Lentiviral Delivery of Transcription Factors to Generate Motor Neurons from Human Induced Pluripotent Stem Cells

This protocol addresses the need for a reliable and scalable method for generating motor neurons (MNs) in vitro from human induced pluripotent stem cells (hiPSCs). This method offers a breakthrough in MN-related disease research as it overcomes the limitations of accessing patient brain tissues. The protocol involves the generation of neural progenitor cells from hiPSCs, followed by the successful preparation of highly pure and functional MNs. To optimize the induction process, the expression of three critical transcription factors, Neurogenin 2, ISL-1, and LHX3, is incorporated into a single lentiviral vector. This novel approach significantly enhances the yield and purity of the generated MNs, providing a robust platform for biochemically identifying dysregulated factors associated with diseased neurons.