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Satoru Yoshida, Hideyuki Miyashita, Miyuki Yasuda, Emi Inagaki, Kazuo Tsubota, Hideyuki Okano, Shigeto Shimmura; Differentiation Of Induced Pluripotent Stem Cells-Derived Neural Crest Cells Into Corneal Keratocytes In Vivo. Invest. Ophthalmol. Vis. Sci. 2013;54(15):4700.
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© ARVO (1962-2015); The Authors (2016-present)
For application of induced pluripotent stem cells (iPSCs) to regenerativemedicine of corneal stroma, we show the ability of iPSCs-derived neural crest cells (NCCs) to differentiate into corneal keratocytes in vivo.
For efficient differentiation of IPSCs into NCCs, we examined modification of NSB method (Lee et al, 2010). To examine the ability of iPS-NCCs to differentiate into corneal keratocytes, the cells were injected into mouse corneal stroma. The injected cells were observed by confocal fluorescence microscopy. Cell morphology was observed by phalloidin staining and the expression of Keratocan, a corneal stroma specific keratan sulfate proteoglycan, was examined by immnohistochmistry using anti-ketatocan antibodies.
By induction with modified-NSB method, over 60% of cells were isolated as NCCs using FACS. Four weeks after intrastromal injection of iPS-NCCs, engraftment of the injected cells was observed. Phalloidin staining revealed dendritic morphology, which is one of the characteristics of keratocytes. In addition, immnohistochmical analysis revealed the expression of Keratocan in the engrafted cells.
iPS-NCCs is useful cell source for regenerative medicine of the corneal stroma by cell transplantation.
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