Figure 4.

In Vitro Self-Renewal Capacity of NSC-like Cells

(A) Photomicrographs of BrdU incorporation into astrocytes transduced with Oct4 in combination with Shh stimulation, which was analyzed to assess the proliferation of NSC-like cells derived from astrocytes at 3 and 7 days. There were significantly more BrdU⁺ cells in Oct4/Shh-induced samples at the indicated time points than in Oct4-induced and control samples. Notably, GFAP expression was also drastically decreased in Oct4/Shh-induced samples. (B) Quantification of BrdU-positive cells among cells subjected to the indicated treatments. All data are reported as the means ± SEM and are representative of three independent experiments with similar findings. **p < 0.01 and ***p < 0.001 versus the corresponding controls. (C) Astrocytes treated under the indicated conditions for 3 and 7 days were analyzed by flow cytometry. G0/G1, G2/M, and S represent different cell phases, and the corresponding data are shown in each panel. (D) Statistical analysis of the percentages of cells in the three phases showed a significant difference between the Oct4/Shh group and the other treatments. The percentage of cell in each phase was calculated by using the cell ModiFIT program. All data are reported as the means ± SEM. *p < 0.05, **p < 0.01, and ***p < 0.001 versus the relevant controls. Scale bars, 100 μm.