This webinar offers a compelling look at how advanced reproductive technology is shifting from human fertility to planetary salvage. It’s a masterclass in using high-level bioengineering to fix the ecological debt we’ve spent centuries accumulating.
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Added:Welcome, welcome. Uh um this is the the last I3 session of the year. It's it's not like the the it's a little bit special. It's going to be about nontraditional species and it'll be it's going to be also about really the advanced uh assisted reproductive technologies that are developed for biomedical science conservation research. And this is very exciting. We have uh three uh very prominent speakers to talk about that. Uh so my name is Pitsoli. I am a research veterinarian at the National Zoo and Smith in Conservation Biology Institute at the Smithsonian in Washington DC. I've been working on uh wildlife reproduction for a long time. And uh I'm joined today to moderate this uh session by Matt Matt Petit who is the chief scientific officer at mature technology limited. Um it's a a barcode system for fertility centers. But importantly enough for this session, Matt serves as a trustee and scientific advisor for uh nature safe uh a conservation initiative preserving life sales for endangered species in collaborations with many many zoos. So uh the the what's really important for all the participants to remember is to please put your questions in the Q&A uh uh section not in the chat. Please all everybody please if you have questions to the speakers please put them in the Q&A. they will be picked up by our our moderator behind the scene and we will be able to as as possible uh to uh to uh to present them to the to the different speakers. So uh without further ado I think um we're going to be starting with our first speaker.
Our first speaker is Andrew. Andrew are you with us? So as you are getting ready Andrew to share your slides let me introduce you. Andrew Pasque, if you don't know him yet, is really the head of the EVO Divo repro group at the University of Melbourne in Australia.
He's a scientist using comparative mamalian genetics to identify critical and conserved networks driving key processes in development. His work has led to the sequencing of several genomes, especially masupial species, including the extinct Tasmanian tiger.
And actually Andrew has established the Tyloin Integrated Genetic Restoration Research Lab, the Tiger Lab, which is a pretty cool acronym to examine really the possibility of having this species back, the the extinction of that species, as well as especially important to develop really the next generation tools to preserve and conserve current endangered massup species. So Andrew, Andrew, uh, welcome. Uh, and please, the floor the floor is yours. You have about 20 minutes, uh, to talk. So what's going to be special with Andrew compared to the other speakers is that we're going to keep him a little bit longer after his talk for Q&A because Andrew unfortunately cannot stay with us for the whole um, webinar. But please, Andrew, welcome.
>> Thank you very much and thank you for the invitation to present at this this incredible uh, conference today. It's great to see so many people online from so many different countries and I'm really excited to tell you about the work that I'm doing here at both the University of Melbourne and with Colossal Biosciences which is a big biotech firm which is really um in exploring the possibility of deextinction and the way that it can really change the conservation landscape. So that's what I'm going to talk to you about today. The reason I've been so passionate about this for so long in my career is because in Australia, unfortunately, we have the highest rate of mammal extinctions of any country in the world, which is crazy when you think of what a wealthy country Australia is. There's really no excuse for being at the forefront of mamalian extinctions. But at the moment, we have over 20% of our land mammals threatened with extinction. And at that rate, we're predicted to still have one to two extinctions every decade moving forward.
And this statistic doesn't take into account adverse weather events which are particularly problematic in Australia.
We're having far far more of these events and there are far greater intensity and these are things like the horrific bush fires that we've had. We had some um about 3 years ago that were really devastating um and big floods as well. We go from one extreme to the other as we're experiencing in so many countries across the world now as a result of global warming. And what this does is any single one of those events can completely, you know, eradicate a species and make a species extinct. Or even when it doesn't do that, it can reduce the number of of animals in that particular species to below a critical mass where they start to enter what we call an extinction vortex where there just simply isn't enough genetic diversity left in that remaining dwindling population to really have a healthy and sustainable population bounce back from that again. So these are things that are really profoundly impacting particularly Australia, but obviously these are massive global issues and adverse weather events are likely going to really push that statistic of how many extinctions we're going to see uh you know a lot faster as well. And I'll talk about how the technology we're developing for deextinction can really help mitigate that as we move forward because I think it's really really important.
So what my lab has been doing then is really trying to think about these next generation conservation tools that we need to develop to try and and stop this from happening basically. So we know that our current conservation toolkit um is not enough. We're still seeing all these animal extinctions. So we need to be able to figure out how we can do some of these things particularly in non-model species. The things that we all work on and the things that are really critical for conservation. So they're things like just being able to do gene editing, being able to then do assisted reproductive technology, something this audience knows an awful lot about. Again, that's perfected for some species, but then completely not at all developed for others. For marsupials, that's been a real challenge for us is surmounting some of those art challenges. And I'll talk a bit about that um today as well. We also need to be able to do things like derive stem cells. And again, that's quite different for different species. We're very good at doing it in mice and humans. And we're not particularly good at doing it in other non-model species. And that requires a lot of time and effort to try and figure out exactly what those cues are to make stem cells in these other species. But if we really want to realize the potential of bioanks and do genetic editing and reintroduce lost genetics back into populations, we've got to figure out this stem cell um problem for uh all species. We really need to figure that out. And then we also run a very large biioanking initiative here in Australia too. Um, we set up Australia's first living cell bio bank, which I'll talk about as well, which is another really critical resource that I'm very passionate about and think, uh, we need more bio banks.
But the main species I'm going to focus on today, the one that my lab is really passionate about bringing back is this one here, the Tasmanian tiger. So, for those of you who have never seen this animal before, it was an incredibly unique massupial. So, it had a pouch and raised joey's in its pouch just like a kangaroo. But I think everybody here can really appreciate just how much it looks like a canid, like a dog. It's got a very, very canided form. In fact, it's the best example of convergent evolution that we know of in vertebrates. And that's when two animals evolve to look incredibly similar. If you look at the skull of a Tasmanian tiger and compare it to the skull of a dog, you'd be really hardressed to see any differences between them. They are incredibly, incredibly similar. But this animal sadly was hunted to extinction in 1936.
The last animal, this one here, died in Hobart Zoo in September. Had a really tragic death. The keeper just simply forgot to put it away one particularly cold night in Tasmania and the temperatures dropped below freezing and this last animal, the last known animal of this particular species froze to death in this pen. So really, really tragic story. But the thing that made this animal so important from a conservation perspective was that this was an apex predatory marsupial. And this was the only single apex predator mammal that we had on the landscape in Australia. And this is really unusual.
If you think about the Americas, if you think about Europe, there are tons of examples of apex predators that you have on the landscape. You have bears and wolves and big cats, like all sorts of things that you have roaming across this landscape. But in Australia, we only had this one. This was our apex predator.
And we know that those species play absolutely critical roles in maintaining ecosystems and ecosystem stability. So when you remove these animals from the ecosystem, we start to see huge destabilization of all of the species within that. And that has profound implications for conservation of all of those animals. And so there's a real imperative that if we have the technology, which we now do, to try and bring back some of these really important cornerstone species and put them back into these ecosystems.
So one of the first things that we can see that's happened as a result of the loss of the Tasmanian tiger, is that this particular species here, the Tasmanian devil, there was a massive upsurge in this horrific disease that this animal gets. And apologies for the graphic images here, but it gets this devil facial tumor disease. It's actually a transmissible cancer, a very unusual disease. And we can see by looking back at historic samples in museum collections that this disease was present in the Tasmanian devil population, but at a very low level historically while the Tasmanian tiger was around. But once the Tasmanian tiger was removed from that population, there's no apex predator anymore.
There's nothing to remove these really severely diseased animals from the population. So instead, these animals persist like the ones you can see up on the screen here to very advanced stages of this disease. And while they're all feeding from the same carcass and also part of their natural behavior is they bite each other's faces when they're all in a den together, they are transmitting that tumor from animal to animal. So when you have your apex predator there, it's removing these animals from the population and it's keeping this disease at a sustainable level. But once the Tasmanian tiger was taken away, we don't have that balance anymore. This disease has run rife. There's nothing to take those animals out of the population that have advanced disease. And so this has spread completely throughout the Tasmanian te population. Now, this would have made that species go extinct already, but we were able to go and isolate uninfected individuals, bring them back into, you know, clean areas, and they're being bred up in captivity, and they are sequentially now released back into Tasmania. But because this disease persists there, they will all eventually contract the disease. And there's no sustainable way to have the population of Tasmanian devils in Tasmania without the Tasmanian tiger present. But they're just one single example. And we can see this imbalance happening completely across the landscape in Tasmania. It's not just the devils. There's been an explosion in kangaroos and walabeees again because there's nothing to predict uh those species. so to control their populations. And so that whole ecosystem is really starting to fall out of balance. And I'm sure many of you are aware of this uh incredible study that was done in Yellowstone where they removed the wolves. The wolf is uh the closest analog to a thyloine actually.
And so it's like removing the exact same ecological role from Yellowstone. And then they tracked so that happened in 1926 and then they tracked it over the the the subsequent years. what was happening in Yellowstone Valley as it's such a, you know, a beautiful pristine national park that people were taking very good uh notes around what was going on in the landscape once the the wolf was removed. And so, as you would expect, you see an explosion of the the deer population. That's similar to our kangaroos and walabeees. So, you have a massive explosion of that. They start eating all of the plants. So, any new plant that's trying to sprout and grow gets eaten by the deer. There are no flowers or berries available anymore. So the bees leave the valley. The birds leave the valley. Eventually the bears left the valley as well. Then the deer start eating all of the fresh growth that's alongside the river because that's the most fertile area of ground.
