Purmorphamine is widely used in stem cell research and regenerative medicine due to its ability to selectively activate the Hedgehog (Hh) signaling pathway. It functions as a potent agonist of Smoothened (SMO), a key protein in the Hh pathway, making it a valuable tool for directing the differentiation of pluripotent stem cells into specific cell lineages, particularly neural and osteogenic cells. In neuroscience research, purmorphamine is employed to induce differentiation of stem cells into dopaminergic neurons, which is relevant for studying and developing therapies for neurodegenerative diseases like Parkinson’s disease. It has also been used in bone tissue engineering studies to promote osteoblast differentiation and bone formation. Due to its small molecule nature and high potency, purmorphamine offers advantages over protein-based Hh activators such as Sonic Hedgehog (Shh), including better cell permeability, stability, and cost-effective use in high-throughput screening assays. It is frequently used in developmental biology to study embryonic patterning and cell fate determination. Additionally, purmorphamine has been explored in cancer research to investigate the role of aberrant Hh signaling in tumor growth, although its use in this area is primarily limited to mechanistic studies rather than therapeutic application.