Webinar

<Tom Allison>
Good morning and good afternoon, depending on where you’re tuning in from. My name is Tom Allison and I’m a Senior Product Manager here at Trailhead Biosystems. So, firstly, thank you very much for joining us on what is our first webinar of 2026. And today’s webinar is going to focus on the custom service segment of Trailhead Buyer Systems.
With a focus on HPC differentiation and protocol development.
The webinar will present information, data and the case study on the capabilities that we’re able to offer at Trailhead, and this will be followed by a Q&A session. So we welcome any and all questions that you have. You can ask them directly during the Q&A portion of the webinar, or you can use the Q&A function that you will see on the toolbar at the top of the screen.
Presented today is our field application scientist Doctor Michael Elias. Dr. Elias is a standard biologist with expertise in regenerative medicine, immunology, and translational research. He earned his PhD in nanoscience from UNC Greensboro, where he focused on stem cell differentiation and cancer therapy.
After completing postdoctoral work at the Cleveland Clinic, where he studied inflammatory disease and fibrosis, he joined Trailhead Biosystems, where he helped develop and scale protocols for iPSC-derived hematopoietic progenitor cells. His work bridges cutting-edge biology with real-world therapeutic application.
Also joining us today is Dr. Angelica Gomes Yocchi. Doctor Gomes Yocchi is a stem cell biologist with expertise spanning IPSC differentiation, hematopoiesis, and process development for scalable cell manufacturing.
She currently serves as scientific director at Trailhead Biosystems, where she leads the mesodermal programs and develops high-dimensional differentiation protocols for cell types, including hematopoietic stem cells, monocytes, macrophages, dendritic cells, red blood cells, as well as endothelial cells.
So with the intros, I will then hand over to Dr. Elias to take us through the presentation.

