New Matter: Inside the Minds of SLAS Scientists

The Ins and Outs of Single Cell Gene Editing (Sponsored by Molecular Devices)

May 08, 2023 SLAS Episode 149
The Ins and Outs of Single Cell Gene Editing (Sponsored by Molecular Devices)
New Matter: Inside the Minds of SLAS Scientists
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New Matter: Inside the Minds of SLAS Scientists
The Ins and Outs of Single Cell Gene Editing (Sponsored by Molecular Devices)
May 08, 2023 Episode 149
SLAS

In this special episode, we're joined by Cytosurge CSO Tobias Beyer, Ph.D., and SEED Biosciences CEO and Co-Founder Georges Muller, Ph.D., for an overview of gene editing with Cytosurge's FluidFM® in combination with DispenCell™ dispensing technologies.

Tobias and Georges explain the FluidFM® technique and how it differs from traditional CRISPR methods along with the advantages the technology has over other methods of gene editing.

For a transcript of this episode, please visit this episode's page on Buzzsprout.

Our Sponsor for this Episode
Molecular Devices makes scientific breakthroughs possible for academia, biopharma and government customers. Dedicated to enabling life science labs of the future, where innovative technology and novel research meet, Molecular Devices empowers scientists to advance discovery, driving earlier diagnoses and safer therapeutics for patients. Spanning cell line development, 3D biology and drug screening, their automated, end-to-end solutions streamline and scale complex workflows while integrated machine learning-enabled analytics allow researchers to mine data easily for insights.

Molecular Devices is the innovation partner that empowers scientists with next-generation technology to advance discoveries, improving the quality of life everywhere.

Learn more about Molecular Devices by visiting:
https://www.moleculardevices.com/

Stay connected with SLAS

About SLAS
SLAS (Society for Laboratory Automation and Screening) is an international professional society of academic, industry and government life sciences researchers and the developers and providers of laboratory automation technology. The SLAS mission is to bring together researchers in academia, industry and government to advance life sciences discovery and technology via education, knowledge exchange and global community building.  For more information about SLAS, visit www.slas.org.

Upcoming SLAS Events:

SLAS Europe 2024 Conference and Exhibition

  • May 27-29, 2024
  • Barcelona, Spain

SLAS 2024 Microscale Innovation in Life Sciences Symposium

  • 11-12 September 2024
  • Cambridge, United Kingdom

SLAS 2024 Sample Management Symposium

  • 16-17 October 2024
  • Toulouse, France

SLAS 2024 Data Sciences and AI Symposium

  • November 12-13, 2024
  • Cambridge, MA, USA

View the full events calendar

Show Notes Transcript

In this special episode, we're joined by Cytosurge CSO Tobias Beyer, Ph.D., and SEED Biosciences CEO and Co-Founder Georges Muller, Ph.D., for an overview of gene editing with Cytosurge's FluidFM® in combination with DispenCell™ dispensing technologies.

Tobias and Georges explain the FluidFM® technique and how it differs from traditional CRISPR methods along with the advantages the technology has over other methods of gene editing.

For a transcript of this episode, please visit this episode's page on Buzzsprout.

Our Sponsor for this Episode
Molecular Devices makes scientific breakthroughs possible for academia, biopharma and government customers. Dedicated to enabling life science labs of the future, where innovative technology and novel research meet, Molecular Devices empowers scientists to advance discovery, driving earlier diagnoses and safer therapeutics for patients. Spanning cell line development, 3D biology and drug screening, their automated, end-to-end solutions streamline and scale complex workflows while integrated machine learning-enabled analytics allow researchers to mine data easily for insights.

Molecular Devices is the innovation partner that empowers scientists with next-generation technology to advance discoveries, improving the quality of life everywhere.

Learn more about Molecular Devices by visiting:
https://www.moleculardevices.com/

Stay connected with SLAS

About SLAS
SLAS (Society for Laboratory Automation and Screening) is an international professional society of academic, industry and government life sciences researchers and the developers and providers of laboratory automation technology. The SLAS mission is to bring together researchers in academia, industry and government to advance life sciences discovery and technology via education, knowledge exchange and global community building.  For more information about SLAS, visit www.slas.org.

