Channel: hPSCreg® clear
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| 1 | hPSCreg® | How to build a European iPSC Collection meeting research needs? | 14 | 31.9 | 15:52 | I'm actually Julie Holder, I'm representing Rosin in South Sciences, which is actually the sort of coordinator of in collaboration with Tim's role in actually coordinating the E-Bisk consortia. And I'm going to talk about building the European IPSC collection to really meet your needs, the researchers needs, and on demand. So what my talk is going to actually be on, the overview, the introduction and the initial hot starts. And we actually really started this collection with a series of well-established IPSC lines that were available from academic collaborators and to really generate the data to allow us to make some of the decisions. Some of the information that I will be giving you is actually on this hot start and how we've actually co-really interrogated these IPSC lines and learned an awful lot from them. In addition, I'm going to talk through some very brief summaries on how we use these cells and then summarize how you can actually obtain them, which will be continued throughout this talk this morning. So it's a consortia consisting of a very large amount of partners and we all integrate together, actually working with each other to facilitate the requirements for depositing the IPSC lines and really expanding and utilizing those IPSC lines in differentiation assays to different lineages. So to build the catalog, as I said, we had these seven IPSC centers that really contributed to the hot start lines. Underline was the Sanger Hipsky lines which are of normal origin. And in addition, the FPA partners actually commissioned particular lines of interest. So we've done a lot of gene editing with Bioneer and this is something that there are a number of posters on throughout this ISSER meeting. In addition, we actually have utilized some IPSC lines from other EU funded projects. And the make-up of these lines has really fallen into some very discreet areas, mostly neurodegenerative and psychiatric cardiovascular, some minor genetic variants, metabolic such as type 2 diabetes, blood and auto-immunity and muscular skeletal disorders. The actual continuum at the bottom on that of this slide actually shows the cell line receipt from the IPSC derivation center and then the foring, we actually register it into Hipsky and we cry a number of particular vials to establish that sort of master cell bank. We then for the cell lines into various plates and culture them on in a feeder-free environment with no antibiotics and continue that throughout the expansion process. So we've had experiences of cell lines that have recovered well or haven't actually recovered well and then we QC and Glyn as Tim alluded to will actually talk a lot more on the QC data that we have established to qualify these cells moving forward. Then when the bank has actually been made of the cells, we actually transfer cells to our mirror facility at Franhoff or IBMT in Germany to actually negate the sort of issues that if one freezer goes down then we have a backup etc and we ship to E-CAC for distribution. So in the course of this project where it's anticipated that we will be establishing 280 new FPIR cell lines there will be cell line panels from diseased patients and also with normal and sibling pairs who have disease or no disease. In addition there is a number of gene edited cell lines that we are establishing and we have brought in a number of collections from Horizon 20 and IMI collaborations. So what we're building is this centralized activity in the EU for the collection testing and distribution of these well-qualified IPS lines to understandidized diverse disease representative lines and panels which are reproducible and will give you a quality of cells to actually allow you to undergo undertake your research we've concentrated because of the FPIR partners interest on most of the neurodegeneration and cardiovascular side so we have representative lines in the catalog from hunting turns from Parkinson's Alzheimer's disease etc and these will actually be expanding as the catalog is built. So it is anticipated that we could or you as a researcher could use the IPS lines to stratify the disease or stratify based on drug response using these differentiated cells in a disease environment and looking at specific disease endpoints. So the current catalog and I've had to put this qualification up that it's as of the 18th of June which is when I prepared this slide. I have had a deluge of emails to say that there's been a number of cells released so this catalog is actually changing dynamically so I would encourage you to go to the website to to ebis.com to actually look at the catalog as available and it will be demonstrated in this session but just to show the diversity of the lines that we have and that this will be building in the future. Also to note specifically in a lot of these lines there are two clones available in the catalog so you can actually build up a very comprehensive understanding of the disease etiology moving forward. So it's been a heroic effort across the whole of the consortia to actually really procure the Hipsky supplying centres to get the information from the donors, the cell history to expand cry preserve and quality control and we're really here now sort of talking about a very well established process that would result in you