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To Empower, Not Haunt Investigators of Extracellular Vesicles (EVs), Exosomes, Microvesicles, & Extracellular Particles

Some in the extracellular vesicle/exosome research community might treat MISEV guidelines like an encyclical, painstakingly scrutinizing the language and examining one’s prior work for shortcomings. Yet, from the very first pages of MISEV2023, [1] its contributors make one thing clear: their goal is to advance good science collaboratively—not to enforce rigid rules. Section 1 introduces the spirit of MISEV, explaining why it exists, what it is (and is not), and how to use it. RoosterBio is proud to have supported and contributed to this effort. [2, 3, 4]

“Spirit” carries many meanings across philosophy, religion, law, psychology, and art, but its Latin root, spiritus, means “breath” or “wind”—something vital, flowing, and sustaining. MISEV’s spirit is not a ghost that haunts researchers but rather is a life-sustaining oxygen flux, helping the field thrive in rarefied scientific air. How does it inspire progress?

MISEV serves multiple roles: an introduction to extracellular vesicle research, a set of recommendations, a tool for reviewers and editors, a rigor and standardization framework, and an indication of broad consensus and areas for growth. [1, 5, 6, 7, 8] Because extracellular vesicles are dynamic—serving as research subjects, technology platforms, diagnostic materials, and therapeutic candidates—MISEV is updated periodically, shaping discussions at local and global conferences. [9, 10, 11, 12, 13]

The need for MISEV stems from the complexity and diversity of extracellular vesicle research. The field spans bacterial, plant, milk, and human stem cell EVs, as well as subtypes like exosomes and microvesicles. Differences in nomenclature, isolation, characterization, and functional analysis create inconsistencies that can hinder progress. Without a unified approach, findings become difficult to compare, slowing the translation of EV-based discoveries into clinical applications. MISEV helps close this gap by offering consensus-driven recommendations, enabling researchers to design robust experiments, interpret results confidently, and ensure reproducibility across studies.

Yet, the latest MISEV2023 aimed to communicate as much about what it is not as what it is. It’s not a one-size-fits-all rulebook, nor does it prescribe absolute checklists of required techniques, markers, or platforms. Researchers must select fit-for-purpose methods and report them transparently. MISEV is also not a barrier to innovation—new approaches might not fit neatly into existing frameworks, but that should not stifle their validation. Likewise, MISEV recommendations should never be used to block publications or funding. And it is not an exhaustive citation catalog; its references serve to illustrate, not to endorse or define the field’s entirety.

The Spirit of MISEV: Asking the Right Questions

Ultimately, MISEV’s spirit is about scientific best practices—not just for extracellular vesicle research, but for science in general. As its primary authors state, the essence of MISEV can be captured in five fundamental questions:

1. What terms do you use, and what do they mean?
2. From what/where did you obtain your EVs?
3. How did you separate, concentrate, characterize, and store them?
4. How confidently can you attribute a function or biomarker to EVs versus other components?
5. Have you shared data and reported methods in sufficient detail to enable replication and reproducibility?

These questions transcend extracellular vesicle research; they define how science should be conducted. Researchers strive for clarity, but there is always room for improvement. Instead of rigid checklists, MISEV2023 prioritizes critical thinking—shifting from a “check-the-box” mentality to a mindset of rigorous inquiry. In doing so, researchers strengthen both their own work and the broader EV field.

Characterization: The Anchor of MISEV

At its core, MISEV2023 revolves around one crucial theme:

Isolation methods in particular can significantly impact extracellular vesicle composition and subsequent analyses. Two groups studying the same EV type but using different isolation techniques may arrive at different conclusions—yet without transparent reporting, these differences remain obscured. MISEV underscores that without careful documentation, meaningful comparisons break down, reproducibility suffers, and progress stalls. While standardization may never be one-size-fits-all, rigorous characterization is what keeps the field moving forward with clarity and credibility. [14, 15, 16]

Looking Ahead

In the end, the spirit of MISEV is about empowering researchers—not haunting them with dogma—so the extracellular vesicle field can advance with both scientific rigor and creative innovation. This philosophy aligns perfectly with RoosterBio’s mission to accelerate extracellular vesicle research by providing scalable, high-quality solutions that bridge the gap from discovery to clinical application. By embracing MISEV’s principles of transparency, reproducibility, and rigorous characterization, the field moves closer to unlocking the full potential of extracellular vesicles as transformative tools in medicine and biotechnology.

