scholarly journals Rapid Fluorescence-based Characterization of Single Extracellular Vesicles in Human Blood with Nanoparticle-tracking Analysis

10.3791/58731 ◽  
2019 ◽  
Author(s):  
Andreas Weber ◽  
Julia Christin Wehmeyer ◽  
Vera Schmidt ◽  
Artur Lichtenberg ◽  
Payam Akhyari
Keyword(s):  
Extracellular Vesicles ◽  
Human Blood ◽  
Nanoparticle Tracking Analysis

scholarly journals Urinary Extracellular Vesicles for Renal Tubular Transporters Expression in Patients With Gitelman Syndrome

2021 ◽  
Vol 8 ◽  
Author(s):  
Chih-Chien Sung ◽  
Min-Hsiu Chen ◽  
Yi-Chang Lin ◽  
Yu-Chun Lin ◽  
Yi-Jia Lin ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Membrane Transporters ◽  
Gitelman Syndrome ◽  
Healthy Controls ◽  
Nanoparticle Tracking Analysis ◽  
Non Invasive ◽  
Transmission Electron ◽  
Renal Tubular ◽  
Inactivating Mutations

Background: The utility of urinary extracellular vesicles (uEVs) to faithfully represent the changes of renal tubular protein expression remains unclear. We aimed to evaluate renal tubular sodium (Na+) or potassium (K+) associated transporters expression from uEVs and kidney tissues in patients with Gitelman syndrome (GS) caused by inactivating mutations in SLC12A3.Methods: uEVs were isolated by ultracentrifugation from 10 genetically-confirmed GS patients. Membrane transporters including Na+-hydrogen exchanger 3 (NHE3), Na+/K+/2Cl− cotransporter (NKCC2), NaCl cotransporter (NCC), phosphorylated NCC (p-NCC), epithelial Na+ channel β (ENaCβ), pendrin, renal outer medullary K1 channel (ROMK), and large-conductance, voltage-activated and Ca2+-sensitive K+ channel (Maxi-K) were examined by immunoblotting of uEVs and immunofluorescence of biopsied kidney tissues. Healthy and disease (bulimic patients) controls were also enrolled.Results: Characterization of uEVs was confirmed by nanoparticle tracking analysis, transmission electron microscopy, and immunoblotting. Compared with healthy controls, uEVs from GS patients showed NCC and p-NCC abundance were markedly attenuated but NHE3, ENaCβ, and pendrin abundance significantly increased. ROMK and Maxi-K abundance were also significantly accentuated. Immunofluorescence of the representative kidney tissues from GS patients also demonstrated the similar findings to uEVs. uEVs from bulimic patients showed an increased abundance of NCC and p-NCC as well as NHE3, NKCC2, ENaCβ, pendrin, ROMK and Maxi-K, akin to that in immunofluorescence of their kidney tissues.Conclusion: uEVs could be a non-invasive tool to diagnose and evaluate renal tubular transporter adaptation in patients with GS and may be applied to other renal tubular diseases.

scholarly journals Characterization of Extracellular Vesicles from Bronchoalveolar Lavage Fluid and Plasma of Patients with Lung Lesions Using Fluorescence Nanoparticle Tracking Analysis

Cells ◽  
2021 ◽  
Vol 10 (12) ◽  
pp. 3473
Author(s):  
Magdalena Dlugolecka ◽  
Jacek Szymanski ◽  
Lukasz Zareba ◽  
Zuzanna Homoncik ◽  
Joanna Domagala-Kulawik ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Lavage Fluid ◽  
Nanoparticle Tracking Analysis ◽  
Characterization Methods ◽  
Sample Composition ◽  
Associated Proteins ◽  
Nsclc Patients ◽  
Further Development ◽  
Reliable Methods

The current lack of reliable methods for quantifying extracellular vesicles (EVs) isolated from complex biofluids significantly hinders translational applications in EV research. The recently developed fluorescence nanoparticle tracking analysis (FL-NTA) allows for the detection of EV-associated proteins, enabling EV content determination. In this study, we present the first comprehensive phenotyping of bronchopulmonary lavage fluid (BALF)-derived EVs from non-small cell lung cancer (NSCLC) patients using classical EV-characterization methods as well as the FL-NTA method. We found that EV immunolabeling for the specific EV marker combined with the use of the fluorescent mode NTA analysis can provide the concentration, size, distribution, and surface phenotype of EVs in a heterogeneous solution. However, by performing FL-NTA analysis of BALF-derived EVs in comparison to plasma-derived EVs, we reveal the limitations of this method, which is suitable only for relatively pure EV isolates. For more complex fluids such as plasma, this method appears to not be sensitive enough and the measurements can be compromised. Our parallel presentation of NTA-based phenotyping of plasma and BALF EVs emphasizes the great impact of sample composition and purity on FL-NTA analysis that has to be taken into account in the further development of FL-NTA toward the detection of EV-associated cancer biomarkers.

scholarly journals 109 Isolation and characterization of myometrium and decidual cell-derived extracellular vesicles

2021 ◽  
Vol 224 (2) ◽  
pp. S75-S76
Author(s):  
Megan Shepherd ◽  
Enkhtuya Radnaa ◽  
Rheanna Urrabaz-Garza ◽  
Talar Kechichian ◽  
Ourlad Alzeus G. Tantengco ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Decidual Cell ◽  
Isolation And Characterization

scholarly journals Prospects and limitations of antibody-mediated clearing of lipoproteins from blood plasma prior to nanoparticle tracking analysis of extracellular vesicles

2017 ◽  
Vol 6 (1) ◽  
pp. 1308779 ◽  
Author(s):  
Morten Mørk ◽  
Aase Handberg ◽  
Shona Pedersen ◽  
Malene M. Jørgensen ◽  
Rikke Bæk ◽  
...  
Keyword(s):  
Blood Plasma ◽  
Extracellular Vesicles ◽  
Nanoparticle Tracking Analysis

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

The Analyst ◽  
2016 ◽  
Vol 141 (2) ◽  
pp. 371-381 ◽  
Author(s):  
Vijaya Sunkara ◽  
Hyun-Kyung Woo ◽  
Yoon-Kyoung Cho
Keyword(s):  
Extracellular Vesicles ◽  
Cancer Diagnostics ◽  
Clinical Implementation ◽  
Characterization Methods ◽  
Cancer Management ◽  
Isolation And Characterization

We present an overview of current isolation, detection, and characterization methods of extracellular vesicles and their applications and limitations as a potential emerging biomarker in cancer management and their clinical implementation.

Analysis of Extracellular Vesicles Using Fluorescence Nanoparticle Tracking Analysis

2017 ◽  
pp. 153-173 ◽  
Author(s):  
Pauline Carnell-Morris ◽  
Dionne Tannetta ◽  
Agnieszka Siupa ◽  
Patrick Hole ◽  
Rebecca Dragovic
Keyword(s):  
Extracellular Vesicles ◽  
Nanoparticle Tracking Analysis

Isolation and Characterization of Extracellular Vesicles in Stem Cell-Related Studies

2017 ◽  
pp. 205-223
Author(s):  
Zezhou Zhao ◽  
Dillon C. Muth ◽  
Vasiliki Mahairaki ◽  
Linzhao Cheng ◽  
Kenneth W. Witwer
Keyword(s):  
Stem Cell ◽  
Extracellular Vesicles ◽  
Isolation And Characterization
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