Complete Growth Medium
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Dulbecco's modified Eagle's medium with 4 mM L-glutamine adjusted to contain 1.5 g/L sodium bicarbonate and 4.5 g/L glucose supplemented with 0.25 mg/ml G-418, 90%; fetal bovine serum, 10%
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Subculturing
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Protocol: 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 37C) until the cells detach. Add fresh culture medium, aspirate and dispense into new culture flasks. Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:10 is recommended Medium Renewal: Twice per week
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Cryopreservation
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Freeze medium: Complete growth medium, 95%; DMSO, 5% Storage temperature: liquid nitrogen vapor temperature
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