The last decade has seen a sharp increase in the number of scientific publications describing physiological and pathological functions of extracellular vesicles (EVs), a collective term covering various subtypes of cell-released, membranous structures, called exosomes, microvesicles, microparticles, ectosomes, oncosomes, apoptotic bodies, and many other names. However, specific issues arise when working with these entities, whose size and amount often make them difficult to obtain as relatively pure preparations, and to characterize properly. The International Society for Extracellular Vesicles (ISEV) proposed Minimal Information for Studies of Extracellular Vesicles ("MISEV") guidelines for the field in 2014. We now update these "MISEV2014" guidelines based on evolution of the collective knowledge in the last four years. An important point to consider is that ascribing a specific function to EVs in general, or to subtypes of EVs, requires reporting of specific information beyond mere description of function in a crude, potentially contaminated, and heterogeneous preparation. For example, claims that exosomes are endowed with exquisite and specific activities remain difficult to support experimentally, given our still limited knowledge of their specific molecular machineries of biogenesis and release, as compared with other biophysically similar EVs. The MISEV2018 guidelines include tables and outlines of suggested protocols and steps to follow to document specific EV-associated functional activities. Finally, a checklist is provided with summaries of key points.
Keywords: ectosomes; exosomes; extracellular vesicles; guidelines; microparticles; microvesicles; minimal information requirements; reproducibility; rigor; standardization.
Minimal experimental requirements for definition of extracellular vesicles and their functions: a position statement from the International Society for Extracellular Vesicles.Lötvall J, Hill AF, Hochberg F, Buzás EI, Di Vizio D, Gardiner C, Gho YS, Kurochkin IV, Mathivanan S, Quesenberry P, Sahoo S, Tahara H, Wauben MH, Witwer KW, Théry C. Lötvall J, et al. J Extracell Vesicles. 2014 Dec 22;3:26913. doi: 10.3402/jev.v3.26913. eCollection 2014. J Extracell Vesicles. 2014. PMID: 25536934 Free PMC article.
Extracellular Vesicles: Recent Developments in Technology and Perspectives for Cancer Liquid Biopsy.Nazarenko I. Nazarenko I. Recent Results Cancer Res. 2020;215:319-344. doi: 10.1007/978-3-030-26439-0_17. Recent Results Cancer Res. 2020. PMID: 31605237 Review.
Towards mechanisms and standardization in extracellular vesicle and extracellular RNA studies: results of a worldwide survey.Soekmadji C, Hill AF, Wauben MH, Buzás EI, Di Vizio D, Gardiner C, Lötvall J, Sahoo S, Witwer KW. Soekmadji C, et al. J Extracell Vesicles. 2018 Oct 24;7(1):1535745. doi: 10.1080/20013078.2018.1535745. eCollection 2018. J Extracell Vesicles. 2018. PMID: 30370018 Free PMC article.
Extracellular vesicles in cancer: exosomes, microvesicles and the emerging role of large oncosomes.Minciacchi VR, Freeman MR, Di Vizio D. Minciacchi VR, et al. Semin Cell Dev Biol. 2015 Apr;40:41-51. doi: 10.1016/j.semcdb.2015.02.010. Epub 2015 Feb 23. Semin Cell Dev Biol. 2015. PMID: 25721812 Free PMC article. Review.
ISEV position paper: extracellular vesicle RNA analysis and bioinformatics.Hill AF, Pegtel DM, Lambertz U, Leonardi T, O'Driscoll L, Pluchino S, Ter-Ovanesyan D, Nolte-'t Hoen EN. Hill AF, et al. J Extracell Vesicles. 2013 Dec 23;2. doi: 10.3402/jev.v2i0.22859. eCollection 2013. J Extracell Vesicles. 2013. PMID: 24376909 Free PMC article.
Cited by 465 articles
ESC-sEVs Rejuvenate Senescent Hippocampal NSCs by Activating Lysosomes to Improve Cognitive Dysfunction in Vascular Dementia.Hu G, Xia Y, Zhang J, Chen Y, Yuan J, Niu X, Zhao B, Li Q, Wang Y, Deng Z. Hu G, et al. Adv Sci (Weinh). 2020 Mar 20;7(10):1903330. doi: 10.1002/advs.201903330. eCollection 2020 May. Adv Sci (Weinh). 2020. PMID: 32440476 Free PMC article.
Biological effects of selective COX-2 inhibitor NS398 on human glioblastoma cell lines.Palumbo P, Lombardi F, Augello FR, Giusti I, Dolo V, Leocata P, Cifone MG, Cinque B. Palumbo P, et al. Cancer Cell Int. 2020 May 13;20:167. doi: 10.1186/s12935-020-01250-7. eCollection 2020. Cancer Cell Int. 2020. PMID: 32435158 Free PMC article.
Synergies in exosomes and autophagy pathways for cellular homeostasis and metastasis of tumor cells.Salimi L, Akbari A, Jabbari N, Mojarad B, Vahhabi A, Szafert S, Kalashani SA, Soraya H, Nawaz M, Rezaie J. Salimi L, et al. Cell Biosci. 2020 May 13;10:64. doi: 10.1186/s13578-020-00426-y. eCollection 2020. Cell Biosci. 2020. PMID: 32426106 Free PMC article. Review.
Bone Marrow Mesenchymal Stem Cell-Derived Exosomes Improves Spinal Cord Function After Injury in Rats by Activating Autophagy.Gu J, Jin ZS, Wang CM, Yan XF, Mao YQ, Chen S. Gu J, et al. Drug Des Devel Ther. 2020 Apr 29;14:1621-1631. doi: 10.2147/DDDT.S237502. eCollection 2020. Drug Des Devel Ther. 2020. PMID: 32425507 Free PMC article.
Deimination Protein Profiles in Alligator mississippiensis Reveal Plasma and Extracellular Vesicle-Specific Signatures Relating to Immunity, Metabolic Function, and Gene Regulation.Criscitiello MF, Kraev I, Petersen LH, Lange S. Criscitiello MF, et al. Front Immunol. 2020 Apr 28;11:651. doi: 10.3389/fimmu.2020.00651. eCollection 2020. Front Immunol. 2020. PMID: 32411128 Free PMC article.
- References, especially those provided to illustrate methods and approaches, are representative only, and are not meant to be a comprehensive review of the literature. Most references were derived from suggestions provided in the MISEV2018 Survey results. Each reference was checked by multiple authors. Citation implies deemed relevance of scientific content and not an endorsement by the authors or ISEV of any particular journal or editorial practice.
- Lotvall J, Hill AF, Hochberg F, et al. Minimal experimental requirements for definition of extracellular vesicles and their functions: a position statement from the international society for extracellular vesicles. J Extracell Vesicles. 2014;3:26913 Available from: http://www.ncbi.nlm.nih.gov/pubmed/25536934 - PMC - PubMed
- Witwer KW, Soekmadji C, Hill AF, et al. Updating the MISEV minimal requirements for extracellular vesicle studies: building bridges to reproducibility. J Extracell Vesicles. 2017;6(1):1396823 Available from: https://www.tandfonline.com/doi/full/10.1080/20013078.2017.1396823 - DOI - PMC - PubMed
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