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Single Extracellular Vesicle Transmembrane Protein Characterization by Nano-Flow Cytometry
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Technical Advances to Study Extracellular Vesicles.

Paula Carpintero-Fernández1, Juan Fafián-Labora1, Ana O'Loghlen1

  • 1Epigenetics and Cellular Senescence Group, Blizard Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, London, United Kingdom.

Frontiers in Molecular Biosciences
|December 14, 2017
PubMed
Summary

Extracellular vesicles (EVs) are nanoparticles from various cells found in body fluids. This review assesses standard and novel techniques for studying EV biology, release, uptake, and cargo.

Keywords:
exosomesextracellular vesicles (EVs)methodsmicrovesicles (MVs)noveltechnique

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Area of Science:

  • Cell Biology
  • Nanotechnology
  • Biochemistry

Background:

  • Extracellular vesicles (EVs) are lipid bilayer nanoparticles originating from various cell types and found in bodily fluids.
  • EVs contain diverse biomolecules (proteins, nucleic acids, lipids) influencing biological and pathological processes.
  • Studying EV biology requires robust methods for isolation, release, uptake, and cargo analysis.

Purpose of the Study:

  • To review and assess standard and emerging techniques for studying extracellular vesicle biology.
  • To provide an overview of methodologies for EV isolation, release, uptake, and cargo characterization.
  • To highlight advancements in EV research techniques across different biological systems.

Main Methods:

  • Review of established techniques including immunoblotting, fluorescent microscopy, and electron microscopy for EV characterization.
  • Discussion of advanced methods like Cre reporter systems and bioluminescence assays for EV uptake and release quantification.
  • Exploration of evolving cargo identification techniques, including mass spectrometry and next-generation sequencing.

Main Results:

  • Standard methods remain valuable for EV visualization and characterization.
  • Emerging techniques offer enhanced precision for studying EV dynamics at the single-cell level.
  • Advances in omics technologies facilitate deeper insights into EV cargo composition.

Conclusions:

  • A comprehensive understanding of EV biology necessitates the application of both standard and cutting-edge methodologies.
  • Technological advancements are crucial for accurate assessment of EV release, uptake, and molecular cargo.
  • This review provides a valuable resource for researchers utilizing diverse techniques to study extracellular vesicles.