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>> I Everyone thank you so much for attending this new you know I check someone are My name is David Welch and I invited Dr Lenz the on Penn State University to present today. But it's a today and I have the pleasure of introducing Dr Lance the I was it was just a professor in the biomedical engineering and biology department at Penn State University Dr Leon received his Ph d. in chemical engineering from the University of Wisconsin Madison and when he was well and during his Ph d. Dr Leon's cardio my as I differentiation from human play stem cells paper was awarded the best by medical paper in t.n.a. s. and it has a really prize of the National Academy of Sciences and 20 Well actually under his post-doctoral training at Harvard University and paralysed Institute for stem cell research I joined Penn State in 2030 and Dr Young developed the world's 1st think reality cell differentiation method for stem cells for shooting I.V.'s only small Monfils which makes its 1st production much more cost effective and efficient Recently Dr Leon received the any stroke was or were the n.s.f. career ward and the Biomedical Engineering Society Rising Star award without further ado I give you a free lance the.

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Thank you. Very well 6 o'clock for this opportunity and today I'm going to talk about some of my walk being the human stem cell area especially using these cells on treating Katic diseases. The 1st of all I want to give your introduction about my Lab Now you might have we have 2 key technologies winds the stem cell technology the other one is ging editing technology so we capture these 2 technologies to generate an in-car clickable human cell types for therapy and we tested these cells in animal models so that's why we have 3 areas listed here to the Co for my lab is we want to use stem cells to understand human embryonic development and we want to develop a cell path to Serapis for treating diseases so 1st of all I want to introduce the type of cells that we are working on.

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So these cells are code Indios to pre-board in the stem cells. So how how do we get these cells to conceive from the left so we can see from our normal human embryo development that we have the so-called embryonic stem cells and these cells have the potential to differentiate into all the symmetric cells in our body such as our neural cells such as our skin cells such as our cardiac muscle cells.

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But the idea of. Human development is. Can be reversed. Onet introduced a by a truck to see I am an actor in 2006 Now previously resort development is a long way process. You cannot reverse the development but in the year 2006 up to the area manakin from Japan he introduced a message that if you can convert for a difference in a cell such as our skin cells back into the embryonic stem cell like cell the he caught or the self induced a proponent of stem cells or i.p.s. east so there is a fascinating idea and in just 6 years later he was awarded a Nobel Prize in Medicine in 2000 jobs.

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To their idea here is we can we can take the skin biopsy proud and have the audience here some of your blood cells all even your toes from your urine samples to also get the cells we introduce for critical genes into the cells to hear these genes are called out to care for stocks to and Simic because this cocktail was 1st discovered by talked the argument aka the they also caught a Yamanaka factor.

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The want to introduce these 4 genes into the in cells the skin cells will be come over to the back into stem cell state so this is more like a turning back the clock in the cells or imagine the cells are coming from a person who's 30 years old and there you can convert at the South back into the.

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Andra like stem cells so that's more like a turning back the clock in a sense that's a pretty amazing technology to watch you got these i.p.s. cells. Their behavior like embryonic stem cells they have the potential to become all the cell types in our board such as well that cells are Excel neurons out and append graphic cells so these cells are very useful for treating diseases for example corrective car exam was a very useful for treating my garden function and the pen graphic cells are very loose for for treating type one diabetes.

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So I'm going to give you one of the example that we use this technology to generate i.p.s. cells so these cells are actually. The cells that we isolated from human urine samples so there are some symmetric cells residual cells from the urine samples and the cells once you culture in a pitcher dish they can expand a lot so we can generate that the human urine samples and then we introduced for human the Kajang being true to your example cells.

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And you can see 2 to 3 weeks later cells completely change their morphology to they look like this before but now they look like a nice cat to say from night colonies and awaiting each colony we have the south end of cells parents of cells become very tiny and the dead come back together so these other human i.p.s. they can see this technology indeed a walked in our lab and we can generate these human i.p.s. cells.

