期刊
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
卷 -, 期 144, 页码 -出版社
JOURNAL OF VISUALIZED EXPERIMENTS
DOI: 10.3791/59143
关键词
Cancer Research; Issue 144; exosomes; extracellular vesicle; tissue; brain; tumor; mass spectrometry; density gradient; ex vivo; oncosomes; biomarkers
资金
- Florida Department of Health Ed and Ethel Moore Alzheimer's Disease Research Program [6AZ11]
- National Cancer Institute of the National Institutes of Health [R01CA204621]
Circulating and interstitial small membrane-bound extracellular vesicles (EVs) represent promising targets for the development of novel diagnostic or prognostic biomarker assays, and likely serve as important players in the progression of a vast spectrum of diseases. Current research is focused on the characterization of vesicles secreted from multiple cell and tissue types in order to better understand the role of EVs in the pathogenesis of conditions including neurodegeneration, inflammation, and cancer. However, globally consistent and reproducible techniques to isolate and purify vesicles remain in progress. Moreover, methods for extraction of EVs from solid tissue ex vivo are scarcely described. Here, we provide a detailed protocol for extracting small EVs of interest from whole fresh or frozen tissues, including brain and tumor specimens, for further characterization. We demonstrate the adaptability of this method for multiple downstream analyses, including electron microscopy and immunophenotypic characterization of vesicles, as well as quantitative mass spectrometry of EV proteins.
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