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Oh hello and we're glad you've joined us for this live webinar prepare for success GMP cell banks as part of a staged standardized platform-style cell production process I'm Suzy Valdez of lab routes and I'll be moderating this session today's educational web seminar is presented by lab roots and brought to you by thermo Fisher manufacturer of GMP give Co cell therapy systems reagents for pluripotent derived cell therapy development for more information on our sponsor please visit them at thermo Fisher comm now let's get started you can submit questions to the speaker during the presentation while they're fresh on your mind to do so simply type them into the drop down box located on the far left of your screen labeled ask a question and click the send button will answer as many questions as we have time for at the end of the presentation if you have experienced technical problems seeing or hearing the presentation simply click on the support tab found at the top right of your presentation window or report your problem by typing it into the ask a question box located on the far left of your screen this is an educational webinar and those offers continuing education credits please click on the continuing education credits tab located on the top right of your presentation window and follow the process for obtaining your credits I now present today's speaker Benjamin fryer Ben is the CEO and co-founder of plura stiix incorporated providing GMP pre expanded pluripotent stem cells contract development services and consultancy for groups and company developing cell and tissue therapies Ben has almost 25 years of experience as a leader and scientist in large pharma academia and startup companies developing combination products and cell therapies for complete biography on our speaker please visit the biography tab at the top of your screen without further ado let me introduce you to our presenter Ben you may now begin your presentation you thank you Miss felt as I'd like to thank miss Valdes from lab roots and also Kyle berry and Amanda summers from thermo Fisher Scientific for the opportunity to present here today to to all of you so today as was mentioned I'll be talking about manufacturing GMP cell banks as part of a staged process and why it is so important and how that's incorporated into cell therapy manufacturing and we'll also talk really about the critical reagents maybe most importantly the quality of the reagents and raw materials also known as ancillary materials that have to go in early in the development process and why it is so important to incorporate quality materials early on so as an overview I think most people that are listening to this webinar know that pluripotent stem cells either from human embryonic stem cells or induced pluripotent stem cells have been described in literature for at least the last 12 years for induced pluripotent stem cells and over 20 years ago from the groundbreaking work from Jamie Thompson in late 90s showing that human embryonic stem cells can be derived and expanded based on that work there's been a lot of proof of concept studies that have been done either with human embryonic stem cells or with induced pluripotent stem cells also just called pscs here pluripotent stem cells make cells or tissues that can be transplanted into animal models of disease and can essentially cure those diseases that includes things like blindness diabetes heart disease spinal cord injuries potentially things like Parkinson's so of course these these cures or treatments are very exciting but translating the promise of those PSC derived therapies to reality or patients requires really a rigorous an often expensive and definitely highly regulated manufacturing that is broken up into unit operations so you don't just start start growing cells make your product and ship it out you have unit operations that define how that process is manufactured and it all starts with making the starting material that is to say the master cell bank and then from that you would make working cell banks and as part of that you need to make sure that those are high quality cell banks that start with high quality starting material that will meet today's regulatory requirements but because the process for development so long you need to consider that they might need to exceed the current regulatory requirements to meet tomorrow's regulatory requirements and as part of that integrating high quality raw materials ancillary materials and in this case we'll call them ancillary raw materials into the product development and manufacturing starting at the very beginning with the cell source and manufacturing of the master cell bank in doing this it reduces process risk regulatory risk and long-term it reduces the economic risk of potentially delayed product release or potentially the company or the sponsors not having enough money to continue so where are we right now in terms of pluripotent stem cell Drive there appease when we look at the the state of the art for other biological manufacturing that is to say antibodies proteins for say enzyme replacement or vaccines or even virus made for other cell therapy manufacturing for things like Kartik there's a host of cell lines that are available and they're already GMP and are ready to go so that would be things like gel where a GK for for protein production Cho for antibodies HEK and vero cells for virus manufacturing and they are essentially plug-and-play meaning you can go to a contract manufacturer and there's a host of them in the United States and in Europe and Asia and they will help you to develop a process that is defined relatively affordable they'll start with mostly if not all animal origin free raw materials ALS and they will make you starting from one of those lines tube is already GMP they will make you a seed bank and you can pick a clone make a master cell bank and then proceed from there and all of this can be routinely done from derivation as a fee-for-service so you could have a virtual company and do it all yourself from basically an office and have a contract manufacturer do it all that is really not the case right now with pluripotent stem cell derived era piece it were much earlier in the development phase for that as an industry right now every trial sponsor whether it's a university and non-profit or for-profit company currently has to develop and use their own process and this can be inconsistent so even if you have the same starting material the same genetic material whether let's for