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Complete Growth Medium
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A 1:1 mixture of Waymouth's MB 752/1 medium and McCoy's 5a medium, 90%; fetal bovine serum, 10%
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Subculturing
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Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:6 is recommended Medium Renewal: Every 2 to 3 days These cells grow as a mixture of floating and adherent cells. Remove the medium with the floating cells, and recover the cells by centrifugation. Rinse the adherent cells with fresh 0.25% trypsin, 0.03% EDTA solution, add an additional 1 to 2 ml of trypsin solution, and let the culture sit at room temperature (or at 37C) until the cells detach. Add fresh medium, aspirate, combine with the floating cells recovered above and dispense into new flasks.
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Cryopreservation
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Culture medium, 95%; DMSO, 5%
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