Upcoming SLAS Events:

SLAS Europe 2024 Conference and Exhibition

  • May 27-29, 2024
  • Barcelona, Spain

SLAS 2024 Microscale Innovation in Life Sciences Symposium

  • 11-12 September 2024
  • Cambridge, United Kingdom

SLAS 2024 Sample Management Symposium

  • 16-17 October 2024
  • Toulouse, France

SLAS 2024 Data Sciences and AI Symposium

  • November 12-13, 2024
  • Cambridge, MA, USA

View the full events calendar

Hannah Rosen: Hello everyone and welcome to New Matter, the SLAS podcast where we interview life science luminaries. I'm your host, Hannah Rosen, and joining us today is Tobias Beyer of Cytosurge and Georges Müller of Seed Biosciences. They're here today to talk about the novel approach Cytosurge is taking in developing CRISPR cell lines with their FluidFM CellEdit service. Welcome to the podcast Tobias and Georges. 

Tobias Beyer: Thank you, Hannah.  

Georges Müller: Hi, Hannah. 

Hannah Rosen: So, to start off with, I would love it if both of you could just kind of give us a brief description of your professional background and expertise. So, Tobias, would you mind going first? 

Tobias Beyer: Yeah, of course, no problem. So, I obtained my PhD from ETH Zürich. The work there was focused on tissue repair and regeneration. And after that I did a postdoc in Canada, Toronto at the Lunenfeld where I studied, made the stem cells. And self-reading, stem cell signaling pathways, and the tools we applied were mainly proteomics and also some genomic approaches. After that I switched back to ETH to the junior group leader, I was working under the umbrellas of Professor Wutz and Professor Corn, where I used my experience like in before to do further research into signaling pathways in stem cells, and that's where I started doing quite a bit of CRISPR work, where I got first into CRISPR and I perfected that my CRISPR skills in the lab of Jacob Clark, usually at the end of six years at ETH. Unfortunately, you get tenure or not I was not, but I had the great luck to join Cytosurge as the Chief Scientific Officer. Maybe of interest to know, I've known the technology well before I started at Cytosurge, one of the pharmacists, we often discussed potential applications over beers and yeah, the CRISPR was one of those so-called bee ideas, I guess. 

Hannah Rosen: Seems like the best ideas are beer ideas sometimes. 

Tobias Beyer: Sometimes, yeah. 

Hannah Rosen: Thanks, Tobias, Georges? 

Georges Müller: Alright, so I'm Georges. I have a PhD in stem cell and bioengineering from EPFL, that's a school in Switzerland. Early in my career I worked on cell and gene therapy projects. It was like, 10 years ago when I started my PhD and in the lab we were developing really cool approach where basically single cells were... were sorted and selected to develop safe and efficient cell and gene therapies. So, that was really, really early in this growing field. Now cell and gene therapy... but we... so, my professor had the idea that at some point, if we want to democratize the field of cell and gene therapy and... and the use of single cell space, we need to develop the tools. So, this is about the time I started my PhD, then five years later I came... I came up with a with a prototype together with my colleague, and then we cofounded Seed Bio. So, that was end of 2018. So now we have a team of 20 people. We are based in Switzerland, the French speaking part as you can hear. We are really big fan of SLAS. I've been there with you guys since the early days of the... of the startup. We've won many awards, Ignite Awards, Innovation AveNEW, I mean all you can imagine. And so, I'm really... I'm really a big fan of SLAS. We won the New Product Award, 2022, that's when we launched the product. Since then, we have sold many units all over, and Cytosurge is one of our customer, and so not only we were able to find customer things to SLAS, we also found, first, partners. In fact, this webinar is sponsored by Molecular Devices, and this is our commercial partner since beginning of the year.So,o I'm very happy to be here representing Seed Bio, but also inviting Tobias Beyer, who's working on a really cool company technology and and... and he, together with his colleague, he's helping, you know, the field to grow and... and... and making it possible. So, we're closing the loop, CRISPR Single cell. Happy to be here. 

