Abstract
Differentiation of the pluripotent neuroepithelium into neurons and glia is accomplished by the interaction of growth factors and cell-type restricted transcription factors. One approach to obtaining a particular neuronal phenotype is by recapitulating the expression of these factors in embryonic stem (ES) cells. Toward the eventual goal of auditory nerve replacement, the aim of the current investigation was to generate auditory nerve-like glutamatergic neurons from ES cells. Transient expression of Neurog1 promoted widespread neuronal differentiation in vitro; when supplemented with brain-derived neurotrophic factor (BDNF) and glial cell line-derived neurotrophic factor (GDNF), 75% of ES cell-derived neurons attained a glutamatergic phenotype after 5 d in vitro. Mouse ES cells were also placed into deafened guinea pig cochleae and Neurog1 expression was induced for 48 h followed by 26 d of BDNF/GDNF infusion. In vivo differentiation resulted in 50-75% of ES cells bearing markers of early neurons, and a majority of these cells had a glutamatergic phenotype. This is the first study to report a high percentage of ES cell differentiation into a glutamatergic phenotype and sets the stage for cell replacement of auditory nerve.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Basic Helix-Loop-Helix Transcription Factors / genetics
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Basic Helix-Loop-Helix Transcription Factors / metabolism*
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Biomarkers / analysis
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Biomarkers / metabolism
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Brain-Derived Neurotrophic Factor / metabolism
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Brain-Derived Neurotrophic Factor / pharmacology
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Cell Differentiation / drug effects
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Cell Differentiation / physiology
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Cell Line
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Cells, Cultured
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Cochlear Nerve / drug effects
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Cochlear Nerve / embryology
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Cochlear Nerve / metabolism
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Deafness / chemically induced
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Deafness / metabolism
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Deafness / surgery
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Doxycycline / pharmacology
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Embryonic Stem Cells / drug effects
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Embryonic Stem Cells / metabolism
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Embryonic Stem Cells / transplantation*
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Glial Cell Line-Derived Neurotrophic Factor / metabolism
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Glial Cell Line-Derived Neurotrophic Factor / pharmacology
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Glutamic Acid / metabolism*
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Guinea Pigs
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Humans
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Mice
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Nerve Growth Factors / metabolism
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Nerve Growth Factors / pharmacology*
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Nerve Tissue Proteins / drug effects
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Nerve Tissue Proteins / genetics
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Nerve Tissue Proteins / metabolism*
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Neurogenesis / drug effects
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Neurogenesis / physiology*
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Phenotype
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Sensory Receptor Cells / drug effects
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Sensory Receptor Cells / metabolism
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Stem Cell Transplantation / methods*
Substances
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Basic Helix-Loop-Helix Transcription Factors
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Biomarkers
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Brain-Derived Neurotrophic Factor
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Glial Cell Line-Derived Neurotrophic Factor
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Nerve Growth Factors
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Nerve Tissue Proteins
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Neurog1 protein, mouse
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Glutamic Acid
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Doxycycline