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This detailed volume presents hands-on technological protocols used to target an array of cell-secreted extracellular vesicles (EVs) in a variety of biological systems. Beginning with methods for EV purification and analysis, the book continues with sections on the study of EV functions as well as specific systems and models allowing for the study of EVs of different origin. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and practical, Cell-Secreted Vesicles: Methods and Protocols serves as an ideal guide to conducting systematic assays in an effort to further our understanding of the mode of assembly, secretion, and targeting of EVs which will serve eventually as new therapeutic openings.
List of contents
Plasmon-Enhanced Characterization of Single Extracellular Vesicles.- Detection of Cell-Derived Exosomes via Surface-Enhanced Raman Scattering Using Aggregated Silver Nanoparticles.- In Vivo Analysis of Heterogeneous Extracellular Vesicles Using a Red-Shifted Bioluminescence Resonance Energy Transfer Reporter Protein.- Characterization of Extracellular Vesicles by Transmission Electron Microscopy and Immunolabelling Electron Microscopy.- Extracellular Vesicles Isolation by a Tangential-Flow-Filtration-Based Large-Scale Purification Method.- Metabolomics Analysis of Urinary Extracellular Vesicles by Nuclear Magnetic Resonance and Liquid Chromatography-Mass Spectrometry.- Methodologies for Scalable Production of High Quality Purified Small Extracellular Vesicles from Conditioned Medium.- Automated Online Isolation and Fractionation Method for Subpopulations of Extracellular Vesicles.- A Novel Assay for Investigating the Role of Exosomes in Tumor Cell-Endothelial Cell Crosstalk.- Nanoparticle (NP) Loading by Direct Incubation with Extracellular Vesicles-Secretor Cells: NP Encapsulation and Exosome Characterization.- Two Complementary Strategies to Quantitate Extracellular Vesicles Uptake Using Bioluminescence and Non-Lipidic Dyes.- Integrin-Mediated Exosomal Homing to Organs.- Methods and Protocols for Using Extracellular Vesicles as Delivery Vehicles in Neuronal Research.- Quantitative Analysis of Extracellular Vesicle Release Using Artificial MicroRNAs.- Purification of Bacterial-Enriched Extracellular Vesicle Samples from Feces by Density Gradient Ultracentrifugation.- Isolation and Characterization of Extracellular Vesicles from Lymphocytes.- Biogenesis of Mesoporous Silica Nanoparticles Enclosed in Extracellular Vesicles by Mouse Renal Adenocarcinoma Cells.- Magnetic Separation of Cell-Secreted Vesicles with Tailored Magnetic Particles and Downstream Applications.- Cilia-Derived Extracellular Vesicles in Caenorhabditis elegans: In Vivo Imaging and Quantification of Extracellular Vesicles Release and Capture.- Exosome-Based COVID-19 Vaccine.