Hannah Rosen: Yeah, it's fantastic. I mean, it's been exciting for us at SLAS to get to see, you know, Seed Bio grow and flourish, and that you're able now to kind of pay it forward and pass it on to other companies. It's just, it's exactly what we hope to see out of that Innovation AveNEW at SLAS. So, it's really quite thrilling for us, I have to say. Well, so, you know, Tobias, I would love it if, you know, you could just kind of start us off here by telling us, you know, what is FluidFM, this technology that Cytosurge is... is producing? 

Tobias Beyer: Yes, I would love to go a bit more into details there. So, it was originally invented at ETH. Basically, what it consists of, the idea is that you combine first microscopy and fluidics. Just maybe a brief introduction to force microscopy. Microscopy is based on cantilever, very small cantilever, that has a tip and it can sense forces when you put it down on cells or any kind of matters. The force is measured by the diffraction of the laser beam, so it's just basically simple physics, so this is nothing new. There has been around for quite a while, but the idea was to use, instead of just cantilevers, to use hollow cantilevers coupled to microfluidic pump. And that invention pair basically enabled us to generate the smallest needle in the world. So just as a comparison, I think it's about 100 or 1000 times smaller than the... the needle of a mosquito. So, it is really tiny.  

And what can we do with this needle or with this whole technology? Because it's all for sensitive, we can go to a single cell, approach it very slowly, and then we have a pyramidal tip at the end of this cantilever with a small opening. What we can see then is that this tip will penetrate a single cell, presumably the nucleus. And once we are in, we can apply slight positive pressure. And through the microfluidics we can inject centiliters of liquid into a single cell. So that's basically the core of the technology. What's also interesting, you cannot only apply pressure, but if you apply negative pressure, you can also sample small amounts from a single cell without destroying it. And that's another application that we're not going to talk to about today, but I will maybe mention it in a side note afterwards. And this is what we call basically the injection mode of the device. There's a second mode instead of a pyramid shape here, of just a cup, you can imagine like a suction cup, and instead of having, for example, RMP in the cantilever, we just have a bit of trypsin in the cantilever. So you have one single cell you really want to have out of your culture. So what you do is you go over the cell, you release trypsin locally, dispense it, the cell will detach and then with the suction cup, you can go down on the single cell that's detached, your apply a bit of negative pressure so it gets sucked on to the cup. Then you move it to another well. So, it's kind of similar to what Georges is doing with the DispenCell. Well, unfortunately it's not that high throughput, so we can only use it for very rare or special cells. So that's basically the... the... the broad view of our technology. 

Hannah Rosen: Wow! Is it difficult to learn how to do this? It sounds like you could very easily destroy all the cells that you're trying to work with. 

Tobias Beyer: Fortunately no, it's not. The whole process is guided by robotics, so it means when we approach a cell, we set the certain threshold of force or maximum force you can use. So as soon as it senses the force and has the force reached, it stops pressing down. So, it's very, very gentle and what we can see is when we just inject regular cells, usually the survival rate is between 90 to 95%. So, the injection process is very, very gentle. Originally these devices were classical AFMs, so atomic force microscope, and honestly, these are really tricky. They're not meant for biologists. Physicists are... are good at those, but it's too complicated. And one of our invention was to transfer net technology and write a program interface that even like, a average talented biologist, biologists like me can understand. OK, so I will take it a lot easier than your average, let's say, spinning disk microscope tool, right.  

Hannah Rosen: Yeah, that’s wonderful. I mean, the fact that you have it all fully automated, I'm sure it just makes everybody's lives so much easier. I'm just thinking back to like, my days in grad school when I was learning patch clamping and just how many cells I destroyed trying to figure that process out and the fact that it's just, you don't have to worry about it. That's just awesome. 

Tobias Beyer: No, you... you... you don't. Sometimes you break your cantilever, or something goes wrong, but usually it is very robust. You said it's automated. I will not say it's 100% automated, it's more a click and point and then you can step away, but it's yeah, semi-automated I would say. 

Hannah Rosen: Very nice. So, you know, you said that you're using this tool for CRISPR applications. Could you describe a little bit, you know, how are you using the FluidFM for gene editing and how is it different from traditional CRISPR trip methods? 

