Real-time imaging of multivesicular body–plasma membrane fusion to quantify exosome release from single cells

Bebelman, Maarten P.; Bun, Philippe; Huveneers, Stephan; Niel, Guillaume van; Pegtel, D. Michiel; Verweij, Frederik J.

doi:10.1038/s41596-019-0245-4

Cited by 104 publications

(120 citation statements)

References 44 publications

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“…Furthermore, Clayton et al published a first roadmap towards collection, handling, and storage of extracellular vesicles from blood, in which the needs included education and quality control parameters [6]. The present survey as well as several recent publications emphasize these needs in EV composition and sample preparation methodology [19][20][21][22].…”

Section: Conclusion and Final Remarksmentioning

confidence: 77%

Methods for Separation and Characterization of Extracellular Vesicles: Results of a Worldwide Survey Performed by the ISEV Rigor and Standardization Subcommittee

Royo

Théry

Falcón‐Pérez

et al. 2020

Cells

254

226

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Research on extracellular vesicles (EVs) is growing exponentially due to an increasing appreciation of EVs as disease biomarkers and therapeutics, an expanding number of EV-containing materials under study, and application of new preparation, detection, and cargo analysis methods. Diversity of both sources and methodologies imposes challenges on the comparison of measurement results between studies and laboratories. While reference guidelines and minimal requirements for EV research have achieved the important objective of assembling community consensus, it is also essential to understand which methodologies and quality controls are currently being applied, and how usage trends are evolving. As an initial response to this need, the International Society for Extracellular Vesicles (ISEV) performed a worldwide survey in 2015 on “Techniques used for the isolation and characterization of extracellular vesicles” and published the results from this survey in 2016. In 2019, a new survey was performed to assess the changing state of the field. The questionnaire received more than 600 full or partial responses, and the present manuscript summarizes the results of this second worldwide survey. The results emphasize that separation methods such as ultracentrifugation and density gradients are still the most commonly used methods, the use of size exclusion chromatography has increased, and techniques based on tangential flow and microfluidics are now being used by more than 10% of respondents. The survey also reveals that most EV researchers still do not perform sample quality controls before or after isolation of EVs. Finally, the majority of EV researchers emphasize that separation and characterization of EVs should receive more attention.

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Section: Conclusion and Final Remarksmentioning

confidence: 77%

Methods for Separation and Characterization of Extracellular Vesicles: Results of a Worldwide Survey Performed by the ISEV Rigor and Standardization Subcommittee

Royo

Théry

Falcón‐Pérez

et al. 2020

Cells

254

226

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“…Most of the time, MVBs are directed to lysosomes containing hydrolase, resulting in degradation of their cargo [23]. Otherwise, MVBs migrate to the cell surface to fuse with PM and release ILVs into the extracellular space, that in turn become an exosome upon cellular exit [24]. Transport of MVBs is directed through accessory proteins: tumor susceptibility gene 101 protein (TSG101), programmed cell death 6-interacting protein Alix, heat shock cognate protein 70 (HSC70), heat shock protein 90β (HSP90β), cluster of differentiation proteins 9 (CD9), CD81, CD63, and involves either the presence of ESCRT protein family, known as exosomal marker family proteins crucial in the ESCRT-dependent formation or alternatively sphingomyelinase enzyme in the ESCRT-independent formation [25].…”

Section: Cellular Origins and Chemical Properties Of Exosomesmentioning

confidence: 99%

Inclusion Biogenesis, Methods of Isolation and Clinical Application of Human Cellular Exosomes

Tschuschke

Kocherova

Bryja

et al. 2020

JCM

130

101

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Exosomes are a heterogenous subpopulation of extracellular vesicles 30–150 nm in range and of endosome-derived origin. We explored the exosome formation through different systems, including the endosomal sorting complex required for transport (ESCRT) and ESCRT-independent system, looking at the mechanisms of release. Different isolation techniques and specificities of exosomes from different tissues and cells are also discussed. Despite more than 30 years of research that followed their definition and indicated their important role in cellular physiology, the exosome biology is still in its infancy with rapidly growing interest. The reasons for the rapid increase in interest with respect to exosome biology is because they provide means of intercellular communication and transmission of macromolecules between cells, with a potential role in the development of diseases. Moreover, they have been investigated as prognostic biomarkers, with a potential for further development as diagnostic tools for neurodegenerative diseases and cancer. The interest grows further with the fact that exosomes were reported as useful vectors for drugs.

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“…Similarly, cells with stemness properties, such as mesenchymal stromal cells (MSCs) [ 86 , 87 , 88 ] and induced pluripotent stem cells (iPSCs) [ 89 , 90 ], have been reported to release exosomes. In addition, exosomes are found in biological fluids including plasma [ 91 , 92 , 93 , 94 ], urine [ 95 , 96 , 97 ] saliva [ 98 ], amniotic fluid [ 99 ] and breast milk [ 100 ]. In the sections below, we provide details of major cells that play a key role in exosome secretion in different metabolic and pathological pathways.…”

Section: Different Types and Functions Of Cells That Release Exosomentioning

confidence: 99%

Exosome: A New Player in Translational Nanomedicine

Aheget

Tristán‐Manzano

Mazini

et al. 2020

JCM

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Summary: Exosomes are extracellular vesicles released by the vast majority of cell types both in vivo and ex vivo, upon the fusion of multivesicular bodies (MVBs) with the cellular plasma membrane. Two main functions have been attributed to exosomes: their capacity to transport proteins, lipids and nucleic acids between cells and organs, as well as their potential to act as natural intercellular communicators in normal biological processes and in pathologies. From a clinical perspective, the majority of applications use exosomes as biomarkers of disease. A new approach uses exosomes as biologically active carriers to provide a platform for the enhanced delivery of cargo in vivo. One of the major limitations in developing exosome-based therapies is the difficulty of producing sufficient amounts of safe and efficient exosomes. The identification of potential proteins involved in exosome biogenesis is expected to directly cause a deliberate increase in exosome production. In this review, we summarize the current state of knowledge regarding exosomes, with particular emphasis on their structural features, biosynthesis pathways, production techniques and potential clinical applications.

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Real-time imaging of multivesicular body–plasma membrane fusion to quantify exosome release from single cells

Cited by 104 publications

References 44 publications

Methods for Separation and Characterization of Extracellular Vesicles: Results of a Worldwide Survey Performed by the ISEV Rigor and Standardization Subcommittee

Methods for Separation and Characterization of Extracellular Vesicles: Results of a Worldwide Survey Performed by the ISEV Rigor and Standardization Subcommittee

Inclusion Biogenesis, Methods of Isolation and Clinical Application of Human Cellular Exosomes

Exosome: A New Player in Translational Nanomedicine

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