fig1

Figure 1. Overview of retinal organoid differentiation protocols. (A) A timeline of each retinal organoid differentiation protocol is shown. The supplements used in each protocol are denoted above the timeline, the extrinsic factors are denoted below the timeline, and the timepoint of excision is indicated with a tungsten needle. “H” indicates the timepoint when the retinal organoids were harvested for the comparative analyses. (B) Each differentiation method successfully produced retinal organoids (shown on day 85). Retinal organoids from each method contained amacrine and ganglion cells (SNCG-positive) and juvenile photoreceptor cells (RCVRN-positive, CRX-positive). Composite images were counterstained with DAPI. IWR-1e: Inhibitor of Wnt response compound-1-endo; SAG: smoothened agonist; DAPT: N-[N-(3,5-difluorophenacetyl-L-alanyl)]-S-phenylglycine t-butyl ester; RA: all-trans retinoic acid; H: harvest; BMP4: bone morphogenic protein 4; SNCG: synuclein gamma; RCVRN: recoverin; CRX: cone-rod homeobox. Black scale bar: 100 µm; white scale bar: 50 µm.