*RoosterBio Note: For thrills and giggles, we had two ChatGPTs discuss this blog with each other in an audio conversation. Listen to it HERE.

 

References

1. Welsh, J. A., et al., Minimal information for studies of extracellular vesicles (MISEV2023): From basic to advanced approaches. J Extracell Vesicles, 2024. 13(2): p. e12404. 10.1002/jev2.12404
2. Stephen, Lenzini. Establishing a Working Range for Effective MSC-EV Dose. RoosterBio Blog 2021; Available from: https://www.roosterbio.com/blog/establishing-a-working-range-for-effective-msc-ev-dose/.
3. Cruz, Crystal. From Benchmark to Airborne: MISEV Guidelines Inspire Fledgling EV/Exosome Analytics Services. RoosterBio Blog 2023; Available from: https://www.roosterbio.com/blog/from-benchmark-to-airborne-misev-guidelines-inspire-fledgling-ev-exosome-analytics-services/.
4. RoosterBio. On the Exosome/Extracellular Vesicle Frontier, Choose Your Own Adventure. RoosterBio Blog 2023; Available from: https://www.roosterbio.com/blog/on-the-exosome-extracellular-vesicle-frontier-choose-your-own-adventure/.
5. Lotvall, J., 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: p. 26913. 10.3402/jev.v3.26913
6. Witwer, K. W., et al., Updating the MISEV minimal requirements for extracellular vesicle studies: building bridges to reproducibility. J Extracell Vesicles, 2017. 6(1): p. 1396823. 10.1080/20013078.2017.1396823
7. Thery, C., et al., Minimal information for studies of extracellular vesicles 2018 (MISEV2018): a position statement of the International Society for Extracellular Vesicles and update of the MISEV2014 guidelines. J Extracell Vesicles, 2018. 7(1): p. 1535750. 10.1080/20013078.2018.1535750
8. Witwer, K. W., et al., Updating MISEV: Evolving the minimal requirements for studies of extracellular vesicles. J Extracell Vesicles, 2021. 10(14): p. e12182. 10.1002/jev2.12182
9. ISEV2020 Abstract Book. Journal of Extracellular Vesicles, 2020. 9(S1): p. 1784511. https://doi.org/10.1080/20013078.2020.1784511
10. ISEV2021 Abstract Book. Journal of Extracellular Vesicles, 2021. 10(S1): p. e12083. https://doi.org/10.1002/jev2.12083
11. ISEV2022 Abstract Book. Journal of Extracellular Vesicles, 2022. 11(S1): p. e12224. https://doi.org/10.1002/jev2.12224
12. ISEV2023 Abstract Book. Journal of Extracellular Vesicles, 2023. 12(S1): p. e12329. https://doi.org/10.1002/jev2.12329
13. ISEV2024 Abstract Book. Journal of Extracellular Vesicles, 2024. 13(S1): p. e12444. https://doi.org/10.1002/jev2.12444
14. Arminan, A., Carson, J., Lenzini, S. miRNA, miRNA in the QIAzol®, Where’s the Fingerprint Among Them All? RoosterBio Blog 2023; Available from: https://www.roosterbio.com/blog/mirna-mirna-in-the-qiazol-wheres-the-fingerprint-among-them-all/.
15. Cramer, Madeline. CD73: A Team Player Caught in the “AKT” of Wound Healing & Cell Survival via MSC Exosomes? RoosterBio Blog 2024; Available from: https://www.roosterbio.com/blog/cd73-a-team-player-caught-in-the-akt-of-wound-healing-cell-survival-via-msc-exosomes/.
16. RoosterBio. Analytics: A Referee for the Tug-of-War Between MSC-EVs’ Nature & Nurture. RoosterBio Blog 2024; Available from: https://www.roosterbio.com/blog/analytics-a-referee-for-the-tug-of-war-between-msc-evs-nature-nurture/.