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Complete Growth Medium
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RPMI 1640 medium with 2 mM L-glutamine adjusted to contain 1.5 g/L sodium bicarbonate, 4.5 g/L glucose, 10 mM HEPES, and 1.0 mM sodium pyruvate, 80%; heat-inactivated fetal bovine serum, 20%
RPMI 1640 medium with 2 mM L-glutamine adjusted to contain 1.5 g/L sodium bicarbonate, 4.5 g/L glucose, 10 mM HEPES, and 1.0 mM sodium pyruvate, 80%; heat-inactivated fetal bovine serum, 20%
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Subculturing
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Protocol: 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 to 2 X 10(5) viable cells/ml. Interval: Maintain cell density between 1 X 10(5) and 1 X 10(6) viable cells/ml. Medium Renewal: Add fresh medium every 2 to 3 days (depending on cell density)
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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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