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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 supplemented with 0.05 mM 2-mercaptoethanol and 100 U/ml recombinant human IL-6, 90%; fetal bovine serum, 10%.
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Subculturing
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Medium Renewal: Every 2 to 3 days Cultures can be maintained by addition or replacement of fresh medium. Alternately, cultures can be established by centrifugation with subsequent resuspension in fresh culture medium. Establish new cultures at 2 X 10 exp5 viable cells/ml and maintain at between 1 X 10 exp5 and 1 X 10 exp6 viable cells/ml.
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Cryopreservation
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Culture medium, 95%; DMSO, 5%
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