Fig. 5.

In vitro BC derivative differentiation into oligodendrocytes. (A–I) BC progeny were grown in EGF/FGF2 medium and plated for 1 d in EGF/FGF2 alone (A–C), followed by sequential Noggin (3 d)/Purmorphamine (1 d) (D–F) or sequential Noggin (3 d)/Purmorphamine (1 d) media followed by glial differentiation medium (10 d) (G–I). (A, D, and G) Phase illustrations of morphological changes after each treatment. (B, C, E, F, H, and I) Immunocytochemistry for oligodendroglial cell stage-specific markers combined with Hoechst staining illustrating the presence of Olig2⁺/nuclear Nkx2.2⁻ cells (B) and the absence of GalC- or O4-expressing cells (C) in EGF/FGF2 medium, acquisition of nuclear Nkx2.2 positivity in Olig2⁺ cells (E) but the absence of O4⁺/GalC⁺ cells (F) after sequential Noggin/Purmorphamine treatment, and induction of CNPase⁺/YFP⁺ (H) and O4⁺/GalC⁺ cells (I) when cells were treated with sequential Noggin/Purmorphamine media followed by glial differentiation medium. (J) RT-PCR on FACS-sorted BC derivatives harvested at each step of the sequential treatment. Arrows point to the magnified cells in the inset.