(1186-C) Rapid and consistent generation of functional motor neurons from reprogrammed human iPSCs using opti-ox technology
Tuesday, February 6, 2024
12:00 PM – 1:00 PM EST
Location: Exhibit Halls AB
Abstract: Motor neurons consist of distinct neuronal subtypes that control the activity of muscles and glands in direct or indirect manners. Motor neurons form a large neuronal network that receives inputs from interneurons, sensory neurons or other motor neurons to control complex behaviours such as locomotion. Pathological perturbation of these motor circuits can lead to the development of motor neuron diseases (MNDs) such as spinal muscular atrophy and amyotrophic lateral sclerosis.
Development of therapies to treat MNDs is hampered by the limited translatability of existing preclinical animal models as well as the lack of reliable and consistent sources of in vitro models. Human induced pluripotent stem cells (hiPSCs) can be used to generate motor neurons for in vitro applications, however current differentiation protocols are often lengthy, inconsistent, and difficult to scale. Our proprietary opti-ox™ (optimised inducible overexpression) technology enables highly controlled expression of transcription factors, which can rapidly reprogram hiPSCs into specific cell types of interest, to provide a robust, consistent, and reliable cell source for in vitro applications.
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 phenotype. ioMotor Neurons show spontaneous neuronal activity with increasing firing rate over 40 days in culture, as shown by multielectrode array activity (MEA). Finally, next generation sequencing methods have shown consistency between three different batches produced through opti-ox mediated reprogramming. opti-ox technology can be utilised for the scalable and consistent production of hiPSC-derived motor neurons. ioMotor Neurons have the potential to advance the development of new therapeutics for MNDs and to further our understanding of motor neuron development and maturation in vitro.