They muddy up the river. The beavers left the valley and then in the end the river actually changed its course through Yellowstone Valley. And they could see this catastrophic collapse of that ecosystem in Yellowstone. And so they made the decision in 1995 to put the wolves back in again. And within 10 years, that entire ecosystem had restored back to that natural balance, including the beavers returning and the river regaining its natural course through Yellowstone Valley. So, it's just an incredible example how the removal of one species can profoundly impact not just the plants and the animals, but even the landscape, the shape of the landscape changes as a result of that. So, we know that these animals, these apex predators in particular, have these absolutely cornerstone roles in maintaining these environments. And that's why it's so important to have them back there. In this case, luckily, you could just go to a neighboring state park, find a pack of wolves, and transllicate them back into the valley. Obviously, we can't do that with the Tasmanian tiger because it's extinct. There are none of them left around. And there's no other marupial that even comes close to the role that this animal played. There's no other animal that could step into that apex predatory role. And so we really have to think about if we want to try and restore that ecosystem, we've got to be able to put that Tasmanian tiger back into Tasmania. So I think in terms of thinking about deextinction and the kinds of species we want to bring back, this is one that really ticks all of those boxes for something that I think we can make a very very strong case for.
This was a completely human-driven extinction. We hunted them to extinction in a very very short time. There's no replacement. There are no other species that look like it that can fulfill that role. It was a cornerstone species in that ecosystem. So it was very very important for sustaining natural balance of everything else. And then importantly for us, it was a relatively recent extinction. So 89 years ago now um since that last animal went extinct. And so because it's relatively recent and Tasmania is quite a pristine incredible place uh in Australia, there is a lot of intact ecosystem left there. and the food sources that it would eat have uh arguably become more abundant now in its absence. So, it certainly has everything it needs not just to survive, but also thrive again back in Tasmania. So, we can tick all of those boxes. And the other unique thing I think about the Tasmanian tiger is because it was such an incredibly unusual and beautiful specimen is there were loads and loads of samples collected for this that were stored in museum collections. And that's given us incredible DNA and even incredible RNA resources for this particular species. And these are some of the specimens that really made this possible. So we sequenced the first genome for the Tasmanian tiger probably about 15 years ago now. And we did it from a joey a baby a pouch young a pup uh that was taken from its mother's pouch which is that specimen you can see on the far left in the in the tall jar.
And then they also um collected that mother's head at the same time. So, her head was removed and placed into a bucket of ethanol in the back of a museum collection. And we've also been able to get unbelievably good DNA resources from teeth. One of the nice things about teeth for us is you can actually drill into the tooth, which is what that movie is showing on the right there, into that soft pulp region, and you can get really pure thyloine, Tasmanian tiger tissue. So, it doesn't have any contamination. Whereas the head that you can see in the middle and then the joey that you can see on the left have been handled by a hundred more people over the last hundred years. And so they have a lot of contaminating DNA all over them as well, which makes the things just a little bit more complicated when it comes to genomics.
But one of the staggering things that we found is that middle specimen, that head in a bucket that has been sitting at room temperature in ethanol for the last 120 years actually has unbelievably uh well preserved not just DNA but RNA in the tissues. So by sampling that head, we've been able to actually look at gene transcription of taste. So we can look at the the taste receptors in the tongue. We can look at the smell receptors in the nose. These are regions of the genome that are very very difficult to annotate unless you have RNA. But we've got RNA from all of them.
Obviously, we can also look at sight because we have the eye, we have the brain, so we can look at brain function and then muscles, bones, all the tissues that you can think of. It's really incredible that they preserved a head because it has so many different tissue types that we can learn so much about thyloine biology from. So really incredible. And so with all of those resources combined, we were able to sequence the first end to end or tie to tie genome for any extinct species that we've done now for the thyloine. So the Tasmanian tiger or thyloine genome is now better than most living animal genomes um just because we put so much effort into trying to rebuild that genetic code. And obviously that's our blueprint for how you recreate a species. So we've got to get that code right. So we've now been able to sequence through every single part of the genome including the ends of the chromosomes, the tieumirs, the middle of the chromosomes, the centromeres, these again notoriously very very difficult parts of genomes to sequence. But because we have so many good resources, we've been actually able to piece that all together. And we can even look at things like the epigenetic profile across that genome as well. It's been so well preserved in these specimens. So really phenomenally able to completely rebuild that genome.
So then how does this process of de-extinction work? Fine, we can sequence our genome of our extinct species and we've done that. But then what happens next? So what we do is we take that genome and we ask once we have that complete genome, what is the closest living relative to our extinct species? So we just compare that genome to living animal genomes. And for us that is a small mouse marupial called a fat tail dunart that you can see at the top there. That is a lot lot smaller in in uh scale to the thyloine. Probably a little bit smaller than its foot. So really really quite small. But when we compare their genomes, they're about 99.85% identical. So only 0015% different. I say only, but it is a 3 billion base pair genome. So that still equates to 4 1.5 million differences. But the way that this technology works is you take living cells from your living animal in this case the fat tail dun art and then you go through using crisper so our really advanced genetic editing tools that we have today to just change all of those nucleotides where they're different going sequentially through and editing all of those eventually 4 and a.5 million edits across that genome.
Once you've done that, you've recreated your thyloine genome and then you can use our standard cloning technologies art to turn that cell back into a whole living animal using things like sematic cell nuclear transfer and their techniques that we're working on currently in marsupials. And right now we're really focused on this DNA editing phase. So we have our established colony of fatal donuts. We've got our thyloine genome and we're going through and making all of those edits to recreate that thyloine genome. But while we're doing that, we're working on all of the assisted reproductive technologies to actually um you know be able to to turn that cell back into a living animal.
Once we've actually got that cell and turning it back into a living animal, obviously we need a surrogate. So in this case, we would actually use the fat tail done up as a surrogate. So some of you might be thinking, how does this work if you've got an animal the size of a Labrador being born from an animal the size of a mouse? But this is one of the incredible things about masupials is they have such tiny tiny babies. And this is a dunartant baby on the day of birth and it's about the size of a grain of rice. A thyloine baby Tasmanian tiger is exactly the same size. So there have been some collected at this early stage.
And so what that means is even our tiny little mouse masubial can actually give birth to a baby thyloine which is really incredible. They have only a 13-day gestation as well. So really short gestation and tiny tiny baby at the end of it. Um it's still a lot of changes to make. So we need to figure out how we sequentially go through and edit all of these genes in the genome. And we've got a few clever ways of doing this now that we can actually scan genomes and identify regions that we think are responsible for giving them their particular form and function. And then using transgenic mice, we can actually test whether these genes are actually driving changes in their body shape and their skull shape. And so this is just an example of one of the regions we found in the Tasmanian tiger genome that looked like it was changing uh skull and limb shape. And so what we did is we just asked what this fragment does in a mouse. And you can see it's turned on everywhere the mouse is blue in the face and in the developing limbs. And then we can actually put that piece of DNA into the mouse genome and ask how it changes shape. And we could actually change the shape of the mouse skull to start to direct it more towards a hyper carnivore. So we can start to learn how we can actually manipulate our genome to uh um to actually change the form of our donut to make out our eventual thyloine.
But one of the things I'm most passionate about is how we can immediately use all of this technology we're developing for deextinction for conservation outcomes for marsupials right now because as I mentioned at the beginning they're in this real desperate need for for more conservation tools. So one of the first things that we needed to do was be able to to harvest oversightes if we want to develop this art. I'm sure everybody on this call appreciates that you really need to have a good supply of oases to do that. We never had protocols for super ovulation of donuts before of any masupial before.
So there was nothing um for masupials in existence. So we have a really talented team of art experts here who worked through uh at the arduous task of trying to figure out what that hormonal regime would be to get them to induce them to ovulate. And we get very good ovulation rates. I wouldn't call it super ovulated. So we call it very good ovulation instead, but we get a really good number of eggs. We probably get about 20 eggs uh per induced ovulation.
They would naturally ovulate about 10, but we can then fertilize those eggs and we can culture them right the way through to Blases' stage. So we know we're able to get really healthy eggs.
And if we induce ovulate and get them to naturally mate, they'll produce live offspring. So we know that our induced ovulated eggs are healthy and good. And so we can now use this as a really viable tool um for helping us develop all of the art that we want to for massupials. This um act of doing induced ovulation also brings animals into heat obviously. Um and so this has been already used for conservation in a lot of our wildlife breeding schemes. So as I mentioned before the Tasmanian devil on the brink of extinction. We really need to do something to try and and save our Tasmanian devil population. But another species that's on the brink of extinction is the northern qual which is that little animal you can see sitting down the bottom of the screen next to the devil. Um but these are both related marsupial species as urids. These are very feisty animals. If you introduce a male to the female and she's not receptive, she will kill the male. This makes them very very difficult to breed in captivity. Particularly when you're trying to maintain any genetic stock that you've got. You cannot run the risk of having one of your males get killed just because the female was not receptive enough. But using this induced ovulation scheme now, we can predictably bring that female into heat, introduce the male at the right time, they can mate and produce a litter and the male doesn't have to die, which is great.
Also, if the female loses her litter for any reason, we can use this to bring her back into heat again and get her to have a second litter within a season. So this is already being used for conservation and has been really pivotal in increasing the output from these captive breeding programs for these animals.
Another thing we spent a lot of time on in my lab was actually perfecting how to make induced pluropotent stem cells. So we figured that out now for marsupials again really complicated. One of the weird things about marsupials back on this slide is you can see that blasty on the far right. There is no inner cell mass. They don't actually form an inner cell mass. The embryo will just form on one side of that ball of cells. And so the signals that are required for making induced pur potent stem cells were slightly different to what we see in mice and humans and other species. So we had to really unpack that. We've done that now. And once we could demonstrate that, Australia was finally um going to get behind us setting up a living cell bio bank, which we've done now at Museums Victoria, where we're trying to collect cells from as many marsupials, endangered and non-endangered, as we possibly can, and bio banking those living cells down now so that we never have to go through this process of de-extinction again. We won't have to re-engineer a genome because we'll have those living cells frozen down, and we can go through that cloning process. And then the last example I'll give you of how we're using this is with the gene editing. So now that we can edit genes so well in marsupials, there are a number of different applications of that. The first one that we're doing is for this northern qual that I said is critically endangered. This is actually the next species that is predicted to go extinct in the wild within 10 years in Australia. And the reason they're going extinct is because they eat cane toads.
