Authors: Shuyan Wang, Bin Wang & Zhiguo Chen
This protocol describes how to differentiate human induced pluripotent stem cells (iPSCs) to Purkinje cells. Human iPSCs are first differentiated to Neph3+ Purkinje progenitors. To promote maturation of Purkinje progenitors in vitro, a co-culture system is used to enhance the maturation of Purkinje precursors on rat and human fetal cerebellar slices. Furthermore, Purkinje progenitor cells are injected into the cerebellum of newborn immunodeficient mice to test the differentiation ability in vivo.
It remains a challenge to differentiate human induced pluripotent stem cells (iPSCs) to Purkinje cells. Purkinje neurons are the only output neurons in cerebellum and often afflicted in spinocerebellar ataxias (SCAs) and other medical indications such as ethanol exposure and autoimmune diseases. Obtaining patient-specific Purkinje neurons would offer a valuable tool to model cerebellar diseases in a culture dish for investigating t