Figure 3.

NPC phenotype is sensitive to EC culture (static or dynamic) and spatiotemporal placement. EC monolayers cultured on a transwell membrane were subjected to static (a, b, e, f) or dynamic (c, d, g, h) culture, while NPCs were cultured below at a distance of ~ 10 μm (a–d) or ~ 1000 μm (e–h) from the EC monolayer. NPCs were immunostained for type A neuroblasts (PSA-NCAM⁺—a, c, e, g: green) and transit amplifying C cells (MASH1⁺EGFR⁺—a, c, e, g: magenta and red) or for active type B cells (GFAP⁺EGFR⁺—b, d, f, h: magenta and red) and quiescent type B cells (GFAP⁺CD133⁺—b, d, f, h: magenta and green). PSA-NCAM⁺ (◂), MASH1⁺EGFR⁺ (*), and GFAP⁺EGFR⁺ (<) are indicated in their respective panels. (I) A significant increase in PSA-NCAM⁺ neuroblasts (* p < 0.05) and OLIG2⁺ transit amplifying C cells (+ p < 0.1) was observed near dynamic EC cultures, compared to cells cultured far from dynamically cultured ECs or at any distance from static ECs. No differences were detected in other NPC subpopulations. Cell types: PSA-NCAM⁺ (A), MASH1⁺EGFR⁺ (C_MASH1), OLIG2⁺ (C_OLIG2), GFAP⁺EGFR⁺ (Bact), GFAP⁺EGFR⁻. Data are represented as mean ± standard deviation. 50 μm scale bar.