Lipidomic and proteomic characterization of platelet extracellular vesicle subfractions from senescent platelets

Transfusion ◽  
2014 ◽  
Vol 55 (3) ◽  
pp. 507-521 ◽  
Author(s):  
Annika Pienimaeki-Roemer ◽  
Katja Kuhlmann ◽  
Alfred Böttcher ◽  
Tatiana Konovalova ◽  
Anne Black ◽  
...  
Keyword(s):  
Extracellular Vesicle

scholarly journals EVQuant; high-throughput quantification and characterization of extracellular vesicle (sub)populations

2020 ◽  
Author(s):  
T.A. Hartjes ◽  
J.A. Slotman ◽  
M.S. Vredenbregt ◽  
N. Dits ◽  
R. Van der Meel ◽  
...  
Keyword(s):  
Protein Content ◽  
Size Distribution ◽  
High Throughput ◽  
Liquid Biopsy ◽  
Ideal Liquid ◽  
Extracellular Vesicle ◽  
Cell Of Origin ◽  
Clinical Implementation ◽  
High Throughput Assay

AbstractExtracellular vesicles (EVs) reflect the cell of origin in terms of nucleic acids and protein content. They are found in biofluids and represent an ideal liquid biopsy biomarker source for many diseases. Unfortunately, clinical implementation is limited by available technologies for EV analysis. We have developed a simple, robust and sensitive microscopy-based high-throughput assay (EVQuant) to overcome these limitations and allow widespread use in the EV community. The EVQuant assay can detect individual immobilized EVs as small as 35 nm and determine their concentration in biofluids without extensive EV isolation or purification procedures. It can also identify specific EV subpopulations based on combinations of biomarkers and is used here to identify prostate-derived urinary EVs as CD9-/CD63+. Moreover, characterization of individual EVs allows analysis of their size distribution. The ability to identify, quantify and characterize EV (sub-)populations in high-throughput substantially extents the applicability of the EVQuant assay over most current EV quantification assays.

scholarly journals Characterization of Sheep Milk Extracellular Vesicle-miRNA by Sequencing and Comparison with Cow Milk

Animals ◽  
2020 ◽  
Vol 10 (2) ◽  
pp. 331 ◽  
Author(s):  
Suyu Quan ◽  
Xuemei Nan ◽  
Kun Wang ◽  
Linshu Jiang ◽  
Junhu Yao ◽  
...  
Keyword(s):  
Small Rnas ◽  
Regulatory Element ◽  
Signal Transmission ◽  
Extracellular Vesicle ◽  
Ovis Aries ◽  
Cow Milk ◽  
Bioactive Components ◽  
Sheep Milk ◽  
Bioactive Component

Milk can mediate maternal-neonatal signal transmission by the bioactive component extracellular vesicles (EVs), which select specific types of miRNA to encapsulate. The miRNA profiling of sheep milk EVs was characterized by sequencing and compared with that of cow milk. Nanoparticle tracking analysis revealed that the concentration of sheep milk EVs was 1.3 ± 0.09 × 1012 particles/mL and the diameter was peaked at 131.2 ± 0.84 nm. Sheep milk EVs contained various small RNAs, including tRNA, Cis-regulatory element, rRNA, snRNA, other Rfam RNA, and miRNA, which held about 36% of all the small RNAs. In total, 84 types of miRNA were annotated with Ovis aries by miRBase (version 22.0) in sheep milk EVs, with 75 shared types of miRNAs in all samples. The miR-26a, miR-191, let-7f, let-7b and miR-10b were highly expressed both in cow and sheep milk EVs, and 14 sheep milk EV-miRNAs in the top 20, occupying 98% of the total expression, were immune-related. Although pathway analysis showed different potential functions of cow and sheep milk EV-miRNAs, there were still some shared points: lipid metabolism (phospholipase D, glycerophospholipid and glycosylphosphatidylinositol), calcium metabolism, and nerve conduction (axon guidance and synapse). This study provides reference for the bioactive components in the milk of different species.

scholarly journals High throughput imaging of nanoscale extracellular vesicles by scanning electron microscopy for accurate size-based profiling and morphological analysis

