March 20, 2022 PAO-02-022-CL-20
A.J. Mellott (AJM): I’m a tissue engineer by trade. My Ph.D. focused on the manipulation of mesenchymal stem cells, intracellularly and extracellularly. During my postdoc, I was culturing literally millions to billions of cells, and, like other researchers out there, was spending a lot of time in a biosafety cabinet filling all kinds of tissue culture flasks day in and day out. It was very tedious, laborious, and frustrating. One day, I had a “eureka idea” moment when I realized that it would be much easier to grow cells from the start in a 3D format rather than grow them first in a 2D format and then transfer them to a 3D scaffold or microfluidics device.
That required determining what primary cells need in order to grow like they would in the body. The first thing that dawned on me was that the substrate needed to be expandable. In 2D cell culture, when cells outgrow the container, they must be disassociated from the substrate and split into multiple containers. That process often disrupts cell division and other behaviors. Substrates that fit together would eliminate the need to disassociate cells. I was able to validate this concept once I obtained my faculty position at the University of Kansas Medical Center. Over time, we added channels and worked on ways to manipulate the mechanics and a few other parameters. Eventually, the core concept was established, and the technology was at a place where it could be commercialized.
Ronawk was created and spun out of the university in 2019. After initial investments from family and friends, sufficient momentum was created to allow me to join the company full time and put all of my energy into working with my cofounder Heather Decker. I focused on refining the technology, and she developed applications. Scott joined us as Chief Revenue Officer and Tom Jantsch as President and COO, both in December 2021.
Scott Leigh (SL): I’ve been involved in biotech sales, marketing, branding, and management for about 25 years. I actually met A.J. and Heather many years ago, when I was selling tissue culture plastics. I am excited to be part of a company that has the potential to not only help alleviate some of the pain associated with cell culture but to revolutionize this crucial process. Scientists will be able to get more cells with considerably less effort. We eliminate subcultures, which will drastically reduce contamination. We are automating cell culture and removing the human element from it. Once an incubator is set up, only the media needs to be changed every few days via pipetting. That’s a huge step forward.
AJM: Even changing the media can be automated, because we have perfusion systems. As a team, we have really grown and come to realize that we can do much more than simplify cell culture. We have a potential solution for fully automating this process and eliminating contamination risks. Researchers can get their data faster and focus on analyzing them for use in the development of next-generation therapies. In addition, because we are using a 3D format, the need for animal studies is reduced, and we can get better answers earlier when testing different drug compounds.
Ronawk’s mission is to accelerate the development of next-generation therapies with the technologies we develop. We want scientists to be the heroes in their labs and help the physicians that ultimately help the patients.
AJM: Our core application is in human healthcare, largely because that is where Heather and I, as well as most of our colleagues, have expertise. We have also been fortunate to be approached by cellular agriculture groups interested in either growing plants or culturing meat. For those applications, we have adapted a few Bio-Blocks and achieved successful results growing plants in preliminary studies. We are now in the process of growing the muscle tissue used for cultured meat products, with some really encouraging results that have demonstrated that there is even more versatility with these Bio-Blocks that we realized. So, while our core focus remains on human healthcare, we are looking at other industries that dovetail with that.
SL: I would clarify that we are being approached as much as we are looking for partners. This technology is agnostic to cell culture and can benefit anyone working with any type of cell, whether in wound-healing drug discovery or for growing meat. The principles are all the same.
AJM: I would also like to stress that what we are doing is not truly a big shift or change, but more a case of mimicking what already happens in biology. We are just better at capturing that behavior because of the way our systems are engineered using blocks for cell growth. Our T-Blocks can be modified to mimic specific environments, while our E-Blocks are standard, ready-to-use options. Our newest X-Blocks are for scientists that want to remove cells in order to conduct a full array of analyses. We are trying to hit all of the points that are important to researchers. What’s especially fun is that we get to collaborate with many different people, and that collaboration really drives the further development of the product and the technology so that it is really useful to all of our end users.
AJM: The eureka moment was when I had a very simple thought: Why can’t we grow cells in 3D from the beginning? When cells start growing in the body, they’re really growing in 3D. Nothing in the body — tissues and organs, etc. — is flat or 2D. It’s all contoured. If we could grow cells in a 3D format from the beginning, maybe we could we get higher-quality cells and create more accurate models.
