Protocol for 3D direct reprogramming of human glia into dopaminergic neurons with gene expression and immunocytochemistry validation

Direct reprogramming of somatic cells into neurons offers a promising strategy for studying neurological disorders and developing cell-based therapies. This protocol describes the 3D direct reprogramming of human glial progenitor cells into subtype-specific dopaminergic neurons. It includes spheroid formation combined with lentiviral transduction, neuronal induction using doxycycline, and validation via gene expression and immunocytochemistry. This versatile approach enables rapid and efficient reprogramming of human cells into specific neuronal subtypes. Overall design: Bulk RNA-seq of human glia progenitors, reprogrammed DA neurons in 3D