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Extracellular vesicles (EVs) hold significant promise as biomarkers and therapeutics, yet their isolation remains challenging due to their low abundance and complex sample matrices. Here, we introduce EV-Osmoprocessor (EVOs), a novel device that leverages osmosis-driven filtration for rapid and efficient EV isolation. EVOs employs a high osmolarity polymer solution to concentrate EVs while simultaneously removing smaller contaminants. Compared to traditional methods such as ultracentrifugation and precipitation, EVOs offers speed and convenience, achieving a 50-fold volume reduction in under 2 h. Our results show that EVOs retained EVs and removed >99% albumin from the cell conditioned culture medium (CCM). The isolated EVs exhibited a particle size distribution centered around 140 nm, which was very similar to EVs isolated via precipitation or ultracentrifugation. The standalone EVOs process achieved a particle:protein ratio (EV purity) of ~107 particles/µg protein. Comprehensive characterization, including cryo-electron microscopy, validation of protein markers and known miRNA cargo confirmed the successful isolation of EVs. Functional assays, based on protection of cardiomyocytes from hypoxia/reoxygenation injury, demonstrated the bioactivity of EVOs-isolated EVs. Furthermore, we show that EVOs can be used to concentrate 30 ml of CCM into a 0.5 ml solution, which was then further processed with size-exclusion chromatography (SEC), improving EV purity to ~109 particles/µg protein. This work establishes EVOs as a promising tool for EV research and clinical applications, offering a streamlined approach to EV isolation with enhanced analytical performance.

権利情報:

• Creative Commons BY Attribution 4.0 International
  

キーワード: Exosome, mesenchymal stromal cell, cardiomyocyte, bioprocessing, polymer

刊行年月日: 2025-12-31

出版者: Taylor & Francis

掲載誌:

• Science and Technology of Advanced Materials (ISSN: 14686996) vol. 26 issue. 1 2485668

研究助成金:

原稿種別: 著者最終稿 (Accepted manuscript)

MDR DOI: https://doi.org/10.48505/nims.5437

公開URL: https://doi.org/10.1080/14686996.2025.2485668

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更新時刻: 2025-07-16 16:14:59 +0900

MDRでの公開時刻: 2025-04-20 17:17:32 +0900