The MISEV is the main international set of guidelines for studies of extracellular vesicles , with recommendations on nomenclature, collection, separation, characterization, experimental controls and reporting of results.
The acronym stands for Minimal Information for Studies of Extracellular Vesicles . The guidelines were developed by the International Society for Extracellular Vesicles (ISEV) to improve the rigor, transparency and reproducibility of research involving extracellular vesicles, also known as EVs.
The current version is MISEV2023 , published in February 2024 in the Journal of Extracellular Vesicles . The document gathered contributions from more than 1,000 researchers across 52 countries and can be accessed through DOI 10.1002/jev2.12404 .
MISEV does not establish a single protocol. It provides guidance on how to select, control, document and communicate the methods used so that conclusions are supported by evidence.
What is MISEV used for?
EV studies use different biological sources and extracellular vesicle separation and analysis techniques . In addition, soluble proteins, lipoproteins, aggregates and other particles may accompany the vesicles during processing.
Without an adequate description of the methods, it becomes difficult to compare results or determine which components were present.
MISEV helps define terminology, identify pre-analytical variables, select methods suited to the study objective, combine characterization techniques and plan functional controls.
How have the MISEV guidelines evolved?
| Version | Main contribution |
|---|---|
| MISEV2014 | Established the first experimental requirements for defining EVs and investigating their functions. |
| MISEV2018 | Expanded the recommendations and reinforced the use of the generic term “extracellular vesicle.” |
| MISEV2023 | Updated production, separation and characterization practices and included guidance on uptake and in vivo studies. |
Nomenclature recommended by MISEV
One of the main recommendations is to use “extracellular vesicle” or “EV” as a generic term when the particle's origin has not been demonstrated. Terms such as “ exosome ” and “ microvesicle ” indicate specific biogenesis pathways and should not be defined solely by size, separation method or the presence of markers such as CD9 , CD63 and CD81 .
When necessary, the term EV may be accompanied by operational qualifiers, such as:
- small EVs,
- large EVs,
- CD63+ EVs,
- EVs derived from a particular cell type, or
- EVs produced under hypoxia.
A small particle is not automatically an exosome. The more specific the nomenclature, the more specific the evidence concerning its origin must be.
EV separation and characterization
MISEV does not establish a universally superior method for separating extracellular vesicles. Ultracentrifugation, density gradients, size-exclusion chromatography, ultrafiltration, precipitation and immunocapture offer different trade-offs among recovery, specificity, processing time and compatibility with downstream analyses.
The method should be selected according to the sample and the study objective. Rotors, processing times, temperatures, columns, membranes, volumes and collected fractions must be described accurately.
Characterization should combine complementary techniques. Nanoparticle tracking analysis (NTA), for example, estimates particle size and concentration within its detection range, but it cannot by itself demonstrate that all detected particles are EVs. Microscopy provides morphological information, while Western blotting, immunochemical assays and omics techniques assess components associated with the preparation.
Expected contaminants must also be investigated, including soluble proteins, lipoproteins, cellular debris and non-vesicular extracellular particles.
Controls for functional studies
Observing an effect after applying an EV-enriched preparation does not demonstrate that the vesicles were responsible. Proteins, cytokines, reagents or non-vesicular particles may also cause the response.
MISEV recommends comparing the EV preparation with the original sample, EV-depleted fractions, non-vesicular fractions and reagent controls. Dose-response and time-response curves strengthen the interpretation. Dose may be reported by particle number, total protein, original sample volume or number of producer cells.
What should be reported in a MISEV-based study?
- sample origin and collection conditions;
- processing and storage variables;
- separation and concentration methods;
- equipment parameters and analysis criteria;
- complementary characterization techniques;
- contaminant assessment;
- functional study controls;
- limitations of the protocol and conclusions.
Experimental details can also be registered with EV-TRACK , a platform created to promote methodological transparency in EV research.
Is MISEV a mandatory checklist?
No. MISEV is a good-practice guide rather than a rigid standard. Not all studies need to use the same techniques, but methodological choices should be justified and limitations clearly reported.
Applying MISEV means aligning the strength of the conclusion with the strength of the evidence. The goal is to make the study clearer, more comparable and more reproducible without preventing adaptations or the development of new approaches.
Conclusion
MISEV2023 is the central reference for researchers working with extracellular vesicles. Its recommendations support appropriate nomenclature, documentation of every stage, the combination of characterization techniques and greater confidence when determining whether a function or biomarker is genuinely associated with EVs.
Rather than simply stating that a study “follows MISEV,” researchers should show which recommendations were applied and how the controls support the results. This transparency strengthens the quality of extracellular vesicle studies and applications.