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Complete Growth Medium
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Iscove's modified Dulbecco's medium with 4 mM L-glutamine adjusted to contain 1.5 g/L sodium bicarbonate, 85%; fetal bovine serum, 15%
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Subculturing
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Protocol: Cultures can be maintained by addition of fresh medium. Alternatively, cultures can be established by centrifugation with subsequent resuspension at 1 X 10(5) viable cells/ml. Do not allow the cell concentration to exceed 6 X 10(5) viable cells/ml.
Interval: Maintain cultures at a cell concentration between 5 X 10(4) and 6 X 10(5) cells/ml. Medium Renewal: Add fresh medium every 2 to 3 days (depending on cell density) Cultures can be maintained by the addition of fresh medium or replacement of medium. Alternatively, cultures can be established by centrifugation with subsequent resuspension at 1 X 10 exp5 viable cells/ml. Maintain cell density between 5 X 10 exp4 and 6 X 10 exp5 viable cells/ml.
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Cryopreservation
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Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO Storage temperature: liquid nitrogen vapor phase
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Culture Conditions
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Atmosphere: air, 95%; carbon dioxide (CO2), 5% Temperature: 37.0°C
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