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Isolating Astrocyte-Derived Extracellular Vesicles From Urine
Xin-Hui Xie1, Mian-Mian Chen1, Shu-Xian Xu1
1Department of Psychiatry, Renmin Hospital of Wuhan University, Wuhan, Hubei, People's Republic of China.
Introduction:
Brain-derived extracellular vesicles (BDEVs) can cross the blood-brain barrier and enter the periphery. Therefore, quantifying and analyzing peripherally circulating BDEVs offer a promising approach to directly obtain a window into central nervous system (CNS) pathobiology in vivo. Rapidly evolving CNS diseases require high-frequency sampling, but daily venipuncture of human subjects is highly invasive and usually unfeasible.
Methods:
To address this challenge, here we present a novel method for isolating astrocyte-derived extracellular vesicles from urine (uADEVs), combining urine concentration, ultracentrifugation to isolate total EVs, and then glutamate-aspartate transporter (GLAST) EV isolation using an anti-GLAST antibody.
Results:
The identity of these GLAST+EVs as uADEVs was confirmed by transmission electron microscopy, nanoparticle tracking analysis, western blotting, and assessment of astrocyte-related neurotrophins.
Conclusions:
Leveraging the convenience and availability of urine samples, the non-invasive uADEV approach provides a novel tool that allows high-frequency sampling to investigate rapidly evolving CNS diseases.
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