BDNF promotes cell proliferation, differentiation, and migration of cultured neurospheres.

Neurospheres were immunostained with Ki67, nestin, DCX and GFAP antibodies to determine the effects of recombinant BDNF (5 pg/ml) on neurogenesis of cultured neurospheres. The groups of neurospheres treated with 0.5% CSF (collected from rats on day 7 post SAH) and without treatment (control) were used for comparison. (A)-(I): photomicrograph showing the distribution of Ki67+ (A, D, and G) and nestin+ (B, E, and H) signals and as merged images (C, F, and I); (J)-(R): photomicrograph showing the distribution of DCX+ (J, M, and P) and GFAP+ (K, N, and Q) signals and as merged images (L, O and R). Scale bar = 50 μm. (S): Percentage of proliferating cells (Ki67+ of Hoechst+ cells) in the neurospheres. (T): Percentage of DCX+ area (DCX+ area/ total area). (U): Percentage of GFAP+ area (GFAP+ area/ total area). Means ± SD, 12 neurospheres were counted for each condition. ns: non-significant; ***, P < 0.001, one way ANOVA with Tukey's multiple comparisons test.