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Complete Growth Medium
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Dulbecco's modified Eagle's medium, 45%; Ham's F12 medium, 45%; fetal bovine serum, 10%; plus the following additives: 10 ng/ml epidermal growth factor; 0.005 mg/ml insulin; 5 ng/ml selenium; 0.005 mg/ml transferrin
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Subculturing
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Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:3 is recommended Medium Renewal: Every 2 to 3 days Remove spent medium, add fresh 0.25% trypsin, 0.03% EDTA solution, rinse and remove trypsin. Allow the flask to sit at room temperature, or incubate at 37C, until the cells detach (about 5 to 10 minutes). Add fresh medium, aspirate and dispense into new flasks.
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