Reprogramming of Adult Retinal Müller Glial Cells into Human-Induced Pluripotent Stem Cells as an Efficient Source of Retinal Cells

Figure 6

Generation of RPE cells from human MGC-derived iPSCs. (a) Phase-contrast images of RPE cells derived from iPSC-5f at passage 1 (P1), four weeks after picking. (b) ZO1 and MITF immunostaining of hiPSC-derived RPE cell monolayer four weeks after picking. (c, d) XZ views after orthogonal reconstruction of confocal stacks showing typical polarized expression of BEST1 (basal) and Ezrin (apical), four weeks after picking. Dash line mark out the apical and basolateral compartments according to ZO1 labeling. (e) qRT-PCR analysis of mature RPE markers in human iPSC-derived RPE cells at P1 and P2. Data are normalized to control RNA isolated from human adult RPE cells. (f) Evaluation of ratio of FITC/DAPI fluorescence in human iPSC-derived RPE cells at P1 and in control RPE-J cell line after 3 h incubation with FITC-labeled POS to determine RPE cell phagocytic activity; binding and uptake of POS were assayed as described Materials and Methods (scale bars: a, b, 50 μm; c, d, 5 μm).

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