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

Figure 2

Induction of U-hiPSCs into retinal fates. (a) Schematic overview of retinal differentiation protocol from U-hiPSCs. (b) U-hiPSCs were dissociated into small clumps and formed EBs under suspension culture on D7. Scale bar = 250 μm. (c) Cells spread out from EBs on adherent surface on D11. Scale bar = 250 μm. (d–h) Immunofluorescence staining revealed that differentiated cells expressed markers specific for anterior neural plate and eye field cells, SOX1 (d), PAX6 (e), OTX2 (f), LHX2 (g), and SIX3 (h). Scale bars = 50 μm. (i) RT-PCR showed that differentiated cells expressed EF transcription factors PAX6, LHX2, RX, SIX3, and SIX6 on D14 with low expression of pluripotency marker OCT4.

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