Basics of Extracellular Vesicles

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  • Introduction to the Course and the Field of Extracellular Vesicles
    • In this module, we will introduce the course and the field of extracellular vesicles. We will discuss the nomenclature and the history of extracellular vesicles as well as the organisms and places that have been shown to release/contain extracellular vesicles.
  • Biogenesis, Cargo and Uptake of Extracellular Vesicles
    • In this module, we will focus on the biogenesis and release of extracellular vesicles as well as the uptake mechanisms of extracellular vesicles when they are encountered by a recipient cell. This module also covers the different RNA, protein and lipid types present in extracellular vesicles as well as a brief overview what the functions of these molecules are. Furthermore, the techniques that are commonly used to detect these molecules as well as databases that can be useful to use when analyzing the cargo of extracellular vesicles will be highlighted.
  • Collection and Processing of Cell Culture Media and Body Fluids Prior to Isolation of Extracellular Vesicles
    • In this module, we will focus on the collection and processing of cell culture media and body fluids prior to isolation of extracellular vesicles. We will discuss some of the considerations and guidelines that are important during the collection of staring material and isolation of extracellular vesicles. This module will help you with the different choices, such as anticoagulants, collection time points and protein inhibitors, which are important for the outcome when working with a particular body fluid compared to conditional media or other body fluids. This module will also illustrate some example of studies on extracellular vesicles from body fluid such as blood, urine, breast milk and cerebrospinal fluid and why it is of interest to analyse extracellular vesicles from these body fluids.
  • Methods for Isolating Extracellular Vesicles
    • This module highlights some of the different methods used to isolate extracellular vesicles. Here the basic concepts for methods such as differential ultracentrifugation, density gradient, size exclusion and kit-based precipitation will be covered and some guidelines will be presented. Furthermore, this module will cover how the techniques are used in the field of extracellular vesicles as well as their limitations and benefits.
  • Techniques for Characterization and Quantification of Extracellular Vesicles
    • This module highlights some of the different techniques used to characterize extracellular vesicles. Here the basic concepts for techniques such as electron microscopy (EM), cryo-TEM, flow cytometry, atomic-force microscopy and nanoparticle tracking analysis will be covered and some guidelines will be presented. Furthermore, this module will cover how the techniques are used in the field of extracellular vesicles as well as their limitations and benefits.

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