Tobias Beyer: Yes, I would love to. So, basically what we do is we take RMP, so Cas9 protein, the guide RNA directed against our target gene. We keep that in mixture and what we do is we fill our cantilever with that mixture. We go to our target cell and gently inject it. And yes, I think the main difference is between like the regular transaction methods is that you do it at the single cell level that you do it, you control the amount of RMP so you know exactly how much you put in. And you were not only just putting it into the cell, but you put it into the cell nucleus. That's where the CRISPR is needed. It's not just somewhere in the cell, it's in the nucleus. So that is the main advantages. Further, what we think is a really important point is that because when you do electroporation. So you electroporate your cells, you never know how much I mean, what ratio of the Cas9 RMP really reached your cell. So, this is one of the issues, so it's a mosaic or not 100% equal transaction method. So, when we do the injections, we know exactly how much is in there. Because we are so gentle and being injected into the nucleus we can use roughly 100 times less Cas9 RMP compared to traditional methods and OK, this is on one hand, it's nice on... on your budget, but on the other hand, it's also very important for the quality of your edit. So, what we also usually say is when you look at the genome editing, it's basically a non-somatic reaction. So, you have an affinity for your DNA. You have your protein, and of course the more protein is in there, the likelier it gets it caught, which is good if you want to have it cut. But you also have the so-called off target effect, so similar sequences in the genome. That have maybe a KD that's in the range of 10 times to 100 times less affinity compared to the on target. But if you swamp the system with so much Cas9, the likeliness that you have off target is quite high. And this is why we also think our instrument has a huge advantage here because if you are 100 times less than our electroporation, so the chance that you get enough storage is also diminished by quite a bit, so that's kind of the basic ideas or approaches. And if you want to go a step further, the other thing is you just don't... don't... you want to have knockouts, but if you want to do knock INS which occur very... the frequency is quite a bit lower. So, what we thought of is if you place the DNA, the DNA repair template, not just somewhere in the cell but directly also where it's needed in the nucleus, you can increase that precise editing by a factor of 5 to 10 fold. That's what we have seen in report assistance so far. So, these are basically our main applications at the moment and the advantages. 

Hannah Rosen: So, it sounds like this is... it's a... you said it's a much more precise way of editing genes and more efficient as well. Is it faster? Do you see results faster than you would compared to traditional CRISPR methods or are they about the same? 

Tobias Beyer: I think that, yeah, it's... the time frame for an app that is basically limited by how fast your cells grow. I mean, they're, maybe they're a bit faster, but I don't think this is the main idea to go with our service, it's comparable right now, but what is definitely different is also the approach. So, what we do is we start from single cell instead from bulk cells, so that cuts down quite a bit on hands on time because you have your single cell right from the start, and then you just grow it up instead of transacting it, sorting it, and then growing it up. So, I would say the time frame is probably the same, but the hands on time is by quite a bit reduced. And this is also kind of where Georges's dispenser comes in. So initially what we did is we used our, like, our pick and place we call it, the... the top shape, the isolation method and yes it works, but we realize the throughput is just not good enough. I mean, this is suitable for if you want to do CRISPR genome engineering. Our device, your own, that is fine. You can do that. But if you want to industrialize the process, the more reliable and more faster method and you're looking around for quite a few devices. Personally, many others, and honestly, the best thing that happened so far to us was a dispenser. So with Georges's DispenCell that we can do is we can go feed our cells in the plates. We know it's right in the middle, we know it's a single cell. And once it has attached, we can go back, we can go and inject it and then let it grow up. So, the combination of these two technologies, they are different, completely different, but combining them really accelerates the whole process. 

Hannah Rosen: Now, I have never done CRISPR myself. I am not by any means a gene editing expert, so forgive me if this is a dumb question, but I'm just wondering, you know, what are the advantages of performing gene editing on a single cell versus a whole batch of cells at once? You know, what is the goal with doing... looking at just a single cell versus, you know, having a whole bunch of cells in a Petri dish throwing, you know, all the... the gene editing at them and then just seeing where it takes. 

