Comparative evaluation of ultracentrifugation and PEG-based precipitation of mesenchymal stem cell‑derived extracellular vesicles: equivalent functionality with enhanced practicality.

AIM: The isolation efficiency of extracellular vesicles (EVs) is essential to ensure their purity and biological properties. Ultracentrifugation is a traditional method which utilizes high centrifugal forces to separate EVs based on their density. The polyethylene glycol (PEG)-based precipitation method is a scalable and cost-efficient alternative to ultracentrifugation. In this study, two popular techniques used for the isolation of EVs from umbilical cord tissue-derived mesenchymal stem cells (UCT-MSCs): ultracentrifugation and polyethylene glycol PEG-based precipitation methods are examined.

MATERIALS AND METHODS: The UCT-MSCs were cultured in a xeno-free environment using human platelet lysate (hPL) to avoid the risk of xenogenic contamination. The MSCs and EVs were characterized to confirm their physical and functional properties. Cytotoxicity was analyzed in both normal and cancer cells, and the cell migration assay was conducted on cancer cells treated with the EVs to examine the biocompatibility and migratory effects.

RESULTS AND CONCLUSIONS: The similarity in physical, molecular, and functional characterization of UC-EVs and PEG-EVs suggests that PEG-based precipitation maintained the integrity and biological properties of EVs. Overall, the PEG-based precipitation method for EVs isolation is safe and efficient, making them suitable for regeneration and cancer research.