example let's say you have an h1 line that you could get as a starting material from from Y sell different expansion processes by different companies or different labs will give different downstream processing and differentiation to a product cell and another thing to keep in mind that's a problem within the field right now is a high cost of raw materials and the raw materials themselves are often research use only there are a couple companies for example thermo Fisher stem cell technologies they have some GMP or clinical grade material that is available for use early on but many other manufacturers don't have that that's something to keep in mind and when you're making these starting material you need to make sure that you're considering the life cycle so for example if you're developing the products can take five to ten years from the time you start the development process the early preclinical studies until the time you get FDA approval from marketing then from there you're looking at tend of potentially up to thirty plus years for your product lifecycle so at the minimum you're looking at 15 years of supporting manufacturing from ups from a single bank and ideally up to if you're lucky 35 40 years or supporting the product life cycle so the underlying master cell bank has to be designed to keep that in mind can you can you make enough without changing the bank and the reason why changing the bank is so problematic is for given the inconsistencies within pluripotent stem cells the early Bank is is the the foundation it's what you build on and you don't want to change it the FDA requires essentially biological manufacturing to be a defined process because the end products are often hard to fully characterize so changing they say the process is the product which means if you change the process you're essentially changing the product and if you do that you need to demonstrate comparability there's three levels of doing that one would be just in vitro assays another could be in vivo assays in an animal model but if the FDA determines that the process has changed sufficiently and they want full proof of comparability you could end up having to do an additional clinical trial to show a comparability so changing a process as its own internal risks and the most expensive outcome could be redoing a clinical trial so starting with a good master cell bank is really important so when we look at the the current state of clinical trials and regenerative medicine right now there are 875 companies most don't just over half of those are located in North America and those companies and other universities nonprofits are running about a total of 977 clinical trials in the regenerative medicine space of those about 60 of them are with clear potent stem cell derived cells or tissues and this is from clinical trials.gov so there's a lot of people that are in the space but it's I'd say still early and most of those trials are in phase one or phase early phase two and they start for the pluripotent stem cell derived therapies they start with either one of two sources on the left you can see embryonic stem cells on the right would be induced pluripotent stem cells embryonic stem cells are come from donated in vitro fertilization material that would be otherwise discarded usually the the donors have elected to to either dispose of the material or donate it and so it's plated out the inner cell mass grows out you get a cell line and that cell line can be expanded and banked all in a laboratory under controlled conditions that's where human embryonic stem cells come from one line comes from one embryo when you're working with induced pluripotent stem cells those can come from any nucleated cell any somatic cell with a nucleus and there are multiple methods to to manufacture them there's you can use virus multiple viral sources Lenti add no retrovirus those are a little bit less likely to be used for or clinical grade product but right now using plasmid or episomes is an alternative and of course direct nodulation with protein or mrna or a mix to induce lower potency in an adult cell back to a higher potent state in the end embryonic stem cells and induced pluripotent stem cells if they are of good quality they should be basically indistinguishable it shouldn't be easy to tell where they came from they should have the same expansion properties the banking processes between them should be the same so therefore the reagents and the raw materials that are used for that process should be applicable to both types of cells the phenotype what you would consider player potency should be the same and they should differentiate relatively similarly knowing that there is differences from line to line but they should be able to form all three lineages of ectoderm endoderm and mesoderm in the end those those pluripotent stem cells are what are used in the potent stem cell field so as you're taking this and you're developing your product starting with these pluripotent stem cells you're going to have to follow if you're working with the FDA they're phased development scheme starts first with proof of concept and this would be the same whether you're working with a drug or a small molecule or a biological or a cell therapy you'd have to show that you've got some proof of concept that they should have some efficacious benefit that you could extrapolate for for a human being then you would move into your preclinical studies which most critically involves pharmacology and toxicology and for cell therapies those are often mixed or biologicals and small molecules they can often be done separately but for preclinical studies in cell therapy they're often combined into the same animal model or multiple animal models that package is then put together along with other work including CMC and your clinical trial plan planning and you file an IND indication for a new drug and if you get the green light and approval from the FDA you can begin a series of escalating clinical trial phases starting with phase one phase two and phase three and at the end of all that you'll have enough data to show that it's most importantly safe but also importantly efficacious and then you can file for a biological licensing approval which gives you a PLA and approval to sell your product which point you'd have to put phase four post-marketing surveillance further for the topic of a master cell bank where it's probably most important to think about this is the first four bullets you have the opportunity to have what used to be called a pre pre-ind meeting I'm now called an interact