Tobias Beyer: I mean both... both approaches are valid from my point of view, but I would say for... for really precise evaluating cells, often that you have to be sure that you have only one initial cell, so it has to be monoclonal. And guaranteeing that from a bulk transaction is just kind of more tedious. So, what people usually do is they... they include, for example, a GFP, mRNA or plasmid that they know, OK, these cells have taken up the CRISPR editing we select for them by fat sorting and then we grow them up. But even that doesn't really guarantee you 100% monoclonal antibody and if you want to do it really, like, for a more clinical application you have to go through rounds and rounds of serial dilution just to be 100% sure that you have one single cell. I mean, that is one of the main differences. Of course, you can go and say, listen, I don't care if I just have maybe a 70% knockout or 80% knockout, I just hope that I will see my knockout. That's fine, but the risk that you miss your phenotype is quite big, because sometimes, like, just hanging out of... of... for serial protein in your cell can cloud outcome. 

Hannah Rosen: So, it maybe depends a little bit on the application, what your end goal is? 

Tobias Beyer: Definitely, yes. 

Hannah Rosen: That makes sense. 

Georges Müller: Good point. In fact, what's really about clinical developments, I mean, the field of cell and gene therapy is not there yet. But in classical biotech and bioproduction, the isolation of a single cell to produce the cell line and the selection of the best clones is a actually a requirement from the FDA. The FDA requires that any clones have to be derived from a single cell progenitor. And then it's up to the companies to, you know, provide evidence and strong statistical data that it is the case. 

Hannah Rosen: I see. So it's not just, this is better for your research, it's actually this is requirement if you want to get FDA approval. 

Tobias Beyer: Yes, that's definitely the case. 

Hannah Rosen: OK, excellent. Thank you. So, you know, what are some of the limitations, then, of this technology and this technique? You know, are there any situations where this is just not an ideal solution for gene editing? 

Tobias Beyer: I would say there definitely are situations. I mean, there are certain cell types, for example, that are just refractory to single cell outgrowth. So, starting from a single cell and then doing gene editing and then realizing that your cell will not grow, that will be painful. So that's the one thing that we test beforehand before we start our service, that each new cell line that we get, we do certain tests to see that we can grow them up. So, if you have one of those cells, of course, bad idea. The second example where you have limits is definitely also cell lines or cells that have limited expansion capacities. So for example, we have to figure out right now a way to grow primary cells that we can edit them and still have enough growth capacity in them left that we can do with the downstream experiments. So that's another limitation. Interestingly, but it's not really limiting, but we just recently found out is that we can also inject into non adherent cells. Of course, we have to trick them a bit, we have to make them adhere just for like, a couple of hours because otherwise, you can imagine, you go with your poking needle onto the cell with quite a bit of speed, and basically if it's not adhering what you end up with this kind of playing cell soccer. So, the cell flies off and it's gone, so that's... it's... it's kind of funny to watch, but it's not so funny for the operator. But we figured a way around to kind of make them stick a bit, and then we can inject. On the other hand what’s definitely and absolute no go is tissue. So, as soon as you have more than two or three cell layers on top of each other the... the geometry of our tip will not allow us to inject. Sure there are other cells like oocytes, there as well they're just simply too big to reach the nucleus of the cell. And the last kind that we're still kind of thinking whether we should attack it or not right now is plant cells. So, as soon as you have a cell wall and not on your cell membrane, it's really difficult to penetrate them. So, these are things that we put aside right now and we're not focusing in it, but I hope in like, mid-term, like in three or four years, we can start tackling these as well. 

Hannah Rosen: Yeah, it's very interesting. So, you know, on your website you... you provide the FluidFM that looks like that people can purchase to use in their own lab, and then you also have the CellEdit service. So, could you maybe just describe, you know, when people are purchasing the FluidFM, you know, what are they... are they using it for? And how is that different from the CellEdit service that you provide? 

