Hello, everyone! My name is Cecilia Lasser, and I'm a researcher at the University of Gothenburg in Sweden, and I'm the coordinator of this course. So I would like to start with welcome all of you very much to this course called Basics of Extracellular Vesicles. I have been working with extracellular vesicle, or EVs, as we also call them, for over nine years, and I think it's a very exciting field. You have already heard Jan Lotvall talk about that extracellular vesicles are small spheres enclosed by a lipid bilayer, and that these vesicles also contain proteins and RNA, which you will hear more about during this course. You also heard Jan Lotvall mention that extracellular vesicle is a rapid expanding field. So you might ask yourself, why are all these people so excited about extracellular vesicles? Well, this is because extracellular vesicles have been shown to have several different biological functions. First of all, they have physiological functions, where they have been demonstrate to be able to induce coagulation and participate during regulation of immune responses and in the communication between the cells in the brain, and also during pregnancy, just to mention a few examples. Then they also have functions during pathological conditions. For example, extracellular vesicles from a tumor cell can suppress immune cells to escape the immune system. They can also promote pre-metastatic niche formation. And they can, together with other factors, facilitate the development of new blood vessel, so that a tumor that is growing and needs more oxygen and nutrition can make sure that it will have the blood supply that it needs to grow. So these are some of the biological functions that extracellular vesicles have been shown to participate and play a role in. It has also been shown that extracellular vesicles that are released by a deceased cell contain different RNA and protein molecules compared to extracellular vesicles released by a healthy cell. Therefore, we are hoping to be able to use extracellular vesicles as a biomarker, so that we can detect different diseases at an early stage. We are also hoping that we can use extracellular vesicles as a vaccine, so to use them to stimulate the immune system in a patient so it will be more effective in the battle against, for example, cancer. We are also interested in trying to load this vesicles with RNA or a drug molecules so that these vesicles can be deliver these molecules to a cell. For example, delivering something to a tumor cell that will make it grow slower or even die. So extracellular vesicles are really participating in several different types of cell-to-cell communication. And you can view them as a message in a bottle that is floating around in our entire body and in the environment, sending these different signals and messages between different cells in an organism. So that's why we are very excited about extracellular vesicles and would like to share that with you. And this is why we made this course that aims to provide the basic knowledge about extracellular vesicles. And as you will see shortly, this course consists of five modules. This course is both for those of you that have never heard about extracellular vesicles before, but you're interested in learning more, and you're interested in cell biology. But it's also for those of you that perhaps have worked in the field for a while, but are interested in learning more about some of the particular techniques that you might not have used before that we will cover in this course. So what we have done is that we have asked some of the leading researcher in the field of extracellular vesicles to give a lecture about their field of expertise. So we have over 20 researcher giving lecture in this course, from North America, Europe, Asia, and Australia. So people all around the world have recorded their lectures to share with you guys. So right now we are in the first week, and you already heard Jan Lotvall introduce the field of extracellular vesicles. You're listening to me right now welcome you, and introducing you to the course. We will also hear from Yong Sung Gho this week about the origin of extracellular vesicles throughout the phylogenetic tree, so all the different organisms that can release extracellular vesicles. And I think you will be quite surprised during this lecture. Then we also have a very exciting talk from Phil Stahl, who was one of the pioneers in the early 80s, when one of the subtypes of extracellular vesicles called exosomes were discovered. He will talk about what the field looked like at that time, and the research questions they had then. And what did they knew when they discover exosomes, and how the field has evolved since then. Then in the second week, Suresh Mathivanan will start by talking about the biogenesis and release of extracellular vesicles, so how they are created. And then David Carter will talk about how these vesicles are taken up by the recipient cells, so that all these messages and molecules within the vesicles can reach another cell after they have been released. Then we have three talks about the cargo of vesicles. So they contain proteins, RNA, and lipids, as I mentioned before. So here, you will hear from Suresh again, but also Andrew Hill and Edit Buzas, and they will talk about what kind of molecules are present in extracellular vesicles. And what can they do, and which techniques can be used to identify them? During the third week, we will discuss what you need to think about when you want to start your research on extracellular vesicles. So there are a couple of things that you have to think about if you're going to use the media from cell cultures to isolate extracellular vesicles from. So I will talk about that. You can also think of how different blood is from breast milk and urine, for example. So depending on which body fluid you want to use to isolate extracellular vesicles from, there are different things that you need to know and consider, because the different body fluid needs to be handled differently. So we will have a talk about the collection and processing of blood by Ken Witwer. Esther Nolte-'t Hoen will talk about breast milk. Lesley Cheng will talk about urine. And Julie Saugstad will give a lecture about cerebrospinal fluid. In the fourth week, we will go through the different methods methods that you can use to isolate extracellular vesicles. So now we have your cell culture media, or you have collected your body fluid of interest, and you have listened to all the lectures in the third week, and you know how to handle them, and now you want to isolate the extracellular vesicles. So I will start by talking about differential ultracentrifugation, which is the method that has been most used so far. Then Su Shul Jang will talk about how you can isolate extracellular vesicles with density gradients. Rienk Nieuwland will give an exciting talk about size exclusion chromatography, which is an old technique, but it's new in the field of extracellular vesicles. So he will talk about how you can apply that technique to isolating extracellular vesicles. Then An Hendrix will talk about kit-based precipitation, which is also a method that has been increasing in popularity in recent years. And then in the end I will do a summary about all the isolating methods that we have talked about and discussed during this fourth week of the course. And then we come to the last week, week number five. So now you have isolated your vesicles. You have picked the isolation method that you think will work really well for your starting material. And now you want to characterize your isolated vesicles. And you want to understand what do they look like, how many are they, what do they contain? So in this week, first Johanna Hoog will talk about several of the different techniques of electron microscopy that can be used in vesicle research. And then Alain Brisson will specifically talk about cryo-transmission electron microscopy. So extracellular vesicles are only a couple of nanometer in size. So they're too small to be observed with a regular microscope. Therefore, we need these techniques to be able to visualize our vesicles. And then Shivani Sharma will give a lecture on atomic-force microscopy. And Marca Wauben will be talk about flow cytometry and how they have adopted that technique so it can be used for the analysis of single vesicles. Since this is a technique that was developed to analyze cells, and as you have heard already many times, vesicles are so much smaller. And then last but not least, Chris Gardiner will give a lecture on nanoparticle tracking analysis, and how it can be used to characterize and quantify extracellular vesicles. So during module four and five, we will discuss several different techniques, and we will describe the principle behind them and how they are used in the field of extracellular vesicles. We will also discuss the limitations and the benefits of each technique, to try to give you a good understanding of the different options that you have. So join us in this exciting world of extracellular vesicles, and we hope that you will enjoy this course. Welcome!
