FIG. 5.

NSC characteristics of FSC/SSC^high and FSC/SSC^low cells. (A) Size categorization of neurospheres derived from the FSC/SSC^high and FSC/SSC^low cells. Neurospheres below 50 μm, 50–100 μm, and above 100 μm in diameter were counted (mean±SD; n=4; **P≤0.01). (B) Secondary NFA done at low density (1000 cells/mL) for the FSC/SSC^high, FSC/SSC^low, and unsorted population (mean±SD; n=4; **P≤0.01, ***P≤0.001). (C) Scoring of the number of unipotent, bipotent, and tripotent neurospheres after single neurosphere differentiation (mean±SD; n=4; *P≤0.05). (D) Immunostaining of a single multipotent neurosphere: O4 (green), βIII-tubulin (red), GFAP (gray). Scale bar=50 μm. Image was taken with a 20× objective. (E) NSC percentage calculated by multiplying NFU at clonal density and percentage of tripotent neurospheres (mean±SD; n=4; **P≤0.01). Passage 2 cells were used for the experiments. NSC, neural stem cell; NS, neurospheres; GFAP, glial fibrillary acidic protein.