Minimal information for studies of extracellular vesicles (MISEV2023): From basic to advanced approaches

Abstract

Extracellular vesicles (EVs), through their complex cargo, can reflect the state of their cell of origin and change the functions and phenotypes of other cells. These features indicate strong biomarker and therapeutic potential and have generated broad interest, as evidenced by the steady year-on-year increase in the numbers of scientific publications about EVs. Important advances have been made in EV metrology and in understanding and applying EV biology. However, hurdles remain to realising the potential of EVs in domains ranging from basic biology to clinical applications due to challenges in EV nomenclature, separation from non-vesicular extracellular particles, characterisation and functional studies. To address the challenges and opportunities in this rapidly evolving field, the International Society for Extracellular Vesicles (ISEV) updates its 'Minimal Information for Studies of Extracellular Vesicles', which was first published in 2014 and then in 2018 as MISEV2014 and MISEV2018, respectively. The goal of the current document, MISEV2023, is to provide researchers with an updated snapshot of available approaches and their advantages and limitations for production, separation and characterisation of EVs from multiple sources, including cell culture, body fluids and solid tissues. In addition to presenting the latest state of the art in basic principles of EV research, this document also covers advanced techniques and approaches that are currently expanding the boundaries of the field. MISEV2023 also includes new sections on EV release and uptake and a brief discussion of in vivo approaches to study EVs. Compiling feedback from ISEV expert task forces and more than 1000 researchers, this document conveys the current state of EV research to facilitate robust scientific discoveries and move the field forward even more rapidly.

Keywords: MISEV; ectosomes; exosomes; extracellular particles; extracellular vesicles; guidelines; microparticles; microvesicles; minimal information requirements; reproducibility; rigor; standardisation.

FIGURE 1

Hierarchy of EP nomenclature. Extracellular particles include vesicular and non‐vesicular particles. This figure presents several distinctions that can be made between classes of EPs, as well as examples of possible nomenclature. EP: extracellular particle; EV: extracellular vesicle; SV: synthetic vesicle; ACDV: artificial cell‐derived vesicle; NVEP: non‐vesicular extracellular particle. See also Section 2 and Table 2.

FIGURE 2

Position of some EV separation and concentration methods on a recovery (yield) versus specificity grid. Dashed blue arrows indicate combinations of methods resulting in increased specificity. Specificity can be of different types: Size exclusion chromatography (SEC) separates EVs by size from many (but not all) NVEPs, but all EV types are recovered together, while differential ultracentrifugation (dUC) separates EV subtypes based on their size/weight, but also co‐isolates NVEPs at high speeds. Note that many ‘exosome purification’ kits use precipitation (P), thus do not isolate pure exosomes or even EVs but a mixture of EPs, while some use affinity precipitation (AP), which may be more specific to EVs but not exosomes. Those who develop new methods should consider positioning their EV outcomes on such a graph.