Methods for Analysis of Extracellular Vesicle Functions

Extracellular vesicles (EVs) are nanoscale vesicles that are secreted by virtually all cells and are involved in numerous biological processes. Studies of EV function have necessitated the development of various in vitro and in vivo methods. Surface modification with specific molecules facilitates targeting of target recipient cells for drug delivery. EV surface modification with nucleic acids, proteins, and lipids can regulate gene expression in recipient cells. Coculture with recipient cells is a typical method for investigating the functions of EVs. EV surface proteins can interact with the membrane proteins of recipient cells to regulate the activation of specific signaling pathways. Furthermore, the EV cargo delivered into recipient cells may modulate gene expression patterns. Animal models of EV treatment are required for functional validation in vivo. Thus, the functions of cell-derived EVs and engineered EVs need to be validated in vivo using animal models to facilitate their application in the clinical setting.