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Qingjian Ou, Caixia Jin, Haibin Tian, Furong Gao, Weiye Li, Lixia Lu, Guo-Tong Xu; High-effect induction of human iPS cells into retinal pigment epithelial cells with small molecules. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):2355.
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© ARVO (1962-2015); The Authors (2016-present)
This study is aimed to establish a more effective method to induce the differentiation of iPS cells into retinal pigment epithelial (RPE) cells with small molecules in serum free medium, in order to facilitate the clinical application of iPS cells derived donor cells.
Human iPS (hiPS) cells were cultured on the matrigel-coated dish in serum free conditioned medium. The iPS cells were first treated with two small molecules, SB431542 and DMH1, for 7days (dual-Smad) and then with other groups of small molecules at the appropriate time. The medium was changed every two days and the cells were separated with accutase before 18 days once the cells were confluence. RPE-like cells derived from hiPS cells (hiPS-RPE) were collected after 24 days induction. The hiPS-RPE cells were confirmed by Q-PCR, immunohistochemistry and western blot for the expression of RPE related genes and proteins.
Combination of different small molecules can induce hiPS cells differentiate into RPE cells after 24 days, and more than 90 percent induced cells differentiate into RPE cells. The hiPS-RPE cells have the typical polygonal morphology and pigments. Meanwhile, they express the RPE cells specific markers.
We can induce hiPS Cells into RPE-like cells under chemically defined conditions with the combination of small molecules in about 3weeks.
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