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Complete Growth Medium
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A 1:1 mixture of Dulbecco's modified Eagle's medium and Ham's F12 medium containing 1.2 g/L sodium bicarbonate, 2.5 mM L-glutamine, 15 mM HEPES and 0.5 mM sodium pyruvate supplemented with 400 ng/ml hydrocortisone, 90%; fetal bovine serum, 10%
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Subculturing
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Remove medium, and rinse with 0.25% trypsin, 0.03% EDTA solution. Remove the solution and add an additional 1 to 2 mL of trypsin-EDTA solution. Allow the flask to sit at room temperature (or at 37°C) until the cells detach. Add fresh culture medium, aspirate and dispense into new culture flasks. Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:6 is recommended Medium Renewal: Every 2 to 3 days
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Cryopreservation
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culture medium 95%; DMSO, 5%
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