meeting with the the FDA where you can get some of their early advice on how to develop your product and a few key questions that you should probably answer in in advance of your pre-ind meeting at the point when you're prepared to do your pre-ind meeting you want to make sure that you've got an idea of what your product is going to look like and you don't necessarily have to have a master cell bank a GMP master cell bank at this point but you need to have a good idea of how you're going to make it because at this point you're going to get approval from the FDA for what you believe you're acceptable toxicology and pharmacology animal models are and this is critical because this is an expensive process it could cost hundreds of thousands of dollars to run these studies and they have something that money can't buy which is time embedded in them so if you're going to use a pluripotent stem cell derived therapy you're going to have to do some sort of a tumor study or Terra teratoma formation study and this is often to the life of the animal so that can be anywhere between six to twelve months of waiting to see if animals develop tumors and that is required to start with you're your master cell bank the FDA as you can see the link below they require the manufacturing process of the cellular product used in the preclinical study should be as similar to the intended clinical product as possible meaning you should start with your master cell bank that you're going to use for your clinical grave manufacturing so you don't want it you can't do those studies until you have that master cell bank and so you can see how early on you're locked in you haven't even filed your int and you need to have locked in one of your manufacturing processes once you've got all those pharmacology and toxicology studies you'll file your IND and you'll be able to move into manufacturing for your Phase one clinical trial the FDA has an explicit and implicit expectation that you will improve your manufacturing process from your Phase one clinical trials through to the end of your phase 2b clinical trials at the beginning before you begin phase three you have to have locked your manufacturing process but that allows you some downstream improvements but the one thing that it's very difficult to improve is the master cell bank although you can improve lots of other things by the end of phase 2 it's really hard to to make any changes to the master cell bank because that essentially introduces comparability risk so when you think about this if you kind of like break it up into what I've called process units or unit operations when you're doing the research work it really is almost seamless you have your starting material might be a cell bank or cell line that's floating around the lab maybe researchers grad students postdocs have traded the line around everybody's got their own so-called bank or master cell bank that they use passage number may not be well tracked the processing is not necessarily consistent but everybody seems to get a pretty good result which is why you're you're moving forward and trying to raise money to to make this into a
linical rate process you expand the cells most you know you might see them a little lighter one day or a little heavier another day to make sure that you don't have to come in and pass cells on a Saturday or a Sunday and at the end you have a differentiation process which works pretty well in a small scale and then those are put into either distant vitro studies or they're put into in vivo studies in your animal model and it's relatively seamless it's often done by one maybe two or three people but it's a small team at a minimum or a maximum I should say when you move into clinical grade manufacturing that's that early part that starting material is broken up really into three unit operations and it's very tightly regulated and ideally it should be locked early in development that's derivation so if it's an embryonic stem cell if that would be anywhere from getting the donation of the blastocyst or the embryo in the case of an induced pluripotent stem cell it would be how you're manufacturing the the IPS lines in the very beginning what are the materials you use to derive that that induced pluripotent stem cell expansion process then can be broke up into a seed material and frozen and you can characterize and pick your your starting material once you've picked it then you make your master cell Bank and all of this is relatively tightly locked and this should all be done with with high quality ancillary materials high quality raw materials so that you don't have to go back and repeat this later on there's any questions about the quality of those materials once you move into the later stages shown in pink on the right which would be making the working cell Bank then thawing a working cell bank which could contain anywhere between one and five million cells in the vial you'll expand that up and the seed train really just means you're growing up lots of pluripotent stem cells for your eventual differentiation process so you might need to make hundreds of millions of cells or even potentially billions of pluripotent stem cells at that point you're ready to differentiate those cells into your product and then you have finish and fill whether it's cold stored and delivered directly to the clinic or you have cryopreservation somewhere in that process and then that's delivered to the to the to the patient or to the point of care all that in pink you could potentially make process changes in optimization prior to process lock in advance of your registration trial which is usually Phase three but if you have to say an orphan designation or an advanced regenerative medicine application that's approved that could potentially be phase 2b or something like that but you need to potentially optimize the process from the point of the master cell bank derivation and you have room in time for that you don't necessarily have it for the the banking process of the master cell bank and so if we look at like really what goes into the workflow that's that's used for making a master cell bank for an intended clinical product it has to be made under GMP good manufacturing process practice and this starts first and foremost by where you're going to do it it has to be done in a clean environment and that's because most of the operations for making a master cell bank are open open would be just kind of your standard 6-well plate or a tea flask you know you've got pipetting that are done open in in more advanced biological