<Michael Elias>
Tom, thank you very much for the introduction. Thank you everybody for joining us this morning. Let’s kick off and just get right into it. So good morning, everyone, and thank you for joining us today to learn about Trailhead Biosystems’ approach to HPC differentiation and custom cell generation.
Here’s a quick overview of what we’ll cover today. First, I’ll briefly introduce Trailhead Biosystems and provide some background on Hematopoiesis as the biological foundation for blood development. Next, we’ll discuss why iPSC-derived hematopoietic progenitor cells are valuable and how they compare to traditional donor sources.
From there, I’ll walk through Trailhead’s approach, including how we use our HD-DoE platform to develop reproducible iPSC-derived HPC populations. We’ll then explore key applications of these cells, including their ability to generate multiple hematopoietic regions.
And finally, I’ll discuss our custom iPSC differentiation capabilities and how we collaborate with researchers to generate HPCs from customer- provided iPSC lines.
Trailhead Biosystems was founded in 2015 and is based in Ohio and focuses on generating specialized cells derived from the mesoderm, endoderm and ectoderm. Pictured here is our facility, which is located in Beachwood, Ohio, just outside of Cleveland.
Since our goal is to generate specialized cells from stem cells, it’s important to understand where the main challenges in the field exist. Over many years, researchers have developed reliable methods to reprogram IPSCS, which can give rise to any cell type. Today, the greater challenge is not reprogramming cells, but directing iPSCs into the specific, specialized lineages researchers need.
This is where Trailhead focuses its efforts. We work on solving the differentiation challenge using iPSC as a customizable starting point to generate functional specialized cells. These cells can be used in a range of applications, including cell therapy, drug discovery, disease modeling, and tissue repair.
Let’s take a moment to look at some of the major challenges facing iPSC-derived cells today. First, the demand for high-quality human cells is extremely enormous, but the available options remain limited. Researchers and companies want to use these cells for therapy, disease modeling and drug discovery if the field continues to encounter the same obstacles.
Second, achieving pure cell populations can be difficult. Many available, many available products don’t reach the level of purity needed for consistent and reliable experiments. Another issue is batch-to-batch variability. Researchers may obtain one result from a particular vial and see very different outcomes for the next.
Supply is also a challenge. These cells are not yet available at scale that research therapeutic markets require. Finally, much of the development production process is still performed manually. This slows progress and introduces variability, making it difficult to scale efficiently.
Together, these challenges represent a major bottleneck in the field, and addressing them is the key to unlocking the full potential of iPSC-derived cells.
Now that we’ve looked at the challenges, let’s talk about how we actually address them. At Trailhead, our approach is to fundamentally rethink how stem cell discovery and manufacturing are performed. Rather than relying on slow, traditional trial-and-error processes, we use machine-enabled experimentation to accelerate discovery.
This allows us to evaluate and optimize conditions at a scale that simply isn’t feasible through manual methods. Using this approach, we explore the high-dimensional space of regulatory inputs, including combination of growth vectors, small molecules, and culture conditions that influence cell fate. By systematically analyzing these variables, we can precisely guide differentiation pathways in.
Improve consistency. The result is higher cell purity and the ability to generate specialized cell subtypes that meet the needs of researchers. Once key process parameters are defined, these cells can be generated consistently and at scale. Because this platform is flexible, it can also be applied across multiple cell types while keeping costs down.
Ultimately, this leads to access to high-quality cells, more reliable experimental results, and greater impact for both research and therapeutic development.
So instead of relying on the trial and error approach typical of traditional protocol development, where one or a handful of vectors are tested at a time, we perform automated, unbiased screening of many compounds and vectors both individually and in combination to determine how they influence cell fate.
Using this experimental data, we then apply high-dimensional modeling to understand the contributions of each compound to cell differentiation both independently and in combination with other factors. We call this approach High-Dimensional Design-of-Experiments, or HD-DoE.
With this strategy, we can generate reproducible subtypes, subtype-specific cells, and subpopulations with impurity, speed, and efficiency compared to traditional methods.
To highlight the advantage of this approach, let’s consider the example shown here. If we want to experimentally test every factor and compound we typically evaluate in one of our studies, including both individual effects and combinations, it would require about 4296 well plates or over 4000 individual wells.
Running an experiment at this scale would demand significant time, materials and cost, making it largely impractical. At Trailhead, however, our HD-DoE approach allows us to capture and model the effects of these conditions using data generated from just a single 96-well plate. Later, I’ll show you how this approach is applied to our HPC cell differentiation.
Let’s talk about the background of Hematopoiesis.
To understand how the blood system develops, we need to begin with the hematopoietic stem cells, or HSCs, which represent a rare population located at the top of the blood hierarchy.
