Journal Article

Directed Differentiation of Human Embryonic Stem Cells Toward Placode‐Derived Spiral Ganglion‐Like Sensory Neurons

Belmadani, Abdelhak, Whitlon, Donna S., Zhang, Chaoying, Edelbrock, Alexandra N., Matsuoka, Akihiro J., Miller, Charles A., Stupp, Samuel I., Chadly, Duncan M., Morrissey, Zachery D., Lyass, Ljuba, McGuire, Tammy L., Homma, Kazuaki, Kobayashi, Shun, Kessler, John A., Tanaka‐Matakatsu, Miho

2017

Overview

The ability to generate spiral ganglion neurons (SGNs) from stem cells is a necessary prerequisite for development of cell‐replacement therapies for sensorineural hearing loss. We present a protocol that directs human embryonic stem cells (hESCs) toward a purified population of otic neuronal progenitors (ONPs) and SGN‐like cells. Between 82% and 95% of these cells express SGN molecular markers, they preferentially extend neurites to the cochlear nucleus rather than nonauditory nuclei, and they generate action potentials. The protocol follows an in vitro stepwise recapitulation of developmental events inherent to normal differentiation of hESCs into SGNs, resulting in efficient sequential generation of nonneuronal ectoderm, preplacodal ectoderm, early prosensory ONPs, late ONPs, and cells with cellular and molecular characteristics of human SGNs. We thus describe the sequential signaling pathways that generate the early and later lineage species in the human SGN lineage, thereby better describing key developmental processes. The results indicate that our protocol generates cells that closely replicate the phenotypic characteristics of human SGNs, advancing the process of guiding hESCs to states serving inner‐ear cell‐replacement therapies and possible next‐generation hybrid auditory prostheses. © Stem Cells Translational Medicine 2017;6:923–936

• Publisher: Oxford University Press,John Wiley and Sons Inc
• Subject: Animals / Axons / Brain Stem - cytology / Cell Differentiation / Cell Line / Cell Lineage / Cell Movement / Cell Survival / Cellular therapy / Cochlea / Cochlear nuclei / Coculture Techniques / Data analysis / Ectoderm / Embryo cells / Embryonic stem cells / Embryos / Growth factors / Hearing loss / Human Embryonic Stem Cells - cytology / Human Embryonic Stem Cells - metabolism / Humans / Mice / Neural differentiation / Neural induction / Neural stem cells / Neural Stem Cells - cytology / Neurogenesis / Physiology / Progenitor cells / Prosthetics / Protocols and Manufacturing for Cell‐Based Therapies / Rats / Sensory neurons / Sensory Receptor Cells - cytology / Spiral ganglion / Spiral Ganglion - cytology / Stem cell / Stem cells / Translational s and Reviews / Transplantation / Writing