Generation of inner ear organoids containing functional hair cells from human pluripotent stem cells

Abstract

Human inner ear tissue derived from pluripotent stem cells could provide a powerful platform for drug discovery or a source of sound-or motion-sensing cells for patients with hearing loss or balance dysfunction. Here we report a method for differentiating human pluripotent stem cells to inner ear organoids that harbor functional hair cells. Using a three-dimensional culture system, we modulate TGF, BMP, FGF, and Wnt signaling to generate multiple otic vesicle-like structures from a single stem-cell aggregate. Over two months, the vesicles develop into inner ear organoids with sensory epithelia that are innervated by sensory neurons. Additionally, using CRISPR/Cas9, we generate an ATOH1-2A-eGFP cell line to detect hair cell induction and demonstrate that derived hair cells exhibit electrophysiological properties similar to those of native sensory hair cells. Our culture system will be useful for elucidating mechanisms of human inner ear development and testing potential inner ear therapies.The human inner ear contains ∼75,000 sensory hair cells that detect sound and movement via mechanosensitive stereocilia bundles 1,2 . Genetic mutations or environmental insults, such as loud noises and ototoxic drugs, can cause irreparable damage to these hair cells, Users may view, print, copy, and download text and data-mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use:

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