What makes HSCs particularly important is their ability to support long-term and secondary engraftment. This means they don’t just restore blood production once, they can sustain hematopoiesis over extended periods of time. HSCs are also multipotent, meaning they can generate every major blood lineage.
This includes red blood cells and platelets, as well as myeloid and lymphoid immune cell populations. Because of their broad potential, HSCs occupy the highest level of the hematopoietic hierarchy. Another defining feature of HSCs is their capacity for self-renewal. This allows them to maintain the stem cell pool within the specialized environment of the bone marrow.
However, despite their importance, HSC presents several challenges. Outside of the bone marrow niche, they’re extremely difficult to expand or maintain while preserving their stem cell properties. This limitation represents one of the major barriers to translating HSC biology into scalable therapeutic applications.
Long-term HSCs, the population residing in a bone marrow with self-renewal capabilities, can be identified by a well-established set of surface markers like CD34 positive, CD38 negative, CD45RA negative, and CD90 positive. These markers are widely used in both research and clinical settings to distinguish true HSCs from progenitor cells.
Overall, HSCs are the foundation of long-term hematopoiesis for this reason. For this reason, developing controlled in vitro differentiation strategies is essential for studying and generating hematopoietic populations.
Now that we’ve discussed the defining features of HSCs, let’s step back and look at how the blood system developed during embryogenesis. Broadly speaking, hematopoiesis occurs in two major waves, primitive hematopoiesis and definitive hematopoiesis. As hematopoiesis begins the first HPCs merge.
These early progenitors are typically identified as CD34 positive and CD43 positive and display multi-lineage potential. Importantly, this population appears during both primitive and definitive ways of hematopoiesis. The earliest wave, known as primitive hematopoiesis, occurs outside the embryo in the yolk sac. During this stage, lineage-committed progenitors are generated.
Primarily producing erythroid cells along with early myeloid progenitors. These cells play an important role in early tissue oxygenation and provide temporary hematic support for the developing embryo. As development progresses, the second wave, known as definitive hemopoiesis, begins within the embryo itself.
This process originates in the aorta, gonad, mesonephros region, or AGM, where specialized hemogenic endothelial cells transition to the first hematopoietic stem cells. These newly formed HSCS enter the circulation and migrate to the fetal liver, which serves as a major site of expansion.
In the fetal liver, the cells proliferate and mature while establishing a more defined transcriptomic profile. Later in development, these HSCS relocate to the bone marrow, which becomes a long-term residence. In the bone marrow niche, they remain largely quiescent under hypoxic conditions, but can become activated when the body requires new blood cells.
So overall, hematopoiesis begins with a short-lived primitive wave in the yolk SAC, while the definitive wave emerging from the AGM establishes long-term HSCs responsible for sustained blood production.
So why iPSC-derived?
On this slide, I want to highlight where hematopoietic regenerative cells are commonly obtained from and why these donor sources can be limiting. The three main sources of primary cells are cord blood, bone marrow, and mobilized peripheral blood. These sources have supported research and clinical work for many years, but each comes with important drawbacks.
Cord blood is limited by the amount that can be collected at birth because the volume is small. The number of cells in each unit is also limited, and there can be significant variation in cell yield and quality between different donors. Even with cord blood banks, the overall supply is still finite. Bone marrow is another traditional source.
But collecting is invasive for donors, and super donor matches are not always easy to find. In addition, the quality and expansion potential of the cells can vary depending on each donor. Immobilized peripheral blood requires treating donors with agents that move stem and progenitor cells from the bone marrow into the bloodstream.
However, donor responses to mobilizations are inconsistent, and the process adds costs, complexity, and more regulatory considerations. Primary donor cells also present compatibility challenges for allergenic use, immune rejection, and the need for HLA matching remains a major concern.
For autologous approaches, obtaining patient-specific cells is not always practical and can limit scalability. It’s also difficult to build large banks of genetically edited primary cells. Together, these limitations make it challenging to achieve a level of consistency, scalability, and reproducibility that modern research and therapeutic development require.
iPSC-derived cells help solve many of the challenges associated with donor-derived hematopoietic progenitor cells. First, they offer consistency. Because IPSC lines can be banked and used repeatedly as starting material, differentiation can be performed under the same control conditions each time.
This greatly reduces lots of lot variation, which is common issue with primary donor material. Another advantage is customization. iPSCs can be generated from individual patients or genetically engineered to model specific diseases. This makes them a powerful tool for studying disease biology and developing targeted research models.
They also provide a renewable source of cells. Unlike cord blood or bone marrow, which are limited in supply, iPSCs can be identified or can be expanded indefinitely in culture. This means HPC production can be sustained and scaled to meet research or industry demand. Standard production also improves reproducibility.
