Neuronal cell differentiation of C17.2 cells cultured in 250 µm microchannels with 12, 24 and 48 hour feeding intervals.

<p>(A) The cell morphological change over 3 weeks. To better demonstrate the positive β-tubulin-III staining in the 24 hour and 48 hour samples, overlays of the β-tubulin-III and nuclei staining were shown in the inset from the boxed areas. In comparison, insets of β-tubulin-III and nuclei staining were also shown for the 12 hour samples, but β-tubulin-III staining was not visible. Red: Nestin. Green: β-tubulin-III. Blue: cell nuclei. Scale bar = 50 µm. The insets were from the boxed region and were at the same scale as the main images. (B) The β-tubulin-III positive cell counts per mm² over time. (C) The neuronal cell counts per mm² over time. (D) The average neurite length per mm² over time. In Figures 4B-4D, the control (with “c”) was C17.2 NSCs seeded at the same surface density in FluoroDishes but without microchannels and fed every 48 hours as in standard subculture protocols. Data were shown as mean ± standard deviation. The * above the bars indicated a statistical difference between the sample and the control by two-tailed Student’s t test (<i>p</i><0.05). The * below the bars indicated a statistical difference in the group by one way ANOVA (<i>p</i><0.05). N≥15.