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

被引：34
|
作者：
Chhoy, Peter [1 ]
Brown, Caitlin W. [1 ]
Amante, John J. [1 ]
Mercurio, Arthur M. [1 ]
机构：
[1] Univ Massachusetts, Dept Mol Cell & Canc Biol, Med Sch, Worcester, MA 01605 USA
来源：
STAR PROTOCOLS | 2021年 / 2卷 / 01期
关键词：
Cancer; Cell biology; Cell culture; Cell membrane;
D O I：
10.1016/j.xpro.2021.100303
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
Extracellular vesicles (EVs) play key roles in transporting key molecular constitu-ents as cargo for extracellular trafficking. While several approaches have been developed to extract EVs from mammalian cells, the specific method of EV isola-tion 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).
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页数：11
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