| Currently, stem cell behavior following injury, inflammation, and viral/bacterial challenge is difficult to study in vivo due to the inability to control variables, limited human tissue resources, the prohibitive costs involved with animal studies, and/or the lack of cross-species infection by species-specific microbes. The goal of this thesis is to develop an ex vivo model of the gut epithelium and underlying mesenchyme to facilitate studies that are not possible due to these technical, ethical, and financial constraints. Microfabricated well arrays that mimic intestinal crypt dimensions demonstrated that PDLA, polystyrene, and 1002F promote proliferation of gut myofibroblasts more rapidly than PDMS. Methods were developed to enhance crypt seeding into microwells and standardize the isolation of epithelial tissue. Advanced 3-D imaging techniques using multi-photon excitation (MPE) and Imaris were also developed. These studies are a stepping stone in the development of an organ-on-a-chip for the intestine, and provide proof of concept for intestinal tissue engineering with microfabricated scaffolds. |
