Department of Molecular, Cell and Cancer Biology, University of Massachusetts Medical School, Worcester, MA 01605, USA.
STAR Protoc. 2021 Jan 29;2(1):100303. doi: 10.1016/j.xpro.2021.100303. eCollection 2021 Mar 19.
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).
细胞外囊泡（EVs）在作为细胞外运输的货物运输关键分子成分方面发挥着关键作用。虽然已经开发了几种从哺乳动物细胞中提取 EVs 的方法，但 EV 分离的具体方法会对膜完整性和产量产生深远影响。在这里，我们描述了使用差速超速离心从贴壁上皮细胞中分离 EVs 的分步程序。分离的 EVs 可以通过免疫印迹、质谱和透射电子显微镜进一步分析，以得出 EV 的产量和形态。有关该方案的使用和执行的完整详细信息，请参阅 Brown 等人。（2019 年）。
