Analysis of Extracellular Vesicles Using Fluorescence Nanoparticle Tracking Analysis

Carnell-Morris, Pauline; Tannetta, Dionne; Siupa, Agnieszka; Hole, Patrick; Dragovic, Rebecca

doi:10.1007/978-1-4939-7253-1_13

Cited by 62 publications

(46 citation statements)

References 7 publications

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“…Particle counting by light scatter, RPS, and similar techniques typically results in overestimation of EV counts since the techniques are not specific to EVs and also register co-isolated particles including lipoproteins and protein aggregates. Possibly, ongoing development of fluorescence capabilities of NTA devices may ultimately allow EV-specific measurement [181], although assay sensitivity and the tendency of labeling antibodies and lipid dyes to form particles pose substantial hurdles to such applications [127,182]. Additionally, particle counting technologies may be biased towards certain particle size ranges (especially 50-150 nm [183,184]) because of pore sizes (RPS), size of calibrator used, sensitivity (for example, smaller particles scatter less light), and ability to cope with multidispersity (DLS versus NTA) [185].…”

Section: ) Intermediate Recovery Intermediate Specificity: Methods mentioning

confidence: 99%

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

Théry

Witwer

Aikawa

et al. 2018

J of Extracellular Vesicle

7,768

212

8,426

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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.

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Section: ) Intermediate Recovery Intermediate Specificity: Methods mentioning

confidence: 99%

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

Théry

Witwer

Aikawa

et al. 2018

J of Extracellular Vesicle

7,768

212

8,426

View full text Add to dashboard Cite

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“…Twelve randomly selected samples from the three different batches were sent to an independent laboratory, Extracellular Vesicle Core at Children's Hospital Los Angeles (California, USA), and were analyzed by nanoparticle tracking analysis for the presence of particles in the extracellular vesicle size range using Malvern Panalytical Nanosight NS300. These samples were also analyzed after staining with a general fluorescent membrane marker, CellMask Orange™ (Thermo Fisher Scientific, Waltham, MA, USA), as previously described [19].…”

Section: Methodsmentioning

confidence: 99%

Umbilical cord-derived Wharton’s jelly for regenerative medicine applications

Gupta

El-Amin²,

Levy

et al. 2020

J Orthop Surg Res

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Background: The last decade has seen an explosion in the interest in using biologics for regenerative medicine applications, including umbilical cord-derived Wharton's Jelly. There is insufficient literature assessing the amount of growth factors, cytokines, hyaluronic acid, and extracellular vesicles including exosomes in these products. The present study reports the development of a novel Wharton's jelly formulation and evaluates the presence of growth factors, cytokines, hyaluronic acid, and extracellular vesicles including exosomes. Methods: Human umbilical cords were obtained from consenting caesarian section donors. The Wharton's jelly was then isolated from the procured umbilical cord and formulated into an injectable form. Randomly selected samples from different batches were analyzed for sterility testing and to quantify the presence of growth factors, cytokines, hyaluronic acid, and extracellular vesicles. Results: All samples passed the sterility test. Growth factors including IGFBP 1, 2, 3, 4, and 6, TGF-α, and PDGF-AA were detected. Several immunomodulatory cytokines, such as RANTES, IL-6R, and IL-16, were also detected. Proinflammatory cytokines MCSFR, MIP-1a; anti-inflammatory cytokines TNF-RI, TNF-RII, and IL-1RA; and homeostatic cytokines TIMP-1 and TIMP-2 were observed. Cytokines associated with wound healing, ICAM-1, G-CSF, GDF-15, and regenerative properties, GH, were also expressed. High concentrations of hyaluronic acid were observed. Particles in the extracellular vesicle size range were also detected and were enclosed by the membrane, indicative of true extracellular vesicles. Conclusion: There are numerous growth factors, cytokines, hyaluronic acid, and extracellular vesicles present in the Wharton's jelly formulation analyzed. The amount of these factors in Wharton's jelly is higher compared with other biologics and may play a role in reducing inflammation and pain and augment healing of musculoskeletal injuries.

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“…In a recent report, Ko et al developed a magnetic nanopore sorting platform that has been used to isolate specific cancer-derived EVs. They EVs expressing a given marker [65]. Despite better size resolution of NTA, a great advantage of FCM is its higher multiplexing capacity.…”

Section: Technologies For Evs Isolation and Analysis And Recent Advancesmentioning

confidence: 99%

Liquid Biopsies in Cancer Diagnosis, Monitoring, and Prognosis

Rubis

Krishnan

Bebawy

2019

Trends in Pharmacological Sciences

420

343

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Liquid biopsy, consisting in the non-invasive analysis of circulating tumor-derived material (the Tumor Circulome), represents an innovative tool in precision oncology to overcome current limitations associated with tissue biopsies. Within the tumor circulome, ctDNA and CTCs are the only components whose clinical application is FDA-cleared. Extracellular vesicles, ctRNA and tumor-educated platelets are relatively novel tumor circulome constituents with promising potential at each stage of cancer management. Here, we discuss the clinical applications of each element of the tumor circulome and the prevailing factors that currently limit implementation in clinical practice. We also detail the most recent technological developments in the field, which demonstrate potential in improving the clinical value of liquid biopsies. Commented [GDR3R2]: Thanks for doing that Commented [MK(2]: Please include all weblinks in a separate section before "References", entitled "Resources" Commented [MK(1]: Please include all weblinks in a separate section before "References", entitled "Resources" Commented [MK(4]: Since this is how it is depicted in the Figure. Commented [MK(5]: I suggest that this be converted into "Key Figure". The Figure represents the contents of the review well. Commented [MK(6]: Are the authors talking about advances in sample isolation? I'd encourage you to give examples to align the readers to what advancements the authors are referring to. Commented [GDR7R6]: Done with some of the innovations discussed in the following chapters.

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Analysis of Extracellular Vesicles Using Fluorescence Nanoparticle Tracking Analysis

Cited by 62 publications

References 7 publications

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

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

Umbilical cord-derived Wharton’s jelly for regenerative medicine applications

Liquid Biopsies in Cancer Diagnosis, Monitoring, and Prognosis

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