 Thank you everyone. Hi, I'm Radhika. I would like to talk to you about how to make a mini-brain tissue in a dish So to do this I will go back a few maybe a hundred years And talk about this Spanish neuroscientist called Santiago Ramonica Hall he is widely known to be the father of neuroscience and apart from making some really beautiful Drawings of the the cells that make up the brain He also came up with some really nice philosophies so he said that as long as our brain is a mystery the universe the reflection of the structure of the brain will also be a mystery now that's a really romantic thought especially for somebody who wants to study the human brain and Me and other neuroscientists and there are many other people who would like to do this How do we do this? How can we learn how the brain works now? Usually you can determine the function of something by looking at the structure of what it's what it's made of what it's Composed of so you can open it up and look inside Can we do this with a human brain? It's possible you can get some brain tissue, but this is really difficult and Human brain tissue is limited Inavailability, so what do we do? Yeah, that was not a joke, but it happened to be a joke, but anyway So we go back to the philosophies of Ramonika Hull and again 100 years ago He said that any man could if he was so inclined be the sculptor of his own brain now to be fair I think he said this more in the context of brain plasticity and how it's adaptable But I think a lot of people try to look at it as why not start from the scratch Okay, we cannot open up a human brain. Let's start making it from scratch bottom up So can you do a do-it-yourself brain? This actually exists by the way there's a Lego brain skull kidney kind of puzzles and So this is sort of the premise. This is what neuroscientists are trying to do and what they're trying to do is make a Brain from the beginning so like trying to put it together like a puzzle like a Lego puzzle So what are the Lego blocks here? What are the building blocks? so we start with something known as human stem cells which I will talk about a bit more in detail and These are the building blocks of what will finally become the brain So think of it like this now you actually have a system where you can follow these building blocks or human stem cells and Build a tissue really sell to sell block to block. So that's really amazing and that's something which has not been done before So to start with this I have to go back to the basics a little bit To introduce you to some concepts and how we started So what are human stem cells? So these are a bunch of cells which have two functions one is to self-renew they constantly keep making more of themselves and Second is to give rise to all different types of cells in the human body So they can give rise to brain cells skin cells muscle cells what you may now where how do we get these cells and Also, how we how do we start like how do these cells come into existence? So we start as a fertilized egg This is when an egg and a sperm come together This is a zygote and this is how all of us start and we go through many cell divisions We are then two cells then four cells and sixteen cells and finally a bunch of hundred cells and this is a very important Stage I must say I mean you go on to become an adult or a baby But this stage is very important also for scientists because this is the stage where you can get stem cells This is the stage where you can get cells which can then become any other cell type in the body so of course scientists realize this and They used cells from the aborted fetuses. So you could get these cells which are Then becoming the stem cells that we can grow in the dish in the lab You can just isolate them and you can grow them in the dish and these cells in the lab Also, just like in the human embryo, they give rise to many different types of cells These cells then organize to become tissues and tissues become organs and organs become a human body This is great, but of course this is coming from an aborted fetus. So there is a lot of ethical issues around it It's not something that scientists can do very freely or easily Is there a better way to get these stem cells so that we can sleep better at night and at lab? so Once again textbook knowledge humans stem cells give rise to all of these specific human cell types These give rise to a human body now This is what we know and we know that these these cells stay as they are So if you are a blood cell you are a blood cell forever But now came a Japanese scientist Shinya Yamanaka who won the Nobel Prize and he told us no This is not the case you can change the identity of a cell which again is a an amazing discovery in my eyes because Imagine that you can take a blood cell or a skin cell and then add a few factors Mumbo jumbo magic and then you can make them stem cells again So this is really like the curious case of Benjamin Button for those of you who have seen the movie You can really take an adult cell put it back into an embryonic stage and that's amazing because Now you have a system in the lab without having to get embryos or fetuses Which you can then use to make any kind of cell which is what happens in the body So this is great. So now that we are equipped with this knowledge that we can make human induced stem cells They are just called induced They are the same as the stem cells from the human but they're called induced because we get them from a different bunch of cells So now that we know this We can now go ahead and look at the recipe for making mini brain brain tissue in a dish It does seem like a lot of voodoo to be honest the first time I read it I'm like I'm I can't do this like what is this? How