Abstract
Exosomes are small vesicles of endosomal origin secreted by most cell types. Recent studies have identified exosomes as important mediators of intercellular communication and as important source materials for many clinical applications, including minimal invasive disease diagnosis. Exosomes have been purified from in vitro cell culture supernatants by many different methods; however the most simple and reliable method involves purification by ultracentrifugation. This chapter describes a detailed protocol for isolating exosomes from cell culture-conditioned medium using ultracentrifugation and their characterization based upon size, number, and protein expression by several complementary methods such as transmission electron microscopy, nanoparticle tracking analysis, western blotting, and flow cytometry.
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Acknowledgments
This work was supported by a New Faculty Development Award from the Young Supporters Board of the UAB Comprehensive Cancer Center to AP.
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Purushothaman, A. (2019). Exosomes from Cell Culture-Conditioned Medium: Isolation by Ultracentrifugation and Characterization. In: Vigetti, D., Theocharis, A.D. (eds) The Extracellular Matrix. Methods in Molecular Biology, vol 1952. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9133-4_19
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DOI: https://doi.org/10.1007/978-1-4939-9133-4_19
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