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The cells were cultured in Dulbeccos Modified Eagles Medium/Nutrient Mixture F-12 (DMEM/F12 = 1:1) supplemented with 5% (v/v) fetal bovine serum, 1 mM pyruvate, 50 g/mL penicillin, 50 g/mL streptomycin, 10 g/mL insulin, 5.5 g/mL transferrin, 5 ng/mL sodium selenite, and 5 ng/mL epidermal growth factor and maintained at 37 C in a sterile, humidified atmosphere of 5% CO 2