title: Higher functionality of extracellular vesicles isolated using size-exclusion chromatography compared to ultracentrifugation.
abstract: Extracellular vesicles (EVs) are nano-sized, lipid bilayer-enclosed particles involved in intercellular communication. EVs are increasingly being considered as drug delivery vehicles or as cell-free approach to regenerative medicine. However, one of the major challenges for their clinical application is finding a scalable EV isolation method that yields functional EVs. Although the golden standard for EV isolation is ultracentrifugation (UC), a recent study suggested that isolation using size-exclusion chromatography (SEC) yielded EVs with more intact biophysical properties. Whether this also leads to differences in functionality remained to be investigated. Therefore, we investigated possible differences in functionality of cardiomyocyte progenitor cell-derived EVs isolated using UC and SEC. Western blot analysis showed higher pERK/ERK ratios in endothelial cells after stimulation with SEC-EVs compared to UC-EVs, indicating that SEC-EVs bear higher functionality. Therefore, we propose to use SEC-EVs for further investigation of EVs' therapeutic potential. Further optimization of isolation protocols may accelerate clinical adoption of therapeutic EVs.
authors.affiliation: Department of Cardiology, Laboratory of Experimental Cardiology, University Medical Center Utrecht, Utrecht, the Netherlands.
authors.forename: Emma A
authors.initials: EA
authors.lastname: Mol
authors.affiliation: Department of Molecular Cell Biology, Leiden University Medical Center, Leiden, the Netherlands.
authors.forename: Marie-José
authors.initials: MJ
authors.lastname: Goumans
authors.affiliation: Department of Cardiology, Laboratory of Experimental Cardiology, University Medical Center Utrecht, Utrecht, the Netherlands; UMC Utrecht Regenerative Medicine Center, University Medical Center, Utrecht, the Netherlands; Netherlands Heart Institute (ICIN), Utrecht, the Netherlands.
authors.forename: Pieter A
authors.initials: PA
authors.lastname: Doevendans
authors.affiliation: Department of Cardiology, Laboratory of Experimental Cardiology, University Medical Center Utrecht, Utrecht, the Netherlands; UMC Utrecht Regenerative Medicine Center, University Medical Center, Utrecht, the Netherlands; Netherlands Heart Institute (ICIN), Utrecht, the Netherlands.
authors.forename: Joost P G
authors.initials: JPG
authors.lastname: Sluijter
authors.affiliation: Laboratory of Clinical Chemistry and Haematology, University Medical Center Utrecht, Utrecht, the Netherlands. Electronic address: pvader@umcutrecht.nl.
authors.forename: Pieter
authors.initials: P
authors.lastname: Vader
mesh_terms: D002478:Cells, Cultured; D002850:Chromatography, Gel / Q000379:methods*; D000067128:Extracellular Vesicles / Q000378:metabolism*; D006801:Humans; D032383:Myocytes, Cardiac / Q000166:cytology / Q000378:metabolism*; D013234:Stem Cells / Q000166:cytology / Q000378:metabolism*; D014461:Ultracentrifugation / Q000379:methods*
keywords: Exosomes; Extracellular vesicles; Functionality; Size-exclusion chromatography; Ultracentrifugation