Updating the MISEV minimal requirements for extracellular vesicle studies: building bridges to reproducibility

Witwer, Kenneth W.; Soekmadji, Carolina; Hill, Andrew F.; Wauben, Marca H. M.; Buzás, Edit I.; Vizio, Dolores Di; Falcón‐Pérez, Juan Manuel; Gardiner, Chris; Hochberg, Fred H.; Kurochkin, Igor V.; Lötvall, Jan; Mathivanan, Suresh; Nieuwland, Rienk; Sahoo, Susmita; Tahara, Hidetoshi; Torrecilhas, Ana Claúdia; Weaver, Alissa M.; Yin, Hang; Zheng, Lei; Gho, Yong Song; Quesenberry, Peter J.; Théry, Clotilde

doi:10.1080/20013078.2017.1396823

Cited by 203 publications

(184 citation statements)

References 11 publications

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“…Given that precipitation is time efficient, inexpensive and demands no specialized equipment, it also seems to conveniently lend itself to integration into clinical usage. However, in a research field as vibrant and international, standardizing reagents and protocols utilized for EV precipitation and characterization are crucial for generating valid and reproducible data across laboratories [22,85,91]. …”

Section: Discussionmentioning

confidence: 99%

Evaluation of serum extracellular vesicle isolation methods for profiling miRNAs by next‐generation sequencing

Buschmann

Kirchner

Hermann

et al. 2018

J of Extracellular Vesicle

198

165

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Extracellular vesicles (EVs) are intercellular communicators with key functions in physiological and pathological processes and have recently garnered interest because of their diagnostic and therapeutic potential. The past decade has brought about the development and commercialization of a wide array of methods to isolate EVs from serum. Which subpopulations of EVs are captured strongly depends on the isolation method, which in turn determines how suitable resulting samples are for various downstream applications. To help clinicians and scientists choose the most appropriate approach for their experiments, isolation methods need to be comparatively characterized. Few attempts have been made to comprehensively analyse vesicular microRNAs (miRNAs) in patient biofluids for biomarker studies. To address this discrepancy, we set out to benchmark the performance of several isolation principles for serum EVs in healthy individuals and critically ill patients. Here, we compared five different methods of EV isolation in combination with two RNA extraction methods regarding their suitability for biomarker discovery-focused miRNA sequencing as well as biological characteristics of captured vesicles. Our findings reveal striking method-specific differences in both the properties of isolated vesicles and the ability of associated miRNAs to serve in biomarker research. While isolation by precipitation and membrane affinity was highly suitable for miRNA-based biomarker discovery, methods based on size-exclusion chromatography failed to separate patients from healthy volunteers. Isolated vesicles differed in size, quantity, purity and composition, indicating that each method captured distinctive populations of EVs as well as additional contaminants. Even though the focus of this work was on transcriptomic profiling of EV-miRNAs, our insights also apply to additional areas of research. We provide guidance for navigating the multitude of EV isolation methods available today and help researchers and clinicians make an informed choice about which strategy to use for experiments involving critically ill patients.

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Section: Discussionmentioning

confidence: 99%

Evaluation of serum extracellular vesicle isolation methods for profiling miRNAs by next‐generation sequencing

Buschmann

Kirchner

Hermann

et al. 2018

J of Extracellular Vesicle

198

165

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“…Efforts to define the minimal standards to support the evaluation and interpretation of experiments have been made in various areas of biomedical research, such as microbiology (47)(48)(49), molecular studies (50), extracellular vesicle studies (51)(52)(53), immunopeptidomics (54), HIV research (55,56), and biobanking (57). Standard definitions have also been shown to significantly improve the reproducibility of clinical flow cytometry experiments.…”

Section: Discussionmentioning

confidence: 99%

MiSet RFC Standards: Defining a Universal Minimum Set of Standards Required for Reproducibility and Rigor in Research Flow Cytometry Experiments

et al. 2019

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Poor adherence to best practices, insufficient training, and pressure to produce data quickly may lead to publications of suboptimal biomedical research flow cytometry data, which contributes to the body of irreproducible research findings. In addition, documentation of compliance with best flow cytometry practices for submission, visualization, and publication of flow cytometry data is currently endorsed by very few scientific journals, which is particularly concerning as numerous peer-reviewed flow cytometry publications emphasize instrumentation, experimental design, and data analysis as important sources of variability. Guidelines and resources for adequate reporting, annotation and deposition of flow cytometry experiments are provided by MIFlowCyt and the FlowRepository database, and comprehensive expert recommendations covering principles and techniques of field-specific flow cytometry applications have been published. To facilitate the integration of quality-defining parameters into manuscript and grant submission and publication requirements across biomedical fields that rely on the use of flow-cytometry-based techniques, a single comprehensive yet easily and universally applicable document is needed. To produce such a list of gold-standard parameters that assess whether a research flow cytometry experiment has been planned, conducted, interpreted, and reported at the highest standard, a new initiative defining the minimum set of standards a robust and rigorous research flow experiment must fulfill (MiSet RFC Standards) was proposed at CYTO 2019. MiSet RFC Standards will integrate and simplify existing resources to provide a universal benchmark a flow cytometry experiment can easily be measured against. The goal of MiSET RFC Standards is its integration into peerreview and publication procedures through partnership with stakeholders, journals and publishers in biomedical and translational research. This article introduces the aims and anticipated timeline and discusses strategies for interdisciplinary consensus and implementation. A single-resource broadly applicable guideline will harmonize standards across different fields of biomedical research and lead to publication of more robust research findings.

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“…In addition, the method of collection, handling and storage conditions of EV can alter the molecular composition of EV [16,17]. …”

Section: Basic and Clinical Aspects Of Extracellular Vesiclesmentioning

confidence: 99%

Essentials of extracellular vesicles: posters on basic and clinical aspects of extracellular vesicles

Nieuwland

Falcón‐Pérez

Soekmadji

et al. 2018

J of Extracellular Vesicle

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The past decade has witnessed an exponential development in the field of extracellular vesicles. Sporadic observations have reached a critical level and the scientific community increasingly recognizes the potential biomedical significance of these subcellular structures present in all body fluids as significant components of the cellular secretome. The Educational Committee of the International Society for Extracellular Vesicles prepared two posters (“Basic aspects of extracellular vesicles” and “Clinical aspects of extracellular vesicles”) to provide essential pieces of information on extracellular vesicles at glance for anyone not familiar with the field.

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Updating the MISEV minimal requirements for extracellular vesicle studies: building bridges to reproducibility

Cited by 203 publications

References 11 publications

Evaluation of serum extracellular vesicle isolation methods for profiling miRNAs by next‐generation sequencing

Evaluation of serum extracellular vesicle isolation methods for profiling miRNAs by next‐generation sequencing

MiSet RFC Standards: Defining a Universal Minimum Set of Standards Required for Reproducibility and Rigor in Research Flow Cytometry Experiments

Essentials of extracellular vesicles: posters on basic and clinical aspects of extracellular vesicles

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