title: Reprogramming of single-cell-derived mesenchymal stem cells into hair cell-like cells.
abstract: OBJECTIVE
  Adult mesenchymal stem cells (MSCs) can be converted into hair cell-like cells by transdetermination.
  
  
  BACKGROUND
  Given the fundamental role sensory hair cells play in sound detection and the irreversibility of their loss in mammals, much research has focused on developing methods to generate new hair cells as a means of treating permanent hearing loss. Although MSCs can differentiate into multiple cell lineages, no efficient means of reprogramming them into sensory hair cells exists. Earlier work has shown that the transcription factor Atoh1 is necessary for early development of hair cells, but it is not clear whether Atoh1 can be used to convert MSCs into hair cells.
  
  
  METHODS
  Clonal MSC cell lines were established and reprogrammed into hair cell-like cells by a combination of protein transfer, adenoviral based gene transfer, and co-culture with neurons. During transdetermination, inner ear molecular markers were analyzed using reverse transcriptase-polymerase chain reaction, and cell structures were examined using immunocytochemistry.
  
  
  RESULTS
  Atoh1 overexpression in MSCs failed to convert MSCs into hair cell-like cells, suggesting that the ability of Atoh1 to induce hair cell differentiation is context dependent. Because Atoh1 overexpression successfully transforms VOT-E36 cells into hair cell-like cells, we modified the cell context of MSCs by performing a total protein transfer from VOT-E36 cells before overexpressing Atoh1. The modified MSCs were transformed into hair cell-like cells and attracted contacts from spiral ganglion neurons in a co-culture model.
  
  
  CONCLUSIONS
  We established a new procedure, consisting of VOT-E36 protein transfer, Atoh1 overexpression, and co-culture with spiral ganglion neurons, which can transform MSCs into hair cell-like cells.
authors.affiliation: Model Animal Research Center of Nanjing University, Nanjing, People's Republic of China.
authors.forename: Zhaoyu
authors.initials: Z
authors.lastname: Lin
authors.forename: Philip
authors.initials: P
authors.lastname: Perez
authors.forename: Zhenyu
authors.initials: Z
authors.lastname: Sun
authors.forename: Jan-Jan
authors.initials: JJ
authors.lastname: Liu
authors.forename: June Ho
authors.initials: JH
authors.lastname: Shin
authors.forename: Krzysztof L
authors.initials: KL
authors.lastname: Hyrc
authors.forename: Damien
authors.initials: D
authors.lastname: Samways
authors.forename: Terry
authors.initials: T
authors.lastname: Egan
authors.forename: Matthew C
authors.initials: MC
authors.lastname: Holley
authors.forename: Jianxin
authors.initials: J
authors.lastname: Bao
mesh_terms: D000199:Actins / Q000378:metabolism; D053687:Adult Stem Cells / Q000502:physiology; D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics / Q000502:physiology; D015415:Biomarkers; D002454:Cell Differentiation; D002460:Cell Line; D002469:Cell Separation; D065150:Cellular Reprogramming / Q000502:physiology*; D018920:Coculture Techniques; D000229:Dependovirus / Q000235:genetics; D005822:Genetic Vectors; D006198:Hair Cells, Auditory / Q000502:physiology*; D007150:Immunohistochemistry; D059630:Mesenchymal Stem Cells / Q000502:physiology*; D051379:Mice; D008810:Mice, Inbred C57BL; D016133:Polymerase Chain Reaction; D013136:Spiral Ganglion / Q000166:cytology
chemical_list: D000199:Actins; D051792:Basic Helix-Loop-Helix Proteins; D015415:Biomarkers; C500943:Atoh1 protein, mouse