title: In vitro differentiation of mouse embryonic stem cells into inner ear hair cell-like cells using stromal cell conditioned medium.
abstract: Hearing loss is mainly caused by loss of sensory hair cells (HCs) in the organ of Corti or cochlea. Although embryonic stem (ES) cells are a promising source for cell therapy, little is known about the efficient generation of HC-like cells from ES cells. In the present study, we developed a single-medium culture method for growing embryoid bodies (EBs), in which conditioned medium (CM) from cultures of ST2 stromal cells (ST2-CM) was used for 14-day cultures of 4-day EBs. At the end of the 14-day cultures, up to 20% of the cells in EB outgrowths expressed HC-related markers, including Math1 (also known as Atoh1), myosin6, myosin7a, calretinin, α9AchR and Brn3c (also known as Pou4f3), and also showed formation of stereocilia-like structures. Further, we found that these cells were incorporated into the developing inner ear after transplantation into chick embryos. The present inner ear HC induction method using ST2-CM (HIST2 method) is quite simple and highly efficient to obtain ES-derived HC-like cells with a relatively short cultivation time.
authors.affiliation: Department of Pathogen, Infection and Immunity, Nara Medical University, Nara, Japan. oujix@naramed-u.ac.jp
authors.forename: Y
authors.initials: Y
authors.lastname: Ouji
authors.forename: S
authors.initials: S
authors.lastname: Ishizaka
authors.forename: F
authors.initials: F
authors.lastname: Nakamura-Uchiyama
authors.forename: M
authors.initials: M
authors.lastname: Yoshikawa
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D002642:Chick Embryo; D017077:Culture Media, Conditioned / Q000378:metabolism; D058732:Embryoid Bodies / Q000166:cytology / Q000378:metabolism; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D006199:Hair Cells, Auditory, Inner / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D017154:Stromal Cells / Q000166:cytology / Q000378:metabolism
chemical_list: D017077:Culture Media, Conditioned