FIG.3.

Transplantation of ES cells into mouse retina explant cultures. (A) EBs were formed from CCE-E, CCE-RX/E, or CCE-CH/E cells, treated with RA, and cocultured on the surface of mouse retina explant cultures. Views of paraformaldehyde-fixed transverse frozen sections immunostained with anti-EGFP antibody (left and middle panels) are shown. Nuclei are visualized by DAPI staining (left and right panels). (B) Appearance of CCE-RX/E cells in the retina explant at 1 and 3 weeks. Views of paraformaldehyde-fixed transverse frozen sections immunostained with anti-EGFP antibody and incubated with DAPI. (C) Immunostaining of markers of various retinal subpopulations in frozen sections of mouse retinal explant cultures containing CCE-RX/E cells. The following antibodies were used: anti-glutamine synthetase (GS) (Müller glia cells), anti-NF160 (horizontal cells), anti-calbindin-D-28k (horizontal cells), anti-PKC (bipolar cells), anti-HU (ganglion and amacrine cells), and anti-Islet-1 (ganglion, amacrine, and bipolar cells). All samples except for the calbindin-D-28k sample were double stained with anti-EGFP antibody. (D) Views of the surface ganglion layer containing CCE-E or CCE-RX/E cells are shown.