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No next to the question is was we generated these human i.p.s. cells out a way for that differentiated the cells into cardiac cells protruding contacted this path to them. The reason why we are interesting product is it is because a vascular disease is the leading cause of death not only in the u.s. but around the world so we can take a look at.

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A typical example of our failure per se so. Let's say there is a healthy person with healthy heart and coronary arteries Ok so everything is fine. But it could be due to some genetic risk factors To the healthy time this person may have plaque formation within the blood vessels now wants this become most severe which were blocking a blood vessel to want you have the patient have a blocked heart or muscle hard a lot of vessels in the heart of muscle cells here because they were not receive enough nutrients and oxygen these cells were dark because of this there would be some remote owning within the heart and eventually this may develop into a heart a failure.

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So this is a very very severe problem. So our idea is things least muscle cells within the heart are dead because of the blockage of the blood vessels here so how about we replace a systemic your region with healthy a lot of vessels out and also healthy Kartik muscle cells the one way we can do this is we can use.

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Them South economics. In order to generate generated human particle muscle cells that we have i.p.s. cells so the idea is they are mimicking embryonic stem cells so then what do we need to do here is you have these cells and how do you give the cells the correct signal to kited them to become the muscle cells.

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So what about our strategy we try to learn from the human Amber development so the question is out how does a mouse make their heart in the in the in animals and so when it will under this animal development from different and. Species those here press you can see a typical example of Amber development here so what we can see here is at different time points these embryos tragedy I'm of knowledge a lot so what we learned here is development is a dynamic process so at different points you had to give different signals to kind of the Predator development and.

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Because of this temper regulation is a very important. So you can see with things here at this stage you already have Peter hot here this is the part that we want to acquire the i.p.s. cells to become and then if you see the mouse it's pretty much similar but seems more complicated and you can see here at different time points the embers were developed.

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So if you look into it and it's extremely complicated there are many many factors involved in this process and if you try to mimic 100 percent of this invisible Amber development you have needed to test a lot of factors at different a concentration at different times points so if you do a simple mass of these combine Asians that are erode more than opinion combined ition So in reality that's not possible for you to test in a lab and no lab can test womp in a combination.

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So my logic would be. Can we identify a single developmental part way out which could to ensure stem cell differentiate into cutter mice so chaotic muscle cells. So when I 1st to do this project and I talk with different p.-i stiffer and ph students no one knows how to go this project because it's completely new and I know has ever differentiated themselves into cutting myself that's being extremely difficult and when I check the Selsey going pas ways.

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I know all of them are important you know in certain ways to guide us and bring it to become the harder region but my logic is this I couldn't offer this a medical I test in all of them How about I choose one of them and a test.

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It's a regulation if that credit card the stem cells to become a muscle cells so I decided to choose this winter so it's going to pass way. So the reason I'm interested in this is because you can see this when pas way the blue region share mocks the activity of this there was one possible way if you see jumper to signal that meanings in this region in the amber is when the pa is all that is active and if you see here it's less pollution here that means we deposit is turned into cats so this tells me.

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That a lot of time ponds in the hotter regions you know when deposit it is all that at a later time point when the Postle is is off so so that means between the other time points and they did I'm going to we have to shut down this possible path to the general idea is it had to turn it all and the some time later you needed to turn it off so we are trying to mimic of this and to see if it's going to work.

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So next I'm going to give you a little bit of background about what is possible. So this is us now this is a human cell and on the cell membrane you have some cell risk receptors so this receptor is called. The receptor so this receptor have the capacity to bang into winter like and this is a parting that can activate when the cars were at the wall so when printing binding to its receptor which we'll.

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Say if one of the keeper team here could have been acting as the sped up to the imprinting once it's safe is it can't enter the nucleus and her own Jing expression. So if we do not have the wind printing. This just gets really paid our kindness will degraded Penticton now in this way there would be no productive available for entering nucleus and they cannot her own gene expression so in that case it's core to when to off Ok.

