Proteomic analysis of small extracellular vesicles purified by three methods

To identify cellular senescence markers using senescent fibroblast-derived extracellular vesicles (EVs), we collected small EVs by three different methods: size exclusion chromatography (SEC), affinity column for phosphatidylserine (PS), and immunoprecipitation (IP) using antibodies against tetraspanin proteins (CD9, CD63 and CD81), from young and senescent fibroblasts to proteomic analysis. In addition, small EVs were collected from culture supernatants of cells derived from patients with Werner's syndrome, a known Progeria, and healthy individuals, and subjected to proteomic analysis.