Pluripotent stem cells (PSCs) possess self-organizing abilities in 3D culture. This property has been demonstrated in recent studies, including the generation of various neuroectodermal and endodermal tissues. For example, PSCs are able to differentiate into specific type of neural tissues, such as the neocortex and the optic cup, in response to local positional information brought about by signals during embryogenesis. In contrast, the generation of cerebellar tissue from PSCs requires a secondary induction by a signaling center, called the isthmic organizer, which first appears in the cell aggregate in 3D culture. Such developmental complexity of cerebellum has hampered establishment of effective differentiation culture system from PSCs, thus far.
We recently reported that cerebellar neurons are generated from human PSCs (hPSCs). In this chapter, we describe an efficient protocol for differentiation of 3D cerebellar neuroepithelium from hPSCs. We also describe the protocols for further differentiation into specific neurons in the cerebellar cortex, such as Purkinje cells and the granule cells.
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Bird MJ, Needham K, Frazier AE et al (2014) Functional characterization of Friedreich ataxia iPS derived neuronal progenitors and their integration on the adult brain. PLoS One 9:e101718CrossRefPubMedPubMedCentralGoogle Scholar
Eigentler A, Boesch S, Schneider R et al (2013) Induced pluripotent stem cells from Friedreich ataxia patients fail to upregulate frataxin during in vitro differentiation to peripheral sensory neurons. Stem Cells Dev 22:3271–3282CrossRefPubMedGoogle Scholar
Hick A, Wattenhofer-Donzé M, Chintawar S et al (2013) Neurons and cardiomyocytes derived from induced pluripotent stem cells as a model for mitochondrial defects in Friedreich’s ataxia. Dis Model Mech 6:608–621CrossRefPubMedGoogle Scholar
Koch P, Breuer P, Peitz M et al (2011) Excitation-induced ataxin-3 aggregation in neurons from patients with Machado-Joseph disease. Nature 480:543–546PubMedGoogle Scholar
Ku S, Soragni E, Campau E et al (2010) Friedreich’s ataxia induced pluripotent stem cells model intergenerational GAA∙TTC triplet repeat instability. Cell Stem Cell 7:631–637CrossRefPubMedPubMedCentralGoogle Scholar