An Innovative Method for Exosome Quantification and Size Measurement

Mehdiani, Arash; Maier, Anatol; Pinto, António Sá; Barth, Mareike; Akhyari, Payam; Lichtenberg, Artur

doi:10.3791/50974-v

Cited by 18 publications

(12 citation statements)

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“…In order to study the impact of liver diseases on the distribution of secreted EVs, NTA was used in this study to monitor and compare EVs characteristics. NTA is a technique based on light-scattering, which enables the rapid sizing and enumeration of nanosized particles in solution [17]. Since its introduction, NTA has become a standard procedure to reproducibility measure EVs size and concentration in their native conditions [18, 19].…”

Section: Resultsmentioning

confidence: 99%

An optimized workflow for analyzing extracellular vesicles as biomarkers in liver diseases

Paluschinski

Loosen

Kordes

et al. 2023

Preprint

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Background & Aims: Extracellular vesicles (EVs) play an important role in intercellular communication, serving as vehicles for the exchange of biological materials and being involved in the regulation of physiological processes. EVs and their associated cargoes are considered a promising source of disease-associated biomarkers. The purpose of this study was to establish an easy-to-use, reproducible, and scalable workflow to efficiently analyze EVs in the context of liver disease. Methods: An optimized workflow was established for the pre-analytical processing and isolation of EVs from plasma and serum. Nanoparticle Tracking Analysis (NTA) was used to characterize circulating EVs in the serum of patients with nonalcoholic fatty liver disease (NAFLD), autoimmune liver disease (AIH), and animal models with impaired liver function. EVs were separated from soluble proteins by an optimized, polyethylene glycol (PEG)-based enrichment protocol. Enriched EVs were either labeled and functionally characterized by monitoring cellular uptake or lysed for biomarker identification. Results: Circulating EVs in the serum of patients with NAFLD or AIH and in different animal models have been characterized by NTA. Here we show that both the quantity and size of EVs in the serum of patients/animal models are significantly different from those of healthy individuals. We show that isolated EVs are functional, and their uptake by acceptor cells can be quantified after fluorescence labelling. Enriched EVs were directly used to analyze RNA biomarkers. Several microRNAs, including miR-15b, -16, -21, -122 and -223, were found to be significantly up-regulated in EVs isolated from the sera of patients with NAFLD and AIH. We show that EVs transport cytokines, and that IL-2, IL-6 and IL-8 were significantly up-regulated in EVs enriched from patients with cholangiocarcinoma (CCA) compared to healthy controls. Conclusions: The workflow presented here represents an accessible and easy-to-use approach that enables the analysis and enrichment of EVs from complex biological fluids and their preparation for functional characterization or downstream analysis. In this study, the levels of several miRNAs were found to be significantly increased in EVs isolated from AIH and NAFLD patients compared with healthy controls.

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Section: Resultsmentioning

confidence: 99%

An optimized workflow for analyzing extracellular vesicles as biomarkers in liver diseases

Paluschinski

Loosen

Kordes

et al. 2023

Preprint

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“…Particle number and size distribution of nanoplastic contaminants were analyzed using a ZetaView PMX-110 light scattering video microscope (Particle Metrix, Meerbusch, Germany) as previously described ( 55 ) and a Titan DynaPro (Wyatt Technology) for DLS analysis, at ~25°C. For DLS, the size distribution was analyzed using the Dyna V6.3.4 software package.…”

Section: Methodsmentioning

confidence: 99%

Anionic nanoplastic contaminants promote Parkinson’s disease–associated α-synuclein aggregation

Liu,

Sokratian,

Duda

et al. 2023

Sci. Adv.

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Recent studies have identified increasing levels of nanoplastic pollution in the environment. Here, we find that anionic nanoplastic contaminants potently precipitate the formation and propagation of α-synuclein protein fibrils through a high-affinity interaction with the amphipathic and non-amyloid component (NAC) domains in α-synuclein. Nanoplastics can internalize in neurons through clathrin-dependent endocytosis, causing a mild lysosomal impairment that slows the degradation of aggregated α-synuclein. In mice, nanoplastics combine with α-synuclein fibrils to exacerbate the spread of α-synuclein pathology across interconnected vulnerable brain regions, including the strong induction of α-synuclein inclusions in dopaminergic neurons in the substantia nigra. These results highlight a potential link for further exploration between nanoplastic pollution and α-synuclein aggregation associated with Parkinson’s disease and related dementias.

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“…One should note that, according to MISEV2018, single-vesicle analysis methods may be difficult to exploit quantitatively, as analysis of an adequate number of EVs is required to increase statistical power [11]. On the other hand, single-particle analysis techniques, such as nanoparticle tracking analysis, measure the biophysical characteristics of single EVs and can be used for the quantification of a large number of EVs, providing a higher statistical power than single-EV high-resolution imaging methods [11, 72, 80, 81].…”

Section: Methods To Isolate and Analyze Evsmentioning

confidence: 99%

Extracellular Vesicles and the Stress System

Makrygianni

Chrousos

2022

Neuroendocrinology

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Extracellular vesicles (EVs) are membrane-enclosed nanoparticles that contain various biomolecules, including nucleic acids, proteins, and lipids, and are manufactured and released by virtually all cell types. There is evidence that EVs are involved in intercellular communication, acting in an autocrine, paracrine, or/and endocrine manner. EVs are released by the cells of the central nervous system (CNS), including neurons, astrocytes, oligodendrocytes and microglia, and have the ability to cross the blood brain barrier (BBB) and enter the systemic circulation. Neuroendocrine cells are specialized neurons that secrete hormones directly into blood vessels, such as the hypophyseal portal system or the systemic circulation, a process that allows neuroendocrine integration to take place. In mammals, neuroendocrine cells are widely distributed throughout various anatomic compartments, with the hypothalamus being a central neuroendocrine integrator. The hypothalamus is a key part of the Stress System (SS), a highly conserved neuronal/neuroendocrine system aiming at maintaining systemic homeostasis, when the latter is threatened by various stressors. The central parts of the SS are the interconnected hypothalamic Corticotropin-releasing Hormone (CRH) and the brainstem Locus Caeruleus-Norepinephrine (LC-NE) systems, while their peripheral parts are respectively the pituitary-adrenal axis, and the sympathetic nervous/sympatho-adrenomedullary systems (SNS-SAM) as well as components of the parasympathetic system (PNS). During stress, multiple CNS loci show plasticity and undergo remodeling, partly mediated by increased glutamatergic and noradrenergic activity, and actions of cytokines and glucocorticoids (GCs), all regulated by the interaction of the hypothalamic-pituitary-adrenal (HPA) axis and the LC-NE/SNS-SAM systems. In addition, there are peripheral changes due to the increased secretion of stress hormones and pro-inflammatory cytokines in the context of stress-related systemic (para)inflammation.

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An Innovative Method for Exosome Quantification and Size Measurement

Cited by 18 publications

References 0 publications

An optimized workflow for analyzing extracellular vesicles as biomarkers in liver diseases

An optimized workflow for analyzing extracellular vesicles as biomarkers in liver diseases

Anionic nanoplastic contaminants promote Parkinson’s disease–associated α-synuclein aggregation

Extracellular Vesicles and the Stress System

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