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Complete Growth Medium
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The base medium for this cell line is ATCC-formulated RPMI-1640 Medium, Catalog No. 30-2001. To make the complete growth medium, add the following components to the base medium: horse serum to a final concentration of 10%.
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Subculturing
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Remove medium, rinse cell sheet twice with 0.25% trypsin, 0.02% EDTA in Hanks' BSS and incubate at 37°C with the trypsin, EDTA for 10 to 15 minutes. Remove trypsin, EDTA, disperse cells in fresh medium, centrifuge, resuspend cells in medium and dispense into new flasks.
Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:8 is recommended
Medium Renewal: Every 2 to 3 days
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Cryopreservation
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Freeze medium: complete growth medium, 95%; DMSO, 5% Storage temperature: liquid nitrogen vapor phase
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