Mild Hypoxia Enhances Proliferation and Multipotency of Human Neural Stem Cells
Figure 4
Mild hypoxia increases the percentage of IhNSC-derived neurons during in vitro differentiation.
IhNSC were differentiated onto an adhesive substrate in medium without mitogens and fixed for immunocytochemical analysis after 3, 10 and 17 days in vitro. (A) Quantification of the percentage of neurons (β-tubIII+) over the total nuclei (DAPI+) number. Values are means±S.E.M (N = 3). Neurons were significantly less represented at 1% O2 with respect to the other O2 concentrations at 10 and 17 days in vitro. The difference among all the values at the different oxygen concentrations was statistically significant (P<0.01) unless indicated (*P<0.05), n.s., = not significant; one-way ANOVA followed by the Student's t-test.(B) Quantification of the percentage of astrocytes (GFAP+) over the total nuclei (DAPI+) number (N = 3). Values are means±S.E.M. At 10 days in vitro the percentage of astrocytes generated in 1% O2 was significantly (*p<0.05) higher with respect to the other conditions (1% O2 vs 2.5% O2, p<0.01). All other values were not significantly different (n.s., = not significant); one-way ANOVA followed by the Student's t-test. (C) Immunocytochemistry of differentiated cells showing the morphology of β-tubIII+ (red) and GFAP+ (green) cells in 2.5% oxygen at 10 days in vitro. Scale bar, 10µm.