The generation of dopamine (DA) neurons from stem cells holds great promise for future biomedical research and in the clinical treatment of neurodegenerative diseases, such as Parkinson's disease. Mesenchymal stem cells (MSCs) derived from the adult human bone marrow (BM) can be easily isolated and expanded in culture while maintaining their immense plasticity. Here, we describe a protocol to generate DA-producing cells from adult human MSCs using a cocktail that includes sonic hedgehog (SHH), fibroblast growth factor 8 (FGF8), and basic fibroblast growth factor (bFGF). Electrophysiological functional DA neurons could be achieved by further treatment with brain-derived neurotrophic factor (BDNF). In summary, a protocol is described for the induction of primary BM-derived human MSCs to specific transdifferentiation; in this case, functional DA neurons. The MSC-derived DA cells express DA-specific markers, synthesize, and secrete dopamine. The described method could be used to generate DA cells for various model systems in which DA-producing cells are implicated in pathophysiological conditions.