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).
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.
What's included
4 videos1 reading1 assignment
Show info about module content
4 videos•Total 69 minutes
Introduction to the Field of Extracellular Vesicles•13 minutes
Introduction to the Course•11 minutes
The Origin of EVs throughout the Phylogenetic Tree•12 minutes
History of Exosomes and Extracellular Vesicles•33 minutes
1 reading•Total 10 minutes
Syllabus•10 minutes
1 assignment•Total 18 minutes
Introduction to Extracellular Vesicles•18 minutes
Biogenesis, Cargo and Uptake of Extracellular Vesicles
Module 2•3 hours to complete
Module details
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.
What's included
6 videos1 assignment
Show info about module content
6 videos•Total 142 minutes
Biogenesis and Release of Extracellular Vesicles•18 minutes
Mechanisms of Extracellular Vesicle Uptake - Part 1•22 minutes
Mechanisms of Extracellular Vesicle Uptake - Part 2•24 minutes
The Protein Content of Extracellular Vesicles•24 minutes
The RNA Content of Extracellular Vesicles•35 minutes
The Lipid Content of Extracellular Vesicles•18 minutes
1 assignment•Total 30 minutes
Biogenesis, Cargo and Uptake of Extracellular Vesicles•30 minutes
Collection and Processing of Cell Culture Media and Body Fluids Prior to Isolation of Extracellular Vesicles
Module 3•2 hours to complete
Module details
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.
What's included
5 videos1 assignment
Show info about module content
5 videos•Total 77 minutes
Cell Culture Media•13 minutes
Blood Plasma and Serum•16 minutes
Breast Milk•9 minutes
Urine•25 minutes
Cerebrospinal Fluid•13 minutes
1 assignment•Total 36 minutes
Collection and Processing of Cell Culture Media and Body Fluids Prior to Isolation of Extracellular Vesicles•36 minutes
Methods for Isolating Extracellular Vesicles
Module 4•2 hours to complete
Module details
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.
What's included
6 videos1 assignment
Show info about module content
6 videos•Total 74 minutes
Differential Ultracentrifugation - Part 1•15 minutes
Differential Ultracentrifugation - Part 2•16 minutes
Density Gradient•12 minutes
Size Exclusion Chromatography•12 minutes
Kit-based Precipitation•12 minutes
Summary of Isolation Methods for Extracellular Vesicles•7 minutes
1 assignment•Total 22 minutes
Methods for Isolating Extracellular Vesicles•22 minutes
Techniques for Characterization and Quantification of Extracellular Vesicles
Module 5•2 hours to complete
Module details
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.
What's included
7 videos1 assignment
Show info about module content
7 videos•Total 108 minutes
Electron Microscopy - Part 1•9 minutes
Electron Microscopy - Part 2•14 minutes
Electron Microscopy - Part 3•7 minutes
Cryo-TEM•14 minutes
AFM•19 minutes
Flow Cytometry•27 minutes
Nanoparticle Tracking Analysis•17 minutes
1 assignment•Total 36 minutes
Techniques for Characterization and Quantification of Extracellular Vesicles•36 minutes
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M
MR
5·
Reviewed on Jan 10, 2019
Awosome course. Lot of information given with paper based data and suggestion. I really love it. I am also working with Exosome. I use cell culture media from PC12 and fibroblast cell line.
A
AA
5·
Reviewed on Jan 17, 2022
This is one of the wonderful courses any scientist or graduates trying to work on EVs must take. I enjoyed every bit of the course.
S
SP
4·
Reviewed on Mar 2, 2018
Very nice nd inormative presentations. But few words in many presentations are not audible.
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