Can toads come from South America. They are not native to Australia. They were transllicated, introduced into Australia. They are toxic. They produce a poison and none of our native wildlife have any natural resistance to that poison. But where can toads naturally exist, there are a few mammals that can easily eat these can toads and survive fine. So by looking at the genomes of those species, we were able to identify that it's just a single amino acid in a single gene. And in fact, it's just a single nucleotide in our three billion base pair genome that makes you either susceptible to kto toxin or resistant.
So what we did is we obtained cells from our northern quals. We genetically edited them to produce that kanto toxin resistance alil. We just changed that single nucleotide and then we expose those cells. We turn them into to stem cells. We expose them to the kanto toxin and we show that when we do that from that single nucleotide edit we can make the the the qual now completely cantoxin resistant. and it had 6,000 times increased resistance to that toxin, which means it's completely resistant to that. Not only does this save the qual from extinction, but it means that they can actually eat canads and help control this spread of this awful pest species across Australia. So, it's a really nice example of how genetic engineering can be used in a conservation space to save our wildlife. So, we're working now on perfecting that art, that cloning process, so we can turn those cells now back into a living qualing them across Australia. So I just want to finish up by going, you know, when I first started my career working in ancient genomes and, you know, sequencing massupial genomes and trying to understand how massupials have evolved over the years, I would go to museum collections and open these drawers and think about how sad it was for all of the species that we had lost and had been collected and, you know, are no longer out there and and in the wild. But now when I see these museum collections, I know that we can sequence whole genomes from these animals. we can understand more about their genetic diversity and we can actually engineer back in that lost diversity to recreate really really healthy populations and resilient populations for our planet moving forward. So this is no longer a sad thing. I'm now so grateful that we collected all of these specimens that we can go back we can re-engineer some of that lost diversity back into animals.
And I actually think that's where deextinction technology will have its biggest hit. I think there'll be a few species that we bring back um through this whole deextinction process, but the majority of the science will be used just to engineer lost diversity back into living animals so that we can make really healthy and sustainable populations of some of these, you know, critically endangered species which are down to just the last few individuals.
There's no way they can sustain really healthy populations again. So hopefully I've shown you today that de-extinction science is really conservation at its core and that we're developing these really new methods to conserve and preserve and then restore biodiversity whether that's restoring biodiversity back into these living populations or bringing back entire extinct species and putting them back into the landscape.
And just this last thought that, you know, we've really changed the planet so quickly and we're changing everything around our poor species and our poor ecosystems at a rate that they just cannot keep up. And we need to be really open-minded about some of these new tools that we need to put into our conservation toolkit to really help build resilient ecosystems again if we want our planet to survive this massive six mass extinction event that we're in currently and to be a sustainable planet moving forward. So with that, I just want to thank the team that you can see there and all of my funding and thank you very much.
>> Okay, thank you so much uh Andrew. That was uh you managed to to deliver a lot of key uh information in a very short amount of time and it was very clear. I loved it. Thank you so much. So we have a lot of questions for you and as we said we're going to spend a little bit more uh time just for you because you cannot really stay for the whole webinar. So there are different types of questions. There are questions that are more technical and then after that there are questions that are a little bit more phys philosophical but not necessarily philosophical but like more like applications and and translation of that knowledge. So let's start with um first some questions that are a little bit more technical and it's all about okay.
So you you talked extensively about uh sequencing the genome and how complicated it was for for the tyloin for example but we have some questions about okay so once you are able to really sequence the genome what about the metilation of the DNA and all the different aspects that where how are you able to trace back really the imprinting of the genes and all the packaging of the the DNA.
>> Yeah. So I mean there is some methylation data from from some of the samples that we had but obviously you've only got limited tissues there. But what we do know thanks to the pioneering work done with sematic cell nuclear transfer is that you can take a fully differentiated cell right put it back into an enucleiated egg and it will you will erase that epigenetic code and then you reset that epigenetic code as those cells go through early embryogenesis and they do acquire the right imprints. So if we take a a massupial imprinted region for example and put it into a mouse uh early embryo and let it go through that process, it will actually acquire the correct methylation. Where histones sit, what those histone marks are and what those methylation marks are across the genome is all encoded in the DNA. So there are certain signatures that tell histones where to bind and then what modifications they should have. It's all there. So going through that process of early cloning can reset that. Obviously, that's one of the biggest challenges with cloning and why we see so many nonviable uh animals from that process is because it's very hard to go through that rewriting process.
But there are methods now that they've really really well perfected in in mice just because it's easy where you would take that cell, create an early embryo, let it get to the blast stage, remove one of the cells from that. So, it's already gone through one round of reprogramming and use that for another round of sematic cell nuclear transfer.
Let it go through again. And you can iteratively do this until you've completely wiped that slate clean and then let that genome reform what its natural imprint should be. So we feel really confident that you can actually get there. I mean we know we can do that with a terminally differentiated cell type that you know a skin cell you can turn back into a whole living embryo by just erasing and resetting that epigenetic code.
>> Okay. Thank you so much. Uh, someone was Jason Angry and was wondering also because you talked about okay DNA recovered from the skull or from the tooth, but if the only source of DNA is hair or fur, is it still possible to reconstruct a genome and are is the information still accurate? Not >> Yeah. So yeah, there's been massive advances in ancient DNA now. So from hair actually re it's actually really good because the hair is there is hair actually I mean DNA in the in the middle of the hair shaft. So they've figured out different um extraction protocols now. So if that person's particularly interested I would say look up Beth Shapiro's work. Um and she has done a lot of work in extracting really good quality DNA from hair of museum specimens that are hundreds of years old. Um the older the DNA is the more fragmented it is. So the shorter the lengths are and the the more difficult it is then to rebuild the genome. So that can make it really challenging with with just hair. But you can definitely sequence a genome with just hair.
Whether you could do a really good genome build or not um would depend on what quality that DNA is.
>> Okay. Thank you. Okay. So let's uh Okay.
Uh now let's let's talk a little bit more about the the gamut. So how do you explain that everything is so complicated from eggs to sperm to embryos in massupules? Why are they so special and so complicated to understand compared to other mammals?
>> I think because they have a different reproductive strategy, right? They really chose to give birth to very small babies and then have this prolonged period of lactation. So everything about them is quite different. But yeah, when you start digging into the the the things about musuples, they have the longest sperm of any mammal. So they have this crazy long sperm that then swim completely differently. The egg gets this uh a shell coat as it's ovulated and travels down the the overduct that is almost impossible to inject a needle through. So we we have the laser. You cannot laser blast through the the the shell coat. It's impossible. We have to get these very special glass pointy nosed pipets and use pzo to even get the needle into the masupial egg. Then the massupial egg is really yolky. So it's really difficult to know exactly where we need to inject things, where we're putting into the egg. But we are working through all of this. You know, again, from a conservation perspective, if we can do art, if we can do AI, if we can do Ixie, like any of these techniques would have profound implications for our management of captive breeding colonies. So there things we need to sort out anyway, but it's just lacks, you know, the investment in really having people work on it. But hopefully we're making a huge amount of headway. I mean just being able to do that induced ovulation has been a gamecher for us because now we can finally get enough eggs to start playing around with all of this work but I hope within the next year or two we'll have sematic cell nuclear transfer sorted we'll have IVF or Ixie sorted um and able to actually produce viable embryos but yeah they're they're complicated in every single aspect nothing is easy in a m >> that's makes them so exciting actually >> it does everything's so interesting I mean the fact that they don't form an inner cell mass is fascinating you know, like it's just a completely different way of forming an early mamalian embryo. But then that made making induced pur potent stem cells a nightmare for us. But you know, like we learned so much when we figure these things out because it is such a different way of approaching development, reproduction, the whole thing.
>> Okay. Okay. So following up on that uh assuming that you can you can produce embryos from tiley can you elaborate a little bit more about the strategy about transferring those embryos to surrogate species and what would be the candidates and what kind of obstacles you are kind of forcing about that.
>> Yeah. So they would gestate within the donut. So, because they're so small when they're born, then they could spend probably about 2 weeks in the mother's pouch before they would outgrow that.
So, they'd get that very, very early milk from the dunart, um, by the time we would transfer them out, they'd be about the size of a bean, like a kidney bean.
Um, that would be about them at 2 weeks old. And then we actually already have the milk that they need to hand raise masupials because we have so many masupials that are hit on the road and are orphaned. And so we actually have this sequential milk formula that you can get from suppliers to actually hand raise uh even very very small marsupials. So we're we're looking at making an artificial pouch that you could put them into. You could put them into a kangaroo surrogate pouch. Um the kangaroos are pretty happy with actually taking surrogate pouch in there, but you that' be complicated. It's more easy just to bottle feed them and put them in a in an artificial pouch. But obviously those first animals you wouldn't release into the wild, right? Like they would be uh quite artificial. whole creation, but you would keep them in captivity in very large areas and then you would hopefully be able to breed from those animals.
That would be the plan is that you would be able to make viable, you know, reproductively viable offspring that you could start to then actually produce then your your thyloines.
>> Yeah. And actually that's kind of the the next question is okay so now assuming that you get the first birth of tyloine a breakthrough in science for sure but how will you I mean not only you but all the people working in this massive effort of the extinction. How will you make sure that you can scale up the effort and then after that where repeat really the the success produce enough animals to really build a sustainable population that could be eventually reintroduced in the wild. So how I mean how are you going to be able to multiply that success and where are the resources going to come from?
>> Yeah. So for the I mean for the thyloine that's a lot easier than it is for a mammoth for example, right? I think we can all appreciate that that's a lot easier in terms of scaling up. You know, you can have a colony of dun arts and have hundreds of dunarts that could be surrogate mums for these particular species. We also sequenced about 50 genomes so far of thyloine. So once you've got your timid tie limit genome, it's very easy to do um you know just basic aluminina sequencing on any crappy sample that we can get from a museum and you can get a really good genome from that because you've got something to map it back to. So we have a very good understanding of population diversity of thyloines right around the time that they went extinct, but then also back hundreds and thousands of years so that we can engineer in you know thousands of years of loss diversity if we need to.