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Because each year involve the. Full protections once a winter like and winds of mishaps are wizened just can't repair a and want to predict in predicting is the most important because we rely on this pretty to turn on the wind possibly was predicted and to the new Chris and I can tell on the pa.

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So based on this cartoon. We can develop several ways to change it all or trade off. So why am I to take it all and if. We can't directly and use a chemical. To inhibit it just gets really bad as long as you can put auk just because we better you know we are not be able to degrade it a bit acting in this way you can turn on when pos way that is.

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This is. Here if you are in combat just because repairer with chemicals now you can do bio or see a trial 9 I own 2 was the later I call this chemical is the edge for the edge in this where we can tell only pos way. If you want to turn tongue off when the pos way why when you can do it and you know this pos Why rely on winter lichens you can directly block with the large and processing that there is no when the like and then you can try to.

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You can't directly a degree day to the conduct in the invite technology there is a technique that you can design a very shot. And then the shuttling it we are binding to bedrock you knowing it and I do have debated about a Christian. If you destroy the doctrine and there is no more productive and to take in all of their fall and I were turned off when possible.

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So there's another approach that we can do as we can construct the product in a binding site in a genome and then in this way we're going to know whether this puzzle is on or off. No I did it to the 1st experiment I did. I inserted a genetic construct into the.

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Stem cell. So if that we deposit where is turned all those were expressed Gippi Ok because we have become green telepathic and I when I couches these cells normally there is no green cells so that means normal human stem cells they do not turn on when possible paths and then I use this chemical to the edge so this is a chemical here to try on one iota to uses chemical to treat a stem cells Indeed I can turn on the when to pass away and some of the cells become going.

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After a trip to the cells with less chemical to activate when possible and if I treated cells with this chemical for 3 days and you can see I can generate more Gippi else if you choose I want to follow only wanted only 5 percent of the cells become green and if you try to force treated as 40 percent of your cells become green and the green cells.

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Fortune to lead the greenhouse indeed. Differentiated and then become addict presented itself so that's a that's a very very good good news for me because our hypothesis is if you activated the way the pos way stem cells will further develop into cardiac cells and indeed we see this phenomena.

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Next we want to test our ideas in the previous year and best of differentiation do we want to see if we capitalize when to activation what that enhanced our ability to generate more cut excel at the here we treated ourselves with this chemical to activate we deposit and then we do the differentiation so the y. axis is a percentage of the beating E.B.'s so p.b.s..

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Themselves clusters tell us we have no printing cells so the beating cells are cut of muscle cells and no other cells in our bodies were spontaneously Pete packed so you can see if we do not treated ourselves with chemical. We only have about 10 percent of the cells.

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They're. Getting cells if we treated ourselves with to Michael Moore or you can see now we have about 25 percent of the cells become pretty prison cells Terry indeed if you treated us as we said chemical. You can increase your ability to generate smoke at Excel. So that we use another protocol to test the idea we use the edge of bio to trigger the cells 1st before we do differentiation you can see in the control you have only have about 5 percent of the cells are caught except hope so that city is a printing own expressed in kind of muscle cells if you treated the cells with the edge you can see now you caught around the 50 percent of the sas becomes cardiac muscle cells so it's a great deal of increased efficiency.

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So what about. We locked up in a container you can see here we use Paetec strategy to. The created. Product in engineering so if you teach you a bit acting and you were shut down when possible who can see if you shut down when to pass when at the time points for example there is arrow you get almost nothing and so there is no kind of much at that you can generate to this child as we deposit it at a very early stage of differentiation is a very very important you should never shut down when the pos went at his or the time point but didn't see it as a little bit a little high Pons you can shut down the pos when and you can increase the efficiency Ok so the idea here is if you want to generate a cut of muscle cells at the other kind parts you should have time on the pos way instead of just on the possible.

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And then add a little bit later time pongs you should have shut down the possible Ok so I did all and then later turned down the about to generate a customizing to test of this idea and then without using the other signals to. Kind of stem cell differentiation we decided to test.

