Figure 1

2D/3D small molecule-based differentiation of hPSC into retinal organoids using animal-derived supplements (xeno). (A) Schematic of the 2D/3D xeno small molecule protocol (B) Bright-field microscopy of 3D optic vesicle-like structures at day 30 and manually picked organoid at day 60 of differentiation (MShef10; scale bars 100 µm) (C) Immunohistochemistry analysis of retinal organoids at week 9 and 17 of differentiation, (MShef4 and MShef10 ESC lines respectively). (D) RT PCR analysis of relative expression of retinal marker genes (mean ± SD; n = 3 independently cultured sets of 10 OVs from N = 1 ESC line, MShef4). (E) Immunohistochemistry analysis of retinal markers at later week 33 stage of differentiation (MShef4 ESC line). ONL, presumptive outer nuclear layer in 3D organoid.