Thanks to a special 3D-printed scaffold tray designed by experts at Cincinnati Children’s Center, researchers can now create larger versions of functional human intestinal organoids twice as fast as traditional methods, and these organoids grow their own nerve cells.
This improved technology could help accelerate the production of human mini-organ tissue large enough to repair damage or restore degraded function in the human small intestine, stomach, and colon. Such tissues may also be valuable for future disease research and for more accurately assessing the risk of organ damage associated with oral drugs.
Project details will be available on May 22, 2026. natural biomedical engineering. The new production system was designed and tested by a team led by researcher Dr. Holly Pauling, senior author Dr. Maxime Mahe, and 17 other scientists from the University of Cincinnati Children’s University and the University of Nantes in France.
The research team used a new “closed culture system” (CCS) to grow small intestine, colon, and stomach organoids from tiny spheres to centimeter-scale tube-like shapes that are nearly 10 times larger than previous methods. Additionally, unlike methods that require complex efforts to introduce neurons, these organoids develop their own nervous systems.
These organoids reach engraftment maturity twice as fast and develop their own functional neurons, demonstrating how engineering principles can drive biological innovation. Our closed culture system is more than just a production method. It’s a scalable, flexible platform for building complex human organizations. ”
Dr. Holly Pauling, Senior Author
New production system drives faster growth
For more than 15 years, experts at the Cincinnati Center for Children’s Stem Cell and Organoid Medicine (CuSTOM) have been leaders in creating miniature versions of digestive system organs, steadily increasing the sophistication of lab-produced tissues. More recently, the team has been working on ways to create enough customized tissue to be transplanted into patients to repair organ damage or restore degraded specialized functions.
The new method uses 3D printing technology to create a tray-shaped scaffold mold from surgical resin, which is then filled with degassed polydimethylsiloxane, a flexible, rubber-like type of silicone.
The new tray contains grooves designed to confine a collection of spherical organoids in a line, which promotes spheroid fusion and maturation. This fusion takes place within a special mixture of nutrients and other components that support the initial growth of induced pluripotent stem cells (iPSCs) into more complex organoids.
By day 6, the individual spheroids have grown into integrated structures along the grooves of the tray. Transfer these to another hydrogel medium and continue growing for an additional 8 days.
By day 14, the organoid constructs had generated all cell types and structures, which previously took 28 days to achieve. These tissues are transplanted into rodents that have been genetically modified to minimize the risk of rejection.
All transplanted tissues successfully engrafted in the rodents, the co-authors said. After growing it in rodents, the research team generated as much as 8 cm of functional small intestine tissue, compared to about 1 cm using the previous protocol. Not only was the structure much larger than previous methods, but its neuromuscular function also became similar to natural human tissue, representing a major advance.
“We can now not only generate complex gut organoids at scale, but also induce their differentiation into functional tissues with integrated enteric neural networks,” Mahe says. “By taking advantage of a defined growth environment, the cells’ innate self-organizing ability facilitates the formation of tissue structures that closely resemble the human gastrointestinal tract.”
Dr. Jim Wells, study co-author and chief scientific officer at CuSTOM, said this new technology overcomes key barriers of scale and capability in organoid research and biomanufacturing.
“The simplicity, reproducibility and versatility of this platform makes it amenable to widespread adoption,” Wells says. “Furthermore, the emergence of a self-organized nervous system within these organoids is particularly important for further studies of neurodevelopmental disorders.”
One step closer to human clinical trials
Michael Helmrath, M.D., a surgeon and scientist at Cincinnati Children’s Hospital and co-director of CuSTOM, has been working for more than a decade to develop intestinal organoids sophisticated enough to be transplanted into human patients.
In 2017, Helmrath and colleagues demonstrated how to combine neural crest cells and intestinal tissue cells in a layered process to create the first human organoids with neural functionality. His team also showed how intestinal organoids can be made to grow larger by transplanting them into mice to provide a blood supply. Since then, intestinal organoids have become more sophisticated, including versions with immune cells in addition to specialized organ cells and nerves.
Now, a new process using rats instead of mice produces even larger amounts of tissue.
“Although it is not yet possible to grow complete, full-size human organs in some kind of aquarium, studies like this have produced large amounts of tissue that are directly matched to individual patients,” Hermlath says. “We believe that once such tissue is transplanted, it will grow further and proliferate as part of the patient’s own organ, restoring its function.”
Further research and development is needed before CCS organoids can be used in human clinical trials, Helmrath said. But if success continues, organoid medicine could allow more infants and children with organ failure to be treated without the need for full organ transplants.
About research
In addition to Pauling, Mahe, Wells, and Helmlath, Cincinnati Children’s co-authors include Akarjot Singh, Garrett Fisher, Conrad Thorner, Praneet Chaturvedi, Kalpana Nattamai, Kalpana Srivastava, Matthew Batty, Nicole Brown, Taylor Hausfeld, Amy Pitstick, Riccardo Barile, Christopher Mayhew, and Takanori Takebe. Three experts from Inserm and Nantes Université were also co-authors.
Funding sources for the study included the National Institute of Diabetes and Digestive and Kidney Diseases (U01 DK103117, P30 DK078392) and the National Agency for Research (ANR-17-CE14-0021, ANR-21-CE14-0017).
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Cincinnati Children’s Hospital Medical Center
Reference magazines:
Pauling, H.M. Others. (2026). Large scale innervated functional human intestinal tissue for transplantation with temporary spheroid confinement. natural biomedical engineering. DOI: 10.1038/s41551-026-01688-6. https://www.nature.com/articles/s41551-026-01688-6
