UMMS Affiliation

Department of Molecular, Cell and Cancer Biology

Publication Date


Document Type



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

Rights and Permissions

Copyright © 2021 The Authors. This is an open access article under the CC BY-NC-ND license (

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

Journal/Book/Conference Title

STAR protocols

Related Resources

Link to Article in PubMed

PubMed ID


Creative Commons License

Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License.