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Overview of Exosomes

Overview of Exosomes

Exosomes are stable, lipid bilayer-enclosed vesicles capable of crossing biological barriers. They can carry a wide range of molecules required for intercellular communication. Once exosomes are released from the cell where they originated, they enter a recipient cell through various pathways such as fusion, receptor-mediated endocytosis, macropinocytosis, and phagocytosis.
Stahl et al. discovered exosomes in 1983, but the exosomes were initially considered waste products released from the...

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Estimation of Urinary Nanocrystals in Humans using Calcium Fluorophore Labeling and Nanoparticle Tracking Analysis

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Published on: February 9, 2021

Quantification of human urinary exosomes by nanoparticle tracking analysis.

Wilna Oosthuyzen¹, Nicole E L Sime, Jessica R Ivy

¹J. W. Dear: University of Edinburgh/British Heart Foundation Centre for Cardiovascular Science, Queen's Medical Research Institute, 47 Little France Crescent, Edinburgh, EH16 4TJ, UK. james.dear@ed.ac.uk.

The Journal of Physiology

|September 25, 2013

Summary

Nanoparticle tracking analysis (NTA) offers a promising method for quantifying urinary exosomes, crucial biomarkers for kidney disease. This technique accurately tracks exosomal changes, improving diagnostic potential.

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

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

Published on: March 28, 2021

Area of Science:

Biochemistry
Nanotechnology
Urology

Background:

Urinary exosomes are valuable biomarkers for kidney disease, but current detection methods are inefficient.
Exosomes contain proteins and RNA from various kidney sections, making them ideal for biomarker discovery.

Purpose of the Study:

To evaluate Nanoparticle Tracking Analysis (NTA) for characterizing and quantifying urinary exosomes.
To assess NTA's ability to detect changes in exosomal biomarkers in response to stimuli and disease.

Main Methods:

Application of NTA to human urine samples to identify and size particles.
Utilized fluorescently labeled antibodies against exosomal proteins (CD24, AQP2) for subpopulation identification.
Investigated the impact of pre-processing steps (ultracentrifugation) and storage conditions on NTA measurements.

Main Results:

NTA identified urinary particles of exosomal size, including a CD24- and AQP2-positive subpopulation.
Ultracentrifugation reduced intra-assay variability in exosome concentration measurements.
NTA successfully tracked exosomal AQP2 changes in response to desmopressin in cell culture, mice, and a patient.

Conclusions:

NTA is a viable tool for characterizing and quantifying urinary extracellular vesicles.
Optimized sample storage, particularly freezing at -80°C with protease inhibitors, is crucial for preserving nanoparticle concentration.
NTA demonstrates potential for non-invasive monitoring of kidney function and biomarker discovery.