Real‐Time Dissection of the Transportation and miRNA‐Release Dynamics of Small Extracellular Vesicles

Extracellular vesicles (EVs) are cell‐derived membrane‐enclosed structures that deliver biomolecules for intercellular communication. Developing visualization methods to elucidate the spatiotemporal dynamics of EVs’ behaviors will facilitate their understanding and translation. With a quantum dot (QD) labeling strategy, a single particle tracking (SPT) platform is proposed here for dissecting the dynamic behaviors of EVs. The interplays between tumor cell‐derived small EVs (T‐sEVs) and endothelial cells (ECs) are specifically investigated based on this platform. It is revealed that, following a clathrin‐mediated endocytosis by ECs, T‐sEVs are transported to the perinuclear region in a typical three‐stage pattern. Importantly, T‐sEVs frequently interact with and finally enter lysosomes, followed by quick release of their carried miRNAs. This study, for the first time, reports the entire process and detailed dynamics of T‐sEV transportation and cargo‐release in ECs, leading to better understanding of their proangiogenic functions. Additionally, the QD‐based SPT technique will help uncover more secrets of sEV‐mediated cell–cell communication.