Purkinje cells are the only output neurons in cerebellum that receive and integrate messages from parallel fibers and climbing fibers. Some familial and idiopathic movement disorders, such as ataxias, are characterized by specific Purkinje cell pathologies. Stem cells-derived Purkinje neurons, especially differentiated from patient-specific induced pluripotent stem cells (iPSCs), offer a useful tool to study the mechanisms of these movement disorders. In this chapter, we describe a method to differentiate human iPSCs into Purkinje cells. Human iPSCs are first differentiated to Neph3+Purkinje progenitors. A co-culture system that employs rat or human fetal cerebellar slices is used to promote further maturation of the Purkinje progenitors in vitro. This method provides a platform to investigate the biology of Purkinje cells in normal and disease conditions. © 2018 Elsevier Inc. All rights reserved.