Tobias Beyer: I mean, I'll have to say the... the FluidFM device dominium is kind of a capital equipment. So, for a smaller lab, it's just not worth the hassle just to buy just to do genome engineering. However, if you have for doing some like a microscopic CORE and you have dedicated personnel that will take care of the device, it will make sense for... for university to go and buy the... the... the Omnium and you're fine with that as long as you don't use it for commercial, like, competing with our cell line editing service. Besides that, what's probably more interesting or what we get more request right now is the... the so-called life seek technology I just mentioned that before it's based on the same tips and the same technology. But instead of injecting into something into a cell, we can aspirate tiny amounts of cytoplasm, for example, and there was a like a hallmark study I would say about the deep branch and by the whole groups, I think it was published in Nature last summer. Basically what they did is they took, like, the AFM equipped with our tips and pumps, and they sampled the same cell over and over again, and took examples and did RNA seq. So, basically what they could do is they could do a temporarily resolved, well, sequencing, transcriptome analysis from the same cell. And that worked very well. So, what we are currently doing is that we kind of take their approach, make it fit to the easier to use... to the Omnium, and we train people on how to do sampling, we train people how to take the, yeah, the biopsies, how to set them down. And then, OK, the downstream analysis we have to take care of them... of that themselves and source it out, but any kind of sequencing service can provide you, after which it's the... the sequencing facilities or sequencing protocols. Kind of, my personal dream is to combine these tools. So, imagine you're going to a CRIPR added in one cell. But you already have taken a sample from that cell and then you see really how the cell reacts to the edit, and then you just go and sample it again. And then you can do a differential gene expression profiling or can detect your editing like in the same platform or in the same platform. But I have to admit I will have to spend some R&D on that to make that happen, but that's kind of where we looking forward to it. 

Hannah Rosen: So, this is more long term plans for this. 

Tobias Beyer: Yeah, I would say like, you know, within the next five years, that's somewhere we want to move on, that we combine things that we can kind of have more, yeah, like a hollow service that. You not only get the edits, but you can also have a glimpse on what's going on in your cells. 

Hannah Rosen: So, what inspired you or interested you in developing the... a technique for single cell CRISPR engineering? 

Tobias Beyer: Honestly, that was during my time at ETH, so I think my... my PhD students at the time they... they tried and tried to do CRISPR genome engineering in human embryonic stem cells, and at one point they got it to work but it was really tedious. And at the same time I was, like, speaking with Mike, the... one of the founders and the brains behind the technology. And it was just like, looking at it, and like ah, we have to try this. Also, I was forced... forced... yeah, I had to make the step from academia to industry, and that was just a fantastic opportunity to start in industry. But the topic I like, CRISPR cells and like, cutting edge technology, that you cannot find anywhere else. And everything came together. And I have to say, Cytosurge is a very special company. It has been around for a while already but we are 35 now, but it still feels like you're working in a small startup, so it's... it's very, yeah, personal. People are welcoming and everybody talks to each other. So, I really enjoy that.  

Hannah Rosen: So, what is the sample prep like for this process? You know, you had mentioned earlier that you were utilizing the Dispencell to isolate the individual cells. Could you go into a little bit more detail of just, you know, how exactly are you doing this whole process from start to finish? 

Tobias Beyer: Yeah, of course I... I can, I'd love to do that so. Usually what we do is what we start off is, we received the cells, we know the... the add in We want to do so we did design the guide RNAs for the customers, we order all the reagents, and then we start off with the so-called optimization round. So, each cell line behaves a bit differently, so we have to figure out how they behave. And, among this sort of optimization round, we optimized seed parameters so that we know, ok, we can see 10 cells, there are single cells, they’re in the middle of the well. So, these are the first parameters. Meanwhile, we go and optimize the injection parameters and, of course, the genotyping and all these things, and at the end of that process, you know, our baselines. And basically what we do then is we just go seed, usually between four and six 12-well plates. And roughly, I would say 90% of the cells are already single cells, we let them adhere. That is of course done with the Dispencell. We let them adhere. We wait between 12 and 24 hours, depending on cell type, that they're adhered nicely.  

And then we put it into the Omnium and we go and inject all these cells. Next day we come back. We usually co-inject the GFP mRNA as a kind of a trace that we know OK, the R&P went in, cell is healthy. We do the statistics and then what we do is basically wait until cells start growing. There we have certain tricks, some cells require specialized medium, some inhibitors, condition medium, just to make them feel more happy that they really grow out. And depending on the cell line, between I would say three weeks is probably the fastest and we had a really difficult one recently and it took us almost two months to get, like, enough cells to do the first genotyping round. There what we do is trypsonize the cell, take a small sample, usually like 200 to 1000 cells per genotyping, leave the rest, grow them up and until they are ready for it to be re split. We already know the genotyping results, so we can select the clones we want to keep, trash the rest, and then expand them further. Cryopreserve them, do some quality controls like mycoplasma testing, see better if there are contaminants in there, and then send it back to our customers. So that's the whole process. 

