Engineering Large-Scale and Innervated Functional Human Gut for Transplantation.

Area of Science:

• Gastroenterology and Regenerative Medicine
• Organoid Technology
• Neurogastroenterology

Background:

• Conventional organoids lack a functional enteric nervous system.
• Generating complex human gastrointestinal tissues is time-consuming.
• Existing methods limit the translational potential for disease modeling.

Purpose of the Study:

• To develop a confined culture system (CCS) for generating complex, functional human gastrointestinal tissues.
• To investigate the development of a de novo enteric nervous system within CCS-derived organoids.
• To compare the maturation speed and engraftment efficiency of CCS organoids with traditional methods.

Main Methods:

• Utilized a confined culture system (CCS) to culture human small intestinal, colonic, and gastric tissues.
• Characterized organoid development using transcriptomic and electrophysiological analyses.
• Assessed organoid maturation, transplantation, engraftment, and in vivo functional augmentation with murine luminal content.

Main Results:

• CCS successfully generated large-scale, innervated, tubular human gastrointestinal organoids.
• Transcriptomic and electrophysiological data confirmed the co-development of a functional enteric nervous system.
• CCS organoids matured in half the time of traditional methods, showing enhanced engraftment and size.
• In vivo exposure to murine luminal content further improved organoid function.

Conclusions:

• The CCS methodology simplifies protocols while enhancing the complexity and generation speed of clinically relevant gut tissues.
• CCS enables the development of functional de novo enteric nervous systems in organoids, a key advancement over conventional methods.
• This system holds significant translational potential for modeling complex intestinal diseases and advancing regenerative medicine.