Generation of Motor Neurons from Human Induced Pluripotent Stem Cells Using Small Chemical Molecules

Motor neurons (MNs) play a critical role in regulating both voluntary and involuntary movements, and their dysfunction underlies various movement-related disorders. The generation of patient-specific MNs provides an unprecedented approach for disease modeling, drug screening, and cell therapy. This chapter presents a comprehensive protocol for the controlled generation and maturation of MNs from human induced pluripotent stem cells (hiPSCs). The protocol details the step-by-step process, including hiPSC culture and expansion, neural induction, conversion to MN progenitors, suspension culture to form neurosphere-like structures, and subsequent maturation of MNs. It also outlines the immunostaining procedure for identifying early and mature MN markers. This robust methodology offers researchers a powerful tool for investigating MN-related diseases and potential therapeutic strategies.