Extracellular vesicles /

Detalles Bibliográficos
Clasificación:Libro Electrónico
Otros Autores: Spada, Sheila, Galluzzi, Lorenzo
Formato: Electrónico eBook
Idioma:Inglés
Publicado: Cambridge, MA : Academic Press, 2020.
Colección:Methods in enzymology ; v. 645.
Temas:
Liposomes.
Extracellular space.
Liposomes
Extracellular Space
Espace extracellulaire.
Extracellular space
Internet Resources.
Index not Present.
Acceso en línea:Texto completo
Tabla de Contenidos:
  • Intro
  • Extracellular Vesicles
  • Copyright
  • Contents
  • Contributors
  • Preface
  • Extracellular vesicles: An exciting and rapidly expanding field of investigation
  • Acknowledgments
  • Disclosures
  • References
  • Chapter One: Genetic labeling of extracellular vesicles for studying biogenesis and uptake in living mammalian cells
  • 1. Introduction
  • 2. Equipment and materials
  • 2.1. Equipment
  • 2.2. Cells and culture medium
  • 2.3. Reagents and chemicals
  • 3. Protocol
  • 3.1. Cell culture
  • 3.2. Generation of transient and stable CD63-GFP/VSVG-GFP expressing cells
  • 3.3. Exosome preparation from conditioned medium
  • 3.4. Cell and exosome imaging
  • 3.5. Cellular uptake of exosomes by confocal microscopy and flow cytometry
  • 4. Concluding remarks
  • 5. Notes
  • Acknowledgments
  • References
  • Chapter Two: Fluorescent labeling of extracellular vesicles
  • 1. Introduction
  • 1.1. Uptake studies
  • 1.2. Biodistribution studies
  • 1.3. Characterization studies
  • 2. Fluorescent labeling
  • 2.1. EV Labeling approaches
  • 2.1.1. Surface marker labeling of EVs
  • 2.1.2. Lipid membrane labeling of EVs
  • 2.1.3. Luminal labeling of EVs
  • 2.2. Post-labeling clean-up
  • 2.2.1. Differential ultracentrifugation
  • 2.2.2. Density gradient centrifugation
  • 2.2.3. Size exclusion chromatography
  • 2.2.4. Filtration
  • 3. Materials, equipment and reagents
  • 4. Protocols
  • 4.1. Sample labeling
  • 4.2. Removing excess dye
  • 4.3. Coverslip preparation
  • 4.4. Fluorescent imaging
  • 4.5. Size assessment using NTA
  • 5. Pros and cons
  • 5.1. Surface marker labeling
  • 5.2. Lipid membrane labeling
  • 5.3. Luminal labeling
  • 6. Conclusion
  • References
  • Chapter Three: Use of antibody arrays to probe exosome and extracellular vesicle mediated functional changes in cells
  • 1. Introduction
  • 1.1. Exosomes and extracellular vesicles
  • 1.2. Antibody arrays
  • 2. Methods
  • 2.1. Cell culture
  • 2.2. Isolation of exosome fraction
  • 2.3. Labeling of exosome fraction
  • 2.4. Treatment of cells
  • 2.5. Antibody arrays
  • 3. Notes
  • 4. Concluding remarks
  • References
  • Chapter Four: Analysis of individual extracellular vesicles by imaging flow cytometry
  • 1. Introduction
  • 2. Single EV analyses
  • 2.1. Equipment
  • 2.2. Materials
  • 2.3. Protocol optimization
  • 2.4. Instrument calibration
  • 3. Protocol
  • 3.1. Preparation of antibody solution
  • 3.2. Sample preparation
  • 3.3. Controls
  • 3.4. Sample acquisition with the ISX
  • 4. Data analysis
  • 4.1. Defining of masks to analyze the raw image file data
  • 4.2. Using the feature manager to determine single events
  • 4.3. Data presentation and calculation of subset concentrations
  • 5. Summary
  • Acknowledgments
  • References
  • Chapter Five: Imaging intercellular interaction and extracellular vesicle exchange in a co-culture model of chronic lymph ...
  • 1. Introduction

Ejemplares similares