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Human induced pluripotent stem cells (hiPSCs) have changed the way human development and disease are studied. It is now increasingly possible to create specific cell types and organ-like structures from an embryonic stem cell state, with improved recapitulation of physiological conditions. Here, we describe a robust and reproducible method to differentiate hiPSCs into cerebellar neurons with several options for downstream applications to address different experimental questions. In addition to conventional dissociated culture conditions, we also present a method for long-term three-dimensional culture of cerebellar organoids for advanced in vitro modelling of the cerebellar microenvironment.

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