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Exosomes from Cell Culture-Conditioned Medium: Isolation by Ultracentrifugation and Characterization

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The Extracellular Matrix

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1952))

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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Authors and Affiliations

  1. Department of Lymphoma and Myeloma, The University of Texas MD Anderson Cancer Center, Houston, TX, USA

    Anurag Purushothaman

Authors
  1. Anurag Purushothaman
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Correspondence to Anurag Purushothaman .

Editor information

Editors and Affiliations

  1. Department of Medicine and Surgery, University of Insubria, Varese, Italy

    Davide Vigetti

  2. Department of Chemistry, University of Patras, Patras, Greece

    Achilleas D. Theocharis

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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

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-9132-7

  • Online ISBN: 978-1-4939-9133-4

  • eBook Packages: Springer Protocols

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