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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 and supplemented with 1 mg/ml G-418, 90%; heat inactivated fetal bovine serum, 10%.
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Subculturing
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Subcultivation Ratio: A subcultivation ratio of 1:6 to 1:10 is recommended Medium Renewal: Every 2 to 3 days 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.
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Cryopreservation
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Culture medium, 95%; DMSO, 5%
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