Generation of Retinal Organoids with Mature Rods and Cones from Urine-Derived Human Induced Pluripotent Stem Cells

Figure 5

Generation of highly mature photoreceptors with rods and cones from U-hiPSCs without addition of RA. (a) An example image of retinal organoids with long and brush-like segments at apical side. Scale bar = 25 μm. (b–h) Representative images of immunofluorescence staining from UE017 U-hiPSCs showed photoreceptors at W21 differentiated into all subtypes, rhodopsin-positive rods, S opsin-positive blue cones, and L/M opsin-positive red/green cones (b–h). Scale bar = 50 μm. (b) An organoid with rich rod photoreceptors occupying the whole outer nuclear layer (ONL). (c–d) An organoid with a patch of S opsin-positive blue cones and rhodopsin + rods. (e) An organoid with rich cone photoreceptors occupying almost the whole ONL. (f–h) Higher magnification of the squared area in (e). (i–j) TEM revealed that U-hiPSC-derived photoreceptors developed typical ultrastructures, outer limiting membrane (arrow), inner segment (IS) with rich mitochondria (), connecting cilium (arrow), and basal body (bb). Scale bar = 1 μm. (k) Higher magnification of the squared area in (j). (l, m) U-hiPSC-derived photoreceptors on W25 expressed functional phototransduction proteins PDE6α and Gtα-1 distributed most in presumptive outer segments (OS). Scale bars = 25 μm.

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