title: Induction of Functional 3D Ciliary Epithelium-Like Structure From Mouse Induced Pluripotent Stem Cells.
abstract: OBJECTIVE
  To generate ciliary epithelium (CE) from mouse induced pluripotent stem (iPS) cells.
  
  
  METHODS
  Recently, a protocol for self-organizing optic cup morphogenesis in three-dimensional culture was reported, and it was suggested that ocular tissue derived from neural ectoderm could be differentiated. We demonstrated that a CE-like double-layered structure could be induced in simple culture by using a modified Eiraku differentiation protocol.
  
  
  RESULTS
  Differentiation of a CE-like double-layered structure could be promoted by glycogen synthase kinase 3β (GSK-3β) inhibitor. Connexin43 and aquaporin1 were expressed in both thin layers, and induced CE-like cells expressed ciliary marker genes, such as cyclinD2, zic1, tgfb2, aldh1a3, wfdc1, otx1, BMP4, and BMP7. Increases in cytoplasmic and nuclear β-catenin in aggregates of the CE-like double-layered structure were confirmed by Western blot analysis. In addition, tankyrase inhibitor prevented the induction of the CE-like double-layered structure by GSK-3β inhibitor. Dye movement from pigmented cells to nonpigmented cells in the mouse iPS cell-derived CE-like structure was observed in a fluid movement experiment, consistent with the physiological function of CE in vivo.
  
  
  CONCLUSIONS
  We could differentiate CE from mouse iPS cells in the present study. In the future, we hope that this CE-like complex will become useful as a graft for transplantation therapy in pathologic ocular hypotension due to CE dysfunction, and as a screening tool for the development of drugs for diseases associated with CE function.
authors.affiliation: Department of Ophthalmology and Visual Sciences Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki, Japan.
authors.forename: Hirofumi
authors.initials: H
authors.lastname: Kinoshita
authors.affiliation: Department of Ophthalmology and Visual Sciences Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki, Japan.
authors.forename: Kiyoshi
authors.initials: K
authors.lastname: Suzuma
authors.affiliation: Laboratory for Retinal Regeneration, Center for Developmental Biology, RIKEN, Kobe, Japan.
authors.forename: Jun
authors.initials: J
authors.lastname: Kaneko
authors.affiliation: Laboratory for Retinal Regeneration, Center for Developmental Biology, RIKEN, Kobe, Japan.
authors.forename: Michiko
authors.initials: M
authors.lastname: Mandai
authors.affiliation: Department of Ophthalmology and Visual Sciences Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Kitaoka
authors.affiliation: Laboratory for Retinal Regeneration, Center for Developmental Biology, RIKEN, Kobe, Japan.
authors.forename: Masayo
authors.initials: M
authors.lastname: Takahashi
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D002924:Ciliary Body / Q000166:cytology*; D004195:Disease Models, Animal; D004847:Epithelial Cells / Q000166:cytology*; D021621:Imaging, Three-Dimensional; D007150:Immunohistochemistry; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D051379:Mice; D008810:Mice, Inbred C57BL