Suchitra Sumitran-Holgersson, Priti Chougule, Gustaf Herlenius, Michael Olausson

University of Gothenburg, Gothenburg, Gothenburg, Sweden

We aimed to produce an acellular human tissue scaffold with a view to test the possibility of recellularization with bone-marrow cells to produce a tissue-engineered small intestine (SI) that can be used for treatment of patients with small bowel disorders. Human small bowel specimens (n=3; each divided into two pieces of 6 cm long segments) were obtained from cadaveric organ donors and treated sequentially with hypotonic buffer, an organic solvent in hypotonic buffer, and a nuclease solution. Each SI piece was recellularized either with a heterogeous population of cells isolated from an allogeneic small intestine sample or CD133+ allogeneic bone-marrow stem cells. Histological and molecular analysis demonstrated that after decellularization, all cellular components and nuclear material were removed. The analysis also showed that the human SI tissue retained its histioarchitecture and major structural proteins. Protein films of common extracellular matrix constituents (collagen IV, collagen I, laminin, and fibronectin) were found in abundance. Immunohistochemical staining found residual angiogenic factors after decellularization. Following recellularization for two weeks with allogenic bone-marrow stem cells we also found viable cells adjacent to an innervated muscularis mucosa and a high repopulation of blood vessels with endothelial cells. In summary, successful decellularization of the human small intestine was achieved producing a biocompatible matrix that retained the major structural components and strength of the native tissue and supported anchorage of newly introduced stem cells.