Department of Molecular, Cell and Cancer Biology
Cancer Biology | Cell Biology
Extracellular vesicles (EVs) play key roles in transporting key molecular constituents as cargo for extracellular trafficking. While several approaches have been developed to extract EVs from mammalian cells, the specific method of EV isolation can have a profound effect on membrane integrity and yield. Here, we describe a step-by-step procedure to separate EVs from adherent epithelial cells using differential ultracentrifugation. Separated EVs can be further analyzed by immunoblotting, mass spectrometry, and transmission electron microscopy to derive EV yield and morphology. For complete details on the use and execution of this protocol, please refer to Brown et al. (2019).
Cell biology, Cell culture, Cell membrane, Cancer
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Copyright © 2021 The Authors. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
DOI of Published Version
Chhoy P, Brown CW, Amante JJ, Mercurio AM. Protocol for the separation of extracellular vesicles by ultracentrifugation from in vitro cell culture models. STAR Protoc. 2021 Jan 29;2(1):100303. doi: 10.1016/j.xpro.2021.100303. PMID: 33554138; PMCID: PMC7848770. Link to article on publisher's site
Chhoy P, Brown CW, Amante JJ, Mercurio AM. (2021). Protocol for the separation of extracellular vesicles by ultracentrifugation from in vitro cell culture models. Open Access Publications by UMass Chan Authors. https://doi.org/10.1016/j.xpro.2021.100303. Retrieved from https://escholarship.umassmed.edu/oapubs/4573
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This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License.