Mouse pdgf aa

OPCs were prepared from whole brains of newborn (P1-P3) ADAMTS4 -/-KO or C57Bl/6J wild-type mice. Meninges were removed, brains were disintegrated with a scalpel, and pooled from animals with the same genotype. Brain tissue was further dissociated with the Neural Tissue Dissociation Kit, and A2B5 + glial-restricted progenitors were isolated by labeling with anti-A2B5 microbeads and magnetic enrichment using MS columns and a MiniMACS Separator according to the manufacturer's instructions (Miltenyi Biotec). 1.5 × 10 6 progenitor cells were seeded on poly-L-lysine coated 10 cm dishes and cultured in proliferation medium (DMEM/F12 without glutamine, 1× B-27 supplement without vitamin A, 1× GlutaMAX, 100 U/ml penicillin/streptomycin, 20 ng/ml each of mouse FGF-b and mouse PDGF-AA, all media components from ThermoFisher Scientific, growth factors from ImmunoTools). Confluency was kept below 50% and cells were passaged approximately every 4 days after dissociation of the cell monolayer with accutase (Sigma-Aldrich). For differentiation of OPCs, cells were switched to differentiation medium (same media components, except 1× B-27 with vitamin A, and without growth factors but instead supplemented with 40 ng/ml each of L-thyroxine (T 4 ) and triiodo-L-thyronine (T 3 ), Sigma-Aldrich) for 1-5 days.