Figure 1 | Scientific Reports

Figure 1

From: Lentiviral mediated RPE65 gene transfer in healthy hiPSCs-derived retinal pigment epithelial cells markedly increased RPE65 mRNA, but modestly protein level

Figure 1

RPE differentiation protocol timeline. (a) Schematic depiction of the differentiation protocol highlighting the time course and media used. The protocol starts with hiPSCs in feeder-free condition that are lifted in clumps using collagenase to form (b) embryoid body-like aggregates (EB) in floating condition in an embryoid body media (EBM). EBs are plated on day 1 on Matrigel Reduced Factor (MRF) in a neural induction media (NIM) and (c) expand. The cells are switched to a retinal differentiation medium with vitamin A (RDMcA) from day 10 to 13, then without vitamin A (RDMsA). (d,e) Pigmented foci (PFs) appear around day 18 to 24 and are collected on day 30. Optic vesicle-like structures can also be observed (e, arrowheads). (f,g) Plated PFs develop pigmented patches (PPs) that are collected, dissociated and seeded to obtain (h) the pure hiPSC-derived RPE (iRPE) cell monolayer.

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