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After the centrifuge, cells were resuspended in 20 mL proliferation medium containing Neural Stem Cell Basal Medium (SCM003, Merck Millipore), B27 without vitamin A (12587010, Thermo Fisher Scientific, Massachusetts, USA), GlutaMAX (35050061, Thermo Fisher Scientific), antibiotics (100 IU/mL penicillin and 100 g/mL streptomycin), 1 mg/mL epidermal growth factor (E9644, Sigma-Aldrich), and 1 mg/mL fibroblast growth factor 2 (100-18B, PeproTech, NJ, USA), and incubated at 37 C with 5% CO 2

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