Figure 1 | Scientific Reports

Figure 1

From: Dual SMAD inhibition and Wnt inhibition enable efficient and reproducible differentiations of induced pluripotent stem cells into retinal ganglion cells

Figure 1

Differentiation of iPSCs into neural retina and retinal ganglion cells (RGCs). (A) Schematic showing iPSCs differentiation in neural retinal differentiation media (DMEM/F12 + 2% B27 + 1% N2 + 10 ng/mL IGF1), including inhibitors of BMP, WNT and TGF-β for 15 days, bFGF2 was included in media from day 4–14. At 16 days in vitro, cells were treated with RGC induction and maturation media. Differentiation using seven different conditions, L, X, LX, LSB, CHIR, LSB and LSBX are shown. (B) Characterization of RPCs using immunocytochemistry with SOX2, RAX and PAX6 along with secondary only control antibodies is shown. (C) FACS analysis demonstrated that 96.7% of RPC cells were positive for RAX, and 98.9% of cells were positive for LHX2, when compared to isotype controls. To normalize for flow cytometry cell counts data is represented as %Max. (D) Gene expression profiles showing RPC markers in three different iPSC lines, showing a high enrichment. Mean ± SEM represented from at least 3 independent experiments per individual.

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