Figure 8.

RPE-OTX2 Cells Secrete Neurotrophic Factors that Promote Cone Rescue

(A) Cone photoreceptor survival by supernatant from AAV-GFP- and OTX2-transduced pig primary RPE cells. CDM represents conditioned non-cell incubated medium. Mean with SEM (n = 6, Holm-Sidak multiple comparison ANOVA). (B) Morphology of the cone cells after incubation with supernatant from RPE-GFP, OTX2, and CDM. Cells are labeled with the viability dye Calcein-AM. (C) Relative qRT-PCR expression of CNTF and BDNF in AAV-OTX2-transduced cells as indicated. (D) Western blot of RPE cells transduced with recombinant AAV vectors as indicated. Pig CNTF was used as positive control. (E) Absence of protection of cones from the cone-enriched cultures by purified CNTF. (F) Quantification of Otx2 and Cntf mRNA in the RPE of the RCS rat at P15. (G) Increase in the expression of CRX mRNA in the eye of RCS rat transplanted by OTX2-modified pig RPE cells. Data were normalized to GAPDH. Mean with SD (n = 3, ANOVA test).