Fig. 4.

Neuronal differentiation of NS12ST cells. a Scheme showing the neural differentiation protocol applied to fetus-derived NS cells. Growth factors were withdrawn, while BDNF concentration was augmented. b After 23 days of differentiation in vitro, NS12ST cells gave rise to βIII-tubulin-, MAP2-, and GFAP-positive cells. GABA- and GAP-43-positive cells were present (×4, magnified after acquisition). c At the end of differentiation, 47.4 ± 4.6% of cells were immunopositive for βIII-tubulin, 47.0 ± 4.0% for MAP2, and 15.2 ± 2.4% for GFAP, while there were no O4-positive cells (n = 1,600 cells for βIII-tubulin; n = 1,163 cells for MAP2 and GFAP; n = 1,585 cells for O4) (columns represent averages, error bars standard deviations). d Gene expression analysis by RT-PCR on NS12ST cells in proliferation (P) and after 15 days of differentiation in the absence (D15-M) or in the presence (D15+M) of morphogens. Mouse fetal brain was used as positive control (+CTRL); -RT was the negative control