But we would start by creating a population that had that more contemporary diversity um that existed around the time when they went extinct.
Um so that's actually quite easy to engineer back in compared to re-engineering your whole thyloine.
That's a much smaller amount of changes that you need to make to incorporate that diversity. So I think we can get diversity back in. I think for something like a massupial it's pretty easy to produce 50 animals you know using surrogates. It's not that challenging.
For some of the larger animals that's definitely going to be a challenge. And then you know for for all of these larger animals too they take decades to become reproductively active themselves.
So, you know, if you want to try and make them to start breeding to make a sustainable population, you have to wait even longer because, you know, they have they take a long long time to become sexually mature and be able to breed themselves. So, yeah, there are challenges. You know, I think this is going to be not a quick process for a lot of species, but I think there are really critical species that we want to get back into those ecosystems before they fall apart.
>> Well, thank you so much, Andrew. Um I think uh we're going to we're going to have to switch to the next speaker. But what we're going to do since you cannot again stay for with us for the whole webinar. We can also kind of try to compile the questions that you were not able to uh to answer with the name of the people and then we can forward that to you because it's very exciting to see how how much people are really kind of eager to know more about all the the the things that you are doing. So thank you so much Andrew.
>> Absolutely pleasure. Thanks for having me.
>> Thank you, Andrew. But it's really evident from the number of questions that have come in in relation to your subject, just how interested the attendees are. So, it would be great if we can keep the conversation going by forwarding some of those questions to you. Thank you very much.
>> Um, unfortunately, we have run out of time with Andrew. Um, but our next speaker is someone who I'm proud to call a friend. Uh, having met nearly 30 years ago when we worked together on an initiative that helped establish what has now become one of the world's leading commercial equin breeding and seaman freezing services, Stallion AI services. Tulis Matson is an entrepreneur. He's a charity founder, a conservation pioneer, a skydiver, and a proud dyslexic. He's dedicated to protecting global biodiversity and he combines cuttingedge scientific technologies with bold advocacy to safeguard endangered species for future generations. In 2020, Tulis founded the charity Nature's Safe with a simple yet ambitious goal to transfer biotechnology from his company to the urgent challenge of halting and even reversing biodiversity loss. Nature Safe is committed to preventing animal extinction by indefinitely pre preserving live selves from endangered species. And this is a mission that could change the future of conservation.
So Tulis, I'm going to hand over to you.
Tulis, would you like to uh >> Yeah.
>> share your slides?
>> Uh yeah, thank you. Um >> can you see that there? Okay, now Matt, >> we can. Yes. Thanks, Tulis.
>> Brilliant. Do you have your sound um enabled? Just a quick sound check. Okay, great. Over to you. Thanks, Tis.
>> Great. Well, thank you, Matt, for that uh amazing introduction and uh yeah, what a fascinating uh first presentation, I have to admit. So, thank you very much for inviting me uh to this webinar and yes, talking about some of the the world that uh I work in and I feel like I do have one of the luckiest jobs in the world and I'm going to give you a bit of an insight to what I do. So um the presentation title is life after life and just showing some of the work but my background basically um I'm a horse person by trade. I've been breeding horses for virtually all my life. I've been freezing horse semen down actually we started with Matt you know nearly 30 years ago. Uh so that's really where it started. Uh so I'm a crybabiologist. Um this horse here is one that went and won the Olympics. This what sort of pays the bills you could say. But my passion is these ones at the bottom here, which are the rare breed horses. And we've had over 1,500 stallions come through our center. And one of my ways is how can we save our rare breed horses and through using new technologies. I love new tech. And how can we develop that? And these are some of the rarest horses in the UK. And that's what we've started off doing. And this is what you're going to see a bit of tonight. Um, we've got several companies uh just all to do with about freezing things down in a way we can bring them back to life. All the cryo side being a crybobiologist and these are just some of them. We have a a logistics company for shipping seaman around the world. We have um uh Gemini Genetics which is a cloning the cloning side stallion AI services the foundation company. Uh we do the same with dogs. So we collect and freeze dog semen and ship that around the world. And we're doing the same with livestock in the sense of freezing their tissue down. We're just about to start that and by banking our livestock, but I'm really passionate about this charity set up called Nature Safe about freezing down species, which you'll see a bit later. I think it's always quite good to go back and look at the history of AI. Um, quite often there's a different type of AI than we're used to talking about nowadays, artificial insemination, uh, rather than artificial intelligence. And the first reference is around the 1300s actually, believe it or not, they actually used to put a sponge inside the mare, let the stallion cover the mare, get the sponge out with a press, they used to press the semen out, extract the seam, and then they used to AI mares that way. So, uh, it's come on a long way since then. And then this fold here is one of the first foss back in 1955 was actually born by using frozen semen. Um, so it's come a long way even since then. This is one of the first videos that was taken in the 1930s when the breeding horses for the German army and this how they used to collect semen from the stallions. They used to stallion cover the mayor. The vet would turn up um to try and extract the seaman looks like followed by the postman. Um but no health and safety there and then and then literally the veterinarian would go inside the soup.
um which is uh yeah luckily nowadays we we have other tech techniques uh of extracting semen but it's always quite good to see how they were used to do it and any of you out there I think you were had a pretty hard deal in life with your job just spare a thought for this person I don't know what they got paid but uh this is the first dummy may uh uh yeah luckily we don't do it this nowadays but if there a great big stallion thundering towards this I would feel sorry for the person sitting on that stall and I hope they got paid a more than the minimum wage. Um, but it's always quite good to see how it started.
And I think this horse says it all. Uh, if you think I'm jumping on there, you've got another thing coming. But, uh, the dummy may now are a little bit more sturdy and a little bit more easier to use. As you'll see in this video, um, this actual breed of horse is called a suffuk punch. Incredibly rare. There's only about, you know, 80 or 90 left in the UK. There's only about three 300 odd around the world. And um we're trying to find ways that we can preserve them and freezing their semen down. And sometimes when you get inbreeding and you get the extinction vortex um quite often it gets more male gender heavy. So we want more females uh on the ground. And with this with these heavy horses, we looked into sperm sexing. We went to the cattle side, which again Matt very much helped us out on, and uh we went and looked into them, see if we could sex some ecquin semen um to see if we can get some more female sperm and it had about a 95% accuracy. And this was the first f uh rare belly f born by this method.
After 11 months of waiting, um, Ruby has produced a Philly fold.
Um, and it's not just any old fo. This is a fold born by sex seaman. We believe it's the first ever in the world for a rare breed animal to use sex semen. And using these technologies can really help this breed. There's only about 70 or 80 females left in the country. There's only 300 left in the world. They need all the tools of the industry. And it's quite incredible. Now what we can do with science and technologies and using this technology maybe just maybe we can help stop these breeds from doing these things but this is hold the future little girl here. So that was quite a a momentous occasion really the there's about 11 months gestation period and I must admit I've never looked between a horse's back legs to make sure it was the right sex before we did that film and it and it was and we've done quite a few more sex seaman foss and they to be fair they've all come out the sex that we wanted uh which is great and you'll see a theme when we go through this now this has been transferred to some more endangered species as well now uh I think they're doing this work in Florida and the sex is being done in in in Texas um but it's it really just shows how it's moving forward and you if you can develop these technologies with the domestic site then you can throw this across onto endangered species. Um we've been working with epidmal sperm taking semen from the testes. We've been doing this for quite some time when a horse had a a tragedy and they asked can we do it and we gave it a go and you see the semen on here how it's totally immotile um or very little and this is sometimes what um uh semen can look like and we add something to it. We call it SSPR, synthetic seminal plasma replacement.
And this wakes the sperm up. And now we do this, you know, we've done about two this week. I've got another two more to do next week. And uh uh we do two or three a week epidmals, not just antiquin on other species as well. Um so this is sort of the last ditch saloon. If they haven't preserved the genetics, there is other ways of doing it. This ball was bought at the sales and after about a week or so went broke its leg and the the guy uh uh vet said, "Oh, there's some guy down in south of England that can actually extract semen." I've never done Bull's testicles. So, I went on to Google and looked at the the anatomy of a ball's testicles. Well, you can imagine everything came up then, but we had a go. And um then we had the first calf on the ground by this method uh back uh I think it's about 2015 and and now this off that one procedure they had 30 offspring uh from that one procedure and now this bull on the right hand side was this calf that really led us on to saying kriy we can use these technologies and we've thrown this across now onto other species as well as you'll see a bit later. So quite often it starts on the domestic side and really throws it over.
Um but travel the world really setting up labs, teaching people how to do this cryo work. Um I've had a very busy few years. I think it's something like 14 or 15 different countries in the last year or so setting up laboratories in India, Japan, Finland, Iceland, and now the Pharaoh Islands. Um, the Pharaoh Islands, if you're not too sure where it is geographically, it's between Scotland and Iceland. It's a population of about 50,000 people. Um, or even maybe even 30,000. And they've got a a horse there called the Pharaoh horse. Uh, there's only 97 of them left, 30 stallions, and they're so inbred. I mean, they got an inbreeding coefficiency of over 30%. So we have we've been been out there last year in 2024 collected and we've been back again this year and we've collected off all 30 stallions and I think this is maybe one of the first times in the world that we've collected a whole breed of of animal the horse and biobanked it basically and we're hoping to go out now and take tissue from each one of these and bank them down as well next year. Um these were quite wild some of these uh these horses but they seem to the beauty about them they had good libido so they didn't they were jumping on the mares and now we've actually trained them to jump on dummy mares as well so which really really did help um but the beauty about them they they used to we used to have to go and quite often catch them from outside um but generally they were very willing to please sometimes the fairways weather isn't the best and the odds standing we actually had to go and uh try and collect outside, which was always interesting. Uh, as you'll see, this uh next next video is right. Our next collection is actually outside. I don't think I've collected in weather like this before outside, but uh we got the stallion there, the mayor's coming up the track here. DJ's in the trucking with the AV. But yeah, pretty amazing surroundings here. Um, but yeah, pretty raw weather out here, but let's see how it goes.
