Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2022:2473:397-403.
doi: 10.1007/978-1-0716-2209-4_25.

Quantitative Measurement of Extracellular Vesicle Content Delivery Within Acceptor Cells

Émeline Bonsergent  1   2 , Shéryl Bui  1   2 , Grégory Lavieu  3   4
Affiliations

Quantitative Measurement of Extracellular Vesicle Content Delivery Within Acceptor Cells

Émeline Bonsergent et al. Methods Mol Biol. 2022.

Abstract

Extracellular vesicles (EVs), including exosomes and microvesicles, are thought to transport bioactive molecules from donor to acceptor cells. Although EV uptake has been qualitatively assessed through subcellular imaging, EV content delivery has been rarely addressed due to a lack of adequate methods. Here we present a sensitive bulk assay to quantitatively measure EV uptake and content delivery in mammalian cell. In this assay, EVs containing a NanoLuc luciferase-tagged cargo are mixed with unlabeled acceptor cells. Cell fractionation separates membrane and cytosolic fractions, and luciferase activity is measured within each fraction to determine the percentage of cytosolic release. This assay can be used to further decipher cellular and molecular mechanisms that regulate the EV delivery process or to quantitatively test specific pairs of donor-acceptor cells.

Keywords: Content release; Exosomes; Extracellular vesicles; Fusion; Microvesicles; Uptake.

PubMed Disclaimer

Similar articles

See all similar articles

Cited by

References

    1. Valadi H, Ekström K, Bossios A et al (2007) Exosome-mediated transfer of mRNAs and microRNAs is a novel mechanism of genetic exchange between cells. Nat Cell Biol 9:654–659 - DOI
    1. Balaj L, Lessard R, Dai L et al (2011) Tumour microvesicles contain retrotransposon elements and amplified oncogene sequences. Nat Commun 2:180 - DOI
    1. Skog J, Würdinger T, van Rijn S et al (2008) Glioblastoma microvesicles transport RNA and proteins that promote tumour growth and provide diagnostic biomarkers. Nat Cell Biol 10:1470–1476 - DOI
    1. Mathieu M, Martin-Jaular L, Lavieu G et al (2019) Specificities of secretion and uptake of exosomes and other extracellular vesicles for cell-to-cell communication. Nat Cell Biol 21:9–17 - DOI
    1. Costa Verdera H, Gitz-Francois JJ, Schiffelers RM et al (2017) Cellular uptake of extracellular vesicles is mediated by clathrin-independent endocytosis and macropinocytosis. J Control Release 266:100–108 - DOI

Publication types

LinkOut - more resources