0:00
So much for that kind of introduction.
0:01
Yeah, the name, I think as we are German, originally we were not aware that it has the British also second meaning, but now it's the company's encounter and now we have to deal with that anyhow, especially on the American market, we get quite nice feedback for that, especially at the networking events.
0:18
But so thank you so much for the introduction.
0:21
The catch phrase is not actually the name, it's more the bionic bioreactor thing, quite bionic because we orient at the principles of nature for the design of our systems.
0:36
So what is behind that?
0:39
We are a small bioengineering company in the West of Germany in Aachen, which is one hour in the West of Cologne, close to the Netherlands, Belgian and Dutch border.
0:52
So and at lunch we can have some nice French Belgian fries and maybe some Dutch beer in the evening.
1:00
So it's very close to that triangular border region.
1:05
We are now 25 employees.
1:07
It's not actually I think we're 28 right now with two product lines.
1:13
I want to show you later on a bit more so and we help especially academics and R&D departments from small other companies to go from 2D into 3D.
1:25
So there's also a service we can offer already.
1:29
We are operational in Europe, the Middle East and North America and we did a lot of different cell type cultivations already.
1:41
And I will show you some more about this later.
1:45
So let's dip a bit into the more the technology.
1:52
So with recent cultivation technologies, we see a lot of draw breaks, otherwise we wouldn't have get any funding or investor money.
2:01
So we see that there are a lot of hurdles still given the bubble aeration technology, which is I mean quite historical to say.
2:14
We see the issues when it comes to IPSCs, all kind of stem cells or sensitive immune cells that even by the occurrence of bubbles, the cells will differentiate in a way we don't want.
2:29
So we're losing the stem cell character of the cells later on in the process or the forces induced by the bubbles by the implosion on the interface between the gas phase and the media phase induces so much force that the cells will erupt.
2:47
And of course we have the foaming issue which form out even cells from the whole batch.
2:54
The foam formation is quite the other, the bad evil twin of the bubble aeration.
3:00
And in the very beginning of our system we try to cultivate micro fermentation, fermentations of microbiologics, for example to produce some green washing detergent.
3:11
And here we start with 200 millilitre biorectors, like a cup of coffee.
3:17
And we needed forty litres to capture the foam of that because while we produce washing detergent, we produced biosurfactants.
3:25
So this was a nightmare in the lab and we came to the idea wouldn't it be nice to circumvent the bubble aeration at all?
3:33
Because normally you could also use just a high surface to volume ratio by surface aeration, for example, in those cells decks here.
3:43
And I mean, this is a picture of a press release from Bayer from the Berkeley Lab where they have hundreds of petri dish stacks.
3:50
I think this [unclear] stacks filled with media.
3:54
And you can imagine that maybe people living in the with that in the loft have some other people cells living in the top part of that skyscraper of having some other cultivation condition than those at the Pateria.
4:14
So the quality of cells is an issue here.
4:17
And to harvest those stacks by hand is another nightmare I don't even want to imagine here.
4:26
So we came up with the idea to decouple the aeration from the surface aeration and the bubble aeration.
4:33
And we do this by membrane aeration.
4:36
And we're not the first one.
4:37
Sartorius had some regards here in the last decades, but our system is way better and I will show you why.
4:45
So this is our 10 litre system.
4:47
I can disclose here the single use system, and we were able to introduce those systems in such a way that it's extremely mild for the condition of cells.
5:00
We have some patrons here also professor Doctor Robert Zweigerdt in the audience.
5:05
Nice that he already tried this system out.
5:07
He will show in the in this evening.
5:09
I think his talk is at 5 maybe a picture of our recent IPSC cultivation and Catalan, for example, is a close partner from us in Germany in the US.
5:25
So how does it look like, as if you could be so nice and push it around here.
5:30
Just be prepared to have something already.
5:33
Yeah, you see that we have two technologies combined here.
5:37
1 is the gassing.
5:39
But before being talking about the gassing, we talk about the flow.
5:42
So the membrane blades are arranged in a way that we have soaking in the media and the cells in suspension on carriers as spheroids as organoids doesn't matter and pushing them quite mildly over the membrane sheets.
5:59
And then we have in here like a fluidic circle and the top of the same one and we have some mixing in the inter space here.
6:08
And this is like nearly 90 percentage of the, just 10 percentage of the shear stress you regularly would have in such a system by a classical stirrer.
6:20
When it comes to those blades, they contain out of dozens of single hollow fibres.
