Mouse OPC Generation from EpiSCs

Mouse protocols were approved by Case Western Reserve University School of Medicine’s Institutional Animal Care and Use Committee. Mouse (Mus musculus) OPCs were generated from epiblast stem cells (EpiSCs) as in ref. 50 (link). In brief, EpiSCs were isolated from 129 S/SvEv male embryos (E3.5; The Jackson Laboratory) and pushed to form neural rosettes^{51 (link)}. Neural rosettes were then passaged into nunclon plates coated with poly-L-ornithine (PO) and laminin, in OPC growth media consisting of DMEM/F12 supplemented with N2 Max (R&D Systems, AR009), B27 (Thermo Fisher, 12587010), 20 ng/mL bFGF (R&D Systems, 23-3FB-01M), and 20 ng/mL PDGFA (R&D Systems, 221-AA). Media was changed every 48 h and cultures were maintained in 37 °C with 5% CO₂.