Isolation and Characterization of Extracellular Vesicles from Cell Culture Conditioned Medium for Immunological Studies.

This article describes protocols for isolation of EVs from cell culture supernatants by differential ultracentrifugation and density gradient centrifugation. It also provides tools and protocols that enable characterization and validation of isolated particles, as well as analysis of interactions between EVs of interest and different subpopulations of human immune cells. © 2020 Wiley Periodicals LLC. Basic Protocol 1: Isolation of extracellular vesicles by differential ultracentrifugation Basic Protocol 2: Isolation of extracellular vesicles by density gradient centrifugation Support Protocol 1: Imaging of extracellular vesicles using transmission electron microscopy Support Protocol 2: Detection of extracellular vesicle protein markers by Western blotting Support Protocol 3: Measurement and counting of extracellular vesicles by nanoparticle tracking analysis Basic Protocol 3: Analysis of extracellular vesicle uptake or association by different subpopulations of lymphocytes in vitro. PMID: 32453501 [PubMed - in process]
Source: Current Protocols in Immunology - Category: Allergy & Immunology Tags: Curr Protoc Immunol Source Type: research