Nanoparticle Tracking Analysis for the Quantification and Size Determination of Extracellular Vesicles

Comfort, Nicole; Cai, Kunheng; Bloomquist, Tessa R.; Strait, Madeleine; Ferrante, Anthony W.; Baccarelli, Andrea A.

doi:10.3791/62447

Cited by 40 publications

(38 citation statements)

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“…NTA can determine the size and concentration of EVs in solution prepared from a wide variety of samples. 36 – 39 This technique monitors the Brownian movement of particles in liquid suspension in real time and can estimate EV sizes ranging from 60 to 1000 nm. 40 Brownian motion of the particles is measured using laser light scattering microscopy, with a camera as the detector, and their hydrodynamic diameter is calculated using the Stokes–Einstein equation.…”

Section: Extracellular Vesicle Characterization Methodsmentioning

confidence: 99%

Overview of extracellular vesicle characterization techniques and introduction to combined reflectance and fluorescence confocal microscopy to distinguish extracellular vesicle subpopulations

et al. 2022

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. Extracellular vesicles (EVs) are nanoparticles (30 to 1000 nm in diameter) surrounded by a lipid-bilayer which carry bioactive molecules between local and distal cells and participate in intercellular communication. Because of their small size and heterogenous nature they are challenging to characterize. Here, we discuss commonly used techniques that have been employed to yield information about EV size, concentration, mechanical properties, and protein content. These include dynamic light scattering, nanoparticle tracking analysis, flow cytometry, transmission electron microscopy, atomic force microscopy, western blotting, and optical methods including super-resolution microscopy. We also introduce an innovative technique for EV characterization which involves immobilizing EVs on a microscope slide before staining them with antibodies targeting EV proteins, then using the reflectance mode on a confocal microscope to locate the EV plane. By then switching to the microscope’s fluorescence mode, immunostained EVs bearing specific proteins can be identified and the heterogeneity of an EV preparation can be determined. This approach does not require specialist equipment beyond the confocal microscopes that are available in many cell biology laboratories, and because of this, it could become a complementary approach alongside the aforementioned techniques to identify molecular heterogeneity in an EV preparation before subsequent analysis requiring specialist apparatus.

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Section: Extracellular Vesicle Characterization Methodsmentioning

confidence: 99%

Overview of extracellular vesicle characterization techniques and introduction to combined reflectance and fluorescence confocal microscopy to distinguish extracellular vesicle subpopulations

et al. 2022

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“…NTA and TRPS can be used for particle size analysis of EVs, and their detection sensitivity is 70-90 nm and 70-100 nm, respectively. NTA technology allows one-time measurement and quantification of EVs, but the equipment is expensive and difficult to operate ( 129 , 130 ). The ELISA technique is greatly flexible and can be modified appropriately for the analyte, but it is also time-consuming.…”

Section: Quantification Methods Of Evsmentioning

confidence: 99%

Extracellular Vesicles and Hepatocellular Carcinoma: Opportunities and Challenges

Wang

Zhang

et al. 2022

Front. Oncol.

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The incidence of hepatocellular carcinoma (HCC) is increasing worldwide. Extracellular vesicles (EVs) contain sufficient bioactive substances and are carriers of intercellular information exchange, as well as delivery vehicles for nucleic acids, proteins and drugs. Although EVs show great potential for the treatment of HCC and their role in HCC progression has been extensively studied, there are still many challenges such as time-consuming extraction, difficult storage, easy contamination, and low drug loading rate. We focus on the biogenesis, morphological characteristics, isolation and extraction of EVs and their significance in the progression of HCC, tumor invasion, immune escape and cancer therapy for a review. EVs may be effective biomarkers for molecular diagnosis of HCC and new targets for tumor-targeted therapy.

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“…It is essential for EVs dose determination and contaminant identification, and can be assessed through biophysical or biochemical methods. Regarding EVs characterization, nanoparticle tracking analysis - a method that tracks individual nanoparticles and derives their size and concentration in suspension– is used for EVs quantification and size determination, while electron-microscopy and cryo-electron microscopy are commonly used for EVs morphological characterization ( Emelyanov et al, 2020 ; Comfort et al, 2021 ; Stam et al, 2021 ). Other methods used for EVs quantification include microfluidics, arrays, and polymerase chain reaction microfluidic systems ( Panagopoulou et al, 2020 ).…”

Section: Extracellular Vesicles As a New Paradigm For Cell-free Therapymentioning

confidence: 99%

Mesenchymal Stromal Cell-Derived Extracellular Vesicles as Biological Carriers for Drug Delivery in Cancer Therapy

Sanmartin

Borzone

Giorello

et al. 2022

Front. Bioeng. Biotechnol.

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Cancer is the second leading cause of death worldwide, with 10.0 million cancer deaths in 2020. Despite advances in targeted therapies, some pharmacological drawbacks associated with anticancer chemo and immunotherapeutic agents include high toxicities, low bioavailability, and drug resistance. In recent years, extracellular vesicles emerged as a new promising platform for drug delivery, with the advantage of their inherent biocompatibility and specific targeting compared to artificial nanocarriers, such as liposomes. Particularly, mesenchymal stem/stromal cells were proposed as a source of extracellular vesicles for cancer therapy because of their intrinsic properties: high in vitro self-renewal and proliferation, regenerative and immunomodulatory capacities, and secretion of extracellular vesicles that mediate most of their paracrine functions. Moreover, extracellular vesicles are static and safer in comparison with mesenchymal stem/stromal cells, which can undergo genetic/epigenetic or phenotypic changes after their administration to patients. In this review, we summarize currently reported information regarding mesenchymal stem/stromal cell-derived extracellular vesicles, their proper isolation and purification techniques - from either naive or engineered mesenchymal stem/stromal cells - for their application in cancer therapy, as well as available downstream modification methods to improve their therapeutic properties. Additionally, we discuss the challenges associated with extracellular vesicles for cancer therapy, and we review some preclinical and clinical data available in the literature.

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Nanoparticle Tracking Analysis for the Quantification and Size Determination of Extracellular Vesicles

Cited by 40 publications

References 0 publications

Overview of extracellular vesicle characterization techniques and introduction to combined reflectance and fluorescence confocal microscopy to distinguish extracellular vesicle subpopulations

Overview of extracellular vesicle characterization techniques and introduction to combined reflectance and fluorescence confocal microscopy to distinguish extracellular vesicle subpopulations

Extracellular Vesicles and Hepatocellular Carcinoma: Opportunities and Challenges

Mesenchymal Stromal Cell-Derived Extracellular Vesicles as Biological Carriers for Drug Delivery in Cancer Therapy

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