SSBD:database

Detail of Fig4A_CE

Hirofumi Kinoshita, Kiyoshi Suzuma, Jun Kaneko, Michiko Mandai, Takashi Kitaoka, Masayo Takahashi (2016) Induction of Functional 3D Ciliary Epithelium-Like Structure From Mouse Induced Pluripotent Stem Cells., Investigative ophthalmology & visual science, Volume 57, Number 1, pp. 153-61

Published in 2016 Jan 1

• doi: 10.1167/iovs.15-17610
• pmid: 26780319
  

(Abstract) PURPOSE: 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 3beta (GSK-3beta) 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 beta-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-3beta 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.

• Animals
• Blotting, Western
• Cell Differentiation
• Cell Proliferation
• Cells, Cultured
• Ciliary Body/cytology[major]
• Disease Models, Animal
• Epithelial Cells/cytology[major]
• Imaging, Three-Dimensional[major]
• Immunohistochemistry
• Induced Pluripotent Stem Cells/cytology[major]
• Mice
• Mice, Inbred C57BL