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A complete a new protocol. So this project is hauled out as early as one point let's say I did pay the arrow we started to travel when the pos went and at a little bit later time point we are going to shut down the pos well but we don't know exactly as a pa to shut down the possible.

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So that's why. We decided to test a different time point. To shut down the pathway to see which time point to give us the best result of. The idea here is we have stem cells here and there we culture the stem cells for 5 days and a 5 day later we call things hero we started to trigger the cells with a chemical called a c h 2 activated the wind pathway and then we use this track talks to shut down wind partway at a different type on so we shut down the pathway on the idea they wanted to say 3 to 4 so we could try all these different compounds and then on to a 3rd we're going to se the percentage of cardiac muscle cells that we have we use a marker part in court to conduct support of the city entity so that's pretty old expressed in.

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Kut excels at the 1st what we find is if you use shutdown with parts where there is 0. You have minimal cut of muscle cells so this is consistent with what we know before you show the never shutdown when to pass when and or at one point if you want to generate a correct myself.

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And then we test different time points to surprising and we find the best time point actually is something here they want point of fact. 36 hours later. So if you add to the docs to shut down with the possible. That they want point 5 you would generate more than 80 percent of the cells to become part of muscle cells so they fissions is extremely high you know at that time point.

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No one can generate of this high efficiency. So this service exile for us is a very tricky time point. It's probably also the reason why no else has being able to discover this you match that's why you do the experiment 2 pm In the after no on their 0 so they want to 5 will be the next day I too am in the morning and you have no one to experiment to am in the morning the reason we discover this is because we test all the time points between days here and there too and we test every hour so you know I by that time I was a Ph d. student at the University of Wisconsin and I have some of the greatest students work with me.

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In chemical engineering and they have a lot of homework to do so they didn't stay overnight there so I just offer them to help me carry this truck every hour you know then you have to go to a lab every hour and it's a little bit shocked to shut down the window possibly So we are very fortunate to discover this unique time point so this is a very very difficult to discover so then we use a more potent.

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To trying it to shut down when to pass when and if we can give a generic to 98 percent of the cells become chronic muscle cells so it's almost all the cells become article muscle cells so this is also for the 1st time many pill a sion of Wong So stick to coding pas when.

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Has your a strategy for the generation of relatively poor population of customizing and so that is an image of the cells that we generate they can see almost all the cells here express the kind of components is. And are these cells functional like a normal human heart of cells so they show up ventricle action potentials and the beating rate is like our human beings can do their bit about.

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The extent to 70 times per minute and then at me show you know one of the videos that we generated so you can see these cells that connect to each other and they're form the speeding waves this just like the ocean waves but remember these are human cardiac muscle cells and if you don't have the stem cell technology it's almost impossible to get this amount of human Kartika muscle cells because no one is willing to donate their heart for research so this is this is for the 1st time we generated a lot of human contact much of that so we did a little bit further calculation of the cell the on the top this is swung single cell Ok so you can see you only have long to see a nucleus here but this is just a want to go to human contact muscles and you can see we have very very nice sock Amir structures here.

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And then this is like a morally ourselves. And there we did scanning electron Michael Graffy so you can see clearly and see the. Structure within the. Article myself the 1st you see these are my father's So these are the my fibers structure and then you see the leaves and you see the deep and so these are the typical structures you can see the ng how to muscle cells and then you see a lot of mitochondria you see a lot of mitochondria.

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More than other cell types because customer cardiac muscle cells and the people all the time they consume a lot of energy so if you check what's going on in this. In this lab stem cell differentiation as compared to the in the wall so what we are. Seeing here although these i.p.s. cells differentiation was down in Alaska but they mimic very well of what's happening still doing the amber development Ok the cells are starting from.

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Stem to our state and then they go through the so called a medical. Cardiac measure the an early cut of my sets and a later cut of mice and. So there is a very interesting night. In beach or lab experiments mimics what's happening in the amber and that's why we can use this stems a motive to start a human embryo development so you know for your information it's extremely difficult to start a human and ember development so you specialize for the 1st 2 mouse wanted to mouse because embers are so tiny and there is.

