Fig. 1

Generation of retinal organoids from human pluripotent stem cells. A Schematic showing the retinal organoid differentiation protocol. Human pluripotent stem cells (H9) were grown to confluence and pushed towards a neuroectoderm lineage with a 3–4 week induction period. Laminated structures (denoted in black triangles in the 3–4 week brightfield insets) were excised and matured in suspension culture. B Gene expression of H9-derived retinal organoids. Fold change was calculated by the ΔΔCt method, with GAPDH as the house keeping gene and normalized to undifferentiated H9 cells. Fold change is presented as mean ± SEM (n = 3 biological replicates using 7–10 pooled organoids per sample), analyzed by one way ANOVA with Tukey’s post-hoc. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001. C Brn3a⁺ retinal ganglion cells (green) in a week 10 retinal organoid. D Week 16 retinal organoid expressing CRX (pink) and NRL (green). E Week 25 retinal organoids expressing recoverin (red), NRL (green), and PKCα white. F Mature L/M cone photoreceptors (red) at week 18 and G S-cone photoreceptors (green) and rod photoreceptors (red) at week 20. Scale bar = 100 μm