Figure 1 | Scientific Reports

Figure 1

From: Retinal Organoids derived from hiPSCs of an AIPL1-LCA Patient Maintain Cytoarchitecture despite Reduced levels of Mutant AIPL1

Figure 1

Generation of 3D AIPL1-LCA ROs from Patient hiPSCs. (A) Schematic of the differentiation protocol. (B) Phase contrast micrographs of differentiation stages: hiPSCs, floating aggregates of hiPSCs following treatment with dispase (W2), and aggregates plated on growth factor reduced (GFR) Matrigel-coated plates reach a typical morphology by week 4 (W4) are dissected manually and are grown in suspension after that (W4’). The typical transparent neuroepithelial domain (*) is formed (W12) with stratified appearance. Inset at larger magnification is shown (W12'). At W20 the projections at the surface begin to emerge (inset, arrowhead). Scale bars: 200 µm. (C) By week 23 (W23) the ROs reached 1–1.5 mm in diameter and displayed dense translucent projections at the apical edge (black arrowhead) that grow after that (W26). The double arrow shows the presumptive ONL. Abbreviations: 3D, three-dimensional; RO, retinal organoids; GFR, growth factor reduced; W, week; Tau, taurine; RA, retinoic acid; FBS, fetal bovine serum; ULA, ultra-low attachment plates; ONL, outer nuclear layer.

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