Abstract
Organoid technology, as an innovative approach in biomedicine, exhibits promising prospects in disease modeling, pharmaceutical screening, regenerative medicine, and oncology research. However, the use of tumor-derived Matrigel as the primary method for culturing organoids has significantly impeded the clinical translation of organoid technology due to concerns about potential risks, batch-to-batch instability, and high costs. To address these challenges, this study innovatively introduced a photo-crosslinkable hydrogel made from a porcine small intestinal submucosa decellularized matrix (SIS), fish collagen (FC), and methacrylate gelatin (GelMA). The cost-effective hydrogel demonstrated excellent biocompatibility, tunable mechanical properties, rapid gelation properties, and low immunogenicity. Importantly, the proliferation and differentiation capacities of small intestinal organoids cultured in hydrogel were comparable to those in Matrigel, with no significant disparity observed. Furthermore, after one week of transplantation in nude mice, the hydrogel-organoid complex exhibited sustained structural and functional stability while preserving the differentiation characteristics of small intestinal organoids. Our study also demonstrated the effective potential of FC/SIS/GelMA hydrogel in accelerating the repair process of small intestinal defects, reducing the area of scar formation, and promoting the regeneration of both intestinal villi and smooth muscle tissue. In summary, this study presents a novel protocol for culturing small intestinal organoids, offering potential implications for future clinical applications and serving as an experimental foundation for the development of tissue-engineered intestines based on small intestinal organoids.