Large-scale and innervated functional human gut tissues for transplantation via transient spheroid confinement.

Confined culture system (CCS) organoids generate large-scale human small intestinal, colonic, and gastric tissues with a de novo enteric nervous system, reaching 8 cm width after 10 weeks.

• Why it matters: Current human gut organoids often mature incompletely and require complex, time-consuming assembloid steps to add a functional nervous system for transplantation.
• What they did: The CCS uses a 3D-printed scaffolding tray to transiently confine spheroid fusion and growth, enabling spontaneous co-development of a functional ENS, assessed by transcriptomics and electrophysiology.
• The result: CCS tissues show selective neuromuscular function with excitatory and inhibitory neurons, mature faster than traditional methods, are up to ten times larger, and engraft better while adapting to a murine luminal environment with preserved barrier integrity.

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