2021 ◽  
Author(s):  
Sara Cavallaro ◽  
Petra Hååg ◽  
Kristina Viktorsson ◽  
Anatol Krozer ◽  
Kristina Fogel ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
High Throughput ◽  
Extracellular Vesicle ◽  
Physical Characterization ◽  
Nsclc Cell Line ◽  
Visual Approach ◽  
High Throughput Imaging ◽  
Scanning Electron

ABSTRACTNanoscale extracellular vesicle (EVs) have been found to play a key role in intercellular communication, offering opportunities for both diagnostics and therapeutics. However, lying below the diffraction limit and also being highly heterogeneous in their size, morphology and abundance, these vesicles pose significant challenges for their physical characterization. Here, we present a direct visual approach for their accurate morphological and size-based profiling by using scanning electron microscopy (SEM). To achieve that, we methodically examined various process steps and developed a protocol to improve the throughput, conformity and image quality while preserving the shape of EVs. The investigation was performed with small EVs (sEVs) isolated from a non-small cell lung cancer (NSCLC) cell line H1975 as well as from a human serum, and the results were compared with those obtained from nanoparticle tracking analysis (NTA). While the comparison of the sEV size distributions showed good agreement between the two methods for large sEVs (diameter >70 nm), the microscopy based approach showed a better capacity for analyses on smaller vesicles, with higher sEV counts compared to NTA. In addition, we demonstrated the possibility of identifying non-EV particles based on size and morphological features. The study also showed process steps that can generate artifacts bearing resemblance with sEVs. The results therefore present a simple way to use a widely available microscopy tool for accurate and high throughput physical characterization of EVs.

Characterization of GPCRs in extracellular vesicle (EV)

2017 ◽  
pp. 119-132 ◽  
Author(s):  
Manoj R. Medapati ◽  
Anula Singh ◽  
Ranjit R. Korupally ◽  
Dana Henderson ◽  
Thomas Klonisch ◽  
...  
Keyword(s):  
Extracellular Vesicle

IDENTIFICATION AND BIOLOGICAL CHARACTERIZATION OF EXTRACELLULAR VESICLE PROTEINS FOR PROPAGATION AND SPREAD OF TAU

2019 ◽  
Vol 15 (7) ◽  
pp. P192
Author(s):  
Satoshi Muraoka ◽  
Annina M. DeLeo ◽  
Manveen K. Sethi ◽  
John D. Hogan ◽  
Zijian Yang ◽  
...  
Keyword(s):  
Extracellular Vesicle ◽  
Biological Characterization ◽  
Vesicle Proteins

scholarly journals Quantitative characterization of extracellular vesicle uptake and content delivery within mammalian cells

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Emeline Bonsergent ◽  
Eleonora Grisard ◽  
Julian Buchrieser ◽  
Olivier Schwartz ◽  
Clotilde Théry ◽  
...  
Keyword(s):  
Mammalian Cells ◽  
Fluorescent Protein ◽  
Extracellular Vesicle ◽  
Content Delivery ◽  
Bafilomycin A1 ◽  
Quantitative Characterization ◽  
Endosomal Acidification ◽  
A Cell ◽  
Dose Responses

AbstractExtracellular vesicles (EVs), including exosomes, are thought to mediate intercellular communication through the transfer of cargoes from donor to acceptor cells. Occurrence of EV-content delivery within acceptor cells has not been unambiguously demonstrated, let alone quantified, and remains debated. Here, we developed a cell-based assay in which EVs containing luciferase- or fluorescent-protein tagged cytosolic cargoes are loaded on unlabeled acceptor cells. Results from dose-responses, kinetics, and temperature-block experiments suggest that EV uptake is a low yield process (~1% spontaneous rate at 1 h). Further characterization of this limited EV uptake, through fractionation of membranes and cytosol, revealed cytosolic release (~30% of the uptaken EVs) in acceptor cells. This release is inhibited by bafilomycin A1 and overexpression of IFITM proteins, which prevent virus entry and fusion. Our results show that EV content release requires endosomal acidification and suggest the involvement of membrane fusion.

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