Georges Müller: So, when... when I listen to... to Tobias and I... I have to say I'm... I'm very excited because what we're talking about today is, this is precision medicine, but it's that's the true definition of... of precision medicine at the single cell level. That's amazing. And we are living in a time where we see more and more applications that like... like that and their, so Cytosurge is... is provides CRISPR as a service and they sell tools. But there are other companies out there doing CRISPR, and we really see now a transition from research projects academia to... to biotech and soon to clinics. So, this is... this is very exciting, exciting time I... I... I believe. And for us as a company that's, you know, give us a chance to... to contribute, obviously with the modest contribution, when you listen to Tobias, the workflow is... is quite long and... and quite complex, but I'm very happy that now, you know, we have a company and we have a product and we can help people like Tobias developing their... their... their workflow.  

So maybe a... a quick word about Dispencell. When we hear Tobias describe it, it's also quite simple. But it's not. The question is, can you put one single cell into a well, at the center of the well, so I can come with my atomic force microscope with the probe and... and come in contact with the single cell. So, I think if you say it like that, that... that... that's not so easy, and the... and the setting of this, the precise setting of the cell at the center of the well was... was critical. So, this is exactly what Dispencell can do. So, Dispencell is a single cell dispenser, single cell seeder. It's a compact pipetting robot that... that can enable the scientists to isolate single cells, and we can do that much faster and with... with high reliability compared to the... to the competition. So, the Dispencell, very briefly, it's a... it's a robot. It's used with kits for stability and with the software. For workflows like that, stability is important, so everything is contained in the chip, as soon as you want to do with the cell, you know, use it for a different project, you just change the chip, use the same machine, but change the chip like the coffee machine, famous one, choose one, and then the software is here to... to provide proof of code unity. So this is the our first... first format and I am very happy to, you know, have customers like... like Tobias that can use them and... and hopefully it's helpful for them at the end of the day. 

Hannah Rosen: Yeah, it's very cool. And I should mention real quick that if people are really interested in the Dispencell and want to learn more, we do have a episode of New Matter dedicated to the new product award that Dispencell won last year, so if you kind of dig back through and we can probably link to it in our show notes if people are interested in learning more, we go a little bit more in depth in that previous podcast episode about the Dispencell and its capabilities. I’m wondering, you know, with this FluidFM and your capabilities, you know, what future advances, if any, do you hope to make with this technology? You mentioned a little bit earlier of some of your long-term goals, but, you know, is there anything else that you're working on? 

Tobias Beyer: I mean, what we also know today is that there is a huge need for larger insertions and larger knock ins. Like, for example, Georges mentioned that the production of biologicals that often requires the insertion of the transgene that the promoter, like the gene of interest and some resistant genes, and these constructs are often, yeah, I would say at least three KBs in length. And that is kind of a limit to do that the... the traditional methods, yeah, you can electroporate it in but as soon as you want to take the like a viral vector, for example, it starts getting difficult to package the whole information into the viral vector plus having the Cas9 present. So what we are really envisioning is that in the future, with optimized conditions from the FluidFM that we can go and place these larger inserts into the cells at a higher efficiency and the greater probability. And that will be definitely kind of the... the next pending stuff that we want to do. And, of course, when you start doing these larger things, I mean, 3 KB's then maybe 10 KBs, and then you basically, are the point where you start writing genomes instead of just only editing them. And that's kind of where the direction we want to go. 

Hannah Rosen: Yeah. Georges, is there any advances that are coming along for Dispencell that are gonna make this process easier for Tobias's team? 