processing often things are closed and considered to be sterile that can be done in a room environment but for any sort of a septic processing it has to be done in a biosafety cabinet which would be an ISO five environment inside of an cleanroom which would be an ISO seven environment that room and that that biosafety cabinet have to have HEPA filtered air and the the environment is monitored by things like settling plates active particle monitors and culture culture plates for monitoring for both just total particle burden and colony forming units any sort of instruments that are used in that environment have to be qualified for use and for work by installation operational performance qualification any of the operators and verifiers who work in that environment have to be gowning qualified qualified to be in the room and also qualified to perform the the unit operations that are going to be done and all of that has to be captured within a batch record that is a their paper and electronic document that verifies and tracks all the work that was done and was so that there's a record of what was done and there also has to be a system to track if any sort of deviations occur if anything that was unplanned or not captured within the batch record to determine on a risk analysis of it if it impacted the quality of the product anything that comes in and is used in that environment has to have traceability so that would be any sort of raw materials or excipient that are going to come in and that means that the product has to come in with a certificate of analysis a CFA and they're held in the warehouse in a quarantine area until they're approved for use and that provides incoming goods control for anything that's coming in and one thing that's not on the slide but it keep in mind is that any sort of supplier is often audited or usually should be audited either with a paper audit indicating that there's some quality supply chain system used by the manufacturing system used by the supplier or potentially depending on on the risk of the incoming product or the phase of the manufacturing an on-site audit by Quality Assurance from the sponsor nothing that keep in mind is that the donor tissue the cell itself whether it's a human embryonic stem cell or an induced pluripotent stem cell you need to make sure that you have fully consented donor release of that material and this is critical because in today's day and age where genome gene sequencing and identity is so easily performed you could quite easily assuming that everything's been anonymized and randomized and you have fully donor consent that donor could be traced and very easily and so you can imagine how family might want to make sure that that they were compensated if that the donor consent wasn't fully in place and then finally so assuming you have all those things you've got a fully consented donor including commercial rights to use those cells and all Rob material traceability cleanroom etc you've made your bank now the bank has to be released based on assays four four four four or more things these are the key for safety identity purity and viability and so when you're making a master cell bank and also working cell bank these these these criteria bold for both both form you'll have to test for phenotype and identity a purity of the of the bank the karyotype meaning that has to be genetically stable it has to have only one donor and that donor has to be human but there's multiple ways to do that but right now for human type 'button stem cells probably the best way to do that would be to use short tendon repeat we test for sterility and that would include mycoplasma fungus and bacterial growth and then also endotoxin which can be either a result of any sort of risk from your your raw materials or bacterial growth within the product then the product before it's frozen and after it's frozen you would check for viability usually by a qualified method which could include things like dye exclusion and some of those are old tested methods like tripe and blue if they didn't bromide are pretty and iodide there are a lot of other methods but in the FDA requirement is that the bank after thaw is greater than 70 percent viable and then finally and maybe most important you need to make sure that the the bank functions as you would want it to in your process and that would be sponsor specific differentiation to whatever it is your fate of interest and so that's for releasing a master a working cell bank the master bank has a deeper dive so when you're releasing it you'll also have to look for any sort of viral potential contaminant or infection in that line there's qpcr assays for the top ones including things like HIV as rt-pcr for RNA viruses cv you have then for and of course because they're PCR they are primer based so you know the sequence but then there's also the risk that there could be viruses that you don't even know about though that includes adventitious viral contaminants for nonspecific reverse transcriptase and then there's in vitro and in vivo assays that are that are intended to determine if there's any virus in the the bank so you would kill culture with MRC 5 euro or a 5 for 9 cells and do it in vitro assay you'd also do an in vivo assay there's direct inoculation of eggs or rodents to look for for viral contaminants also if your product has ever come in contact with any sort of bovine material you'd also have to potentially do bovine virus detection and and if you have used anything that was porcine like trip trypsin drive from from pig gut or you've told cultured them with Mouse feeders you'd have to do porcine or murine virus panels so it's really important that you have if you can get animal origin free recombinant and human materials only then you will reduce the risk of having to do any additional viral testing it's also really important that you understand the material supply chain of your suppliers how did they make those materials because that's where additional risk comes in any of that raw material contact puts that that master cell Bank and therefore your entire process and potentially company at risk if you don't have high quality materials coming in so I've given you a lot of tables give you a picture just to kind of describe this so like I said we're looking at unit operations cell banking process you start at unit one and that really making your your seed materially seed bank and with the with the embryonic stem cell it starts with a with an embryo or a blastocyst it's cultured out and in the case of induced pluripotent stem cells would