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
. 2022:2504:207-217.
doi: 10.1007/978-1-0716-2341-1_15.

Generation, Characterization, and Count of Fluorescent Extracellular Vesicles

Flavia Ferrantelli  1 , Valentina Tirelli  2 , Valeria Barreca  1 , Francesco Manfredi  3
Affiliations

Generation, Characterization, and Count of Fluorescent Extracellular Vesicles

Flavia Ferrantelli et al. Methods Mol Biol. 2022.

Abstract

Extracellular vesicles (EVs) are membranous particles released by all cells in the external milieu. Depending on their origin, they are given different names: exosomes are nanovesicles that originate from the endosomal compartment, whereas microvesicles bud from plasma membrane. Both contain molecules that are crucial for the onset and spreading of different pathologies, from neurodegenerative diseases to cancer, and are considered promising disease markers. On the other hand, EVs are often used as therapeutic tools, and can be engineered to carry drugs and chemicals. This chapter describes a method to produce EVs, mainly exosomes, containing the green fluorescent protein (GFP) linked to an exosome anchoring protein (Nefmut). This enables counting and tracing of fluorescent EVs by different methods, including conventional flow cytometry.

Keywords: Exosome count; Extracellular vesicles; Fluorescent exosomes; Fusion protein; Nef.

PubMed Disclaimer

References

    1. Harding C, Heuser J, Stahl P (1983) Receptor-mediated endocytosis of transferrin and recycling of the transferrin receptor in rat reticulocytes. J Cell Biol 97:329–339 - DOI
    1. Wolfers J, Lozier A, Raposo G et al (2001) Tumor-derived exosomes are a source of shared tumor rejection antigens for CTL cross-priming. Nat Med 7:297–303 - DOI
    1. Aline F, Bout D, Amigorena S et al (2004) Toxoplasma gondii antigen-pulsed-dendritic cell-derived exosomes induce a protective immune response against T. gondii infection. Infect Immun 72:4127–4137 - DOI
    1. Li S, Yao J, Xie M et al (2018) Exosomal miRNAs in hepatocellular carcinoma development and clinical responses. J Hematol Oncol 11:54 - DOI
    1. Liu S, Hossinger A, Göbbels S et al (2017) Prions on the run: how extracellular vesicles serve as delivery vehicles for self-templating protein aggregates. Prion 11:98–112 - DOI

MeSH terms

LinkOut - more resources