Figure 1: Neural stem cell-derived oligodendrocytes. (A) Schematic representation of the culture protocol for NSC-derived OL culture generation. Following isolation, cells grow as suspension neurosphere cultures exposed to bFGF/EGF to keep them proliferating and preserving their multipotency. After the first splitting, the spheres are exposed to bFGF/PDGF-AA growth factor, to start driving lineage specification (oligospheres). After the second splitting, dissociated spheres are seeded on laminin-coated supports in the same medium. To start the differentiation process, after 3 days cells are exposed to the differentiation medium, containing thyroid hormone as the differentiation trigger. (B) The panel illustrates the changing morphology of the cells, from multipotent 3D structure (neurospheres) to the fully differentiated OL. Immunofluorescence reactions were performed as already described, using lineage-specific markers and epifluorescence microscope. Scale bar for neurosphere: 100 μm; for immunocytochemistry pictures: 20 μm. bFGF: Basic fibroblast growth factor; CNTF: ciliary neurotrophic factor; CNPase: 2′,3′-cyclic-nucleotide 3′-phosphodiesterase; DIV: day in vitro; EGF: epidermal growth factor; MBP: myelin basic protein; NAC: N-actetyl cystheine; NG2: neural/glial 2 chondroitin sulfate proteoglycan; NSCs: neural stem cells; OLs: oligodendrocytes; OPCs: oligodendrocyte precursor cells; PDGF: platelet derived growth factor; T3: triiodothyronine. Figure 1B is sourced from our unpublished data.