be your donors somatic cells that are induced back to player potency you might either bulk passage or single cell clone and then you would have potentially one or more seed banks if you could take those banks and before you move on to unit operation to which is the actual master cell bank would take that material and you would test it you'd want to make sure that it was genetically stable if it's an IPS cell you didn't have any say Sima core for incorporation into the wrong places you'd want to make sure that it was had a nice population doubling time it didn't grow too fast it didn't grow too slow it was a nice stable line and probably most importantly if you're making a product that it differentiates to the fate that you want so now you've got a seed bank and let's say that's somewhere between five and 20 vials you're going to take one of those vials and you're going to return to the GMB facility and you're going to make your master cell bank and that's going to take somewhere between say two to five passages you start in a small either a 6-well plate or a small tea flask you can grow that out into cell factory or a cell stack and or multiple tea flasks and you'll have enough cells to make 100 or more files of the master cell bank that can then be used thought and used to make a working cell bank and nearly the same process as the master cell bank and in this case when we talk about the life line at a lifetime life cycle of the product i'm a hundred vials of a master cell bank can make about 100,000 total product Lots because each vial of master cell bank can make about a hundred miles or more of working cell bank so you just multiply that out assuming that each each working cell bank vial can make a product lot that's ten thousand possible product Lots from relatively small banks and assuming you do two production runs a month or just a little bit more and it's about twenty-five production runs a year which would be a really high number normally you'd want to do a lot less than that but let's say you're you're really running full gun and full bore and you've got that 25 product runs product Lots being manufactured you're off of this simple scheme that would be enough for 40 years of your product lifecycle so it's enough to support a long long term manufacturing process so just in summary you've got your see material process starting either with an embryo or a small amount of donor cells you're going to make somewhere less than say 20 vials you're going to check those those vials in that Bank for do they work in your process are they either the right material then you're going to have to release them you're going to want to know in the case of an IPS cell if you have any sort of residual virus or plasma and corporation or episomes left in the cytoplasm you're going to check to make sure are they still are they Player potent are the genetically stable and of course are they clean are they sterile and do they function in your process then you pick one you move into your master cell Bank and now you're doing this much larger release specification Compendium release for sterility and the toxin mycoplasma checking for their pluripotency are they are they the right identity are they genetically stable do they function and in the case of IPS cells do they have any sort of insertions that you don't want and then a working cell Bank is a sub subset of those relief specifications and in each case you don't have to make a lot of vials I mean this isn't a large production process the thing about it is it takes a lot of time and if you can see here on the third or fourth line time to completion you know it could take 14 to 21 days or more for the C material you know 20 20 plus days potentially you're around 20 days for a master cell bank and another say 15 to 20 days for your working cell bank if you add that all up you're looking at just in time alone spent of a GMP facility of about 7 to 8 weeks and so this is expensive GMP facilities are not cheap operations are not cheap and so you need to click plan these out very carefully in advance because you don't want to change the master cell bank the cost even if you do it early on is quite expensive GMP suite with reagents and labor and I think these are on low-end numbers there are tens of thousands of dollars the actual bank release testing for the the tables I showed obviously those are hundreds of thousands of dollars so they're quite expensive and the time is something that money just can't buy time just to generate in test those cell banks can take nine months or longer including the the tumor studies they're required for your toxicology studies to to sponsor an hour support an IND filing and so the risk to the sponsor at the cell bank isn't done properly are if you change it cell banks by the nature are inherently variable in performance you can end up having to go back and reoptimize your manufacturing for your differentiation and that cost could be millions of dollars the additional cost could be incomparability requirements that that any regulatory agency might require and so if you imagine that you've got a product that could be a blockbuster drug meaning makes a billion dollars a year or more and you end up taking a nine or twelve month delay because you had to remake a master cell bank that's a billion dollars in cost it's money that actually has been actually realized but it could be the difference between success and failure of a company especially if you have a competitor that your neck and neck with and they get there first your your you could end up losing completely so just to review the risks to the master cell bank and I'm the slide after this is really how to mitigate these risks you want to make sure you have proper donor consent for the incoming raw material but that is the cell itself you want to make sure that any sort of raw materials or ancillary materials they're used in you're making your bank are of high quality so if you're going to use feeder cells I'd recommend GMP human feeder cells if you're going to use well I would recommend against Mouse feeder cells you can use them for example H 1 and H 9 have been used for human clinical trials but they are not there they're considered xeno genic lines
