Isolation and Characterization of Exosomes from Cultures of Tissue Explants and Cell Lines
| Autor: | Vladimir Baranov, Eva Dehlin, Olga Nagaeva, Lucia Mincheva-Nilsson |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Endosome Immunology Biology Cell Fractionation Exosomes Exosome Cell Line Immunophenotyping Flow cytometry Tissue Culture Techniques 03 medical and health sciences 0302 clinical medicine Centrifugation Density Gradient medicine Extracellular Animals Humans Cells Cultured Staining and Labeling medicine.diagnostic_test Flow Cytometry Microvesicles Cell biology 030104 developmental biology Cell culture Culture Media Conditioned 030220 oncology & carcinogenesis Density gradient ultracentrifugation Ultracentrifugation Biomarkers Function (biology) |
| Zdroj: | Current Protocols in Immunology |
| ISSN: | 1934-368X 1934-3671 |
| DOI: | 10.1002/cpim.17 |
| Popis: | Exosomes are specialized, nanometer-sized extracellular vesicles of endosomal origin actively secreted into the extracellular space by a variety of cells under normal and pathological conditions. Exosomes have recently emerged as important intercellular communicators and modulators of diverse mechanisms and cellular responses. Characterization of their composition and function will open possibilities for new diagnostic methods and promising therapeutic approaches based on nanobiology. This unit provides a standard isolation procedure for purification of exosomes based on density gradient ultracentrifugation with sucrose. The process of isolating exosomes relies on obtaining proper source fluids/supernatants as well as qualitative and quantitative assessment of the isolated vesicles. The methodological procedures here can be divided in three parts: (1) pre-isolation procedures aiming to obtain fluids containing exosomes, with a focus on protocols for organ explants and cell cultures; (2) a procedure for exosome isolation with several gradient alternatives; and (3) post-isolation procedures for estimating the purity and yield of the exosomal fraction. © 2016 by John Wiley & Sons, Inc. |
| Databáze: | OpenAIRE |
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