They were quite brilliant. I mean, as you can see, the libido is great.
There's not many animals out there that you could do it in a gale force winds and they still perform. So, but it was incredible. And we then we got the um seaman uh from one. They've never been aed. Everything's been natural covering.
So, we aed our first mayor um back in 2024 um which was uh just which was one of the first ones well it was the first one that's ever been done and then this year we've the the mayor fold. So, this is the first fold born by artificial insemination.
This really showed what these technologies can actually do to um a breed of horse that is is you know could be fascely on the brink of extinction.
And then we can translate that across to other species. Other species that we've actually worked on as well with camels.
This wasn't necessarily on the rare breed side, but this is just trying to further technologies. Uh camel seaman is very difficult to freeze. Uh and they've got the beauty camel and the racing camel. I usually ask question which do you think is more expensive? But usually comes back to the quite often the beauty camels are very very expensive out there. So we're doing some work as well in how we can freeze camel semen as well.
Other work we do is uh genetic preservation, freezing uh tissue down in a way that can be used in the future for cloning purpose on a domestic side. So um we've got a company called Gemini Genetics and this has been running for seven or eight years now. And uh we do cats, dogs, and horses. Uh and the the work is actually done in America with Vioen, but we actually freeze all the tissue down in the UK and then ship it out there. This is one of the original uh clones here. You can see Gem Twist.
He was a geling. So, in other words, he couldn't breed. So, they decided to clone him. And the one on the right is the clone. And that's the one we have in our yard behind me. Um and he is a stallion in his own right. And he's done exceptionally well. And he's basically, you could say, a sperm bank for the for the original. Uh that just shows what these technologies can actually do.
Uh this is uh oh one of the ladies in my life. She's amazing. She's called Jem.
She's a clone dog and she's actually by my feet now. So when we maybe finish this presentation if we can you can all have a look at her. But she's a clone dog. She's one of the top used to be clone one of the top field trial dogs in in the country. Uh and this we went up snowed in one of the highest peaks in in in the country. So it just shows what is possible uh out there. We thought well if we can do these technologies to all these equids why can't we transfer this over onto other species.
So this is where nature safe was born.
We're facing the sixth mass extinction of all time between 50 and 100 species a day and sometimes nearly going extinct.
It's quite frightening. By the time we've all gone to bed tonight or some of you getting up, you know, in these next couple of hours, another 15 maybe 20 more species would have gone extinct, which is really could be really really frightening. So, our mission is to save uh by collecting and definitely storing reproductive uh and cells and cell lines from species um from all over the UK and the world. And this, if I get rid of all the slides, this says it all really.
This is from Revive and Restore. It just shows that you know all the the species going into the tank and some of the work we can do on it which we heard a bit before in the previous presentation about gene editing or reproductive technologies but a lot of the technologies haven't actually arrived yet. But what we do know if we don't put them in the tank in this end we might have nothing out the other end. So, I think it's imperative that we bank as many species we can before it's too late because they're just some of these species are going at an alarming rate.
And you'll see at the bottom here all you know when these species were preserved. But we collect the samples and we got up to 5 days from when an animal passes away to get those samples in. And we get the reproductive cells and the skin cells and the ears and the hang on um and and we tend to get ovarian or test good tissue uh eggs and sperm um and skin samples. I'd say 85% of the samples we get in are skin um just because it's easier to get to with sperm and eggs. We really need those samples within 12 to 24 hours uh when we're freezing them down. Um one of the fastest declining species on the planet is coral. Uh believe it or not, um we've lost 50% of our coral since 1950. They are the lifeblood of our seas. And I was literally at the Horn Museum only last week uh for the second time. This was the first time we went there. Uh corals spawn uh most for once a year roughly 2 weeks after or before moon 2 hours after sunset. And um you can see the coral spawning here and they release them in bundles. There's sperm and egg together and then they go to the top uh and then we separate them out. The sperm um come away from from the eggs and then we can cut the sperm at the bottom.
>> It's absolutely incredible.
>> This coral sperm. Look at this uh under the eye sperm. Look at those sperm go. I wish some of the other species would have stuff as good as this. Um and we can actually And that just shows it amazed me uh how good the semen was on some of this coral. There's a 3D printed device here that um we did um and we've frozen cryovials um in filter sea water with DMSO and going down at 20° a minute. Um and then a few days later they thawed that out and mixed the uh frozen semen into the eggs and you can see the sperm this side. Look at the tail on that. I mean it's absolutely incredible. um uh looking at these uh semen samples and we got the first cryo coral babies in Europe which is absolutely amazing. So we feel like it uh it's absolutely amazing what this science can actually do. Uh and again coral is very very important. They're the lifeblood of our seas.
Yeah, I've watched this video maybe about I don't know 100 200 times. It always gets the hairs at the back of my neck going up. These are the last two northern white rhinos on the planet.
There's none left. uh apart from effectively they're they are still alive but they're ex uh virtually on the brink of extinction but this is what Richard Vine had to say.
Hello everybody, my name is Richard Vine and I'm talking to you from the old Peda Consery in Kenya.
Behind us you can see the last two remaining northern white rhinos on the planet and we are doing our best to recover the species from the brink of extinction. both are infertile and therefore artificial reproductive techniques are the only options that are open to us. Nature Safe is also playing a critical role uh in saving the world's biodiversity by creating one of the first living tissue banks on the planet.
Their work is incredibly important and we look forward to working with them in the future.
I mean, that's just such a sad video and it's, you know, us as a mankind, we've, you know, produced that scenario and we've got to try and make sure, you know, we uh try and hold the extinction process as much as we can for many of our species. It really is very sad to see that video. So, the science behind it, as you may know, there's lots of bio banks all over the world, but there's very few what's called living bio banks.
There's lots of dead bio banks where they're freezing at minus 80 and those cells uh with no cryop protection those cells rupture and break. Uh so the cells are nonfunctional but the DNA is still there intact but where our cells are all all alive. So we're free freezing in a living bio bank and that is the difference of what we do. I say we take the skin samples usually the ear because those cells regenerate much easier for us. U but there are other parts that we are freezing down. The beauty about this, you know, if an animal dies on a Friday, Saturday, Sunday, even Monday, they can ship those skin samples to us and it's got the whole DNA of that animal in, we can freeze that down and for 10, 20, even a thousand years and help bring that species back if it's on the brink of extinction.
Uh, our data and record keeping are absolutely paramount. You can only it's only as good as obviously storing these samples if you've got the data package behind it. And we use the matcher system. I know Matt's involved with this, but it is this is something that's really showed uh when people look into the charity we run some of the big benefits of that we've got a really good barcoded system uh with the whole identity of that particular animal on those cryobials and um you know when you've got a few hundred or thousands of samples you can get away with other system but when you've got thousands and millions maybe hundreds of thousands samples is absolutely imperative a system like this is is abs can actually make such a huge difference in how you store your samples down.
And this just shows some of the, you know, this is on the new scientists last year saying the northern white rhino could it be saved from extinction literally by frozen skin cells. So this is just showing, you know, some of this work and what it can be done. This was a rhino that passed away in 2019.
We just froze the ear down, part of the ear of the rhino. And then this year we got the ear out of the uh to the minus 196 uh nitrogen tanks. We thored it out and starting to get the cell lines uh growing. After a week, a few days we had a few hundred and after a week we had millions of cells. And the beauty about this is you can replicate. You can replicate when you have sperm samples or in straws. Once those straws have been used, that's it. But you have an infinite supply when you're uh regrowing these cells up. And this was highlighted this year uh on the BBC uh as well, just showing how so important this technologies are to helping save some of these species that are on the brink. And as it was alluded before, it's called the extinction vortex. when you get these really low numbers of species uh and they're virtually in this vortex, nature safe could come uh and actually using that genetic line and bring that species and to help it climb back out of that extinction vortex when that gene pool is so small that you can bring back those lost genetics and it really can make a difference when it comes to effidal sperm. I say it's not just the horses and the cows. Yeah, we do it on all these other species now. And this was a cheetah very inbred don't normally have the best po uh quality semen again because the inbreeding but this is just shows what it can be done it passed away they sent the testes into us and we froze it down I don't know whether you've heard of something called the the mouse deer uh one of the smallest deer in the world the testicle bay is 1.3 g incredibly difficult to extract the semen but we did and we actually managed to get some good quality semen from that the mountain chicken frog incredibly rare there. Um I think there's only a few hundred left in the world. Um and this we we do a lot of uh electron microscope with the with the pictures and get these amazing pictures to see and we're trying to build something called a sperm atlas to try and get some really good pictures uh with this. Uh we don't know whether this is a normal good sperm or not. So we need to build up their data.
A voner and when you do this work a lot of time it doesn't always work. Uh sometimes someone has maybe never even frozen from a bonera for instance. So it's we have these spreadsheets that are building more and more and more. We got hundreds of different species on our spreadsheets and we're learning all the time. So when it doesn't work, we alter it. But this is what one of the main successes. We froze it down, thaw the semen the next day and the semen was incredible. So not only have we preserved that animal's genetics, we've we've got a protocol which is just as important if maybe even more important.
So if this work needs to be carried on in the future, those protocols in place and building up this sperm atlas can really give us a good intel whether this semen is good, bad, how it relates to that particular uh species as well. And so this is something that we're building on and we're just about to send a load uh from a snow leopard I think and some other species we're about to send away and look at their um uh semen quality and looking at h how that semen looks as well. Uh work that's even been done on snakes collecting semen from snakes. Uh um so there's no species that we don't want to be preserving and this is working with the African Lion Safar in Canada. um and they perfected this collection technique and we've been helping them with the crowd preservation side when there's going to carry on trying to do some more work with this because there's very little done on this side of things.
I say preser preserving skin samples. Uh the other technology is cloning we talked about and obviously using nuclear cell transfer. Uh and this is one of the uh first rare breed you know uh horses to be produced by this uh nearly 40 years ago. They've saved the tissue cell from 40 years ago to produce this fault.
