Untangling mechanisms for cerebellar neural specification using human pluripotent stem cell-derived organoids

The cerebellum is one of the most complex structures of the brain composed of a high diversity of GABAergic and glutamatergic neurons. Whereas cerebellar biogenesis has been extensively studied in the mouse, an in-depth characterization of genes and pathways involved in cerebellar specification and maturation in the humans remains overlooked. Here, we used human pluripotent stem cells (hPSC)-derived cerebellar organoids (CRBOs) to study the temporal biogenesis of neuronal subtypes. Our results show that CRBOs acquire caudal neural tube identity at an early stage followed by a time-dependent expression of mature cerebellar neuronal markers in vitro , mimicking human neurodevelopment. CRBOs show the generation of both cerebellar excitatory and inhibitory neurons and the expression of glial cell markers, suggesting the generation of a high variety of cerebellar cell types in vitro . Further, in vitro CRBOs show expression of cerebellar disease associated genes, such as those related to ataxia. Our results establish CRBOs as a valuable platform to explore the mechanisms of human cerebellar development and related disorders.