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Decelluarization and Reseeding of Porcine Colon with Human CaCo-2 Cells for Whole-Scale Organ Engineering Applications

Abstract

A new approach to whole-scale organ engineering is the seeding of stem cells upon a decellularized scaffold of a desired organ. Current efforts seek to use this method for whole-scale organ engineering. Here, we report a novel decellularizationrecellularization approach to engineering a human colonic graft with human enterocytes and xenogeneic scaffold. Porcine colon tissue was decellularized with protein-solubilizing agents and recellularized by incubation with Caco-2. Immunohistochemistry, DNA staining, and SEM were used to study the effects upon the scaffold. Exposure to 2%SDS-0.5%Triton solution for four hours resulted in the best decellularization and preservation of microstructure, while tissues treated with 2%SDS-0.5%Triton and 0.05%Trypsin/EDTA supported the best recellularization. Our method provides a potential strategy for streamlined regeneration of a viable colonic lumen from xenogeneic and allogeneic sources while maintaining colon microstructure and potential for recellularization.

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