did people come up with it? And it's really pretty cool because the scientists there's a Japanese scientist Yoshiki Sasai and two scientists who are currently in Austria Madeleine Lancaster and you're getting you get no blush who came up with this so-called cocktail or the Mechanisms or a perfect protocol to make mini brains in a dish. So let me run you through this This is their work and I would like to simplify this the best I can So there are some things that you need of course so the first thing is a cell and tissue growth medium So think of it like an energy drink that the ones that we drink like Red Bull or something Which has many supplements and these help the cells to grow and reproduce and then there are Growth factors for specific parts so you can really take some supplements to augment certain parts which you want Sorry, and then of course infrastructure. You need to do this in a lab. This is not something you should try at home So, okay, how do we start like I said we have these stem cells and these Stem cells are usually they are stuck to the dish. That's how they grow They are attached to the dish so think of them as a bunch of people who are sitting in a bar They're sitting they are relaxed. They're drinking beer and they're sitting so now How about we remove them? So if we remove these cells from the dish, what do they do? Automatically they come together they clump up Just like people if you ask them to stand up if I asked you all to stand up now You will automatically form groups to people you like or you make interactions with that's exactly what these cells are doing They once they are apart from the dish. They start talking to each other They have interactions and they come together and they form a group or a ball of cells To this we now add all the energy drink which has the nutrients We add the growth factors and then it becomes a Well-organized ball of cells. Okay, so now it has some sort of organization and it's growing we need to make it grow further and To do this we try to mimic the way it grows within a human brain So we try to give it some sort of a structure or a scaffold so it can grow better and You can imagine this as jello shorts. I guess everybody has done jello shorts So if you put like a cherry or a plum or an apple into gel into the jelly it can set and it really It's exactly what we are doing We put these bunch of cells into a jelly like substance a hydro gel and we let them grow further So this helps them get nutrition and it also helps them grow better So now they grow and they become even bigger floating balls and then finally they become this which is what I call the mini brain balls and They grow up to a size of up to three to five millimeters and this entire protocol from day zero to the day We get this is approximately a month of course you can keep them in culture longer in the lab we also keep it for up to four to six months and There are labs who also keep it for up to a year. So they grow and mature much more So this is what they look like live in action. It's slightly skewed the video. I'm not Greater doing this video stuff. I can grow these things, but I can't take videos Of it. So this is what it looks like So now of course now that I've shown you what it looks like and how they have made you will be like This is not a human brain. Yes, of course. It's not a human brain So again, I have to introduce this and as a cautionary approach that I need to explain what There is a difference between an organ and an organoid like there's a reason why they are called organoids So this is the scientific term for many brains. They're called brain organoids There have been of course not just with the brain people have been curious and they've made many organs of many other organs So there are many hearts many kidneys many eyes. There is a boom in this industry. Yeah, so To just to Bring home this message an organoid or a mini organ is basically a miniature and a simplified version of an organ It is produced in vitro vitro is glass so in the lab and of course, it's three-dimensional So it is useful. It's much more useful than a two-dimensional culture and it shows the microstructure of the organ that you're trying to mimic and Since I'm interested in this and we are talking about this just another picture of these brain organoids So once again, they always a brain organoid is a mini brain. This is what we're referring to here All right, so how do we know that this is really a good system? We need to what are the similarities between a mini brain or an organoid and an actual human brain? So there are many similarities. I'm just going to touch upon a few. So this is An image of a section of the human brain and this is an image of a section of an organoid You don't need to worry about the colors or whatever But I think anybody who looks at this can see that the structure the way the colors are organized a very similar in both Of them which goes to show that these colors basically represent some proteins which are marking these these structures and these proteins are very similar or the Components of these both structures are very similar in the human brain and the human organoids and there is a lot of data with respect to genetic genetics which is also similar and the timeline of development and the kinds of cells produced so Basically, they're very similar to the human brain in its embryonic development So of course if we study the function of something it helps us study the dysfunction of the of this organ or the structure So to study the dysfunction of the human brain. There have been many different ways to do this So absolutely worth a mention is animal models there is no way that we can get anywhere in neuroscience if we didn't