Fig. 7

Therapeutic effects of hESC-derived RPE cells for the treatment of corneal endothelial dysfunction. a Corneal transparency of rabbits was measured by a slit lamp microscope at days 1, 3, 7, and 14 after cell injection. b OCT image-based central corneal thickness analysis. c Immunofluorescence staining images of α-SMA (green) and Vimentin (green) in the transplanted area 7 days after surgery. Nuclei were stained with DAPI (blue) (scale bar: 25 μm). d qRT-PCR analysis of CEC related genes, RPE related genes, and antioxidant genes between cultured hESC-derived RPE cells in vitro and transplanted hESC-derived RPE cells. Quantification represented the levels of relative mRNA expressions normalized to GAPDH. e ZO1 (green), ATP1A1 (green) and MITF (red) were stained in the central area of the cornea 14 days after surgery. The transplanted cells were stained with human specific antibody human nuclei (HuNu) (red). Nuclei were stained with DAPI (blue) (scale bar: 50 μm). f Slit-lamp photograph of a rabbit cornea injected with hESC-derived RPE cells at day 30 postoperatively. The white dotted circle indicates the range of transplanted cells with slight pigmentation. g Immunofluorescence staining images of ZO1 (green), ATP1A1 (green), and MITF (red) in the transplanted area 30 days after surgery. The transplanted cells were stained with human specific antibody human nuclei (HuNu) (red). Nuclei were stained with DAPI (blue) (scale bar: 50 μm). h Concentrations of VEGF and PEDF at day 30 after surgery. Data are mean ± SEM. In vivo experiments were performed using three independent animals per group. Significance (*P < 0.05, **P < 0.01, ns: nonsignificant) relative to control. hESC, human embryonic stem cell; RPE, retinal pigment epithelium; OCT, optical coherence tomography; hESC-RPE, hESC-derived RPE cells; qRT-PCR, quantitative real-time reverse transcription polymerase chain reaction; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; Sham, non-injected corneal endothelial damaged group. VEGF, vascular endothelial growth factor; PEDF, pigment epithelium-derived factor