eaning they've been in contact with another species cells and so it's not something that you'd want to do if you can avoid it but it's not it's not a game stopper if you're using any sort of bovine component in your process FBS or bovine albumin want to make sure it comes from a qualified herd meaning a usda-approved heard that has a certificate of origin and this is because of prion disease there is no assay for it the only way to prevent it in your manufacturing is to is to start with high quality material and if you can't prove that later on that could be a game stopper so TSE has a real risks here you're processing your product the media I'm depending on how it's made there's warehoused actual mice in the warehouse can can transmit virus to the components and so you want to use media from a reputable vendor who actually tests for that your growth factor should be all Deeley human recombinant and if possible animal origin free and that includes for pluripotent media things like insulin bf/gf and tgf-beta and other growth factors that can be in the media your small molecules if possible should be filtered and your DMSO should be a pharma grade cell culture plastic usually if you're working from something like Nalgene dunk or Corning you're going to get a very high quality product but some people use cheaper sources I'd recommend against it go with a with a with a reputable old and trusted vendor if you're using a coating which most clear potent stem cells require a coating avoid Maitre gel if you can that's that's a really are you--oh product you want to go with thing that is manufactured fit for purpose and that would be a recombinant product or a synthetic product like RG d or recombinant laminin and for any sort of human donor materials you want to make sure that you have a certificate of analysis that they are clean for any sort of viruses and just keep in mind that the real risk here is is two things transmissible spongiform encephalopathy the prion disease and viral contamination and you can test for viral contamination later on but you can't test for TSE so you need to make sure that anything that's made on any product that's made with bovine source in your supply chain or that is a bovine source material that you're using directly has come from a USDA certified herd or New Zealand Australia herd so in terms of how do you mitigate this the USP has a very clear guidance on this that 10:43 was recently issued a couple years back and it talks about how to incorporate ancillary materials or Celgene and tissue engineered products into your manufacturing and they do this by providing a four tier table system so Tier one is the lowest risk tier 4 is the highest risk and tier 4 really includes a lot of the things I'm talking about previously like bovine derived materials any sort of xeno product anything that's not intended for potentially incorporation into an infusible or directly treatable product things that are for Diagnostics a or for research would be in Tier three or Tier four and they provide guidance for qualifying the use of materials so if a material is say research use only but you have to use it because there is no GMP or self-therapy grade product they provide guidance for qualifying that material to use in manufacturing your cell gene or tissue engineered product and that is key because you know early on the FDA and other regulatory agencies wreckin recognize there is a risk you can't always get a GMP reagent if you're if you're pushing the envelope and trying to make a new therapy they do expect that you work with a supplier that is committed to improving their quality systems and parallel as you advance through your phased development to to commercial manufacturing for for human use products and that means you really want to make sure from the very beginning you work to integrate high quality raw materials and ancillary materials into your manufacturing process from the very first seed bank onward and that means if you can use GMP or an animal origin free products as soon as possible especially for your cell banking and source all your raw materials if possible from suppliers experienced in supporting cell therapy development ideally they should be stable long-term partners because they're going to have to commit to to supplying that product to you as long as you're making your product and if they do so that will reduce your risk of potential disruptions things like back orders I know if you're in a lab and something's backwards six weeks it's you know it's a hassle if you've got a GMP suite set up and you've got operators and you've paid for that time and you're back ordered on a critical reagent you have to pay for that time that can cost a lot of money product changes should be clearly communicated to you if you're using them and you don't want to have to worry about any sort of discontinued products because that can of course create great risk in your in your manufacturing system and you want to identify if possible more than one source for your raw materials ideally at least two suppliers avoid sole supplier products if possible I know that's not always possible but but that's something to keep in mind it just reduces your risk as a manufacturer or as a sponsor so in summary I think I've hopefully communicated how important it is from seed material on through your master cell bank all the way through to end product manufacturing how important it is especially is beginning to use high quality raw materials from an established reputable vendor as early as possible because those raw material risks extend throughout the supply chain and they put the entire project and product at risk if they're not dealt with you want to book and future forward planning you want to know where you're going to be where you want to be knowing that's going to take many years and you want to anticipate increased regulatory stringency over time or your product and so you don't just want to meet today's two you want to know that tomorrow standards are going to be a little higher and you want to try and meet those in advance so that you can at least pass the bar for tomorrow even though you greatly exceed it today and all of this if you do this and put this in place now you can avoid the loss of time money and potentially catastrophic risk to your product or your company by generating a high quality seed and master cell banks in the beginning that can sustain the product throughout its lifecycle so my last slide just a little bit about plura sticks I won't talk too much about it but I'd like to thank everybody who has come today and watched and listen to this talk and if if you have any questions I'm happy to take any questions you have at this time thank you and Thank You Ben for your presentation a quick reminder for our audience on how to submit questions just click on the ask a question box located on the far left of your screen we'll