have animal models and Then people started using some 2d culture models where they took cells from the human brain and tried to culture it And this is a bit difficult and it's possible, but of course it it's not as knowledgeable it doesn't give as much knowledge as More in vitro or in vivo systems And then now we have arrived at this 3d culture models, which is what I'm talking about the mini organoids with mini brains So here we really until now like I said We've always been studying a human disease or human neurological disease in the in a mouse model or a primate model But now we have this unique technique where we can study human brain diseases in a uniquely human system and Of course since we can do that we can also reduce the number of animal experiments We cannot do away with it, of course, but we can reduce them Which is great So I would like to give you some examples of how human brain diseases are being studied with with these mini brains So just one of the many examples is I guess you've heard of the Zika virus which Caused a lot of problems in Brazil in Asia and Africa This is a virus that infects cells within the embryo so in a pregnant woman and It causes neurological damage and even death So what people have now done is use the Zika virus and they have infected Mini brains with the Zika virus and what they found out was that there is a specific kind of cell which is damaged because of this this virus infects a certain kind of cell and they also found that in one of the Symptoms of this disease is that you have a smaller brain some microcephaly compared to a normal head So the children have smaller head sizes and they also could show this with the organoids So what they did was they made these mini brains and they with the they took cells from the patient and this This saw that the mini brains were even more mini So they could really actually recapitulate what was happening in the human brain and also study a very human specific Mechanism to see that there are certain kinds of cells which are killed specifically by the Zika virus Which is really good for if you want to try out therapy to know these kind of cells and also they found out that there There were some DNA damages etc. So this is obviously a very useful technology to study something specifically human and Just to give you an idea there are many other diseases like Alzheimer's brain tumors schizophrenia, etc. Which are currently being studied with these with this technology and This is also to emphasize the fact that the rise of the organoids it sort of happened around 2012 2013 and already by within five years There's a lot of data with respect to a lot of diseases So what is the future of this technology? Of course people are trying to do some more cool stuff with it and since I spoke about disease modeling and studying of Human diseases what people are also now doing is that they can actually now put These small structures onto chips and they can do drug testing so that you can really put the the brain organoids from a Normal person and a patient and you can compare how the drug works on both of them. So this is really cool this is stuff which is happening in some companies also in universities and People are also trying to do some other cool stuff where they try to put make two different types of brain tissue So brain tissue from maybe a right side of the brain or the left side of the brain and they try to put it together And see how they interact with each other. This is again something which is pretty awesome But yes, okay. This is a slide full of references. Just just saying I love Shakespeare and I have to mention him somewhere As scientists we have to always call a spade a spade. It's really important to not oversell what we are doing We're not making a human brain. So when I keep using these this word mini brains It's something I do to make it easier for everyone to understand But I don't know if anybody knows this. This is a really cool movie. Watch it. It's called Princess Bride. Princess Bride Yeah, so yeah, we have to be very cautious when we use this word Because just to give you an example, this is a fly brain. This is a mouse brain and this is a human brain Where are we now? We are somewhere here with the organoids just to highlight this point but We can make it better. Of course every technology. I mean, this is really this technology is in its infancy It has just been born. So can you imagine what can be done with this? One of the things which is being already done is that so if you have seen the structure of a human brain if you just go Back here, you see that they have all these folds and you know, these are it's a very peculiar structure And this is you don't see that in the mini brain organoids So what people are now doing is really doing a lot of experiments to make this a more folded structure Which is really more authentic so to say and what they're also doing is right now Trying to put some other cell types like there are cells in the brain Which are called glial cells or the cells that keep the other the neurons together Which also have a lot of functions and the blood cells. So they're really trying to make it more complete in terms of a brain tissue So what motivates us just to come to the end I Think so there are a lot of different theories to how to interpret this Painting and I think Michelangelo was trying to for me. He was trying to show a human brain here. There are many other Theories of course, but of course as a neuroscientist I see a human brain here and I feel that it's really Really important that stem cell biologists and neuroscientists coming together can really give rise to what I call the creation of the perfect organoid So These okay, by the