Maturation of Human Pluripotent Stem Cell-Derived Cerebellar Neurons in the Absence of Co-culture
Teresa P. Silva, Evguenia Bekman, Tiago G. Fernandes, Sandra H. Vaz, Carlos A. V. Rodrigues, Maria Margarida Diogo, Joaquim M. S. Cabral, Maria Carmo‐Fonseca
Abstract
The cerebellum plays a critical role in all vertebrates, and many neurological disorders are associated with cerebellum dysfunction. A major limitation in cerebellar research has been the lack of adequate disease models. As an alternative to animal models, cerebellar neurons have been differentiated from pluripotent stem cells. However, previous studies were able to produce only limited amounts of Purkinje cells. Moreover, in vitro generation of Purkinje cells so far required co-culture systems, which may introduce unknown components to the system. Here we describe a novel differentiation strategy that uses defined medium to generate Purkinje cells, granule cells, interneurons, and deep cerebellar nuclei projection neurons, that self-formed and matured into electrically active cells. Using a defined basal medium optimized for neuronal cell culture, we successfully promoted the maturation of cerebellar precursors without the need for co-culturing. We anticipate that our findings can be translated into better models for the study of cerebellar dysfunctions, and represent a valuable advance towards the development of autologous replacement strategies for treating cerebellar degenerative diseases.