When cells are generated using defined protocols, the resulting populations are more consistent, allowing experiments to be compared more reliably across studies, labs and institutions. iPSC platforms are also highly scalable. These cells can be adapted to larger manufacturing systems, making them useful not only for research but also for high-throughput screening and industrial applications.
Finally, the range of applications is broad. iPSC-derived HPCs can be used in drug discovery, toxicology studies, immune system modeling, and regenerative medicine, as well as regenerating downstream immune and blood cell types. Overall, iPSC-derived HPCs provide a consistent, reproducible, and scalable alternative to traditional donor-derived HPCs.
Sources.
Let’s move on to the trails approach and how we tackle these.
Now that we’ve discussed the biology and background of HPCs, let’s look at how Trailhead generates hematopoietic progenitor cells from iPSCs. Our process follows A three-stage workflow. Starting with pluripotent stem cells, we first drive them towards the mesoderm, then guide those cells into the hemogenic endothelium.
And then finally direct them into the hemogenic or hematopoietic progenitor cells.
The most important step in this workflow is stage two, the formation of the hemogenic endothelium. This stage represents the key developmental gateway to blood, and achieving it correctly is critical for generating functional HPCs. To optimize this transition, we apply our proprietary HD-DoE platform.
This approach systematically evaluates combinations of the small molecules and proteins to identify conditions that precisely control signaling pathways during differentiation. By adjusting the time and concentration of these signals, we can carefully guide the cells into the hemogenic endothelial state.
In total, more than 250 factors were screened during this development process. The result is a highly robust and reproducible differentiation workflow. It reflects the steps of embryonic development while still providing the scalability and control needed for research and translational applications.
This ultimately enables A reliable production of high quality HPC’s in our stage 3.
Before reaching stage two, we first verified the cells had successfully committed to the mesoderm. This is confirmed by immunocytochemistry using marker CD 309, also known as VEGF receptor 2, along with the transcription factor brachiuri. The data isn’t shown here, but it’s confirmed that stage one differentiation had occurred properly.
The next step is confirming the transition to stage two, the hemogenic endothelium. This slide focuses on stage two, which as mentioned earlier, represents the developmental gateway to blood formation. At this point, the mesodermal cells are cultured in our optimized stage two media, which is promoting induction of genes and surface markers characteristic of the hemogenic endothelium.
Key markers expressed at this stage included runx 1, CD31, Fly one and CD34. On this slide you can see representative muta-sided chemistry images highlighting several of these markers, including SUC 17C309, runx 1.
Cadet to CD31 and CD144. These images confirm protein expression across the cell population.
In addition, we assessed the population by flow cytometry, which shows strong marker expression in the majority of the cells. For example, 82% of the cells expressed CD31, 43 expressed RUNX 1 and 86% expressed CD34. Together, these results show the cells are not just initiating the transition, but are forming a robust and reproducing.
Hemogenic endothelial population.
To generate HPCs from the stage 2 hemogenic endothelium, we developed a stage 3 differentiation condition designed to promote expression of key HSC-associated markers.
In this analysis, we evaluated unsorted thawed TrailBio HPCs, and the population shows a highly pure early-stage hematopoietic phenotype. More than 70% of the cells coexpress CD34 and CD43, which are well-established markers of the hemopoietic progenitor cells or the HPCS.
At the same time, CD 235, a marker associated with erythroid differentiation, remains low, indicating the cells are not prematurely committed to downstream lineages.
Another important characteristic is the retention of CD90. Approximately 60% of the HBC population is CD90 positive and CD43 positive, and these cells do not co-express CD45R A, which is typically associated with a more differentiated cell state. The pattern is consistent with an early HSC-like phenotype.
CD 90 in particular is known to mark early hematopoietic stem cells, and its expression is usually lost as cells begin to differentiate. The fact that a large portion of our population retains CD90 suggests the cells remain in a more stem-like state.
When we examine the classical HSC phenotype, CD34 positive, CD38 negative, CD45 RA negative and CD90 negative, we see that roughly 60% of the cells meet this profile. This represents A substantial enrichment of stem-like cells in higher proportion than we typically observe with other commercial HPC products.
So overall, these cells not only express the expected progenitor markers, but they also display a class early HSC phenotype with strong enrichment compared to alternative HPC sources.
Here we have a brief overview of the TrailBio HPC product. On the right, you can see a brightfield image taken after thaw showing a healthy population of viable cells in culture. Flow cytometry analysis confirms strong post-cell survival, with viability consistently higher than what we observed with comparable competitor products.
This is reflected in the bar graph, where TrailBio HPCs show strong recovery post-thaw. In addition to high viability, the cells maintain an HSC-like marker profile. They express CD34 and CD90 while remaining CD44C45RA and CD38 negative, which is consistent with an early stem-like hematopoietic population.