way, these are the organoiders. I would say so this is Marisa Karoo I work with her and this is Benedict. Unfortunately. He didn't come to the school event So we don't have a picture with him But we work here at the LMU at the biomedical center in Martin's Reed And you can always drop by if you want to have a chat or ask any questions or see some organoids Yeah, thank you So, thank you very much very interesting. I love the idea of floating mini brain balls I think someone's used that as an insult to me on more than one occasion Throw the floor open to any questions, please as a hand shot up the very very back was the first No, we can't say that and I don't claim it of course you're studying something in isolation This is one of the biggest limitations of having in vitro cultures or anything which you're doing in the lab because you you cannot get I mean the same kind of results that you do when You study something which is a complete functioning human being or a complete functioning organism But considering that we don't have another option to do this in this is really the best we can do right now Okay, thank you next question Okay, so questions to people try and investigate brain behaviorism in the organelles I would say the technology is quite naive right now for this kind of Application so first of all yes, these structures do have connections so they do make electrochemical connections there are The unit that builds the brain so there are neurons inside them and they do fire action potential So they they talk to each other there are connections, but It's you have to still remember that it's a five millimeter sized tissue It's really small and it does not have all the connections that is required to what you are saying, you know to study behavior You know consciousness, etc. It's really far away from that really far. Okay. Thank you What is the group factor? Oh, maybe can you get it to induce the same to be Okay, so where do we get these growth factors that make the stem cells turn into brain cells? there are many different growth factors and it really depends on the kind of Brain structure that you want so for example if you need In the beginning you have to give them a certain set of growth factors which are common so you give them things like BDNF GDNF which is like brain derived neural growth factor and there are different. I don't know I can name these things but The after which you have to give them more specific growth factors depending on what kind of brain region you're looking for there are people who make like hippocampal organoids a very specific region of the brain or mid brain organoids or hind brain organoids So then depending on that you have to give them different factors It's a difficult to get them This growth factor, you know, it's not difficult. They're kind of expensive But yeah, it's not difficult as long as it's known what the growth factor is you can get them Yeah in a lab environment. Yes, you can get anything with money. Yeah It's question here to your opinion or to your Knowledge doesn't require more than growth factors to make a Functioning brain that is able to process information. So do we need more than just growth factors to make this thing work? Absolutely Yeah, absolutely. I think this I I mean I had this on a slide But I think it's very important to know that to make this tiny five millimeter structure You need you need to know neuroscience. You need to know stem cell biology You need to know bio physics. You need to know the material material science because there's so much dependent on the kind of dish that it grows in you need to really understand a lot of these Sciences to make something even as small as that so it's not just a addition of growth factors or giving them whatever energy drinks There's really how these cells interact with each other I mean, of course, I haven't sat and done all this This has all been done by scientists before and I think it's really important to To emphasize the fact that it's really so many different sciences coming together to be able to make such a Small system even so you have to understand many different aspects of it How these cells interact how they interact with the dish how they interact with each other and Many other things like that. I think I think one last question then we'll have a break question Are there any other applications of these? Or no, it's for example, like using it as a computing power Questions can be used the other noise and anything else in robotics Consciousness, I don't know if you see any application in like your future Consciousness I Would say it's kind of like the answer would be similar to her the answer to her question because I don't think the organoids are there yet To be able to study some things so complicated as consciousness. I mean, we don't even know what consciousness is with the human brain Completely in its, you know full capacity. So I think that's really really naive at this point But maybe yes, there's a point where you can really mix computational systems and Stem cell biology and some get somewhere close But right now it's really difficult to and unless you really get a structure, which is so complete It's very difficult to study behavior and consciousness We are really looking at this system as something which you can study very early stages like very early stages of growth When the embryos developing and there's so many diseases which happened during that stage Which cannot be studied really well and the system helps more for this Rather than for studying something like consciousness to my to the best of my knowledge and of course Maybe there are many people working on this right now. So Great what I know. So thank you very much indeed. Thank you