The answer to the question was yes, but that then led to other questions, such as whether cells grown in a 3D format can expand, how nutrient diffusion and gas exchange could be handled, and so on, in order to provide an environment that closely mimics the original, natural cell environment. Answering those questions has involved some trial and error and optimization over the last seven years. Even though Ronawk has only been around since 2019, Heather and I have been working on many aspects of the technology for much longer. That’s where the layering has come in as we continue to refine the product.
Distilling it down, we wanted to create something that would allow cells to grow in 3D that was also modular in the sense that the substrate could mimic the environment in which cells grow naturally and support continuous propagation. And, just as importantly, make it so that it is useable for researchers so that they can get the data they need from their processes. That’s been our main motivation driving the development of our products.
AJM: Much of this comes from my bias as a tissue engineer. From a tissue engineering perspective, having the macrostructure designed like a puzzle piece gives the end user the ability to control how the cells actually grow and expand. In addition, the microchannels inside the Bio-Blocks align when the Bio-Blocks are put together, so that the cells on each Bio-Block come in contact with one another after they have been growing in 3D.
With our growth plate, the Bio-Blocks can be connected in a line to study spinal cord regeneration or blood vessel construction. There are also options for making simple square formats, as well as branched constructs using intersecting Bio-Blocks. As a result, the end user has more control over how the cells grow in actual 3D environments.
In addition, creating these discrete substrates that can be connected together offers the potential for the co-culturing of different types of cells. The hope is that this design allows researchers to combine multiple different cell types to make more complex organoids or sophisticated tissues. One example might be growing bone cells or osteoblasts in one Bio-Block and cartilage or chondrocytes in another and then combining them to observe how the cells interact in 3D.
SL: I would add that for contract research organizations (CROs) and contract development and manufacturing organizations (CDMOs), using our X-Blocks allows for much higher productivity. For instance, lining the bottom of one regular roller bottle with X-Blocks enables the production of as many cells as would be produced in 10.6 roller bottles with a surface area of 850 cm2. Similarly, putting X-Blocks in cell stacks and large bioreactors would also supersize them.
Of course, some engineering may be needed, but the potential is there to increase production by orders of magnitude. One CRO we spoke to explained that projects are often delayed by about 36 weeks while a sufficient quantity of cells is grown to seed the bioreactor. With our technology, that timeframe can be compressed to a few weeks. That level of time savings would have a huge impact for their clients.
AJM: That gets back to our goal of accelerating drug development by enhancing our clients’ capabilities through the use of the 3D culture technology. It always comes back to how we can make our clients the heroes in what they are doing.
Our technology accelerates cell culture across multiple applications. And we continue to make improvements and increase its versatility. We are talking about growing cells, and when those cells are used to produce exosomes, extracellular vesicles, recombinant antibodies, growth factors, and myriad other biomolecules, this technology simplifies things.
AJM: With stem cells, it is well known that, as they are subcultured to passage repeatedly, they eventually reach a point where they senesce –– stop dividing. When put in our Bio-Blocks, however, there is a dramatic reduction in senescence. This behavior was discovered when one of our researchers subcultured adipose-derived stem cells, for which a significant portion of the population was senescent, in one of our Bio-Blocks, and then stained them. Surprisingly, the senescent expression was reduced by nearly 80%.
That result suggests that there is something about the environment in the Bio-Blocks that is waking these cells up and causing them to start dividing again. There are many questions to explore here, and we are working on them. Are the cells dividing again because they are in a structured 3D environment with multiple touchpoints within the 3D Bio-Blocks? Is it because of greater stiffness or the cell signaling that occurs in the Bio-Blocks? It is going to take a while to tease all of that out. We continue to replicate the initial result and are excited about the implications for researchers in multiple fields. If there is a way to essentially wake up senescent cells or reinvigorate them from a regenerative medicine perspective, that’s huge.
AJM: First and foremost, our Bio-Block technology affords economy of scale. As Scott said, it’s now possible to supersize what can be done in even basic plasticware containers. A lot of the times in the lab, cell-related experiments are done on samples that are pretty small, which can make it difficult to really infer results. Now, more cells can be grown in the same small-scale equipment, affording larger quantities of samples to work with.
