmann
authors.affiliation: Department of Hematology/Medical Oncology, University Medical Center Göttingen.
authors.forename: Matthias
authors.initials: M
authors.lastname: Schulz
authors.affiliation: Department of Hematology/Medical Oncology, University Medical Center Göttingen.
authors.forename: Lena
authors.initials: L
authors.lastname: Ries
authors.affiliation: Department of Hematology/Medical Oncology, University Medical Center Göttingen.
authors.forename: Claudia
authors.initials: C
authors.lastname: Binder
mesh_terms: D000072283:A549 Cells; D015415:Biomarkers / Q000378:metabolism; D001773:Blood Cells / Q000166:cytology; D002462:Cell Membrane / Q000378:metabolism; D055252:Cell-Derived Microparticles / Q000737:chemistry / Q000378:metabolism*; D019040:E-Selectin / Q000378:metabolism; D055354:Exosomes / Q000378:metabolism; D000067128:Extracellular Vesicles / Q000737:chemistry / Q000378:metabolism*; D005434:Flow Cytometry; D006801:Humans; D017493:Leukocyte Common Antigens / Q000378:metabolism; D046529:Microscopy, Electron, Transmission; D019007:P-Selectin / Q000378:metabolism; D010316:Particle Size; D060245:Tetraspanin 29 / Q000378:metabolism
chemical_list: D015415:Biomarkers; D019040:E-Selectin; D019007:P-Selectin; D060245:Tetraspanin 29; D017493:Leukocyte Common Antigens