Rapid and consistent generation of functional motor neurons from reprogrammed human iPSCs using opti-ox technology

We have used opti-ox technology to rapidly reprogram hiPSCs into motor neurons, termed ioMotor Neurons, which are a homogenous population of cells with classical neuronal morphology and neurite outgrowth. As early as 4 days in culture, cells express the pan-neuronal markers MAP2 and TUBB3, the cholinergic markers ChAT and VAChT and the motor neuron-specific markers MNX1 and ISL1/2, as assessed by both ICC and RT-qPCR. Bulk RNA sequencing of ioMotor Neurons demonstrates a rapid acquisition of a motor neuron signature. ioMotor Neurons show spontaneous neuronal activity with increasing firing rate over 40 days in culture, as shown by multielectrode array activity (MEA).