Second, we can actually grow organoids within a Bio-Block for up to 14 days. Most organoids only survive for about 72 hours, depending on the cell line and other factors. Having access to organoids for a much longer period makes it possible to study the pharmacokinetics and drug metabolism much longer — and in a more 3D model. Third, our Bio-Block technology creates a standardized format, not only for growing cells in 3D but also for performing 3D coculture.
These three things weren’t possible before without a lot of work and consideration. Now, we’ve simplified those processes so researchers can easily run those experiments in their labs and get their data faster.
AJM: We’ve conducted a variety of pilot studies with different end users trying different cell types that either we don’t readily have access to or are just not in our specialty areas. Those partners provide us with data on how those cells perform, which helps us.
SL: I would break the partnerships into two groups. In one group are CDMOs and CROs that are conducting experiments but have not informed Ronawk about the specifics, because their work is proprietary with their clients. These companies are doing beta testing and finding that our Bio-Blocks are accelerating their projects, because they can now perform culture runs in existing equipment but with much greater numbers of cells.
The second group comprises research funding. They are using our Bio-Blocks to create libraries of specific types of cells that they then provide to all of the research groups that they are funding for their studies. This approach is advantageous, because it is possible to control the phenotype and thus have more uniformity across all of the various research groups participating in a study. In addition, by only requiring one passage, we are minimizing the risk of cross-contamination.
Ronawk's Bio-Blocks offer more consistency for all downstream applications. This technology will accelerate drug discovery and wound healing therapies. It will ultimately enable personalized medicine. Libraries of cells could also be taken into the field for surgical wound healing and plastic surgery. All of these things are now possible. Instead of working with one cell line, researchers can just as easily grow six, 10, or more cell lines, making it easier to test their hypothesis on a wider range of cell types with less genetic drift and minimal changes in phenotype. It is staggering to consider what people might be able to do with this.
AJM: What we’re finding when we’re talking with different clients and collaborators is that they’re not changing any of the equipment or the plasticware or anything else that they’re doing; we’re just simplifying their work. They really like the idea, but they have a difficult time believing that the protocol is as simple as it is. We address that uncertainty by performing demonstrations, which provides a level of assurance.
SL: Switching to our Bio-Blocks can be viewed as a paradigm shift like others in the past, such as going from flask to 96- to 384-well plates. It really doesn’t take researchers too long to accept the concept once it is explained and demonstrated. We are helping people scale up without having to go to larger equipment.
AJM: With our Bio-Blocks, we are growing cells across the X, Y, and Z axes. Going from 2D by adding the Z dimension allows the jump from surface area to volume, and we are guiding cells — just like in the body — to grow throughout that volume in all three dimensions.
SL: Unfortunately, the current terminology is limited. Typically, when people think of 3D cell culture today, they think of organoids. But our Bio-Blocks go well beyond organoids. Essentially, they allow vertical pharming for cell culture. It is like putting a ten-story building on a one-acre plot and expanding the work area to 10 acres. Where people worked with 1 billion cells, now they can work with 10 billion or more in the same equipment. Once they understand what our blocks enable, there is usually an “aha moment” which is really fun to be a part of.
AJM: What is great is that, because we positioned them to grow cells, our Bio-Blocks are essentially comparable to other plasticware. As a result, they fall into the Class I Device category. For the U.S. FDA, that means taking the 510k route.
If we move to producing biologic drugs from cells grown in our Bio-Blocks or possibly the cells themselves being released from the Bio-Block for use as a cellular therapy in the future, we will need to follow a drug approval pathway. Starting with the lower-risk applications allows us to have conversations with the FDA and allow the agency to become familiar with our product. We can also discuss what we need to do to approach those drug-related applications: what data to collect, how to ensure safety and efficacy at the clinical stage, and what the development of new drugs, devices, and/or other therapies should look like.
SL: I think polymerase chain reaction (PCR) is a phenomenal model. It all started with Watson and Crick naming DNA and winning the Nobel Prize (1962). Karl Mullis introduced PCR in 1985. It took a while to be adopted, though. Even in the 1990s, many labs weren’t doing PCR. Then quantitative PCR (qPCR) was introduced, and it took several years before it was widely adopted. Today in pharma, it is a foundational assay. It takes time for new technologies to be accepted. But we see researchers experiencing this “aha” moment almost every day, which is just amazing. And the number of people, institutions, and companies reaching out to us is also quite something.
