This thesis presents the Rotating Organoid Culture Kinetics Insert (ROCKI), a novel 3D-printed device designed to combine the cellular complexity of ALI organoid culture with pump-free, unidirectional fluid flow to support multi-media, multi organ coculture systems within standard 6-well culture plates. Using mouse liver organoids, ROCKI cultures were compared with conventional submerged and ALI organoid methods. Fluid flow was found to influence altered organoid morphology and growth rate in both submerged and ROCKI cultures, and ROCKI organoids achieved a level of morphological complexity and hepatocyte-, endothelial-, and macrophage-specific marker expression comparable to ALI cultures, more closely aligning with primary tissue than static submerged organoids. These findings demonstrate that ROCKI can support complex, multi cell type organoid cultures under physiologically relevant fluid flow without the need for pumps or specialized microfluidic equipment.