Taken together, these results show that our HPCs not only survive the thawing process well, but also retain the stem-like characteristics needed for dependable research applications.
Let’s talk about the multilineage capacity and applications of these cells.
To begin this section, we first looked at one of the most widely used functional assays for HPCs: the colony-forming unit (CFU) assay. In this experiment, true bio HPCs were seen in methylcellulose-based media in culture for 14 days. During this period, the cells generated multiple distinct colonies.
On the side, you can see representative examples, including granulocyte colonies, CFUG, macrophage colonies, CFUM, and erythroid colonies such as BFU and CFUE. These colonies represent both myeloid and erythroid lineages, demonstrating the cells maintain broad multilineage differentiation potential.
We next evaluated the ability of these HPCs to commit to specific and medial lineages, the monocyte and macrophage lineages. After stage 4 differentiation, cells are analyzed by flow cytometry. Here, you can see a clear population expressing the monocyte and macrophage markers CD14 together with CD33.
A similar pattern is observed when looking at CD14 together with CD16.
We also examined additional immune markers, including CD163 and CD45, which further support differentiation towards the monocyte and macrophage lineages. Importantly, these cells can be further differentiated into functional macrophages and functional dendritic cells, demonstrating that these cells can progress into downstream innate immune populations.
These results show that trail bio HPCs are capable of both broad multilineage differentiation in CFU assays and direct differentiation into immune lineages such as monocytes, supporting a range of hematopoietic and immune biology applications.
On this slide, we’re highlighting more linear specific differentiation capabilities to TrailBio HPCs. Together on the left, you can see differentiation towards the myeloid lineage. In this workflow, our HPCs were first guided into myeloid progenitors during stage 4 and then further differentiated into neutrophil-like cells at stage 5.
Float cytometry analysis shows a strong population expressing CD11B and CD15, which are well established markers associated with associated with neutrophil like identity.
On the right we show erythroid differentiation. In this case, TrailBio HPCs were directed into erythroid progenitors and then further matured into erythroid cells. This differentiation was carried out in the presence of dexamethasone. Flow cytometry confirms clear co-expression of CD71 and CD235, which are characteristics of characteristic markers of the erythroid.
It’s important to note that both these differentiation protocols are standard approaches commonly used in literature. They were not developed using your HDDOE platform.
Taken together, this data demonstrates that TrailBio HBCs can be guided into multiple hematopoietic lineages, including both myeloid and erythroid pathways, providing strong functional evidence of the differentiation capacity. This level of flexibility is important because it shows that the cells retain their multiple potent nature while also allowing us to steer differentiation towards specific lineages to.
Pending on the application.
Here we show differentiation of our HPCs into microglia performed independently by the Gaskill lab at Drexel University. Microglia are the resident immune cells of the central nervous system and arise from early hematopoietic progenitors during development because of its developmental relationship.
HPCs provide a biological, biologically relevant starting point for generating microglia in vitro. On this slide, you see TrailBio HPCs at the beginning of the differentiation process at day zero, where the cells display an expected progenitor morphology following thaw after directed differentiation.
By day 11, the cells develop a more ramified morphology, which is characteristic of a microglia-like state.
Day zero and day 11.
On the right, immunocyte chemistry confirms expression of the microglia-specific marker P2RY12, shown in green with DAPI labeling the nuclei.
This slide focuses on functional validation of the microglia generated from our HVC platform. In this experiment, the cells were stimulated with increasing concentrations of bacterial endotoxin, commonly used to trigger an inflammatory response. The assay measures activation of NF Kappa B signaling pathway, which is considered a master regulator of inflammation.
NFCAPB controls the expression of many genes involved in immune activation and inflammatory signaling. Under resting conditions, NFCAPB remains inactive in the cytoplasm. When the cells are stimulated with LPS, the pathway becomes activated and NFCAPB translocates to the nucleus where it drives its transcription of inflammatory genes.
In these images, NF Kappa B is shown in green. DAPI blue labels the nuclei and the red outlines the cell body, revealing NF Kappa B translocation into the nucleus.
Here as LPS concentrates increase, you can see more NF Kappa be localized to the nuclei. This increase in nuclear translocation is quantified in the bar graph showing a clear dose dependent activation response. Again, this analysis was performed by the Gaskill Lab at Drexel University providing independent validation of the functional response.
This data shows that microglia derived from TrailBio HBCs are not phenotypically correct, but also functionally responsive to inflammatory stimulation.
Now that we’ve looked at how these microglia can be generated and functionally validated, let’s briefly discuss how they can be used. One major area is neuroinflammation research. Because microglia are primary immune cells of the brain, they play a central role in cytokine signaling and inflammatory responses within the CNS.
They’re also valuable for drug discovery and screening, where researchers can evaluate compounds that modulate microglial activation or immune signaling pathways. Another important application is in co-culture systems. Microglia can be studied alongside neurons, astrocytes, or brain organoids to better understand cellular interactions within the neural environment.
