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S. Yoshida, S. Shimmura, H. Okano, K. Tsubota; Differentiation of Induced Pluripotent Stem Cells Into Corneal Epithelial Cells. Invest. Ophthalmol. Vis. Sci. 2009;50(13):1780.
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Regenerative medicine using donor-derived cells and tissues, somatic stem cells, and ES cells include some problems such as rejection and ethical objection. Application of induced pluripotent stem (iPS) cells to regenerative medicine is expected to bypass these problems. To apply iPS cells to corneal epithelial disorders, we examined differentiation of iPS cells into corneal epithelial cells.
For differentiation of iPS cells into epithelial cells, dissociated cells were seeded onto mitomycin C-treated feeder cells. After several weeks of culture with FBS, in order to confirm the phenotype of epithelial cells had been induced, expression of cytokeratin 18 (K18) as a non-squamousepithelial marker, cytoleratin 14 (K14) and p63 as the stratified epithelial marker, and cytokeratin 12 (K12) as a corneal epithelial marker, were examined by immunohistochemistry.
Immunohistochemistry revealed K18-positive colonies and K14-positive both mouse and human iPS cells-derived colonies, which cultured on feeder cells for 2 weeks. In addition, K14-positive cells were p63-positive but not K18-positive cells. Furthermore, we found some K12-positive cells in K14-positive colonies. In the case of mouse iPS-derived cells, most colonies expressing K14 at high levels did not overlap with the colonies expressing K18 at a high level. In contrast, K14-positive cells from human iPS cells were dispersed among K18-positive colonies.
These results suggested that the culture method using mitomycin C-treated feeder cells is available to produce K12-positive corneal epithelial cells from iPS cells.
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