Figure 2.

Efficient differentiation of outer segment-bearing photoreceptors in 26-week hESC-derived retinal organoids

(A) Schematic of retinal differentiation protocol with bright field images of retinal organoids.

(B–D) Representative bright field images showing brush borders (white arrowheads).

(E–G) Semithin images of brush border region.

(H–J) Retinal neuroepithelium showing phalloidin positive OLM (pink), mitochondria-rich ISs (green), and elongated PRPH2+ OSs (red).

(K) RT-qPCR analysis showing the relative expression of RHO and ABCA4 in RDM90 and ALT media (±SEM; n = 15 pooled NRVs, N = 3 experiments).

(L) Histogram showing the ratio of OS-like to IS structures as a percentage (±SD; ∗∗p < 0.01, ∗∗∗∗p < 0.0001; n = 30 images, N = 3 experiments).

(M) Graph showing the length of segment structures for all conditions (±SD; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗∗p < 0.0001; n = 40 images, N = 4 experiments). Nuclei were stained with DAPI (blue).

Scale bars, 20 μm (E–G), 25 μm (H–J), 50 μm (B–D), 70 μm (A). OLM, outer limiting membrane; IS, inner segment; ONL, outer nuclear layer; OS, outer segment.