Differentiation of human induced pluripotent stem cells into dorsal and ventral spinal neural progenitor cells for application in spinal cord therapeutics

Abstract

Traumatic CNS injury can attenuate cognitive, motor, and sensory function with minimal potential for full recovery. Research utilizing human induced pluripotent stem cell (hiPSC) -derived neural cell types for in vivo remodeling and neuromodulation after injury has grown substantially in recent years. In the case of spinal cord injury, however, most researchers currently utilize hiPSC -derived ventral motor neurons for cell transplantation after injury, and there is very little exploration of dorsal sensory neuron transplantation, despite the devastating impact of sensory dysfunction. The lack of studies that utilize sensory populations may be due in part to the relative scarcity of dorsal horn differentiation protocols. Building upon our previously published work that demonstrated the rapid establishment of a primitive ectoderm population from hiPSCs, our lab has worked to define the process of producing a diverse population of both ventral spinal and dorsal horn progenitor cells. Our work creates a novel system allowing dorsal and ventral spinal neurons to be differentiated from the same intermediate ectoderm population, making it possible to construct the dorsal and ventral domains of the spinal cord while decreasing variability. This twenty day protocol produces fully mature dorsal and ventral spinal neurons, allowing reconstruction of both sensory and motor pathways that are critical to functional recovery after injury. This technology can be used in tandem with biomaterials and pharmacology to improve in vitro modeling of sensorimotor circuits and in vivo cell transplantation for spinal cord injury, increasing the potential for translation of spinal regeneration therapeutics.