A Monolayer System for the Efficient Generation of Motor Neuron Progenitors and Functional Motor Neurons from Human Pluripotent Stem Cells

Methods for the conversion of human induced pluripotent stem cells (hiPSCs) into motorneurons (MNs) have opened to the generation of patient-derivedin vitrosystems that can be exploitedfor MN disease modelling. However, the lack of simplified and consistent protocols and the factthat hiPSC-derived MNs are often functionally immature yet limit the opportunity to fully takeadvantage of this technology, especially in research aimed at revealing the disease phenotypes thatare manifested in functionally mature cells. In this study, we present a robust, optimized monolayerprocedure to rapidly convert hiPSCs into enriched populations of motor neuron progenitor cells(MNPCs) that can be further amplified to produce a large number of cells to cover many experimentalneeds. These MNPCs can be efficiently differentiated towards mature MNs exhibiting functionalelectrical and pharmacological neuronal properties. Finally, we report that MN cultures can belong-term maintained, thus offering the opportunity to study degenerative phenomena associatedwith pathologies involving MNs and their functional, networked activity. These results indicatethat our optimized procedure enables the efficient and robust generation of large quantities ofMNPCs and functional MNs, providing a valid tool for MNs disease modelling and for drugdiscovery applications .