Quite incredible. And back then they may may never even thought that it'd be used for cloning. So preserving tissue can have so many different uh pluses. The blackfooted ferret virtually went extinct in the 1970s um and was cloned and was brought back because the genetic pool was so small.
Somebody had the foresight to preserve uh skin samples from 30 years ago. So this really just shows how important it is to preserve these animals before it's too late, before they they're gone.
I think was briefly spoken about in the last presentation, but obviously with the skin samples, you saw the cloning, but there's so other future technologies that will be coming around in turning a skin cell into an egg or into a sperm.
And obviously, this has been done on uh on on the mice and a few other uh species, but it's it's something those future technologies that we should be really looking at.
Our vision uh is to build up hubs and this is we would love to hear from anybody out there on this uh on this webinar. If you're interested in um partnering up or learning more about this, we're about knowledge transfer. We cannot do this alone. It's all about coming together with these technologies.
Um this is a crisis we're finding uh with many of our species and we've got to leave this legacy uh behind for future generations. I think if we don't, they'll look back at us and say, "You have the ability to do this, but you did nothing." So, it's so important that we actually forge ahead and preserve these these species. This is something I love and enjoy and setting up hubs and bio banks around the world. And yeah, we're calling out for your help and we'd love to hear more from you if you're willing.
Um, this is some other work that we did in South Africa. This is me. Part of my job was hanging on to an elephant's penis for about an hour a day. Um, but it sounded a bit weird, but actually it was quite incredible. We did seven bull elephants over a week. U, we collected semen from them, tissue samples, and again, we've lost so many elephants over the years, and they're in big game reserves, which is amazing. Protects them. But again, they can inbreed again.
So, we need to be collecting.
So far to date uh we have collected from 366 different species just in the five years. But that is obviously a drop in the ocean of what we got to do. Um so but we are this is our cryo store here.
As you can see here we have over you know nearly a million straws in store of ecquin semen but also it's where we store all these other species in these uh cryo vaults.
We've been just been nominated for the Earthshot Prize, which is quite fantastic. We just learned about this literally two weeks ago. Um, so if anybody don't know, this is one of the big Earthshop prize. Uh, it's once a year. It's5 million pounds given out to five great causes. So fingers crossed uh for that. But we need your help. Uh, it doesn't obviously we we we need funding, but we're always looking for people to come on board. So it's something you're interested, please reach out. These are some of the people that really do help us. We are a charity at the end of the day. And and I think we all know this person, the great Sir David Atra, and he um in his last one of his last programs called the mating game. I've spoken to you now for just under 25 minutes or something. And what he's summing up in these three sentences or four sentences is what sums up the whole lot. By using these current technologies, if we so wish, we have the ability to save any animal or at least save its tissue until we discover how. And I think that's a a very sort of fitting ending uh to to us.
And I thank you for listening. I'd love to hear some questions uh as well. And thank you, Matt, for and there's some details if anybody wants to take a quick picture of that before it disappears. Uh and you can reach out to me on any of those social handles as well. you that's uh that was a fascinating presentation.
It's great to see how you're endeavoring to transfer uh assisted reproductive technologies already developed in domestic species to um to conserve more of the critically endangered species. Um in the interest of time, we're going to move on to the next speaker to list, but there will be uh there will be there are lots of questions coming in. Um so there will be time at the end uh to to get more involved in a Q&A session. So I'm going to hand back over to Pierre. Thank you again, Tis. Pierre is going to introduce the next speaker. Thank you.
>> Yeah, thank you. Uh, so last but not least, Andres. Andres Gambini. Ciao, Andres, how are you doing?
>> Good to good to you.
>> Okay, so Andres is right now he's in Australia, but of course he's not from Australia. He's a lecturer at the University of Queensland, but he comes from Argentina where he did his PhD in 200 he finished his PhD in 2008. And actually the title of his presentation is kind of a a brief summary of his uh CV because he's been working on all those different species. He's made he's been made he's been the one really making you know producing the first clone horse you know in South America in Australia. So that's uh there is he has a lot of things to say but now that he's austral in Australia he's also working he's not going to talk about that because he's also working on on marupials and uh last year is they produced the first IVF embryos of a kangaroo which is also a huge achievement. So, Andres, uh, you use multi many different advanced assisted reproductive technologies and the floor is yours and welcome.
Well, thank you so much uh Pierre for the for the invitation to to the to the webinar and thank you everyone that is joining us today and I think my my talk today will cope really well to the previous two amazing uh talks that we just had because I'm going to be talking about horses and other members of the family uh such as teras and donkeys and mule and how we can apply a sister reproductive technologies and more specifically um different ways of producing embryos for animal production but also for for conservation and if I have time as Pierre mentioned but only if I have time and and Pierre allows me I will briefly comment uh and give you a snapshot of the work that we have done in in marsupials this year and also in in the salt war crocodile so why am here today talking about this and and and here mention a little bit um on it but I'm a veterinarian originally from Argentina and I had the opportunity to do my PhD in fourth cloning back in in in 2010 um and we were able to produce the first clone PS in South America that were born in in Argentina and and I've been working with this technology pretty much since then another technology that I've been working on is Ixie uh and OPU so cap combined with Ixie and I'm very sure that all the the audience is very familiar with this technology and we were able in our lab again in Argentina back in 2018 to produce the first in our country from produced by OPU followed by And at that time I also started collaborating with Australia where I'm based now. I have a position at the University of Queensland now and I'm doing teaching and research in in Brisbane. But uh we started a collaboration with Melbourne University and we were able to produce the first clone pole in Australia. So as you can see I've been producing embryos in horses for a very long time now. Um, and I just wanted to show you how this technology that when I started was not really well established, but now OPU and Ixie, they are both established technologies in domestic courses. Opu and Ixie um is it's now uh we probably have more than 40 um laboratories around the world providing the service of OPU and Ixie in horses. In Europe the number of embryos that are being produced in the lab through Ixie is higher than the numbers of embers that we are conventionally flashing from the mares.
And in sense of in in terms of cloning we pretty pretty much the same. We we have now at least seven commercial companies that have established many of these companies are actually in Argentina and in Brazil because these are the countries that we can have access to a lot of ovaries from from slur houses and and that facilitates cloning and there are at least 3,000 clone folds that have been born. the the the numbers are of course really hard to estimate because you know this is commercial work and the numbers are not always um uh told. So just to just to show you the work the workflow of what we are doing right now in Australia in Queensland we have our embryo lab in Gaton campus and we have connection with industry partners that they collect all sites from Mars and of course semen from stallions and they send the gamuts to the lab. In the lab, we can produce embryos and these embryos are then frozen and sent back to the industry partners that they will perform embryo transfer. And this is not new, right?
And you're probably very familiar with this if you're a human embryologist or you know an embryologist in general. But what I want to take one minute today is to have this amazing opportunity to to reach a lot of people that is working in the human IVF world to show that I am seeing a huge opportunity on using the course as a translational model for understanding more the impact of human assisted reproductive technologies. We have now a commercial ongoing opex programs all over the world and we have the opportunity to use the mayor as the model to understand more female infertility.
Uh and there are many things that you can see in the slide that are just you know ideas where we can explore and do research. But in my mind, aging for example is one of those things that we can collect a lot of data from the OPU exe program that we have in courses that we can potentially take messages from the human IVF uh world and that's because we are collecting all from may across all different ages.
Very similarly, the stallion can potentially become our model for understanding uh male infertility. And there are a lot of uh similarities and we are using and trying to apply this technology to stallions that have some um degree of infertility and and and looking at OPU and Ixie as a solution for that stallion. And again, plenty of ideas that we can combine. Something that I believe is truly amazing as an opportunity is that we we we want we we we are creating athletes. We are producing offerings and we are monitoring these offerings. So again a huge opportunity to contribute on how producing embryos in the lab is impacting that um long-term and the multigenerational uh impact of a sister to reproductive technologies.
And and lastly, something that is amazing for me as an embryologist as well is that most of the things that we are using nowadays in an ecoin lab are from the human IVF lab. So we are using the same vitrification protocols that are being used for human. We're using the same culture media, same incubators, same needles and so on. So again um just I wanted to take that one minute to show how the opportunity that we have now here with forces and just to give a few examples of what we're doing uh in the lab. Not not not super uh rock science but we're using laser technology for example to select and immobilize sperm as you can see here in the video. Um this has been done in humans already but there's little there's not a lot of um studies looking at potentially the impact of this method of of sperm selection and if if it is something that we should be implementing more or not.
Another very beautiful example on how we can contribute from from the horse into into the human as if the reproductive technology is the fact that this year was formed the first course through maternal spindle transfer. This is a technology that it's already applying human but there's a lot of debate around it and we have now an opportunity to use the course as a model where where we have a lot of mares that are infertile that we cannot get embryos through opu and Ixie and we can potentially innovate using this type of other nuclear transfer technologies but I'm not going to uh keep talking too much about forces today because I want to spend spend most of my time in the other members of the family. Uh and one of these members is donkey. I'm I'm fascinating about donkeys myself and and a lot of the people will you know think that a donkey is pretty much very similar to to a horse and yes they do belong to the same family but the common ancestor was more than 4 million years ago. So they are two very distinct species with very different um um behaviors and characteristics. But we have these two domestic species as part of members of the um equits. But they are not the only members. We have more uh species that belong to this fascinating family.
And this is where the zebras come in place and the wild donkeys come in place. So we have three different uh species of zebras and and three different species of wild donkeys. And this is uh just to make it easier um for the audience today. But what is really concerning is that as an example one of one of the species of don of donkeys the African walley is considered critically endangered right now. there are less than 200 mature individuals in in the wild and there's a clearly a need of uh taking action and this is where we believe as it was said in the previous talk that applying assist reproductive technologies that we have developed for domestic animals into into wildlife it's is what the contribution that that we can do similarly in in the group of the sebras there's one species zebra that is also endangered right now listed as endangered that's the greatest zebras and I'm going to show you the the the work that we have done to try to contribute to develop assisted reproductive technologies for for zebras and lastly as it was said um again in the last talk the shioki force another member of this family also considered endanger and in fact cloning was already applied for for this species But what it makes this group of animals super interesting is that we can mix and match everything really. So probably you are very familiar with the mule which is the the cross with a the mayor with a J.
