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Invest Ophthalmol Vis Sci. 2016 Jan 1;57(1):153-61. doi: 10.1167/iovs.15-17610.

Induction of Functional 3D Ciliary Epithelium-Like Structure From Mouse Induced Pluripotent Stem Cells.

Author information

1
Department of Ophthalmology and Visual Sciences Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki, Japan.
2
Laboratory for Retinal Regeneration, Center for Developmental Biology, RIKEN, Kobe, Japan.

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 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.

PMID:
26780319
DOI:
10.1167/iovs.15-17610
[Indexed for MEDLINE]

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