Figure 1.

Human EGFR⁺ GM Cells Isolated by FACS Display Stem Cell Properties In Vitro

(A) Immunofluorescence in human GM tissue shows many EGFR⁺OLIG2⁺ (^∗∗), scattered EGFR⁺GFAP⁺OLIG2⁺ (^∗∗∗), and exclusively EGFR⁺ (^∗) cells, some of which show radial morphology (arrows) next to the developing ependyma (dashes) (see also Figures S1A–S1F and S2A).

(B) Representative FACS isolation of EGFR⁺/EGFR⁻ cells using EGF-APC for positive selection, and CD24/CD34/CD45-PE and DAPI for exclusion (GM, 21gw).

(C) Acute immunofluorescence of sorted GM EGFR^+/− cells (2 hr after FACS) shows predominant distribution of EGFR in the positive fraction (93%) (^∗∗p = 0.002), and comparable expression of SOX2 and Ki67 in both fractions (n = 3 independent experiments) (see also Figure S2G).

(D) Representative primary NS growth at 6 days.

(E and F) Quantification of primary NS growth (n = 12 independent experiments; ^∗∗∗p = 2.9 × 10⁻⁵) and (F) NS size (n = 5 independent experiments; ^∗∗p = 0.01) at 6 days (EGF + FGF).

(G) Under differentiating conditions, EGFR⁺-derived cells show tri-lineage differentiation toward astrocytic (GFAP⁺), oligodendroglial (O4⁺), and neuronal (TUJ1⁺) fates (representative example of three independent samples).

Scale bars, 50 μm. Magnification of NS images, 10×. Bar graphs show mean ± SEM.