Scaffold-Assisted Generation of Innervated Gastrointestinal Organoids for Transplantation

The field of human pluripotent stem cell (hPSC) based organoid science has rapidly evolved to generate more complex structures resembling functional human tissues. Here, focusing on gastrointestinal organs, we developed a novel methodology for generating large-scale innervated organoids for transplantation providing tissue for both in vivo and ex vivo studies. By implementing a scaffolding tray, we generated small intestinal, colonic, and gastric organoids with an elongated tubular shape compared to conventionally generated organoids. Transcriptomics and electrophysiology data demonstrated that this method fostered the co-development of additional cell types absent in conventional organoids, namely a de novo functional enteric nervous system. When compared to traditional methods, intestinal (HIO), colonic (HCO) and gastric (HaGO) organoids grown in a scaffolding tray reached in vitro maturation in half the time, supporting engraftment with enhanced efficiency, resulting in tissues several centimeters in size, significantly larger than their conventionally made counterparts. This novel methodology represents a step forward in the field, simplifying current protocols and expediting the generation of clinically relevant transplantable functional gut tissues.