Figure 4.

In vitro analysis of SC properties of purified Nestin-CFPnuc⁺ and Nestin-CFPnuc⁻ cells

(A–G) Ten days after in vitro expansion under subclonal conditions (<5 000 cells/mL) in EGF/FGF, FACS-purified adult CFPnuc⁺ spinal cord cells (SpCs) (A and B) or CFPnuc⁺ DRG cells (F and G) gave rise to CFP⁺ spheres.

(C–I) After growth factor removal and adhesion, spinal cord primary spheres differentiated into typical CNPase⁺ multi-branched oligodendrocytes (C), GFAP⁺ ramified astrocytes (D), and MAP2–MAP5⁺ process-extending neurons (E), whereas DRG primary spheres differentiated into GFAP⁺ bipolar spindle-shaped Schwann cells (H) and MAP2–MAP5⁺ process-extending neurons (I).

(J and K) Phase illumination shows that, after FACS purification, CFPnuc⁻ SpCs died over time (J), whereas CFPnuc− DRG cells generated adhering bipolar cells (K).

(L and M) After growth factor removal, CFPnuc− DRG primary spheres differentiated into αSMA⁺ flat fibroblasts, IBA1⁺ pancake-shaped microglia/macrophages (L), and GFAP⁺ spindle-shaped PNS glia but not into MAP2–MAP5⁺ neurons (M).

Scale bars: 20 μm (A–G, J, and K) and 100 μm (C–E, H, I, L, and M). See also Figure S4.