That's that's a male donkey. But we can also do the other way around and and produce a kiny. So this will be the offspring of a jenny that we cross with a a domestic horse with the stallion.
And this is just if you think about it an amazing opportunity to understand and to study genetics and epigenetics and the maternal environment and and so many other stuff uh that offer us this the members of this family. And similarly with the sebret, we can mix seats with horses and we we're going to have live offspring. That's a source and we can mix severs with donkeys and we're gonna have a son. So yeah, it's it's just fascinating that these intercross are producing viable offspring. Although most of the offspring will be sterile, infertile.
But what is even more interesting is that within this within the members of this family the the genetic plasticity that we have is just amazing.
researchers some yeah few years back they demonstrated that if we took for example a zebra embryo from a zebra so it's a 100% zebra embryo and we transfer that zebra embryo into a mare into a synchronized mare that mare can get pregnant and can deliver a healthy zebra hole and the same thing we can do with donkeys and mules and zebras and horses So it's a a fascinating uh aspect of the family that allow us to to to have the opportunity to have and use domestic equit as uh recipient animals for wild equit.
So I'll now go straight into applying the technologies that we have been hearing about and and and talking about in the last two two lectures. So, we had the opportunity to partner with one of the of the Sue here in Argentina when I was working here as a researcher and I got a call from a friend that was working there and she she mentioned that Deborah have just died and I said, you know, uh just send me the testice and send me a piece of skin. And that's what we did. We were able to isolate several sperm from the tail of the epidmus and freeze that sperm using our knowledge on on on force semen freezing. And with this with the skin cells we were uh with a piece of skin we were of course able to isolate fibroblast skin fibroblast and freeze those fibroblast.
What we did next was say used the zebra sperm and injected that zebra sperm into a horse egg.
And this uh will be producing source embryos in the lab. And with the skin cells, what we did, we used a horse egg.
We inuculated that horse egg. We remove the DNA from the course and we introduce and we use the stomatic cell from a zebra as a DNA donor for for that oite to then produce an embryo that will have the genetic of a zebra.
So let me tell you uh a little bit of the findings of of this research that we did and the first thing that we do in our lab. We like a lot to do Ixie Pier know this and one of the things that we do is we can of course with Ixie as you know you bypass all the barriers of fertilization and you can inject any sperm into any site pretty much and and that's what we did at the beginning we we used pig eggs um because that was uh this easy to have access to pig eggs we invitro mature this eggs and then we injected pore sperm and we knew that uh When we inject the horse sperm into a pig, we will trigger that initial activation and initial steps of embryo development. So we can see the two peruclei here for example and the two polar bodies. And when we injected zebra sperm in pig eggs, we were able again to see that we can still have peruclear formation. Although the rates were a little bit lower compared to horses, they were still happening. So we decided to keep moving and then inject that sperm now that several sperm into a horse egg and and this is a bit of our table with with the results and I'm not going to spend too much explaining the numbers but if you look pretty much of course we have a horse control here. So this is injecting horseer into horse eggs and and this is the blastosis rate when we inject a zest sperm into a horse eggs. So it seems to be a a bit low but of course this was just one animal and we pretty much demonstrated that what we are seeing in vivo can be replicated in vitro and we can produce embryos in the lab um that these hybrid embryos in the lab.
What happened with with the cloning? So with the cloning um one of the technologies or or the um innovations that we developed during my PhD was embryo aggregation. So what we did we produce horse uh sorry zebra embryos. All these zebra embryos are coming from the same cell line the same animal. So they are all genetically identical. But after doing the cloning we remove the son of palucida. So what you can see here are three embryos that they don't have sona palucida and they are under a time-lapse machine and we culture the embryos together in the microwwell and we do aggregation of three embryos. So basically this embryo aggregation technique allow us to create an embryo one single blastosis that it's coming from three embryos officially.
So this we we publish a lot around this technology and we demonstrated that embryo aggregation improves embryo quality, pregnancy rates and and and cell differentiation and so on. I can I can give a a a full talk on on embryo aggregation but yeah I'm going to stop the video because basically this will reach to uh the blastoy stage and uh but the video is is a bit long to to spend all the time on it.
So what we saw is that compared to horse cloning um that we were using a cell line from from from a horse as a as a as a control group. We were able also to produce zebra blastosis uh in that cloning rounds. And it looks like cloning zebras it's easier or more efficient than cloning horses. It might be but if you work in cloning you know that there's a strong impact of on the cell line that you're using. So there it's not something that we can that we can officially say but you can you can look at the blastosis rate and you can we were amazed on how uh easy was to obtain blastosis when we were using the a horse um inucleated egg to reprogram that ferbra skin fibroblast. But we decided also to to look at the embryos more in detail and understand whether uh that or plasm was doing a a proper reprogramming of that cell and we did that by looking at and on some markers at the blastoy stage and you're probably familiar with this two which is a marker of the inner cell mass and then yap one which is a marker of the tropo ectoms that of course are going to be more placenta and then we compared clone horse embryos with zebra clone embryos and we pretty much didn't find any difference suggesting that a horse or plasm can reprogram a zebra skin cell and and contributing to understand even more the genetic plasticity that we have within the members of the family.
Um so now just just for the sake of time I'm going to um um move on. with with donkeys. We were able to achieve the first donkey IVF embryo in 2022. We reported that doing embryos in donkeys is not as easy as it is in courses. We had to set up the oven pickup technique. Um this is a work of one of my PhD student here in Argentina at the University of Rio Puerto. Um but long story short we were able to successfully collect all sides and in animals compared to humans we always collect immature oid. So we we then need to do in vitro in vitro maturation.
There are a lot of things that we learn uh when we move from one species to another specy. We were we're looking at how opu impacts the estro cycle the the fertility of the donkeys and so on. But what I want to spend maybe a little bit more of time it's in this next slide because we are now thinking on a domestic mo donkey as a potential model to understand more about aging and assisted reproductive technologies because donkeys compared to horses are more resilient animals. They live longer.
So we are looking at for example mitochondrial activity in in in donkey oides that are young and donkey o sites that are old and we're also of course comparing that with horses and we're using different dice to look at the mitochondria and whether that mitochondria is active or not. Um and just to summarize a bit of of our finding is what we are what we are seeing is that all donkey also seems to to have a total number of of of mitochondrial within the egg. Um and of course that kind of give us a little bit of more total number of active mitochondrials. So the ones that are healthy which were the red before. But when we look of course at the ratio of the you know what out of total mitochondria which one are active the still all donkeys um animals are not oh sorry eggs are are not able to compensate from from a healthy young donkey also. Anyways this is the contribution ongoing that that we have in donkeys. We we needed to set up in vitro maturation. We realize that in vitro maturation in donkeys is way longer than in horses. We still need to know if this is because the media that we have developed is not yet 100% um fine-tuned for donkeys or if it is that just naturally donkeys will take more time to extrude that first ba body as you can see there. Um and then we decided to do some ixie some intracytoplasmic sperm injection into donkey oytes and uh we were able to to obtain the first donkey blastosis produced in vitro and we're now producing more blastosis.
Um we are doing that comparation between young and old donkeys and of course we're also having our mule control group. So if you think about it, producing a mule in the lab, it's injecting a a donkey sperm into a horse egg. And and this is this uh of course for us is very useful to to have as a control for that uh sperms uh to to be able to measure the sperm uh impact on development and and so far we are not having like huge difference across all these different hybrids and and species. We're also doing some timelapse imaging work and and looking at the morphoginetics of the emerald, but I don't have time to go into the details of that today. So, just to kind of wrap up my talk, I think um I'm close to the time. Um I I'm hope I'm hoping that I have provided some evidence that we now have a beautiful model that we can use to understand more about human reproductive technologies and that domestic force as reproductive technologies that we now have shown that the technology can be moved from other members within the same family and that we can pretty much apply this technology to those other members of the family that are critically endangered as a as I show towards the beginning. And I don't know, Pierre, if I have time to show something about the kangaroos.
>> Okay. You have you have one minute. Go ahead.
>> Okay. I'm going to skip acknowledgement then and then I'm going to just jump into the work that we did and we um show we published this early this year and and that was producing the first um kangaroo IVF but if it's not officially an IVF it's an exil. I'm just showing you here a beautiful slide with some of the of the pictures of of how a kangaroo egg look like. This is this is this is an ovary. First we are collecting ovaries from from dead kangaroos. Um and then we collect the oates from the ovaries very in a very similar way that you will do this with cats or with mice.
And what you can see here these are cumulus all site complexes from kangaroos.
And then we had to track down in vitro maturation for these species because as as Andrew mentioned, marsupials are complete different and but we did it. We were able to invitro mature kangaroo sperm. You can nicely see there the first polar body and then we injected a kangaroo sperm that we collected from the epidmus and then we freeze and and we were successfully on producing embryos that cleave and then we um we just fix and look at DNA quality of the embryos.
We're also doing something similar in koalas. These pictures right here, these are uh uh immunofllororesence on koalas sperm. Koalas sperm is fascinating. They have 12 different type of heads. There's a lot that we can uh that we can learn from from understanding biology of marsupial. This is a koala ovary and one of my PhD student is developing um koala oite recovery and we just published a paper now where we um recovery for for the first time all sites from from koalas and we are looking at uh basic stuff just as the you know how the normal site in Koala should look like and just you know to have fun and because I'm in Australia I couldn't help myself and we did some crazy uh research on far north Queensland in Australia with saltwater crocodiles. I'm talking about 3.5 meters animals and we were able to collect semen from cloak massage from these animals and we are also now working on um preservation technologies and and chilling media for uh developing and helping assisted artificial insemination in these species. All right, that's it. I'm going to go back to my acknowledgement questions, but I think yeah, thank you so much for for everything.
