FIG. 2.

SGN-NSCs differentiated into glutamatergic neurons. (A) Diagram showed ScN differentiation in adherent culture: BDNF was added to differentiation medium on day 0 whereas control (CTRL) group only used the differentiation medium. Cells were maintained for the same time period for 3–5 days in both groups and then fixed and proceeded immunostaining. ScNs expressed neural stem cells markers Sox2 and nestin whereas nestin expression was decreased when ScNs differentiated. (B) Cells derived from SGN spheres expressed neuron markers βIII-tubulin (TUJI), NF, NeuroD (NeuD), NeuN, and glutamatergic neuron marker VGLUT1. (C) Passage 3 SGN spheres were cultured in adherent cultures for 3 days. TUJ1-positive cells were found in the absence (upper panels, CTRL) and presence of BDNF (lower panels, BDNF). (D) In quantitative analysis, ScNs treated with BDNF had significantly more TUJ1-positive cells than the control group (**P < 0.01; Student's t-test). (E) Quantitative analysis indicated no significant differences on VGLUT1-positive neurons between the control and BDNF-treated groups (n = 6, P > 0.05, Mann–Whitney U test). Scale bar: 50 μm in (A), 20 μm in (B); 50 μm in (C). BDNF, brain-derived neurotrophic factor; NF, neurofilament; ScNs, SGN-NSC-derived neurons. Color images available online at www.liebertpub.com/scd