DMEM/F-12

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Publication protocol

Neural Stem Cell (NSC) Cultures: Cortices of E12.5 or E13.5 mouse embryos were dissected in cold DMEM/F12. Each culture was derived from an independent embryo. Cells were dissociated with StemPro Accutase (Thermo Fisher Scientific A1110501) and plated at 50,000 cells/ml in petri dishes and grown in NSC maintenance medium composed of DMEM/F12 (Thermo Fisher Scientific 11320033), N2 (Thermo Fisher Scientific 17502048), B27 (Thermo Fisher Scientific 17504044), 20 ng/ml FGF2 (Miltenyi Biotec 130–093-841) and 20 ng/ml EGF (Miltenyi Biotec 130–093-825) at 37°C, 5% CO2. After 3 days, neurospheres were dissociated with StemPro Accutase into single cell suspensions and plated at 50,000 cells/ml in NSC maintenance medium either in petri dishes for neurosphere cultures or on poly-L-Lysine (30 μg/ml) and laminin (3 μg/ml) coated coverslips for adherent cultures. These cells are considered passage-1 NSCs.

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