Medium-sized spiny neurons (MSNs) are the only neostriatum- projection neurons, and their degeneration underlies some of clinical features of Huntington's disease. Here we used human developmental biology and exposure to key neurodevelopmental molecules to inform human pluripotent stem (hPS) cells differentiation into MSNs. In a feeder-free adherent culture, ventraltelencephalic neuroectodermal specification is induced by BMP/TGF-β inhibition and subsequent morphogens treatment. The emerging FOXG1+/GSX2+ telencephalic progenitors are terminally differentiated, resulting in the systematic line-independent generation of CTIP2+/ DARPP-32+MSNs. Similarly to mature MSNs, these neurons show in vitro inhibitory, spontaneous, and repeated spikes and dopamine neuromodulation, and synaptic integration ability in vivo. When transplanted into the striatum of quinolinic acid-lesioned rats, hPS-derived neurons survive, integrate, and are capable of differentiating into DARPP-32-expressing neurons, leading to a restoration of amphetamine-induced rotation behaviour. In summary, hPS cells can be efficiently driven to acquire a functional striatal fate using a stepwise method representing a platform for in vitro developmental neurobiology studies and drug screenings approaches.