AJM: That’s been really exciting for us. Initially, we were the ones reaching out. Now we have people contacting us on a regular basis asking for more information and wanting to set up meetings. The number of organizations that we’re engaged with just keeps growing by the day.
AJM: We do. We aren’t discussing them yet, but there is a list of ideas we want to explore and that will hopefully lead to the development of new Bio-Blocks that can be commercialized. We have collaborators who ask if we can develop products that serve specific functions. Many of them are already on our list, and it is nice to get that confirmation. It is also helping us to understand the pain points for them and to figure out what the demand is and what direction we should go in — where we should focus our energy to be the most helpful.
AJM: The core team was Heather and me initially. We are scientists and are fortunate to have gone through a number of programs that have helped us with the business vernacular. Our initial success helped us to attract individuals like Scott and our new President and COO Tom Jantsch, as well as out new Chief Marketing Officer, Trice Alford. Our core leadership team is mostly complete.
Now we are looking at the next hires with respect to production, sales, marketing, and day-to-day operations. That is where Tom, Scott, Heather, and Trice will play big roles. I will also be looking at all the data that are being generated to determine our strategy going forward and address the next immediate need.
Our ability to pair science and technology with business has been really beneficial. We are building on that with the collaboration and synergy between our team members, which allows us to work together across all different positions and get a strong understanding of what we need to do next.
SL: I agree — it isn’t just the technical expertise. We are also looking for a certain type of person. We want believers, not only in what we’re doing but in the potential science. And we want them to get the technology out in front of as many people as possible in order to give researchers access to this impactful technology. The COVID-19 vaccines are a great example of that kind of thing. It took teamwork and passion to get those new vaccines developed and manufactured in 10 months. We’re looking for the same.
I also would like to say that the group at Ronawk is the nicest, most collaborative group of people I’ve ever worked with. Everyone really believes in the technology and its promise. It is just an incredible place of ideas.
AJM: I think you hit the nail on the head. We are very much team-oriented and collaborative. It is definitely not a one-person show; it’s us collectively building this technology and company and helping others. That type of diversity and all of these different ideas allows us to accelerate our efforts because we get to see so many points of view, which makes us better as a team.
AJM: Our technology enables so many new applications and new experiments on multiple fronts. In the future, I see Ronawk spinning out some subsidiaries that focus on very unique technologies for specific end-use applications that are enabled by our core technology. The mission will remain to help researchers and ultimately patients, farmers, and many others.
As a teaser, I will mention that we see real potential for our technology in aerospace, with possible customers like NASA, Space-X, Blue Origin, Virgin Galactic, and so on. Now that the space industry includes private tourism, there is a need for a way to easily grow food onboard those spacecrafts. This will be a bit down the line, but we believe our cell culture blocks could help deliver that type of solution.
I believe Ronawk will have a few spinouts in the next two to five years. We will have our hands in various ventures, some that involve partnering with large companies — because for so many life science applications, everything comes back once again to the cell. By making it easier to grow cells, we just make it easier for our customers to enhance their own value propositions.
SL: Our success will be defined by our customers’ successes. They are the ones doing the really great work. We are just helping them do more, faster. Look at what researchers will do and what pharma companies will do with our products in a few years. They become the heroes because we’re enabling them to achieve their vision. That’s the real key here.
AJM: Ronawk was founded by scientists with a vision of helping and enabling others. We are always open to hearing from people who want to learn more. because like Scott said, the success of our clients is our success. And that’s really why we’re here.
Dr. A.J. Mellott earned his doctorate in Bioengineering from the University of Kansas in 2014, where he focused on genetically reprogramming human umbilical cord mesenchymal stem cells to become mechanosensory hair cells using non-viral methodologies. He went on to build a successful basic research program for the Department of Plastic Surgery at the University of Kansas Medical Center. In 2019, Dr. Mellott co-founded Ronawk to create 3D technologies that accelerate research for the development of next-generation therapies.