Figure 2
Effect of NGF supplementation on axonal growth of hESC- and hiPSC-derived RGCs. (a) Axonal growth of hESC- and hiPSC-derived RGCs was investigated by immunostaining of neurofilament protein L (NFL). Axonal growth of hESC- and hiPSC-derived RGCs is promoted by 50 and 200 ng/ml NGF supplementation from D24-30 (centre and right panels, respectively). The assessment from D24 is performed in retinal maturation medium (RMM). From D24 to D27, the embryoid bodies (EBs) are cultured in floating state and the EBs are attached to the dishes on D27, following 3 days’ adhesion culture. (b) Real-time PCR analysis of mRNA expression of axonal markers, TAU, NFL, and TUJ1 (n = 5). Expression of NFL is significantly upregulated at all concentrations of NGF supplementation (ANOVA, Dunnett’s T3 test, p ≤ 0.05). TAU expression is significantly increased only for 200 ng/ml NGF supplementation (ANOVA, Dunnett’s T3 test, p ≤ 0.05). No significant increase of TUJ1 expression is detected for all concentrations of NGF supplementation. There is no different reaction between hESC- and hiPSC-derived RGCs. Scale bar, 100 μm. Error bars indicate ± standard deviation (SD). Each column shows an average value for the studied samples. The sample size for all mRNA data is five (n = 5). NS, not significant.
