FIGURE 2.

Time-dependent effect of blood in VZ cell cultures. After 5 days of differentiation samples were analyzed at 3, 24, and 48 hours of control (Cont) or blood exposure. Controls exhibit normal differentiation from NSC to ependymal cells (A, C), with significant increases in the percentage of βIV-tubulin multiciliated cells (C). Blood exposure significantly reduces both the total number of cells (DAPI-positive cells/0.0225 mm²) (B) and the percentage of multiciliated cells at 24 hours and 48 hours (C). *p < 0.05; **p < 0.01; ***p < 0.001. (D) Double-labeling with active caspase 3 (green) and βIV-tubulin (red) showed no difference in the number of apoptotic cells. (E) Blood exposure-dependent proportional change of total cells versus ependymal cells (EC). EC decrease was twice as high after 24 hours and even greater after 48 hours than the cell loss, showing a significant discrepancy between total cell loss and the lack of EC. *p < 0.05; ***p < 0.001; Student t-test was used to analyze the total cells (B) and the blood-dependent variation (E); one-way ANOVA and a Tukey posthoc test were used to analyze the mean percentage of ciliated cells (C). Scale bars: 50 μm in all panels.