Pitfalls associated with lipophilic fluorophore staining of extracellular vesicles for uptake studies

Journal of Extracellular Vesicles
Jens B Simonsen

Abstract

Post-staining of extracellular vesicles (EVs) with lipid-anchored fluorophores (LAFs) such as PKH67 is a widely used strategy for studying EVs but it is associated with several pitfalls. The pitfalls discussed in this commentary are related to LAF labelling of non-EV species due to (1) lipoprotein contamination in EV samples, (2) desorption of the LAF reporters from vesicles into proteins and lipoproteins in blood and serum, and (3) the capability of the amphiphilic LAF compounds to form EV-like particles. Awareness of these challenges and developing solutions to overcome these are important to ensure that we make relevant interpretations when using LAFs to track EVs.

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Citations

Dec 5, 2019·Biomaterials Science·S BusattoP Bergese
Jun 10, 2020·Future Medicinal Chemistry·Johann Mar Gudbergsson
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Jan 15, 2022·Journal of Extracellular Vesicles·Xiaoyu YuHonghong Yao

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Methods Mentioned

BETA
confocal microscopy
density gradient centrifugation
transfection
flow cytometry

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