Figure 4.

Effects of glial cells pretreated with glutamate or H₂O₂ on NSC neurospheres in a Transwell co-culture system.

(A) Phase-bright photomicrographs showing adherent neurospheres after NSCs were plated on uncoated wells in growth medium for 7 days with astrocytes, microglia or mixed glia with or without glutamate or H₂O₂ pretreatment. In NSC cultures, adherent neurospheres displayed a spherical shape. In NSC/microglia co-cultures, most adherent neurospheres displayed a spherical shape. In NSC/astrocyte and NSC/mixed glia co-cultures, adherent neurospheres showed irregular shapes and had a few of cells migrating away. Scale bar: 50 μm. (B–D) Quantification of the ratio of differentiated neurospheres in NSC co-cultures with microglia (B), astrocytes (C), and mixed glial cells (D). Data are expressed as the mean ± SEM from three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001 (one-way analysis of variance followed by a least significant difference test). Glu: Glutamate; NSC: neural stem cell.