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Methodological Guidelines to Study Extracellular Vesicles.
Frank A W Coumans1, Alain R Brisson1, Edit I Buzas1
1From the Biomedical Engineering and Physics (F.A.W.C., T.G.v.L., E.v.d.P.), Vesicle Observation Centre (F.A.W.C., A.G., T.G.v.L., E.v.d.P., G.S., R.N.), and Laboratory of Experimental Clinical Chemistry (A.G., G.S., R.N.), Academic Medical Center, University of Amsterdam, The Netherlands; Extracellular Vesicles and Membrane Repair, UMR-5248-CBMN CNRS, University of Bordeaux, IPB, Pessac, France (A.R.B.); Department of Genetics, Cell- and Immunobology, Semmelweis University, Budapest, Hungary (E.I.B.); VRCM, UMRS-1076, INSERM, Aix-Marseille University, UFR de Pharmacie, Marseille, France (F.D.-G., R.L.); Haematology and vascular biology department, CHU La Conception, APHM, Marseille, France (F.D.-G., R.L.); Exosomes Research Group, Department of Pathology, VU University Medical Center, De Boelelaan 1117, Amsterdam, The Netherlands (E.E.E.D., D.M.P.); Department of Laboratory Medicine, Karolinska Institutet, Stockholm, Sweden (S.E.-A., Y.L.); Department of Physiology, Anatomy and Genetics, University of Oxford, United Kingdom (S.E.-A., I.M.); Bristol Heart Institute, University of Bristol, United Kingdom (C.E.); National Heart & Lung Institute, Imperial College London, United Kingdom (C.E.); 1st Chair and Department of Cardiology, Medical University of Warsaw, Poland (A.G.); Laboratory of Experimental Cancer Research, Department of Radiation Oncology and Experimental Cancer Research, Ghent University, Belgium (A.H., O.d.W.); Cancer Research Institute Ghent, Belgium (A.H., O.d.W.); Department of Biochemistry and Genetics, La Trobe Institute for Molecular Science, La Trobe University, Bundoora, Victoria, Australia (A.F.H.); Department of Medicine, University of North Carolina at Chapel Hill (N.M.); Institute of Technology, University of Tartu, Estonia (I.M.); Scintillon Institute, San Diego, CA (J.P.N.); Cardiovascular Research Center, Icahn School of Medicine at Mount Sinai, New York, NY (S.S.); and EV Core Facility, University of Helsinki and EV-Group, Division of Biochemistry and Biotechnology, Department of Biosciences, University of Helsinki, Finland (P.R.M.S.).
Extracellular vesicles (EVs) are crucial in health and disease. This review updates methods for studying EVs, focusing on blood, and highlights common pitfalls for accurate research.
Area of Science:
- Biomedical research
- Molecular biology
- Cardiovascular research
Background:
- Extracellular vesicles (EVs) are increasingly recognized for their roles in physiological and pathological processes.
- The growing interest in EVs stems from their potential for diagnostic and translational applications.
- EVs are present in all bodily fluids, with human blood being a key focus for cardiovascular research.
Purpose of the Study:
- To provide an expert-based update on recent advances in methods for studying extracellular vesicles (EVs).
- To summarize current recommendations for EV analysis, from sample collection to characterization.
- To highlight common misconceptions and methodological challenges in EV research.
Main Methods:
- Review of recent literature and expert consensus on EV research methodologies.
- Focus on techniques for isolation, detection, and characterization of EVs.
- Emphasis on protocols relevant to EVs derived from human blood.
Main Results:
- Advances in EV study methods offer new diagnostic and translational possibilities.
- Standardized considerations and recommendations are crucial for reliable EV research.
- Identification of common misconceptions and pitfalls to improve experimental rigor.
Conclusions:
- Extracellular vesicles (EVs) are vital biomarkers with significant potential in diagnostics and therapeutics.
- Methodological standardization and awareness of pitfalls are essential for advancing EV research, particularly in cardiovascular studies.
- This review provides critical insights for researchers studying EVs in human blood.

