Development and Functional Applications of Human iPSC-Derived Spinal Motor Neurons

The aim of this study was to produce spinal motor neurons from human iPSCs with sufficient purity for use in a multitude of downstream assays. In particular we wanted to produce motor neurons that could be cultured in defined conditions over long periods of time, without being hampered by outgrowth from proliferative cell types. Using an optimized differentiation protocol that improves upon published methods, we were able to produce motor neurons from iPSCs at greater than 60% purity as measured by Isl 1/2 and Tuj1 positive staining. These cells can be stored frozen, thawed, and cultured in media without glia for extended periods. We collected ICC, qPCR and MEA data to characterize the motor neuron cells and used iPSC lines from multiple donors to demonstrate a robust protocol that produces motor neurons independent of donor iPSC line. These data show the characteristics and utilization of motor neurons produced from iPSC.