Key techniques for studying extracellular vesicles

Extracellular vesicles (EVs) are heterogeneous nanoparticles (30–500 nm) found in all biological fluids. The composition of EVs largely reflects the composition of the cells that secreted them. At present, EVs are widely used to search for diagnostic and therapeutic markers, and therefore studies of EVs composition and properties are becoming more and more in demand today. Despite the obvious potential of EVs in biomedical applications, the lack of unified methods for their isolation, analysis and quantitative measurement significantly limits scientific progress in this field. In scientific literature there are descriptions of dozens of methods for isolation and study of EVs, and often researchers do not present comparative advantages and disadvantages of the methods used. In this regard, a review of methods for the study of EVs becomes relevant. This review presents a comprehensive analysis of modern methods for the study of EVs. Each method has its own advantages and limitations affecting the yield, purity and characteristics of isolated vesicles, as well as the accuracy of the results obtained. The choice of the method for the isolation and analysis of EVs should be made considering the purpose and specificity of the study, since there is no universal approach at present. A thorough understanding of methodological nuances is critical for optimizing reproducibility and reliability in EV research.