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This volume examines established methods and protocols to isolate and characterize extracellular vesicles (EVs) and their composition, among other techniques including purification, imaging, biofluid-specific and cell-specific isolation and downstream genomic and proteomic profiling. The international group of expert scientists who have contributed to this collection provide a variety of different techniques related to the growing assortment of EV applications, given that at times using only one technique or two is insufficient to address the question at hand. 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, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Comprehensive and practical, Extracellular Vesicles: Methods and Protocols serves as an ideal guide for researchers seeking to expand our knowledge of EV functions and applications.
Table des matières
Extracellular Vesicles: A Brief Overview and Its Role in Precision Medicine.- Red Blood Cells: A Source of Extracellular Vesicles.- Isolation of Extracelullar Vesicles by Ultracentrifugation.- Sequential Filtration: A Gentle Method for the Isolation of Functional Extracellular Vesicles.- Paper-Based Devices for Isolation of Extracellular Vesicles.- Filter-Based Extracellular Vesicle mRNA Isolation and High-Throughput Gene Expression Analysis.- Specific and Generic Isolation of Extracellular Vesicles with Magnetic Beads.- Polymer-Based Purification of Extracellular Vesicles.- Size Exclusion Chromatography: A Simple and Reliable Method for Exosome Purification.- Purification Protocols for Extracellular Vesicles.- Characterization of Extracellular Vesicles by Surface Plasmon Resonance.- Extracellular Vesicle Isolation and Analysis by Western Blotting.- Analysis of Extracellular Vesicles Using Fluorescence Nanoparticle Tracking Analysis.- Characterization of Extracellular Vesicles by FlowCytometry.- Characterization of Extracellular Vesicles by Size-Exclusion High-Performance Liquid Chromatography (HPLC).- Multi-Surface Antigen Staining of Larger Extracellular Vesicles.- Microcapillary Chip-Based Extracellular Vesicle Profiling System.- Detection and Characterization of Extracellular Vesicles by Transmission and Cryo-Transmission Electron Microscopy.- Imaging of Isolated Extracellular Vesicles Using Fluorescence Microscopy.- In Vivo Tracking of Extracellular Vesicles in Mice Using Fusion Protein Comprising Lactadherin and Gaussia Luciferase.- Tracking Extracellular Vesicles Delivery and RNA Translation Using Multiplexed Reporters.- Extraction and Analysis of Extracellular Vesicle-Associated miRNAs Following Antibody-Based Extracellular Vesicle Capture from Plasma Samples.- Extracellular Vesicle miRNA Detection Using Molecular Beacons.- Rapid Isolation of Extracellular Vesicles from Blood Plasma with Size-Exclusion Chromatography Followed by Mass Spectrometry-Based Proteomic Profiling.- An Adaptable Polyethylene Glycol-Based Workflow for Proteomic Analysis of Extracellar Vesicles.- Isolation of Extracellular Vesicles from Ovarian Follicular Fluid: Evaluation of Ultracentrifugation and Commercial Kits.- Isolation of Extracellular Vesicles in Saliva Using Density Gradient Ultracentrifugation.- Isolation of Extracellular Vesicles from Breast Milk.- An Integrated Double-Filtration Microfluidic Device for Detection of Extracellular Vesicles from Urine for Bladder Cancer Diagnosis.- Electric Field-Induced Disruption and Releasing Viable Content from Extracellular Vesicles.- Production and Characterization of Extracellular Vesicles in Malaria.- Isolation of Extracellular Vesicles from Stem Cells.- The Use of Peripheral Extracellular Vesicles for Identification of Molecular Biomarkers in a Solid Tumor Mouse Model.- Therapeutic Applications of Extracellular Vesicles: Perspectives from Newborn Medicine.- Therapeutic Use of Tumor Cell-Derived Extracellular Vesicles.
Résumé
This volume examines established methods and protocols to isolate and characterize extracellular vesicles (EVs) and their composition, among other techniques including purification, imaging, biofluid-specific and cell-specific isolation and downstream genomic and proteomic profiling.