6:26
And you will see this a bit closer when you get the prototype in your, the vessel here, not in your hands.
6:32
Those hollow fibres are well like plastic macaronis you can imagine.
6:38
And in into the lumen of the macaroni of the hollow fibre, we flush the gas.
6:44
So the process gas comes from top, from the stirrer, here inside it's split in the headspace, and from the headspace it's distributed to each single hollow fibre, to hundreds of those hollow fibres.
6:58
And here the magic happens because we have a dense membrane, a dense hollow fibre.
7:03
So it has no pores, it's completely smooth on the surface.
7:07
And the molecules of the oxygen diffusing alongside the concentration gradient from the membrane into the media, in the media they are already solved.
7:17
They can be consumed directly by the cells and the cells producing more cells product and carbon dioxide.
7:25
Normally the carbon dioxide would reduce or heighten or change the pH.
7:31
But here also alongside the concentration gradient, the carbon dioxide diffuses backwards and as we have a constant airstream inside the stirrer, we flush out the carbon dioxide.
7:43
The rest is collected at the bottom and through a different a channel we can flush that out of the system and this is really the function of the lung or fish gills inside the bioreactor.
8:00
With that technology we are able to control the pH and the dissolved oxygen level way closer than all other technologies which we are aware about because we have a very high surface area with the membrane.
8:12
In regards to all other technologies, it's like 20 fold the headspace area realised in the stirrer and this by very mild conditions.
8:22
So we can mimic several parts of the human body.
8:25
With that we can mimic hypoxic regions or acidic or basic regions of a body.
8:37
With that close control. When you normally need like 3 hours to calibrate your sensors, with the O and pH with your classical bioreactor system, here you go, after 20 minutes, you can directly start your cultivation.
8:53
Maybe something to announce.
8:56
As you may see, we have the bottom part of the bioreactor and in the next version of that or in let's say an upgrade of that, we will exchange that bottom part with a cell retention filter.
9:08
And here we can tailor made to the regards of our partner and customers the pore size to keeping back the cells, microcarriers, whatever it is inside the vessel that they're having the same cultivation conditions for the whole condition, for the whole process.
9:27
But we can nicely exchange the media over the whole surface area.
9:32
Also by design of that we can backflush so that we control biofilm part biofilm building up at that part of the vessel quite nicely.
9:44
So that was quite some bold statements going to the facts.
9:48
If you compare the tip speed we use, we and others use in the system to reach KLA under classical conditions, you'll see quite fast that by given tip speed, we have like 10 times higher KLA input in the system.
10:08
That means at the same time we can only use 10 times less energy we're reaching the same KLA so that we can for extremely sensitive cells have very mild conditions here for the cultivation and thriving of those cultures.
10:26
Of course the tip speed is not directly the energy dissipation rate, which would be the benchmark number, but it's quite hard to get those numbers from others.
10:36
But in publications we at least can see the tip speed and the rough number.
10:39
I think this is quite good for estimates some effects.
10:48
As you see the 300 millilitre single use vessel is already on the market.
10:53
We have at our booth also the two litre and the 10 litre stirrer which can be used as an upgrade in classical glass vessels.
11:03
We will launch the two litre single use vessel in January next year and the 10 litre vessel in Q3 next year and single use bioreactors.
11:14
And we're having working prototypes for 50 and 200 litre at our facility in Aachen.
11:21
The upper limit what we envision for this technology because we still have the issue that the surface area we can offer at the membrane is always slower growing than the volume we can add in the bioreactor would be around 1000 to 2000 litres.
11:40
Talking about two product lines we have on the left side our yeah, we call it the vanilla cigarettes addition or menthol cigarette addition, which is to tease people to use that system.
11:51
It's a quite cheap and a nice system to use just into your incubator.
11:56
And we rely here on the atmosphere of the incubator and the gas concentration of the incubator.
12:02
And this is like a sophisticated stove flask.
12:04
Yeah.
12:05
So you have still the active aeration part, we have the temperature, and you can just use with low amount of effort and the technology.
12:16
But of course you don't have per se online monitoring and online control of the values for that.
12:23
Unfortunately, as everywhere we need a complete controller system with gas control, gas mixing and pumps for sophisticated glucose feed and stuff like that.
12:35
But if we use this one, we can get quite comprehensive data sets for the scalability.
12:43
And then we can use the small vessel here in this nice stand, which can also be used for the perfusion system.
12:49
I introduced, because it's actually nearly the same vessel.
12:54
Yeah, just with the exchanging of the bottom part.
