Emerging techniques in the isolation and characterization of extracellular vesicles and their roles in cancer diagnostics and prognostics

Sunkara, Vijaya; Woo, Hyun‐Kyung; Cho, Yoon‐Kyoung

doi:10.1039/c5an01775k

Cited by 93 publications

(95 citation statements)

References 98 publications

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“…We found significantly higher protein content in CFS/ME patients than in controls (mean ± SD, 65.49 ± 10.75 mg/ml vs. 43.62 ± 5.10 mg/ml; p = 0.001, respectively). The Bradford assay is commonly used to determine the protein content of isolated EVs and to estimate the amount of circulating EVs [13]. Moreover, analysis of the EV protein content may differentiate between CFS/ME individuals and controls.…”

Section: Resultsmentioning

confidence: 99%

“…Detection of purified EVs from CFS/ME patients and HCs by single-targeted LFIA was performed as previously described [13], using the following monoclonal antibodies: anti-CD9 (clone VJ1/20) and anti-CD63 (clone Tea3/18) (both from Immunostep, Madrid, Spain). Briefly, EV samples were homogenised with the detection antibody conjugated to gold nanoparticles (AuNP-anti-CD63).…”

Section: Methodsmentioning

confidence: 99%

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Circulating extracellular vesicles as potential biomarkers in chronic fatigue syndrome/myalgic encephalomyelitis: an exploratory pilot study

Castro-Marrero

Serrano-Pertierra

Oliveira-Rodríguez

et al. 2018

J of Extracellular Vesicle

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Chronic Fatigue Syndrome (CFS), also known as Myalgic Encephalomyelitis (ME) is an acquired, complex and multisystem condition of unknown etiology, no established diagnostic lab tests and no universally FDA-approved drugs for treatment. CFS/ME is characterised by unexplicable disabling fatigue and is often also associated with numerous core symptoms. A growing body of evidence suggests that extracellular vesicles (EVs) play a role in cell-to-cell communication, and are involved in both physiological and pathological processes. To date, no data on EV biology in CFS/ME are as yet available. The aim of this study was to isolate and characterise blood-derived EVs in CFS/ME. Blood samples were collected from 10 Spanish CFS/ME patients and 5 matched healthy controls (HCs), and EVs were isolated from the serum using a polymer-based method. Their protein cargo, size distribution and concentration were measured by Western blot and nanoparticle tracking analysis. Furthermore, EVs were detected using a lateral flow immunoassay based on biomarkers CD9 and CD63. We found that the amount of EV-enriched fraction was significantly higher in CFS/ME subjects than in HCs (p = 0.007) and that EVs were significantly smaller in CFS/ME patients (p = 0.014). Circulating EVs could be an emerging tool for biomedical research in CFS/ME. These findings provide preliminary evidence that blood-derived EVs may distinguish CFS/ME patients from HCs. This will allow offer new opportunities and also may open a new door to identifying novel potential biomarkers and therapeutic approaches for the condition.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Circulating extracellular vesicles as potential biomarkers in chronic fatigue syndrome/myalgic encephalomyelitis: an exploratory pilot study

Castro-Marrero

Serrano-Pertierra

Oliveira-Rodríguez

et al. 2018

J of Extracellular Vesicle

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“…A large number of excellent reviews have been published recently reporting the different EV isolation and characterization techniques. [15][16][17][18][19][20][21][22][23] However, our goal in this review is Extracellular vesicles (EVs) have recently been at the center of attention of cellular biologists and physicians as their role in intercellular communications has become progressively revealed. EVs display a huge diversity concerning their biogenesis and functions, leading to a still evolving classification comprising exosomes, microvesicles and apoptotic bodies.…”

Section: Doi: 101002/adbi201700040mentioning

confidence: 99%

Extracellular Vesicles: Isolation Methods

2017

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Extracellular vesicles (EVs) have recently been at the center of attention of cellular biologists and physicians as their role in intercellular communications has become progressively revealed. EVs display a huge diversity concerning their biogenesis and functions, leading to a still evolving classification comprising exosomes, microvesicles and apoptotic bodies. One of the main technical challenges to studying EVs is to isolate them without interfering with their structure, in order to be able to reveal their functions and to use them as biomarkers. Moreover, the new area of therapeutically using EVs needs clinical grade methods of isolation. In this review, different methods disposable to researchers and clinicians to isolate EVs are described, focusing on the physical principles that allow understanding the advantages and limitations of each technique. The new growing field of microfluidic systems which offers the opportunity to associate isolation and characterization on a single chip will be presented highlighting its potential in the field of EV studies.

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“…Simultaneously, detection techniques for analyzing and quantifying exosomes have been developed for both research and clinical applications. These widely used detection techniques include scanning electron microscopy (SEM), transmission electron microscopy (TEM), atomic force microscopy (AFM), enzyme‐linked immunosorbent assay (ELISA), dynamic light scattering (DLS), and nanoparticle tracking analysis (NTA) . Despite the great advances in exosome detection techniques, further development is needed to meet the requirements of rapid, sensitive, and low‐cost detections.…”

Section: Introductionmentioning

confidence: 99%

Progress in Microfluidics‐Based Exosome Separation and Detection Technologies for Diagnostic Applications

Lin

Chen

et al. 2019

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232

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Exosomes are secreted by most cell types and circulate in body fluids. Recent studies have revealed that exosomes play a significant role in intercellular communication and are closely associated with the pathogenesis of disease. Therefore, exosomes are considered promising biomarkers for disease diagnosis. However, exosomes are always mixed with other components of body fluids. Consequently, separation methods for exosomes that allow high‐purity and high‐throughput separation with a high recovery rate and detection techniques for exosomes that are rapid, highly sensitive, highly specific, and have a low detection limit are indispensable for diagnostic applications. For decades, many exosome separation and detection techniques have been developed to achieve the aforementioned goals. However, in most cases, these two techniques are performed separately, which increases operation complexity, time consumption, and cost. The emergence of microfluidics offers a promising way to integrate exosome separation and detection functions into a single chip. Herein, an overview of conventional and microfluidics‐based techniques for exosome separation and detection is presented. Moreover, the advantages and drawbacks of these techniques are compared.

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Emerging techniques in the isolation and characterization of extracellular vesicles and their roles in cancer diagnostics and prognostics

Cited by 93 publications

References 98 publications

Circulating extracellular vesicles as potential biomarkers in chronic fatigue syndrome/myalgic encephalomyelitis: an exploratory pilot study

Circulating extracellular vesicles as potential biomarkers in chronic fatigue syndrome/myalgic encephalomyelitis: an exploratory pilot study

Extracellular Vesicles: Isolation Methods

Progress in Microfluidics‐Based Exosome Separation and Detection Technologies for Diagnostic Applications

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