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. 2020 Oct 27;8:585675. doi: 10.3389/fcell.2020.585675

FIGURE 2.

FIGURE 2

Optimization of hiPSC-derived RGCs differentiation. (A) Schematic diagram illustrating the protocol for RGC differentiation from hiPSC-derived retinal organoids. (B) Phase-contrast micrograph of retinal cells 1 week after plating of dissociated cells derived from D56 organoids; scale bar, 50 μm. (C) Immunostaining on dissociated cells derived from D56 retinal organoids, showing the co-expression of BRN3A and THY1, or PAX6 with RBPMS, or βIII-tubulin allowing the identification of RGCs 1 week after plating. Photoreceptors are also identified according to CRX and recoverin (RCVRN) expression. Nuclei staining with DAPI in blue; scale bars, 50 μm. (D) RT-qPCR analysis of THY1, BRN3A, and RBPMS in D56 organoids and in retinal cells 1 week after plating (mean ± SEM; N = 3 differentiations per time point; n ≥ 10 organoids/differentiations). Gene expression at each time point is indicated relative to D56 organoids. *p < 0.05, Mann–Whitney test).