Differentiation of hAEC to progenitors of cortical neurons.

(A) Representative micrographs at 20x from hAEC differentiated to Nestin-positive cells (green) after different treatments; the nuclei were stained with DAPI (blue). When the hAEC were treated with SB431542 + Noggin + EGF + bFGF the proportion of Nestin-positive cells increased. (B) Percentage of Nestin-positive cells obtained from hAEC during the proliferation stage with different treatments. Results are expressed as percentages of means ± S.E.M. from 9 fields counted in duplicate from three independent experiments. (C) Representative micrographs of hAEC treated with SB431542 + Noggin + EGF + bFGF. On the 14th day, we found cells that were positive for OTX2 (red), TBR2 (green), NeuN (red), or β-III-tubulin (green); the nuclei were stained with DAPI (blue). Arrow indicate neurite prolongation. Scale bar 50 μm. * p < 0.05 as compared with basal medium (CM).