Protocol for combining BMP, MEK, WNT inhibition and iNGN2 to enable rapid neuronal differentiation and regional...

Neurogenin 2 (iNGN2) overexpression induces human induced pluripotent stem cells (hiPSCs) into neurons but results in a mixed population of peripheral and central nervous system neurons. Pre-differentiation using BMP, MEK, and WNT inhibition ("BMWi") prior to iNGN2 promotes telencephalic neuron differentiation. We outline neural induction, neural progenitor replating/freezing/thawing, and telencephalic neuron maturation and describe protocols for patterning motor neurons, ventral midbrain, and sensory neurons (sensBMi). For complete details on the use and execution of this protocol, please refer to Habich et al.¹.