Effect of different scaffolds on neural differentiation. (A) Neurite length of SH-SY5Y cells grown on different scaffolds, as measured by conventional quantitative microscopy and HCS. (B) Neurite length of SH-SY5Y cell line, as measured by HCS; (C) Neurite length of primary cortical neurons, as measured by HCS; (D) Neurite length of neurons derived from NSCs, as measured by HCS. In all cell systems, conventional 2D, semi3D-random and semi3D-aligned scaffolds having different coatings are compared. Data are expressed as mean ± SD, and represent the mean of three independent experiments. Statistical analysis: two-way ANOVA (SH-SY5Y F[9, 100] = 5.165; primary cortical neurons F[14,45] = 6.317; NSCs F[18,60] = 15.40) followed by Tukey's post hoc test. Asterisks represent the difference between the semi3D group versus the relative 2D control group within the same DIV (*p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001), letters indicate the difference between coated semi3D groups versus the relative uncoated semi3D control group (a = p < 0.05; b = p < 0.01; d = p < 0.0001). 2D, two-dimensional seeding; SD, standard deviation.