Scale Up Spheroid Production: FibroBiologics' Path to Clinical Scale

Rather than using two-dimensional or suspension cell cultures, as most cell therapeutic companies do, FibroBiologics' products are based on 3D fibroblast spheroids. Dr. Khoja explained why. "I don't think [2D fibroblast cultures] represent, and publications have also indicated, they don't represent the natural response and characteristics of how these cells actually work in organs and tissues," he said. "In spheroid form, their secretion profile, their metabolic profile, and their transcriptomic profile are significantly different as compared to 2D cells, and they resemble more closely the natural response role of these cells in the human body."

Early on, FibroBiologics tried several traditional methods for spheroid production, but the team encountered difficulties, including high variability in spheroid size, high labor requirements, contamination risks, and lack of scalability. Dr. Khoja described some of these challenges and why the team looked for alternatives.

"Being able to produce these spheroids to a specific size reproducibly was difficult with other methods. For example, let's take into account the hanging drop format, which is one of the oldest ways of making spheroids. It works, but it's incredibly labor-intensive and low-scale. It's difficult, it takes a lot of time, and it's error-prone. For small-scale projects, that's how we initially started doing our testing. But in terms of scaling up, it was just not possible," Dr. Khoja said.

The team also tried another method, using stirred suspension bioreactors for spheroid production. "We looked at the spinning wheel method of making these spheroids, and yes, they generated the spheroids. It could be scaled up, but the sizing variability was extremely wide. You could get spheroids in the micron size all the way up to several hundred microns. So, the variability was way too great. And it would not have been useful for us, even though it could be scaled up," Dr. Khoja said.

Another limitation of these methods is that they require significant hands-on labor, making them both prone to contamination and challenging to scale. Dr. Khoja explained: "Since they're so labor-intensive, there are multiple... potential points of failure and contamination with those [methods]. For example, with hanging drop, you always have to pick up the plate cover, dispense and put it back."

Dr. Khoja said his team found success when they began using Corning^® Elplasia^® 12K flasks during preclinical studies with fibroblast spheroids. Elplasia products are spheroid production systems designed for reproducible, scalable, scaffold-free 3D cell culture and uniform spheroid growth. "When we did find Elplasia flasks, we were just amazed how well it worked right from the beginning, very little optimization on our part to get it working."

Two significant advantages were the uniformity of spheroid size as well as the simplicity of adjusting spheroid size in Elplasia vessels.

"We were very easily able to adjust the size of the spheroids based on the cell number. We've done those experiments. We know how many cells we can put in to get a specific size range and get that size range with a very tight distribution. So, we're very happy with that," Dr. Khoja said.

Uniformity in spheroid size is critical for some of FibroBiologics’ products. Dr. Khoja explained that this is partly due to safety concerns: "For a topically administered product, which is our product for wound healing, sizing accuracy is not that important. However, for a systemically administered product, like the drug products that we're going to be using for MS and psoriasis, sizing is very important because you don't want your drug product to be so large that it will clog capillaries or induce a certain reaction."

The Corning Elplasia technology is part of Corning's family of products with a hydrophilic, neutrally charged Ultra-Low Attachment (ULA) surface that inhibits specific and nonspecific immobilization, forcing cells into a suspended state. The ULA surface, in combination with the rounded microcavity geometry, enables cells to aggregate and form 3D spheroids of uniform size and shape. Elplasia plates and flasks have found uses in drug screening, cancer research, stem cell research, and now, clinical cell therapy work. Elplasia 12K flasks, introduced in 2022, generate approximately 12,000 spheroids in a T-75 flask footprint.

The process of getting up and running with Elplasia 12K flasks and optimizing the protocol was "smooth and very timely," Dr. Khoja said. "And of course, that was with the help of Corning Applications." The support that Corning Applications scientists provided was "phenomenal" and helped the FibroBiologics team optimize spheroid production in the Elplasia 12K flasks fairly quickly, he said.

