title: Differentiation of an auditory neuronal cell line suitable for cell transplantation.
abstract: The auditory neuroblast cell line US/VOT-N33 (N33), which is conditionally immortal, was studied as an in vitro model for the differentiation of spiral ganglion neurons (SGNs) and as a candidate for cell transplantation in rodents. It expresses numerous molecular markers characteristic of auditory neuroblasts, including the transcription factors GATA3, NeuroD, Brn3a and Islet1, as well as the neuronal cytoskeletal protein beta3-tubulin. It displays active migratory behaviour in vitro and in vivo. In the presence of the fibroblast growth factors FGF1 or FGF2 it differentiates bipolar morphologies similar to those of native SGNs. In coculture with neonatal cochlear tissue it is repelled from epithelial surfaces but not from native SGNs, alongside which it extends parallel neuronal processes. When injected into the retina in vivo, EGFP-labelled N33 cells were traced for 1-2 weeks and migrated rapidly within the subretinal space. Cells that found their way into the retinal ganglion cell layer extended multiple processes but did not express beta3-tubulin. The ability of N33 to migrate, to differentiate, to localize with native SGNs in vitro and to survive in vivo suggests that they provide an effective model for SGN differentiation and for cell transplantation into the ear.
authors.affiliation: Department of Biomedical Sciences, Addison Building, Western Bank, Sheffield, S10 2TN, UK.
authors.forename: A J
authors.initials: AJ
authors.lastname: Nicholl
authors.forename: A
authors.initials: A
authors.lastname: Kneebone
authors.forename: D
authors.initials: D
authors.lastname: Davies
authors.forename: D I
authors.initials: DI
authors.lastname: Cacciabue-Rivolta
authors.forename: M N
authors.initials: MN
authors.lastname: Rivolta
authors.forename: P
authors.initials: P
authors.lastname: Coffey
authors.forename: M C
authors.initials: MC
authors.lastname: Holley
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D051792:Basic Helix-Loop-Helix Proteins; D019208:Brain-Derived Neurotrophic Factor / Q000494:pharmacology; D002452:Cell Count / Q000379:methods; D002454:Cell Differentiation / Q000502:physiology*; D002465:Cell Movement / Q000502:physiology; D017690:Cell Transplantation; D002478:Cells, Cultured; D003051:Cochlea / Q000502:physiology; D018920:Coculture Techniques / Q000379:methods; D004268:DNA-Binding Proteins / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004338:Drug Combinations; D016220:Fibroblast Growth Factor 1 / Q000494:pharmacology; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D005346:Fibroblast Growth Factors / Q000494:pharmacology; D050990:GATA3 Transcription Factor; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects / Q000502:physiology*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D007150:Immunohistochemistry / Q000379:methods; D051379:Mice; D008810:Mice, Inbred C57BL; D009419:Nerve Tissue Proteins / Q000378:metabolism; D009474:Neurons / Q000187:drug effects / Q000502:physiology* / Q000637:transplantation; D020933:Neurotrophin 3 / Q000494:pharmacology; D009925:Organ of Corti / Q000166:cytology* / Q000254:growth & development / Q000502:physiology; D051381:Rats; D012160:Retina / Q000637:transplantation; D013997:Time Factors; D015534:Trans-Activators / Q000378:metabolism; D050818:Transcription Factor Brn-3; D050819:Transcription Factor Brn-3A; D014157:Transcription Factors / Q000378:metabolism; D014162:Transfection / Q000379:methods; D014404:Tubulin / Q000378:metabolism; D014947:Wounds and Injuries / Q000503:physiopathology
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; D019208:Brain-Derived Neurotrophic Factor; D004268:DNA-Binding Proteins; D004338:Drug Combinations; D016220:Fibroblast Growth Factor 1; D016222:Fibroblast Growth Factor 2; D005346:Fibroblast Growth Factors; D050990:GATA3 Transcription Factor; D049452:Green Fluorescent Proteins; D009419:Nerve Tissue Proteins; D020933:Neurotrophin 3; D015534:Trans-Activators; D050818:Transcription Factor Brn-3; D050819:Transcription Factor Brn-3A; D014157:Transcription Factors; D014404:Tubulin; C494748:Gata3 protein, mouse; C494189:Pou4f1 protein, mouse; C462941:beta3 tubulin, mouse; C485184:enhanced green fluorescent protein; C093564:Neurogenic differentiation factor 1