OP‐4 ADVANCES IN TISSUE ENGINEERING TO REGENERATE THE INTESTINAL EPITHELIUM

Abstract

Introduction:Irreversible intestinal failure (IF) due to anatomical or functional loss is a devastating condition is associated with significant morbidity and mortality. Patients referred for intestinal transplantation suffer low survival rates due to the high graft‐rejection rate and sepsis as a consequence long‐term immunosuppression. These clinical challenges reveal an urgent need for research into new treatments for IF. Here, we present recent advances in our work on intestinal tissue engineering.Methods:Ethical approval is in place to collect intestinal tissue from patients at Great Ormond Street Hospital. Intestinal crypts were isolated and organoids were cultured as previously described. Decellularization of whole intestinal sections was performed using 2 cycles of the Detergent Enzymatic Treatment (DET) as previously reported. Organoids were cultured on the epithelial surface of the acellular matrix in vitro for 10 days. Cell survival was analysed by histology, immunofluorescence, immunohistochemistry and electron microscopy techniques.Results:Intestinal crypts from the small intestine and colon were successfully isolated from endoscopic biopsies and cultured in order to generate 3 dimensional organoids. The presence of goblet cells and enterocyte cells were confirmed on immunohistochemistry. Whole sections of intestine of varying lengths were successfully decellularized and assessed using H&E staining and scanning electron microscopy. Organoids were seeded onto the epithelial surface of the acellular intestinal matrices and allowed to engraft for a period of 10 days in vitro. The formation of a neo‐epithelium with evidence of a microvillus brush border was evident after 10 days in culture. Furthermore, the cells had successfully migrated into the crypt domains of the matrix.Conclusions:We demonstrate the potential of tissue engineering for intestinal failure by combining organoid cultures with decellularization expertise. Our results confirm human intestinal organoids to be the ideal source of progenitor cells to regenerate the intestinal epithelium, with potential clinical applications for transplantation.