answer as many questions as we have time for before I ask the final first question we have a question for you have you worked in a GMP environment before this question will now pop up on your screen and thanks for your participation look at our first questions coming in from our audience then can you make a cell line GMP if it wasn't originally generated in a GMP environment excellent question so this is a common question that's asked and I think the answer right now I think is yes so for example the 89 line that is available from YSL was made under under research conditions so that would have been considered a research master cell bank that that question that that bank has been used essentially as the seed bank so as a unit operation and that that research master cell bank becomes the so-called seed material a vial of that is then transferred into a GMP environment and entire batch record and process is developed around that and then it's manufactured and released and there's a essentially a risk assessment that has to be done on all the materials that were used to make that line in the beginning so it's kind of a way of kind of figuring out what other additional viral panels you should you should look for I guess the biggest risk with any sort of with doing something in that manner is that you need to make sure you have early passage cells and it's actually not given that the FDA or other regulatory agencies will will accept it by the time you get to filing your PLA so you might be able to make your product early on but you might be essentially pigeon-holed into a very small indication so for example if you wanted to go for show that you were making with a neurons and you wanted to treat a specific type of blindness you might only be able to use that product for that specific type of blindness to be able to add on indications or other therapeutic areas you may not be able to do it because they would consider that product to be too high of risk and so you end up basically having to restart the process so I guess the question the simple answer is yes it's possible to take a research bank and make it into a GMP master cell bank right now but I think if if you can afford it if it's possible it's better to make the master cell bank from a GMP source in the very very beginning Thank You Ben and is there a difference between a research master cell bank and a GMP master cell bank and can our research bank be converted to GMP bank during development I would say probably you can I like I said it's possible to convert from one bank to another I just I think that it the cost is really the issue in terms of risk down down down down the line and so I would recommend if possible especially if you're working with an IPS cell start with a GMP seed material and a GMP master cell bank embryonic stem cells a little bit harder but it is possible to take a research master cell bank make that your seed material and make a GMP master cell bank later on Thank You ban and one audience member would like to know to your slide 18 point how important is it to you quality by design so quality by design is essentially used asking the question beforehand how you know testing the parameters of your of your process so it's a little bit easier to do with a master cell bank really what you're looking for is you're looking for population doublings and essentially quality you're controlling for four risks and stresses in the process early on I think it's it's important to use but I don't know exactly how how much work is required to do quality by design early on I think having the good master cell bank probably feeds more directly into quality by design later on in the process so that you have a starting consistent process and product that you can a bank that you can use for manufacturing later on Thank You Ben and why do you need a GMP cell trying to make study material for pre ind-enabling studies so the risk well so the request in a pre-ind well in an IND submission is that you have to have shown that your material is number one and most importantly it's safe and so the safety question for stem cell any sort of cell which has the ability to grow and form all the other tissues is that it could form a tumor and so the question is that you know you have a master cell bank that master cell bank is where it's essentially the starting point for all things so if you generate different master cell banks the potential is that one of them could have a different risk for sorts or tumors or for ectopic tissue formations so if you do your combination pharmacology toxicology studies with your master cell banks and they're not there they're not the one that you're going to use in patients the FDA or some other regulatory is going to wonder why or what the other risk is and so this is probably the biggest problem with with trying to change master cell banks you'll end up having to redo that study so you could potentially use one master cell Bank of this GMP for your pre ind-enabling studies but if you want to switch to another bank you'll probably end up having to do those those teratoma toxicology studies again and that's just cuts down it's just it's a time cost so we could end up having to repeat human clinical trial studies so it's I think it's really critical to get that that done first thank you and thank you to our audience members for their questions that are coming in then our next question is is it important to have a MCB if the cells being used are from the source they are going back into from for example from patient a back to patient a and no one else yeah so this would be essentially almost like the autologous model of of cells so if there's this question of minimal manipulation so for example if you're pulling cells out and you're going to put them right back in so for example if your banking blood from somebody for a surgery and you know if you want to make sure that they have a blood supply that you could infuse back into them that's considered minimal manipulation and you won't have to to make a master cell bank of that but if you're going to expand those cells and you're going to change them you're going to end up having to do a proper release on those on that product and that release panel is is expensive I guess it really depends on the product so for example if you're making a pluripotent stem cell derived product you're going to have to make a master cell bank and you're going to have to release the master cell bank and the real question there is the risk to the patient and the risk comes from the raw material so virus and sterility