Protocol for the separation of extracellular vesicles by ultracentrifugation from in vitro cell culture models

STAR Protocols

Caitlin W. Brown¹  Peter Chhoy²  John J. Amante²  Arthur M. Mercurio² 

[1] Corresponding author;Department of Molecular, Cell and Cancer Biology, University of Massachusetts Medical School, Worcester, MA 01605, USA;

关键词: Cell biology;    Cell culture;    Cell membrane;    Cancer;   

DOI  :  

来源: DOAJ

【 摘 要 】

Summary: 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).

【 授权许可】

Unknown