Directed differentiation of human induced pluripotent stem cell to sensory neurons by combined small molecule inhibitors

Abstract

Human induced pluripotent stem cells (iPSCs) hold great promise for cell therapies and tissue engineering. iPSC-derived neural cells provide prospects for studying neurodevelopment and modeling neurological diseases. A prerequisite for these studies is a reproducible protocol that efficiently yields an abundant number of neural cell types. However, differentiation of iPSCs with extrinsic factors is a slow, step-wise process, mimicking the complex timing of human development. Progress has been made in identifying signaling pathways that direct the differentiation of iPSCs into specific lineages. Using a combination of small molecule inhibitors, we attempt to develop a new protocol to generate sensory neurons from human iPSCs. Human iPSCs were exposed to small molecule inhibitors. The iPSC-derived sensory neurons were maintained for two weeks in growth factors required for their survival. After differentiation, over 90% of the total cell population expressed the neuron-specific protein, Tuj-1 and neurofillament. Nearly 80% of the total cell population co-expressed peripherin and Brn3a, the specific markers for sensory neuron. Therefore, we demonstrated that a combination of small-molecule inhibitors of signaling pathways promotes highly efficient peripheral neural induction from human iPSCs. Our in vitro model of iPSC-derived sensory neuron provides a promising strategy for controlled production of sensory neurons and may serve as a useful tool for studying human neurodevelopment and modeling neurological diseases.