(191v) Spray Delivery of Organoids to Reconstitute Intestinal Epithelium on Decellularized Native Extracellular Matrix

The native extracellular matrix (ECM) serves as a unique platform for tissue engineering because it provides an organ-specific scaffold in terms of both matrix composition and tissue architecture. However, efficacious cell-seeding techniques that would allow for recellularization of these ECM constructs with appropriate cell types to enable biologic function remain under development. In this study, the impact of spraying as a seeding technique for repopulation of decellularized small intestine was investigated. The effects of spray parameters on CaCo-2 cell viability and distribution were analyzed via fluorescent microscope. High cell viability and homogeneous cell distribution were obtained when cell suspensions were sprayed via airbrush at low pressure. The effect of spray pressure on the length and number of organoid clusters were evaluated by image analysis and monolayer formation was explored by H&E staining after 1 week of culture. While organoids seeded via pipetting onto the decellularized scaffold demonstrated clumping, organoids airbrush spray-seeded at 0.35 bar onto decellularized scaffold demonstrated monolayer formation. The results suggest that seeding cells via airbrush spraying holds great potential, especially for the recellularization of tubular organs.