12:57
And we can go with the same controller from 300 millilitre to two litres to 10 litres.
13:02
So scalability here is directly given.
13:05
And we were able to prove that for example, with the Catalent trials.
13:11
So talking a bit more about the data maybe, yeah.
13:17
So we in the last year we were quite busy by using a different cells and enforcing our cooperations with partners all over the globe to achieving some biological data here.
13:30
And you'll see we have some results.
13:33
We can publish some of course regarding the partnership can be disclosed only under NDA.
13:40
But if there's something in your interest, please ask me later after the corporation or yes, and we would love to get in touch with you.
13:50
So for example, one of our first results was with Swiss University to cultivate IPSCs on microcarriers.
14:00
And for sure this is not the most fancy way to cultivate IPSCs, but it was the 1st result and we were able to show that with classical bubble aeration cells are quite fastly dead because the even by low aeration.
14:16
I think this was 0.1 volume per volume.
14:22
The cells cannot withstand those bubbles. In a classical T flask, which is the T of the black line.
14:29
Here we got quite a decent amount of cells in cultivation within classical bioreactor it was a bit lower, but I mean regarding the error bars not significantly.
14:40
And with our reactor we got the best results.
14:43
I mean, otherwise I wouldn't show the slide, but we are quite proud on this and has enforced us to go further in this way.
14:53
TreeFrog which will have also a presentation later on use our technology or in the cooperation.
14:59
We use that to cultivate their IPSCs and they are very nice encapsulation technology and very able in regard to their classical I think 110 millilitre system to show superior cell expansion and also longer expiration of pluripotency markers.
15:24
Here with a rooster bio, we were able to cultivate human bone marrow derived MSCs on carriers as well.
15:37
And we reached quite a nice viable cell density of around 49 million cells in the 300 millilitre batch of us.
15:52
This one of my favourites.
15:53
This was with Cologne Hospital in Germany.
15:57
And regarding the resolution of the Pima, you have to trust me here that those particles to see you're floating, you may see floating around are not bubbles but organoids.
16:09
These are cardiomyocytic organoids more than 200,000.
16:15
We see that in our two litre system here.
16:17
And after the, I mean, we did the formation of the partners at the formation Petri dish, then we seeded them into this bioreactor.
16:25
We did the differentiation here in our system.
16:28
And after seven days, the more than 90 percentage of the cardiomyocytes were beating, which is quite Frankenstein science.
16:37
Yeah, this is absolutely astounding to see those mini heart tissues starting to beat.
16:47
As mentioned with Catalent, we did what we would call an on the fly scalability or scaling here.
16:54
We used the iNK technology, or Catalent used their own iNK technology also some results which they also presented in Boston this year and we started with 100 millilitre media and then we see that they self drive so nicely in our system.
17:12
We directly put the same cells on the fly in our two litre system and we were able to achieve from far more than 1000 fold expansion factor, some more classical cells.
17:30
Let's see if the video.
17:32
No.
17:35
Could you be just so kind and check if the video would play. Well normally it spins not so fancy, but yeah.
17:48
Anyhow, what we did here is CHO cell cultivation with Rentschler Biopharma from Germany for example, but also with some academic partners.
17:58
In that batch we reached CHO numbers of 35 million cells per millilitre in perfusion, we reached two 248 million cells per millilitre in perfusion, which is a really thick soup.
18:14
So as we can now state that a ration of the gas transfer is no longer the bottleneck of cell cultivation, especially for sensitive cells.
18:27
We did something similar with HEK cells.
18:29
I don't have the slide here that was yes, 150 million cells per millilitre.
18:36
I believe as well around 150 million cells per millilitre with HEK cells with which we are also fancy to use for production of viral vectors, for example, or vaccines.
18:50
Looking for partners right now to do that.
18:52
So if you have some ideas here, we would love to cooperate on this.
18:56
OK, 5 minutes.
18:57
Luckily it's one of my last slides.
19:02
Another topic which is quite en vogue is mean is exosome or EV production.
19:08
Of course, we tried something here with the Universal Hospital of Aachen.
19:13
We used THP-1 cells for those production and we figured out that with our system the EV production was quite higher than in classical shaking flask or T flask.
19:28
Yeah, like yeah, double 83rd percentage higher than in classical T flask.
19:36
I also think we were able to reproduce with MSCs, yeah, which are also a classical system for EV production.
19:44
With that, I thank you so much for your attention and I would love to chat with you about that topic.
19:51
Thank you so much.