As FibroBiologics progressed from early to later preclinical research and began planning for human clinical trials, they required additional spheroids, which exceeded the capacity of the Elplasia 12K flasks.

In pursuit of further scalability, the team at FibroBiologics collaborated with Corning in testing a new product, the Corning Elplasia 48K vessel. This product utilizes the same ULA surface and rounded microcavity geometry at a larger scale to generate approximately 48,000 spheroids per vessel.

"We worked very closely, and they were extremely helpful in helping us optimize and develop a really nice workflow for utilizing the 48K Elplasia vessel," Dr. Khoja said.

As one of the initial testers of the product, FibroBiologics was able to offer feedback to Corning to optimize features and performance characteristics based on their workflow. "Of course, when you're developing a product, a new product, there's always going to be some challenges, and we expected that, and I think Corning expects that as well. And I think, to be honest with you, the process was fairly smooth from iteration to iteration. Corning was very quick at turning around suggested changes based on what we've seen and sending us another iteration to test."

For Dr. Khoja, the Elplasia 48K vessel is essential for FibroBiologics' work. "It's the scalability. For example, we have a 120-patient clinical trial coming up in Australia in early 2026, for which we have to make four and a half million spheroids. There's no other way we could have made those without the availability of the 48K vessel. Yes, we could have used the 12K vessel, which is available, but again, we would have had to use four times as much."

Corning Elplasia products have multiple features that support uniformity and health of spheroids. Media can be exchanged without disturbing the spheroids in the microcavities in the Elplasia 12K and 48K vessels. Additionally, because Corning ULA products for spheroid production are now available in a range of sizes and formats, users can scale output while utilizing the same surface coating and microcavity geometry. This allows the spheroids to grow in a similar environment, providing consistency from the R&D and preclinical phases through process development for clinical trials and manufacturing phases.

Elplasia 12K and 48K vessels achieve high uniformity through their microcavity design and shared culture media across microcavities. "Even for our wound care product, which is topically administered, we consistently get very good sizing. We're about plus or minus 15% of the size we're looking for. So, we're very happy with the consistency and viability, and the yield has been very good as well," Dr. Khoja said.

Dr. Khoja noted that another advantage of moving up to the Elplasia 48K is that the larger-scale production per vessel reduces the risk of contamination because there are fewer points of human contact.

"Of all the training runs that we've done so far and all the Elplasia plates that we have utilized internally for our work, we have not yet had a—knock on wood—single case of contamination. So, it's worked beautifully so far."

FibroBiologics' clinical trials include the 120-patient trial in Australia, planned for the first quarter of 2026. The Phase 1/2 trial will test the company's treatment for diabetic foot ulcers using spheroids grown in Elplasia 48K vessels. This will provide the first clinical data from spheroids grown in Elplasia products, as well as the first-ever clinical data on spheroids used for a therapeutic purpose.

In addition, FibroBiologics is using the Elplasia 48K to produce spheroids in preparation for upcoming Phase 1/2 trials in MS and psoriasis.

Beyond the clinical trials in 2026, Dr. Khoja sees ahead an even greater need for high-volume spheroid production at FibroBiologics. As the company looks ahead to Phase 3 trials in larger patient populations that will enroll about 200 to 250 patients, and to clinical product manufacturing, it will turn to Corning for further product development.

"If this product gets approved, then we need to figure out how to manufacture it to scale for clinical use," Khoja said. "So hopefully Corning will come up with something bigger for us to accommodate the yields."

The Elplasia 48K vessel exemplifies the benefits of customer-driven innovation at Corning, where scientists and engineers responded to FibroBiologics’ need for a larger-scale spheroid production solution, with the team at FibroBiologics providing feedback throughout the process.

In addition to the Corning Elplasia 12K flask and 48K vessel, other Elplasia products include Elplasia microplates and Elplasia open-well plates. Learn more about these products and find Elplasia application notes, protocols, and other resources on spheroid production.