These cells are also useful for disease modeling, particularly when derived from patient-specific or CRISPR-edited IPSC lines, enabling the study of disease-relevant genetic backgrounds. In addition, researchers can perform functional assays such as measuring phagocytosis, synaptic pruning, and immune cell surveillance activities that are central to microglial biology.
Overall, TrailBio HPCs provides a scalable and reproducible starting population for generating microglia, enabling a wide range of downstream applications in CNS research.
Let’s talk about a service that Trailhead offers, our custom IPSC differentiation service.
I want to briefly highlight our custom IPSC differentiation capabilities. In addition to our off-the-shelf product, TrailBio also partners with researchers who want to generate cells from their own iPSC lines. One of the major advantages of working with us is access to our HD-DoE platform, which allows us to rapidly identify and optimize the conditions needed to produce specific cell types in a scalable and reproduceable way.
Another key benefit is access to our internal expertise across all the three germ layers, the mesoderm, the endoderm and the ectoderm. Our team has extensive experience designing differentiation strategies for a wide range of specialized cell types. This approach allows researchers to work with patient-derived or gene-edited iPSC lines, enabling disease-relevant models while maintaining the.
Maintaining the consistency that comes from standardized differentiation workflows. Altogether, this provides a flexible way to generate customized cell populations while leveraging TrailBio’s platform and expertise.
Here’s a simplified overview of how a typical custom differentiation project works. First is a project consultation where we discuss the research goals, starting IPSC line and desired cell type.
Next, the client line, the client provided line undergoes sourcing and expansion to generate a stable starting population.
In the third step, our team performs the optimization and differentiation, applying our HD-DoE platform and optimized protocol to guide the cells into the desired lineage.
Finally, the cells undergo characterization and quality control before delivery with supporting data, with the option for additional functional testing based on the project goals. This workflow allows researchers to move from a starting IPSC line to a validated specialized cell population in a reproducible way.
To illustrate how this workflow translates into real-world impact, I’ll now walk through a recent project example.
This slide highlights the internal quality control we performed during our differentiation process. As stage two, we confirm the generation of the hemogenic endothelium by measuring expression of key markers such as CD34, CD31, KDR and RUNX 1, which indicates successful progression into the hemogenic endothelial state.
At stage three, we characterize the resulting hematopoietic progenitor cells. Here we expect a CD34 positive population with appropriate CD43 and CD90 expression consistent with an early hematopoietic progenitor phenotype. It’s also worth noting that we often observe higher CD34 expression when using our own internal IPSC lines, which were optimized with this protocol.
However, this data shows the protocol still performs well with other IPAC lines, demonstrating that is that is a robust and applicable across different genetic backgrounds.
We also evaluated the post thaw performance and in this example the cells show about 87% viability after cryo preservation indicating strong recovery following thaw. Together these stage specific QC checks ensure the cells meet our quality and standards before shipment.
To further validate performance, the client independently characterizes cells after delivery. Here we see evaluation of key hematopoietic stem and progenitor markers, including CD34 positive and CD38 negative populations. Importantly, the CD34 positive, CD38 negative, CD45RA negative and CD90 positive population was at a level similar to our off-the-shelf.
Bio HBC product.
Post off viability, post off viability remained high, demonstrating stability through the crowd preservation. In this case, the client also compared different differentiation approaches across protocols, and the TrailBio workflow produced the strongest HPC HSC-associated phenotype.
Together, this data provides independent confirmation that our custom differentiation workflow can generate HPCS with an HSC profile consistent with our established product standards.
Before we end this webinar, I want to briefly highlight a few key takeaways. First, our HDDOE platform combines software, robotics, and high-dimensional modeling to accelerate IPSC differentiation and optimize cell development. Using this approach, we generate IPSC-derived HPCS that are scalable, customizable, and show reduced lot-to-lot.
Variability compared to donor sources. These cells retain key stem-associated markers such as CD90 displaying HSC-like phenotype, maintain low CD235 and show strong post-thaw viability. Functionally, they support multi-linear differentiation, including CFU formation and downstream immune cell generation such as microglia, monocytes, neutrophils, and erythroid cells. Our platform also enables custom iPSC differentiation projects, allowing researchers to apply these workflows to their own iPSC lines. If you’re interested in learning more or starting a project, you can reach out to support@trailbio.com.
If you have any questions, please reach out to our sales team or myself. We’ll now move into the Q&A portion of the webinar. I’m joined by my colleague, Angelica, the scientific director at Trailhead, who led much of the development behind these cells and brings deep expertise in immune biology. With that, we’re happy to take any questions.

