Fig. 6.

Differentiated EGFP-rESC-RPC2 integrate with host retina and preserve the retinal structure 4w after transplantation. a-d Differentiation of rESC-RPC2 into various retinal cells. Antibody against EGFP (green) and antibodies against retinal cell markers (red) were used for immunofluorescence staining. DAPI (blue) was used to highlight the nuclei. GCL ganglion cell layer, INL inner nuclear layer, ONL outer nuclear layer. Scale bars: 50 μm. e and f Integration of EGFP-rESC-RPC2 with retinal neurons. Colocalization of presynaptic markers Bassoon and Synaptophysin with EGFP expressed by donor cells in OPL was confirmed with immunostaining. GCL ganglion cell layer, INL inner nuclear layer, ONL outer nuclear layer. Scale bars: 50 μm. e' and f'. The magnified images of the rectangles in e and f. Arrow heads indicate the colocalization of EGFP and Bassoon or Synaptophysin. GCL ganglion cell layer, INL inner nuclear layer, OPL outer plexiform layer, ONL outer nuclear layer. g Preservation of the retinal structure by EGFP-rESC-RPC2 transplantation. The thickness of ONL of rESC-RPC2 treated eye was compared with that of vehicle (PBS) injected retina. Data are shown as mean ± SEM, t test, *P < 0.05 and **P < 0.01, n = 6 (rats) for each group. EGFP enhanced green fluorescent protein, rESC rat embryonic stem cell, RPC retinal progenitor cell, DAPI 4′,6-diamidino-2-phenylindole