Primary mouse OPCs were isolated from cortices of P4-P8 mouse brains. Cerebral cortexes were dissected and then dissociated by mechanical trituration until the cell suspension has no small clumps. Then dissociated mouse cortices cells were resuspended in panning buffer. Suspension was sequentially immunopanned on anti-GalC and anti-O4 antibody coated plates. The adherent OPCs were trypsinized and plated onto poly-D-lysine coated plates. The cultures were maintained under proliferating condition which is DMEM/F-12 (GIBCO, Cat#11330032) with addition of 2% B-27 (GIBCO, Cat#A3353501), 1% N2 (GIBCO, Cat#A1370701), 20 ng/ml PDGF-AA (Peprotech, Cat#100-13 A), 10 ng/ml CNTF (Peprotech, Cat#450-13), 20 ng/ml bFGF (Sino Biological, Cat#10014-HNAE), and 1 ng/ml NT3 (Peprotech, Cat#450-03). For OPCs differentiation, 60 nM L-triiodothyronine (T3; Sigma-Aldrich, Cat#T6397) was added to the media and PDGF-AA, bFGF and NT3 were removed. For iOLs, we harvest cells at 24 h after T3 induction. And for mOLs, we induce OPCs differentiation for 72 h. Rat OPCs were obtained from P2–P4 rat brains as mouse OPCs with slight modifications. Briefly, dissociated rat cortices cells were sequentially immunopanned on anti-GalC and anti-A2B5 antibody coated plates to harvest
