title: Directed Differentiation of Human Embryonic Stem Cells Toward Placode-Derived Spiral Ganglion-Like Sensory Neurons.
abstract: 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.
authors.affiliation: Department of Otolaryngology and Head and Neck Surgery, Chicago, IL, USA.|Department of Communication Sciences and Disorders, Chicago, IL, USA.|Knowles Hearing Center, Chicago, IL, USA.
authors.forename: Akihiro J
authors.initials: AJ
authors.lastname: Matsuoka
authors.affiliation: Department of Otolaryngology and Head and Neck Surgery, Chicago, IL, USA.
authors.forename: Zachery D
authors.initials: ZD
authors.lastname: Morrissey
authors.affiliation: Department of Otolaryngology and Head and Neck Surgery, Chicago, IL, USA.
authors.forename: Chaoying
authors.initials: C
authors.lastname: Zhang
authors.affiliation: Department of Otolaryngology and Head and Neck Surgery, Chicago, IL, USA.|Knowles Hearing Center, Chicago, IL, USA.
authors.forename: Kazuaki
authors.initials: K
authors.lastname: Homma
authors.affiliation: Department of Molecular Pharmacology and Biological Chemistry, Chicago, IL, USA.
authors.forename: Abdelhak
authors.initials: A
authors.lastname: Belmadani
authors.affiliation: Department of Otolaryngology and Head and Neck Surgery, Chicago, IL, USA.
authors.forename: Charles A
authors.initials: CA
authors.lastname: Miller
authors.affiliation: Department of Otolaryngology and Head and Neck Surgery, Chicago, IL, USA.
authors.forename: Duncan M
authors.initials: DM
authors.lastname: Chadly
authors.affiliation: Department of Otolaryngology and Head and Neck Surgery, Chicago, IL, USA.
authors.forename: Shun
authors.initials: S
authors.lastname: Kobayashi
authors.affiliation: Department of Biomedical Engineering, Chicago, IL, USA.
authors.forename: Alexandra N
authors.initials: AN
authors.lastname: Edelbrock
authors.affiliation: Department of Otolaryngology and Head and Neck Surgery, Chicago, IL, USA.
authors.forename: Miho
authors.initials: M
authors.lastname: Tanaka-Matakatsu
authors.affiliation: Department of Otolaryngology and Head and Neck Surgery, Chicago, IL, USA.|Knowles Hearing Center, Chicago, IL, USA.
authors.forename: Donna S
authors.initials: DS
authors.lastname: Whitlon
authors.affiliation: Department of Biomedical Engineering, Chicago, IL, USA.
authors.forename: Ljuba
authors.initials: L
authors.lastname: Lyass
authors.affiliation: Department of Biomedical Engineering, Chicago, IL, USA.
authors.forename: Tammy L
authors.initials: TL
authors.lastname: McGuire
authors.affiliation: Department of Medicine, Chicago, IL, USA.|Department of Biomedical Engineering, Chicago, IL, USA.|Simpson Querrey Institute for BioNanotechnology, Chicago, IL, USA.|Department of Chemistry, Northwestern University.|Department of Materials Science & Engineering, Northwestern University.
authors.forename: Samuel I
authors.initials: SI
authors.lastname: Stupp
authors.affiliation: Department of Neurology, Feinberg School of Medicine, Northwestern University, Chicago, IL, USA.
authors.forename: John A
authors.initials: JA
authors.lastname: Kessler
mesh_terms: D000818:Animals; D001933:Brain Stem / Q000166:cytology; D002454:Cell Differentiation; D002460:Cell Line; D019070:Cell Lineage; D002465:Cell Movement; D002470:Cell Survival; D018920:Coculture Techniques; D000066449:Human Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D051379:Mice; D058953:Neural Stem Cells / Q000166:cytology; D051381:Rats; D011984:Sensory Receptor Cells / Q000166:cytology*; D013136:Spiral Ganglion / Q000166:cytology*
keywords: Cellular therapy; Embryonic stem cells; Neural differentiation; Neural induction; Progenitor cells; Stem cell; Transplantation