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We have a little to us Paul. Imaging of the own Amber development that you just cannot see it by the time part but with the stem cell technology you can use our cells and account of the cells in the lab and then you can start it what's going on in embers.

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Another important thing I want to mention is the I p s l technologies these cells look exactly like embryonic stem cell part we never use an amber materials there is no Amber materials at all here so you know then there's no ethical concerns here because what do we do is we isolated as all persons human cells cells from you example there's no Amber at all we never use any human ambers Ok there is no ethical concerns here.

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And I was with generators these. Are my sides and we cultured them for a long period of time so very very interesting that these cells can maintain their spontaneously beating activity for more than one year we can culture cells and they are over a spit in the air and even after one year of this stupid.

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But I stopped my experiments after one year it's not because they don't you know that I don't something it's because. I have to graduate from my Ph d. degree and I I'm not going to feed them anymore and so that's why I say you know I stopped the experiments but they looked very nice even after one year of college in the lab Ok you know if you call yourself for a longer period of time the cardiac muscle cells become more mature it's just like you know say I was hearing the amber development Peter kind of my eyes I saw him mature and then once a baby was gone and then become an adult person muscle cells become more mature the 17th year if you culture cells.

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In the lab for a longer period of time tell us who have become more mature the technical stuff at the how do we know they become more mature cells it's because the mature cells expressed of this maturity are pretty in court a message would be you can see the percentage of the cells expressed this and message of in the gray areas are increasing during the time.

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And then the other feature is to help lose the most must act to become more mature you can see the percentage is also increasing doing. So there were no cells become more mature. And another feature of mature cells is much much house where Stop Pearl information and you can see the appeal and I can 67 as the marker will turn to freighting cells you can see they are losing these proliferating markers that means they become less proliferating and that means they are more mature.

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So then next we decided to develop a technology that we can use. A chemical to provoke when possible and instead of using. The modeling it to do it because a small Einar requires you had a commodification of the cells from just you know off a screening with several chemicals Wayfinder that's a chemical called of the piece this chemical.

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Can be used to think if it's something to pass when you can see if you use this chemical. This is use a chemical approach to a fission that is generating the percentage of customizes generated as pretty much similar like a used Jing approach path now it depression of the winter production is as effective.

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As an act in a knockdown for generating kind of mice and the other to think about now the protocol has simplified with just the 2 chemicals the 1st chemical as courtesy edge which were activated when pos way the 2nd chemical is called either of the p. which will inhibit it when possible and if you use these 2 chemicals you can make good customizing.

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And is a cottage much of the house that we generated can also be used to generate products to show the name is showing a lot of the videos about the tissue that we generated. Though this is correct issues that we generated they can see there are a lot of cells here and post and what do we have here is I'm sure you are more.

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On burst and this to show it's connect to most transmitter so they can sense how much force this kind of tissue can generate. The ones we generate of these tissue want to generate as a to show we test how much post that they can generate and then we treated additional always a drug called to beautifying So this track actually is a drug to use in kinetic and so they use this truck to treat the heart of failure patients so this deal beach rock increases the force that the heart can generate it think did it all was that growing harder to show you can see the raid.

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Is from the correct issue with a drug you can see it clearly it generates more forces Ok compared to the tissue without the drug treatment and so what we see here is we can use them to drive the car to to show to make car to mimic what's really happening in can in its path.

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Ok So in summary. This is the approach that we do so we call it Jewish project because we just can't string has been and winning it but tests so you can see you start a ways human stem cells and the goal is to generate a cut on my side and all you need to do is you have to buy 2 chemicals and then you need to trigger the cells with the 1st chemical at what it wants and a 2nd a chemical a little bit a later time points and then you can generate.

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The efficient to customize at more than 90 percent of the cells become customising and the cells can be used to for. Therapy Ok So that is all for my talk today I would like to a cannot achieve my funding. From Penn State and also from my age thank you all for attending this lecture now it's time for me to take you know question.

