Rho-associated coiled-coil containing protein kinase (ROCK) signaling is a primary regulator of cell behavior in stem cell biology and organoid systems. This review examines the diverse roles of Rho/ROCK signaling in cytoskeletal remodeling, cell adhesion, differentiation, and mechanotransduction. Special emphasis is placed on its role in organoid formation, patterning, and functional maturation. An understanding of how this pathway integrates with other signaling networks, such as Ras, Hippo, and PI3K-AKT, is crucial for optimizing organoid fidelity. We propose future research directions that enhance organoid models through targeted modulation of ROCK activity, thereby opening new opportunities for regenerative medicine and disease modeling.