<Tom Allison>
Thank you very much, Mike. That was a great presentation on what we can do at Trailhead. We do have a few questions that have come in, but just a reminder, if you do have any burning questions that you want to ask, the Q&A button is on the toolbar at the top of the teams here. So again, please feel free to ask any questions that you have.
So moving on to the questions that we have received, um, the first one, Angelica, then how do custom projects differ from your off-the-shelf product?

<Angelica Gomes Ueltschy>
Hi, everyone. Can you hear me well?

<Tom Allison>
Yeah, all good.

<Angelica Gomes Ueltschy>
Good, good. So, coastal projects use our established differentiation and manufacturing protocols to generate sales from a customer-supplied IPSC line. Sometimes minor modifications are necessary to adapt to the protocol, but in general, these protocols are seamless to the ones we use to produce our off-the-shelf product.

<Tom Allison>
Thank you. Another question that’s come in. Do you offer customization for customer-provided disease-specific iPSC lines?

<Angelica Gomes Ueltschy>
Yes, we do. We, of course, customize through our customer differentiation services. Customers can provide their own iPSC lines. These lines could be disease-specific or gene-edited lines, and we apply our established differentiation protocols to generate the desired cell type.

<Tom Allison>
Right.
Um, for the HPCs, and I think we’ve got a similar question in the chat that’s come through. What media or coating is required?

<Angelica Gomes Ueltschy>
OK, good question. So, there are multiple commercially available based on metas or hematopoietic cultures and those can be used. The cytokines and growth factors that you need to use will depend on the differentiation conditions that you’re focusing on.
In general, regarding coating, lower adhesion plates are used to culture the HPCs, but this can vary depend on the differentiation protocol that you are interested.

<Tom Allison>
OK.
Do you think so part the second part of the question that that came in, do you think there is opportunity for improvement with respect to the cell culture services that that services that are used within the differentiation protocols?

<Angelica Gomes Ueltschy>
I’m sorry, what was the question Tom?

<Tom Allison>
So the question, the second part of the question that came in, was whether you feel that there is any significant opportunity for improvement in terms of the surface self-culture services that are used within the protocol. Can you improve the protocol further?
By looking at the matrices used.

<Angelica Gomes Ueltschy>
Yeah, yeah. I think some downstream lineages derived from the HPCs, they might need special coating conditions. For instance, for microglia, there are multiple different coatings that are required.
Some people use job track, some people use fibronectin, so there is definitely room there for improvement.

<Tom Allison>
Um, another question. Are all the protocols that you used at Trailhead developed using HD-DoE?

<Angelica Gomes Ueltschy>
Yes, the HD-DoE is the core, the heart of our platform, and is used to develop more robust protocols, and it speeds up optimization of our protocols.

<Tom Allison>
OK.
And then once, um, you know, once sales are purchased or a project has been finished, what kind of support is provided after that?

<Angelica Gomes Ueltschy>
Once you buy ourselves, we provide technical and application-focused support to ensure that you achieve success, successful differentiation, and successful outcome of the cells. We have our field application scientists who can help with that, and of course.
Me as well, if it’s necessary.

<Tom Allison>
Another question has come in. How long does custom iPSC differentiation take from start to delivery? A difficult question, I guess, to answer. Maybe I’ll jump in here and say it really depends on.
The data package that you require first and foremost, and so the QC metrics that you want to see. So that is customizable according to the project that you lay out, and so that can change the timing.
From start to delivery, in terms of Angelica, can you talk about the length of time for the actual differentiation itself once the iPSC line has been established and it works, and we’ve optimized that?

<Angelica Gomes Ueltschy>
Oh, you mean like how long the differentiation protocol lasts for the HPCs? It’s normally around 12 days. And yeah, normally it doesn’t change. If you change the cell line, the protocols we have.
Conduct our protocols in multiple lines, including the one that Mike showed here, and the differentiation we know was pretty similar in the cell lines tested.

<Tom Allison>
Yes.

<Angelica Gomes Ueltschy>
So, of course, the scale of the project will also impact the timeline, but yeah.

<Tom Allison>
So then the question on the microglia, in terms of microglia, do you have any data for growth in organoid conditions? And once differentiated, do we know how long these can be cultured or passage for?

<Angelica Gomes Ueltschy>
So we have conducted a few studies with microglia.
But I’ve seen microglia in the literature being cultured for a long time regarding organoid conditions. I’m not familiar with that. We haven’t tested that.
But I would assume the growth in organoid conditions would be maybe better than the cell alone because the microbes have, you know, friends and the other cell types to support them to have a talk back and forth. So, I’d say that would be better. But yeah, I think the cells can be cultured for a good amount of time if they are culturing the appropriate conditions.

<Tom Allison>
Yeah. And so, the data that Michael showed was not done by us, a group that took our HPCs and did the differentiation themselves. The protocol is published. And so, if you are interested in seeing that publication, we’re very happy to help with that too.
OK. It looks like that’s the end of the questions that have come in. I just want to thank you again for joining us today. You know if you do have an interest in Trailhead’s custom capabilities or off-the-shelf products, we’d love to hear from you. Please reach out to your representative. You can email us at support@trailbio.com.
And finally, the webinar will be available digitally in the coming days on the website. So do look out for that. And again, thank you. We hope that you have a great rest of the day.