Generation of Human iPSC-Derived Cerebellar Organoids

Human induced pluripotent stem cell (hiPSC)-derived brain organoids are a novel tool to model human-specific aspects of brain development. Here, we describe a reproducible protocol for generating hiPSC-derived cerebellar organoids that have been experimentally validated to contain typical cerebellar cell populations. Generated three-dimensional cerebellar organoids can be kept in long-term culture or further be maintained in dissociated culture settings to allow for single-cell resolution in downstream applications. The current protocol is an improvement on earlier methods that require co-culture of hiPSC-derived neurons with murine cerebellar cell populations.