>> Well, thank you so much, Andres. That was really great. So, we're going to we're going to >> we're going to switch directly to the everybody's going to come up on on camera and uh we're going to switch to the Q&A session and uh we're going to request our uh speakers, our guests to uh to really provide some uh quick and snappy answers to a question because uh we don't want to to to be too long. But again the most important is that we have questions that are that really show that you know there is a huge interest from from the the audience. Um I would say that um first of all Andres I'm going to start with you. Um you you you demonstrated pretty well that the the the horse could be a very good model for for human reproductive medicine. And uh there is a question uh about actually that is related to that is because um apparently it's very difficult to superovulate uh aquids in general. So is that true by the way and uh does it create kind of a potential challenge if you we you when for your work in IVF or ixing?
>> Yeah that's that's a good question.
Yeah, it's super ovulation in forces has it's it's challenging because of the anatomy and hisystologology of the of the ovary in equits. Um, so it's it's it's being tried many times and it's not it's not something that it's uh commercially viable right now and the efficiency of the superobulation is just not going anywhere. So that's that's a big difference with with the human IVF um workflow in in in domestic species in pretty much all domestic species we need to uh if we want to collect a lot of oates we need to collect those oates from immature follicles or antron follicles that are growing. That being said we can still recover an invivo mature oite. We can inject hormones in the mare to induce that pre-ovulatory follicle to ovulate but we will only recover one. Uh and that's why the technology has now moved into just aspirating all the folics all the follicles that we can see in the ovary and doing vitro maturation instead of trying to collect invivo mature all sites.
>> Thank you.
>> Thank you. I have a question for Tullis actually. Um this is a question that relates to conservation of species and it's fairly evident from your talk that you know there it's almost a race against time to proactively cryopreserve um as much biodiversity as possible. But the question is if the world fails to act fast enough what class of animals or species lost do you fear will have the greatest ripple effect on ecosystems and why?
>> Thank you Matt. Um, I mean to me it's there's one answer here. It's insects at the end of the day. Yes, we're doing all these other the mammals and that side of things, but the insects, the pollinators, we need them and you know, uh, without them, you know, our our whole ecosystems and the biodiversity of our planet can can fall apart. So, the insects and they're very difficult to cry reserve and and so we're actually going to be actually looking at butterflies crow trying to crow reserve butterfly lavi for the f, you know, I think one of the first times. Uh that's a PhD student we're looking at. We've lost 80% of our butterflies in the last I think it's 40 years which is frightening. So our insects are a challenge. There's no doubt about it.
And uh I think if we to lose them uh we're in serious trouble. I suppose it's less glamorous as a as a class of animals because they're not, you know, fluffy and cuddly and large, but they're they're probably Yeah. As you say, you know, we often forget the importance and of them and how they are really integral to the uh stability of ecosystems.
>> Absolutely. Absolutely. And and the rhinos, the elephants, and all these amazing other species out there, there's a lot of work focused on them, but less fluffy animals, as you say, there's less work on them, but they're vitally important.
>> We better get on with it. Thanks, Tis.
>> Okay. Um, a question for for you, Andreas, and it's going back to more like about the >> Can you hear me? I'm sorry. Looks like my internet is getting slow. Okay. Uh, it's more about the cloning and it's a question from Valeria Canya. Um so how genetically identical are cloned animals to their nuclear donors particularly considering denovo mutations and mitochondrial differences.
So I don't know if you can get a snappy answer on that one but >> yeah yeah I I'll try but that's a very very very good point. when we are doing cloning it we we need to be considering that we are putting a a somatic cell that has its own you know mitochondrial DNA within a that will have a complete different pool of of mitochondrial DNA so we are different definitely changing the mitochondrial DNA to that nucleus and there's a lot of evidence that there's a you know strong connection and communication within the a nucle the nucleus of the cell with a with the mitochondrial. So that's definitely impacting the the cloning and more we we really need to pay attention to this when we when we think on cloning as a strategy for conservation, right?
Because we we don't want to mess up things. we want to try to to create embryos and and future animals that we know uh that are genetically valuable for for for the population and and and just changing mitochondrials is is not the way to go. So there's a lot of other technologies that we can applies to replace the mitochondria later on. So it's not impossible but that's a really good point.
>> Thank you.
>> Thank you. Another question relating to cloning um which we've had from a number of uh of attendees in relation to biodiversity. So if if biodiversity depends on genetic variation, can cloning ever truly support it or does it merely by time in a race against extinction? Do and that's a question for either of you as you're both involved in um the practice of cloning.
>> I think the classic case in this is the Blackfoot. We we we saw that you know some people say cloning how does that help biodiversity uh um you know with with you know if you clone lots of uh of one particular animal no it doesn't but if you bringing a lost genetic line back that has been lost for history and that biodiversity of the genetic pool is very very small that clone coming back can really help climb out that extinction vortex and the and and the blackfooted fair was a classic example that of bringing a a species back that died nearly 40 years ago to help a species that's here now. Um and and that just showed what it can do >> integration with with other species. Um >> yeah. Okay. Thank you.
>> Yeah. Just to comment on that. I think that's one of the beauty of cloning, right? It's it's a it's a machine to copy genetics. But if the genetics that you are coping is a is a very valuable genetic that is that that was lost or that a genetic that didn't have time enough to contribute to that population then you can put it back and then you can officially increase genetic v variability within your population and that's what you need to save a species that is a brink of extinction. You need genetic variability.
>> Thank you. There's another question from Valeria actually regarding um another form of assisted reproductive technology which is liophilization um freeze drying I think so have have either of you worked with sperm liophilization particularly regarding DNA integrity preservation and subsequent embryo development. Uh do you see any potential for this technology?
>> Yes, we I mean I can I can quickly comment on that. We we are indeed working with that. We I have one of my PhD students his his PhD is based on uh reinforce and donkey sperm. We are looking at DNA fragmentation levels and we're looking at how other important proteins within the sperm changes with um free ring. We're doing proteomics on that. So we will soon give you a lot of a lot of more information about it. But what we can see and of course be has a lot of background on this as well. Um but there it's a it's a very good way to keep the DNA intact. Of course the sperm will die but the sperm uh will remain remain intact. Yeah.
>> Yeah. Pier do you have a view on that?
Uh as you have some knowledge of that.
>> Well yes I mean we we don't we don't necessarily work on liophilization. We we work on desiccation or dehydration here. uh we don't use the same techniques because liophilization uh is supposed to have first freezing process and then sublimation of the ice but what we do is we dry directly and actually Andres came spent some time in in our lab earlier this year um to to work on domestic cats and and other types of endangered species and but I agree with what Andre is saying it's a it's a very intriguing and and cool options But we still need to do a lot of basic research because of course there are obvious applications to human uh reproductive medicine. But we need really to be careful about you know the the consequences of the the preservation and the stress of the cells you know during dehydration and all this kind of stuff.
>> Wouldn't it be wonderful though if we could end up in in a world where there was there was no um low ultra low temperature storage and we just add water like coffee. That would be fantastic.
Thank you.
>> Okay, Matt, you had a you had a question. So again about the the success rates of cloning.
>> Yes, it's a very quick question. Um, you know, could you give us an indication as to to what are the success rates that you could expect from cloning in domestic species?
And that's again I to either of you >> perhaps to list do you have any experience of that?
>> Um just with the it depends what you call success rate. Uh when the samples tissue samples come in yes uh if we get them within usually 5 days the success rate is incredibly high. I mean it's over 95%. It's very high when we get those cell lines to grow and freeze them back down. uh when we ship them off to to to Biogen um I know the success rates been incredibly high if you're looking obviously the embryo failure rate that's different I don't know those figures to be fair off the top of my head so yeah Andreas you might be able to >> yeah yeah I can comment a bit more on on that side of course now that we have many commercial companies that are not necessarily publishing you know the new changes on the technology the the technology is officially a little bit higher potentially in a commercial world than than we have in science and the efficiency will change in in different species. So different species cloning technology has different efficiency. In one of the species that we're doing a lot of cloning which is horses I can comment on that and we are probably having nowadays a 30% blastosis rate with cloning. So the the blastosis rate efficiencies is is quite high. um the pregnancy rates are the a bit of the problem. Not the early pregnancy rates, but those pregnancy that are able to to keep going until till till the term.
There's a lot of pregnancy losses within within within the within the process.
And the main issue is the placenta. So what we really know is that in clone animals, the placenta does not develop properly. Though there's a lot of work trying to improve the trophy or the placenta development in clone animals that's uh will definitely improve the efficiency of the of the technology and something that is important for for the audience is to understand that cloning relies a lot from the somatic cell. So there are some cell lines that are very good and you create a lot of blastosis, a lot of pregnancy, a lot of live healthy falls and some other cell lines are just not giving you anything ever.
So there's there's something that we don't really necessarily know in depth that is making that huge difference.
>> Yeah. And this is living proof.
>> Yeah. Of what is possible.
>> Oh, so it does work.
>> It does work. so much opportunity to to develop and devolve the technology. It's the prospects and the future is looking very bright and the the proof of the pudding is in the eating as we can see in the arms of Tulis.
>> Okay. So, I think it's time to uh to thank everyone uh and to close our webinar. So, of course, I'm going to start by thanking you uh Tullis and you Andres, and we thank also um Andrew even if he's not with us. Uh thank you to the audience and um I would like to thank of course Matt for his uh his help to to co-pilot this uh this webinar. So thank you so much.
>> Likewise. Thank you Pierre. And and we'd also like to recognize the the I3 initiative and the organizing committee uh Jacqu Cohen, Peter Naji or Peter Nars, sorry, Maryanne uh Sevates, Thomas Elliot, Fran Farley, Violet Sura, and of course Giles Palmer. Thank you very much guys. It's a wonderful initiative and we really look forward to what's to come in 2026.
So on that note, please do join us again next year to talk about many new hot topics and the latest from the IVF world. If you are celebrating, then please have a wonderful uh happy and safe festive season and we wish you health, wealth, and happiness in 2026.
Thank you everybody and thanks again to the presenters.
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