This course aims to provide the basic knowledge about extracellular vesicles (EV) a generic term including exosomes, microvesicles, microparticles, ectosomes, oncosomes, prostasomes, and many others. It covers areas such as EV history, nomenclature, biogenesis, EV cargo as well as the release and uptake mechanisms, collection and processing prior to isolation, different isolation methods, characterization and quantification techniques.
This course is divided into five modules. Module 1 is an introduction to the field and will cover the nomenclature and the history of EVs. Module 2 will focus on the biogenesis, release and uptake mechanisms of EVs as well as the different EV cargos (RNA, protein, lipids). In Module 3, we will focus on the collection and processing of cell culture media and body fluids such as blood, breast milk, cerebrospinal fluid and urine prior to isolation of EVs. Module 4 and 5 will present different isolation methods and characterization/quantification techniques, respectively. Here differential ultracentrifugation, size exclusion chromatography, density gradient, kit based precipitation, electron microscopy (EM), cryo-TEM, flow cytometry, atomic-force microscopy and nanoparticle tracking analysis will be presented.
The recommended prerequisites are college-freshman-level biology and biochemistry.
After a completed course you should be able to:
+ Discuss the nomenclature and subgroups of extracellular vesicles.
+ Describe the RNA, protein and lipid content of extracellular vesicles.
+ Describe the basic concepts about the most common isolation and characterization techniques and how these techniques are used in the EV field.
+ State the benefits and limitations of the most common isolation and characterization techniques for extracellular vesicles. + Explain the considerations that are important during the collection and isolation of EVs from different body fluid.
+ Describe the release and uptake mechanisms of extracellular vesicles
All lectures are given in English.
Each of the five modules will be followed by an exam. All exams will be in the format of multiple choice questions.
The course is organized in collaboration between the International Society for Extracellular Vesicles (ISEV), University of California Irvine (USA), University of Gothenburg (Sweden) and Pohang University of Science and Technology (South Korea).
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.