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I think is an issue the presentation of for those in the audience if you have any questions please put them into the you're in a. Chat I'll start as I have a couple questions in regards to you gentlemen in general what's more important getting a high purity or higher you.

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Yeah that's a great question I have both of them are important. Because for example for the context. The neuron and our cell want to generate these cells will lose proliferation paragraph says so in this case you want to. You want to have both purity and heart. But the 4 cell type Let's say you want to generate some a liver cells better sites from stem cells so in that case.

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High yield a May not that important because the reason is because you want to generate pure of these liver cells they can proliferate So even a small amount of you can culture the cells and they can proliferate and they can generate a lot of these cells but the for newer or contacts out most of them i important.

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Ok then my 2nd question is and then this is your talk that you grow yourself on a special substrate so that enables them to eat the way they were eating. Yes So that's a good question so we just. Alter the Skutnik a muscle cells on the fiber Nekton coated in culture dishes.

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Kartik cells are per for fiber Nekton if you do measure g.i. of. Carnage and they also walk walks it's not as good as 5 connect. That's all my question I guess. To see is going on for some sleep tonight I. Might have a question just feel free to ask questions.

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Yes The question is are there other pack ways you consider before diabetes when signalling for how you might say you know. That's a quick question that's a great question that's an and before my approach. You know there are. You know there are already some other people try to different apos ways to for example there is a possible record of the m.p. part way though the palm of a generous is hurting pos way these part ways are also very very critical for stem cell differentiation and the people tried that and I also tried it.

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At the very beginning so I combined at the Winter pos where we said b.m.p. parts might be what happens indeed when the pa modulation enhanced the b.m.p. pos way effect and but later what I find is all I need it is modulating the when possible even I do not model it b.m.p. possibly it still worked so you know tried to make it as simple as possible so that's why my final differentiation project go is only involved in modulation when the pos way was just to chemical.

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The let's cover the check completely changes everything Ok there's another question yes it is about the 3 great issues with the cells Yes that's possible as a matter of fact. We are collaborating with leather lab they are experts in 3 d. printing and they are using our human kind of muscle cells to print streety kind of Titian's Yes it is visible.

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Doing of the question What are some future directions freezing is carrying my sick cells leashes are they are you know crazy **** they're organized they're being developed or some other method to 3 maybe 3 tissue structures and the cells yet so there are there are several directions you know for use in the cells I mean this is of course you know we have the ability to make these cells the 1st direction with a b. especially in the pig from typical companies so they develop a lot of you know drugs but only a few of them can find it you know and to the market so one of the big problem is for whatever drug you need to consider the side effects this president that track has you know that's ripe for to be developed to treat a cancer etc But then it has a doctor today facts on human heart than that that you are quick witted but then how do you test that you know you don't want to wait until you.

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Finish the kinetic offense while in effect to and then you see you see that side effects on the human heart your eye to penalize $500000000.00 She doesn't want to that so what they want to do now is they want to use human Kartik muscle cells also Reidy Katic tissues and then they want to test to that laptop today using these human cells so previously they use the mouse to house because they just cannot have human contact cells but the mouse cell.

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You know given that you have shown no toxicity from out character that may still have the prison for humans or because you know mouse card Excel is different from human kind except to now many of the big pharmaceutical companies they really want to have human contact muscle cells of 3 d. human kind of tissues for drugs up to testing so I think a definite That's one of the projects and at the other one picture action with a b. use these cells for clinical trials as a cell therapy and it's being.

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I know there are some labs in Japan they already did it for using these cells to treat a real human heart patients so they already did the test experiment for 3 to 5 patients so it's already been going are so but we haven't to try or you know the large scale clinical phase 3 trial using these cells to so far looks pretty promising so I like both of the directions you know why I suppose you have to look for therapy wise for drug testing most of them are pretty promising of using these cells Yes we.

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Didn't see that is a new questions to we just ravenous here Ok yeah Kami that sounds good if you think you're going to everyone. Who is one of the socks and if you actually answer to this information. My pleasure.