Figure 6

NSCs on ONAS show a distinct proliferation pattern when cultured at clonal density (3.3 cells μl⁻¹). (A) Time-lapse digital images of NSCs grown in the presence of mitogens on either the flat surface (a–f, a’–f’) or ONAS (g–l, g’–l’) at 3.3 cells μl⁻¹. Boxed area in a-l is enlarged in a’–l’. The each scale bar is 50 μm in a–l and 25 μm in a’–l’. (B) Time-lapse digital images of NSCs cultured in the presence of EGF and FGF2 on flat surface or ONAS at 6.7 cells μl⁻¹. Scale bar, 50 μm. (C) NSCs grown in the presence of mitogens for 4 days were fixed and stained with phalloidin (green) and anti-vinculin antibody (red). Nuclei were stained with DAPI (blue). Boxed areas in a and b were enlarged in a’ and b’. Arrows and arrowheads in a’ indicate lamellipodia and filopodia, respectively. Scale bar, 25 μm (in a and b) and 10 μm (in a’ and b’). (D) Field emission scanning microscopy photos of NSCs cultured on Flat or ONAS. NSCs grown in the presence of mitogens for 4 days were fixed and photos were taken. Boxed areas (a and b) were enlarged in a’ and b’. Arrows and arrowheads in a’ indicate lamellipodia and filopodia, respectively. Scale bar, 10 μm (in a and b) and 2.5 μm (in a’ and b’).