can be a problem based on whatever you're doing to expand those cells and to change them so you'll need a master cell bank the the full release panel may not be it may not be to be so extensive but you will need it a master cell bank thank you and then does the master cell bank strategy change if the pluripotent stem cell is genetically modified example expresses a transgene or how the gene edit this is a is that why I love this question because it's question I've asked before so this I think that let me just preface my answer by saying I don't think my answer is the only answer and I'm not sure that the FDA would agree but I'll give you my opinion on it and that is you'd want to start with a master cell bank that was not edited and then you would make essentially working banks that would be genetically modified but all that then has to be done in the GMP environment and you have to fully characterize that the working cell Bank is genetically modified so you may not have to redo all the the viral testing but you will have to do two netic stability on that working cell bank our next question is we have a cleanroom that is not GMP can we manufacture our material there and use it for clinically representative pre-ind studies so for for a cleanroom that's not GMP I think if you're making your your working cell banks or so called GLP material that it's going to that follows the supply chain and the batch records in the process that you're used for making the end product you can you can manufacture material for your pre-ind studies it's not in a GMP environment however the master cell bank that does have to be GMP so at some point you're going to have to make a break and make that GMT master cell bank knowing that you can take some of those vials back to a research lab which is clean but not GMP and make essentially development or research working cell banks and under GLP or sufficient quality to to meet regulatory requirements you can make your end material but you can't you can't skip the GMT master cell bank so I guess when you get into final or actual phase one manufacturing for patients everything soup-to-nuts has to be done in the GMP environment but if you're making material for pre ind-enabling studies preclinical and animal studies that it has to start from a GMP master cell bank but you could make it under GLP in a less clean or less less controlled environment for ind-enabling studies Thank You Ben and we appreciate a live participation from our audience we have time for a few more questions our next question is what is the difference in the cost of cell culture for GMP products and for laboratory purposes the probably the biggest cost is oftentimes research use only or are you oh I didn't spell that out in the tal
are you oh just means research use only and a GMP product often there's there's very little maybe no difference in how they're made the difference is in the the amount of labor and time and documentation that's required to differentiate between between a GMP product or a cell therapy grade product and a research use only product so that's that's why there's so much more cost it's in the human side and the liability that is involved in that for the for the supplier so GMP reagents are just by nature more expensive because there's a lot more traceability that goes into that testings more extensive things like that so when you're working in a GMP environment the same kind of thing it's the lab will look the same it'll have an incubator will have a biosafety cabinet you know it's it doesn't it operates like any other lab would except for the fact that you have to have controlled entry controlled exit you have to have traceability and the environment you have to have traceability on operators traceability on all these machines and so just it's expensive of keeping that record and keeping that cleanroom is what causes the GMP prices to go up and it's so in the short end the difference is really traceability and human labor that's required for that and therefore that is what increases the cost between GMP and non GMP thank you for that answer our next question which molecule molecular assays do you suggest for testing genetics ability of the MGB right now the the assay which is and not molecular it's it's a it's gross morphology essentially is dban G band is considered the the the gold standard and it's what's used to test for genetic stability so you'll look at say 30 to 50 cells and you should see the carry type should be normal stable a lot of groups are now starting to add in new or technologies whole genome sequencing we've got integrations looking to see if there's any sort of especially for type one stem cells if there's integration of your of your your clear potent genes into the the genome of the cell of interest i I mean there's I think there's a fair amount of assays that you can use to give yourself comfort to give your investors comfort to give your patients and your physicians comfort that you have a stable line but right now the required requirement would be would be karyotype Thank You Ben and we have time for one more question our final question is what are the consequences to not using a GMP master cell bank your IND won't be approved so to do a lot of work and you won't get you won't you won't be able to begin clinical trials then do you have any closing remarks that you'd like to share with the audience Dale no I well I guess a yes I do so I just like to thank everybody who came and and participated listened to our talk if you have any questions you'd like to follow up with you can find us at our website laura sticks comm and I look forward to hopefully meeting some of you in person and if we can do anything to help you we'd love to help and I'd like to also thank thermo Fisher and lab roots for for the time and attention today and for inviting me to present this has really been fun thank you and before I go out I tell also thank the audience for joining us today and for their interesting questions questions we did not have time for today and those submitted during the on demand period will be addressed by the speaker via the contact information you provided at the time of registration the webcast can be viewed on-demand through August 2019 leverage will alert you via email when it's available for replay we encourage you to share this email with your colleagues who may have missed today's live event a poll will now pop up on your screen and your answers and your suggestions will help improve our webinar program and they are greatly appreciated until next time bye bye
