Extracellular vesicles are small membrane-bound vesicles secreted by cells that play an important role in intercellular communication. They are categorized into small vesicles, microvesicles, and apoptotic vesicles, depending on their origin and size. They contain various biomolecules, including lipids, proteins, RNA, and DNA, which enable them to play a significant role in signal transmission between cells. They influence brain and immune system function and are involved in the pathophysiology of disease. They are potential biomarkers for various disease conditions, including cancer, neurodegenerative diseases, and psychsiatric disorders, including depression. To standardize the isolation and characterization of vesicles, we followed minimal information for studies of extracellular vesicles to identify the best isolation method for clinical use. Based on the obtained results, both ultracentrifugation through a sucrose cushion and isolation of vesicles with a reagent kit based on percipitation (Total Exosome Isolation Kit (from plasma) (InvitrogenTM, Thermo Fisher Scientific, ZDA)) proved to be comparable. Both methods successfully facilitated all analyses for the characterization and confirmation of the presence of extracellular vesicles, as well as the determination of miRNA expression. For clinical use, the reagent kit for precipitation is most suitable due to its speed and simplicity.
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