being able to order a particular cell from a well characterised cell bank to allow you to do the research. This gives a feel for the process steps that are involved in the expansion of the cells and routinely we would have approximately 50 within a working cell bank but the availability to go back to our master cell bank to generate passage, similar passage numbers to the initial working cell bank if required when the cells have actually the vials have been exhausted at eCAC. The QC is a this is a very general snapshot to show that we have cell phenotype genetic identity, the pluripotent potential, chromosome stability and sterility of all the the cell lines within the catalog and this actually shows a vial that is actually being distributed from eCAC with the code what passage number and it links it back to the data sheet that actually gives you all of the information on these cells. So we had these hot start lines and we had 47 in total and I just wanted to share with you how they performed during the process so remember that this is seven IPSC derivation centres. We actually had a training program so that all of the researchers supplying the cells into the bank were actually trained in all the processes that we wanted to adopt. So what media to use, how to passage them, this was actually a very well regarded workshop and as actually one of the highlights that people actually said in the whole process when it was initiated. So as you can see here there were some individual results on the recovery of the cells actually showing that there were 12.8% of the cells, there were issues recovering the cells either didn't stick down or they didn't expand, they didn't perform as expected. There was quite a high percentage of spontaneous differentiation in these cells but they were very clean with regards to viral screening but a little bit of fungal contamination across these 47 lines. However what was worrying but what does agree with literature out there was that 17% of the cells that we received in the central facility were actually not what we expected so there was some mix-ups at the actual derivation centre so a female actually was a male in one instance so you know this is something for other banking initiatives to be aware of that you do need to look at your cell line identity quite carefully. But on the positive side sorry it should be N equals 38 not 38% 38% of the cells sorry 38 of the cell lines that we received actually went on to 100% to go on to trial in each differentiation of forming mesoderm and endoderm and ectoderm. So ECAC distribution this is actually just the process showing that the files are stored in a unique ebisc freezer so there's no contamination with other cell lines of unknown microplasma levels and they're easily traceable and they're shipped within the appropriate packaging. The examples that I've got are three so this is actually a slide from one of the partners to Finnegin who actually have a hepatocyte and also pancreatic cell differentiation protocol that we have been using to actually look at the ability of the IPSC lines to differentiate into hepatocytes and pancreatic cells and this actually shows very nicely that the IPS lines that have been banked within ebisc are actually have the ability to differentiate into very well characterized hepatocytes as shown by the albumin levels and by the presence of the alpha-1 antitripsin. So this is actually a key metric to show that these cells have the ability to differentiate. This panel is actually from a bioneer line which is in the catalog showing that they differentiate to a neural lineage the map 2 are actually present on dendritic cells and total tau showing that you've actually got a good network and beta 3 beta 3 tubulin as well. So this is an example from the University of Bonn, Oliver Brussell's lab actually with a disease line and I wanted to show you the actual differentiation capabilities of a control line on the right hand side of the left hand side of the right hand panel. You've got the control line showing that you've got an increased new right length but in this hereditary spastic paraplegia or HSP you can actually see that there is a reduced new right length in the IPSCs once they've been differentiated into neurons and this is actually something that is being looked at in a drug screening campaign to see if you can recover this new right length in the disease lines. So just showing that they have the ability to to really give very good results in a differentiation capability. So just to say when summarising that we're actually building this centralized activity we're at the first stage of of really delivering the initial catalog and this is constantly changing and we will be putting in a huge amount of IPS lines from specific diseases and it's an opportunity for you to if you have an interest in generating a particular line from a rare disease then this is something please come and talk to us but we are really I think delivering to a very high standard. So just to say this is not a single entity this is a consortia and the next slide just shows the breadth of the individuals who have been involved in this and this is actually taken from the last year's general annual meeting which was held in Granta Park but we've had a more recent one in Berlin and I'm sure that if you'd like to see the slides we can actually present you with them. So thank you very much for your attention and I'll take any questions. | ↗ |