Isolation and detection strategies for decoding the heterogeneity of extracellular vesicles

Extracellular vesicles (EVs) are membrane-bound particles released by cells, playing crucial roles in various pathophysiological processes. A comprehensive understanding of EV diversity offers valuable insights into disease mechanisms and may lead to novel diagnostic and therapeutic approaches. However, the heterogeneity of EVs in terms of size, content, and function presents significant challenges in elucidating their precise roles and applications. The study of EV heterogeneity begins with effective isolation and detection methods, which are essential for characterizing different EV subpopulations. This review provides an in-depth discussion of various isolation techniques, evaluating the strengths and limitations of each. The importance of selecting appropriate methods based on research objectives, as well as the desired purity and yield of EVs, is emphasized. Furthermore, the review examines detection and characterization methods, particularly those based on EV size and surface markers. Emerging technologies and innovative approaches to improve the precision and efficiency of EV isolation and detection are also explored. In conclusion, the review underscores the critical need for continued advancements in isolation and detection strategies to fully understand the complexity of EV heterogeneity. Such progress is essential for realizing the potential of EVs in clinical applications, including diagnostics, drug delivery, and personalized medicine.