The linear measurements and 3D maps may prove useful when attempting more anatomically accurate reconstructions.

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Background Short bowel syndrome (SBS) is a life-threatening condition with few solutions. Tissue engineered intestine (TEI) is a potential treatment, but donor intestine is a limiting factor. Expanded epithelial surrogates termed enteroids may serve as a potential donor source. Materials and Methods To produce TEI from enteroids, crypts were harvested from mice and enteroid cultures established. Enteroids were seeded onto polymer scaffolds using Matrigel or culture medium, and implanted in immunosuppressed mice for 4 weeks. Histology was analyzed using Periodic Acid Schiff (PAS) staining and immunofluorescence (IF). Neomucosa was quantified using ImageJ software. To determine if TEI could be produced from enteroids established from small intestinal biopsies, 2×2mm pieces of jejunum were processed for enteroid culture, enteroids were expanded and seeded onto scaffolds, and scaffolds implanted for 4 weeks. Results Enteroids in Matrigel produced TEI in 15/15 scaffolds, whereas enteroids in medium produced TEI in 9/15 scaffolds. Use of Matrigel led to more neomucosal surface area compared to media (10,520±2,905 μm vs. 450±127 μm, p<0.05). Histologic examination confirmed the presence of crypts and blunted villi, normal intestinal epithelial lineages, intestinal subepithelial myofibroblasts, and smooth muscle cells. Crypts obtained from biopsies produced an average of 192±71 enteroids. A single passage produced 685±58 enteroids, which was adequate for scaffold seeding. TEI was produced in 8/9 scaffolds seeded with expanded enteroids. Conclusions Enteroids can be obtained from minimal starting material, expanded ex vivo, and implanted to produce TEI. This method shows promise as a solution to the limited donor intestine available for TEI production in patients with SBS.

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