Department of Biology and Chemistry, Changwon National University, Changwon 51140, South Korea.
Environmental Disease Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon 34141, Korea; Department of Functional Genomics, University of Science and Technology (UST), Daejeon 34113, Korea.
STAR Protoc. 2024 Sep 20;5(3):103183. doi: 10.1016/j.xpro.2024.103183. Epub 2024 Aug 1.
Extracellular vesicles (EVs) are membranous nanoparticles classified based on their size and surface markers, which can be specific to various cell origins. Here, we present a protocol for the isolation of pulmonary-specific EVs in mice. We describe steps for differential centrifugation, density gradient centrifugation, and commercially available polyethylene glycol(PEG)-based precipitation, employing pulmonary-specific EV-bound chemicals and antibodies. We then detail procedures for the characterization of these EVs through nanoparticle tracking analysis, flow cytometry, scanning electron microscopy, and transmission electron microscopy. For complete details on the use and execution of this protocol, please refer to Lee et al..
细胞外囊泡（EVs）是根据其大小和表面标志物分类的膜性纳米颗粒，可以特异性地来自于各种细胞来源。在这里，我们提供了一种从小鼠中分离肺特异性 EVs 的方案。我们描述了差速离心、密度梯度离心和商业上可用的基于聚乙二醇（PEG）沉淀的步骤，采用了肺特异性 EV 结合的化学物质和抗体。然后，我们详细介绍了通过纳米颗粒跟踪分析、流式细胞术、扫描电子显微镜和透射电子显微镜来表征这些 EV 的步骤。有关此方案的使用和执行的完整详细信息，请参见 Lee 等人的研究。