Georges Müller: Well, for Dispencell, of course there are... there are many, many new products to come and new product awards to... to win. But we just want to jump in here, Tobias mentioned about, you know, the need to industrial... industrialize the process, and we also mentioned about the FDA. So down the line there is a need for automated solution. This is the way to expand the process for I would say affordable cost because the companies we're talking about, the ones that are developing those workflows, they're usually small to mid-size companies, so they don't, I mean, correct me if I'm wrong Tobias, but they don't necessary extended unlimited budget, so costs, those are important. So, I would say automation and... and cost is is important and... and I regard industrialization, one thing that we mentioned is the clone selection, because we created the cells then you need to select the clones. And one of the things that, you know, can be implemented in such a workflow is a... is an automated imager like Net imager and there are quite a few in the field here. Although because this webinar is sponsored by Mol Dev, I should mention the... the... the clone select imager, which is a great product which you can use to image tens of plates at the same time and get what's the so-called monoclonality assessment, proof of clonality. The proof that one cell has been isolated.  

Tobias Beyer: I mean, this will be definitely of great interest for us. And if you, I don't know if that's possible, but one of the things we struggle with is, like, that we don't have to look too long for the cells before we want to inject. We kind of need to coordinate. Could you imagine that you can write an algorithm that kind of detects the single cell and then measures the... its relative position to the rest of the well and then gives you the coordinates that we could feed into our Omnium? 

Georges Müller: You see that, Hannah? I told you he will have an application.  

Tobias Beyer: I mean, that is... that is exactly the things that we are working on. And we try to optimize the whole process because the poor R&D associate that sits at the microscope and has to look for one cell in the in the total plate can be kind of frustrating, so something like that, that would be very much helpful. 

Georges Müller: Well, we should reach out to the people at... at my desk, see what they think. But I'm sure they would be... they would be interested. And... and just to... to comment on that, so now Dispencell is available through Molecular Devices, in particular in the in the US and in Asia. In Switzerland, you can also buy it through Seed Bio, but... but, you know, for a startup like... like Seed Bio, having such a... a deal with Molecular Devices is... is very important. In particular, if we want to now help, you know, companies to industrialize their process, having a partner like Mol Dev which is, you know, very famous in the field and which has high credibility, which is known by the FDA as a provider, that's important, which also provides support overseas because we're now based in Switzerland, so it's good to have such a partner. So yeah. In... in... in... this... in this vision of having big companies to industrialize, it's... it's great to have companies like... like Mol Dev, and... and again I'm... I'm very thankful to SLAS putting us... for connecting the dots. 

Hannah Rosen: That’s what we're here for! So, if there is a researcher out there listening, who has heard everything that you have to say and they're on board, they want to get started using your CellEdit service, what do they need to do or to know in order to get started? 

Tobias Beyer: It's basically simple. The only thing you really need to know is the target you want to target preferentially with like, a unique symbol, because otherwise it's a bit hard to untangle. Sometimes in the case there are multiple isoforms from the... the same gene, maybe also specify... know which splice variant you want to target, know which cell line you want to use and then just, I think, on our web page there is even a kind of a quote that gives you an estimate of how much that will cost, so you can use that and then contact our sales department and usually within 24 hours you will be in touch with one of our representatives, and as soon as it gets a bit more complicated, someone of our team will be there too to advise you and try to find the best solution to do your knockout or your small knock in. 

Hannah Rosen: Right. Well, we are almost out of time for today, but before we go, I just wanted to ask both of you if there's anything that we didn't get a chance to discuss today that you really want our listeners to know either about the FluidFM, the CellEdit service, the Dispencell, anything that we've discussed so far? 

Tobias Beyer: I think from my side that was very... I think we touched all the... the main points. And please, if you have further questions, do not hesitate to... to reach out to us and I'm happy to take the time also to talk to you personally or if you have particular questions, very difficult edits, we’re here to help. 

Georges Müller: And from Seed Bio, yeah, we are happy to... to... to listen to the needs of scientists and see how we can... we can help. In fact, we're gonna be at SLAS Europe in a few days, and here I'm biased because I'm also co-chairing the scientific conference. So, if you listen to this podcast, please come to the... come to the conference, listen to the greats talks, invited speakers, and feel free to come to the booth, Molecular Devices, Seed Bio and Dispencell will be at the booth, happy to connect there and also put you in touch with the people at Cytosurge. 

Tobias Beyer: Right. 

Hannah Rosen: Yeah, thank you both so much for coming today and for telling us all about this cool technology, and we look forward to seeing you at our European conference in a few weeks, or days depending on when this airs. 

 

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