Cargo Stability of Extracellular Vesicles Derived From Non-Diseased Sources: Effects of Storage and Handling Conditions

Extracellular vesicles (EVs) are nanoscale, membrane-enclosed particles that transfer proteins, nucleic acids, lipids, and other bioactive molecules between cells. Their cargo-bearing capacity supports applications in drug delivery, regenerative medicine, and biomarker development, but ex vivo handling can alter vesicle recovery and cargo readouts. Storage temperature, pH, freeze-thaw exposure, storage duration, and buffer composition differentially affect EV-associated proteins and nucleic acids. This review evaluates preservation studies using EVs from non-diseased cells, tissues, and biological fluids, with emphasis on the distinction between EV recovery, cargo abundance, molecular integrity, and functional preservation. Across the available literature, optimal conditions depend on EV source, biological matrix, isolation method, storage duration, cargo class, and analytical endpoint. Standardized protocols that separately measure vesicle recovery, cargo abundance, molecular integrity, and biological activity are required to define source- and application-specific preservation strategies and resolve the mechanisms of storage-associated cargo alteration.