The contractions of the intestinal wall and the movement of luminal chyme exert a diverse range of physical forces on the epithelial and mesenchymal cells of the intestinal mucosa. These forces, along with other features of the mucosal environment, such as the composition of the extracellular matrix, the topography of the lamina propria, and epithelial-stromal crosstalk contribute to establishing distinct proliferative and differentiated epithelial compartments in the crypts and villi, respectively. The homeostasis of this crypt-villus axis is critical for normal intestinal function, and is deranged in many diseases. However, there is currently a lack of experimental models that can replicate these important epithelial dynamics. The objective of this research is to develop biomaterial and biomechanical techniques to provide physiological inputs to co-cultures of human intestinal enteroids (HIEs) and primary intestinal subepithelial myofibroblasts (ISEMFs). My hypothesis is that these scaffolds will better recapitulate the organization and epithelial homeostasis than currently available models. I will construct these models in two specific aims. First, I will grow monolayers of HIEs on top of villus-shaped, synthetic hydrogel mimics of the intestinal lamina propria that will simulate both the stiffness and the extracellular matrix profile of the intestina basement membrane and contain a subepithelial population of ISEMFs. These features will organize the HIEs into distinct villus and crypt regions with differentiated epithelium coverin the gel villi, and proliferating epithelium located in the intervillar regions. Second, I will expoe HIEs, ISEMFs, and HIE/ISEMF co-cultures to cyclic stretch that mimics the effects of peristalsis. This mechanical stimulation will promote HIE proliferation and differentiation through effects on both the epithelial and the mesenchymal cells. The models developed in this research are expected to yield fundamental insights into intestinal physiology, inform efforts in intestinal tissue engineering, and provide a framework for studying the pathogenesis of intestinal disease.