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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:
- 10 Units/ml human recombinant insulin
- fetal bovine serum to a final concentration of 15%
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Subculturing
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Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:4 is recommended
Medium Renewal: Every 2 to 3 days
Remove medium, and rinse with 0.25% trypsin, 0.03% EDTA solution. Remove the solution and add an additional 1 to 2 mL of trypsin-EDTA solution. Allow the flask to sit at room temperature (or at 37°C) until the cells detach.
Add fresh culture medium, aspirate and dispense into new culture flasks.
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Cryopreservation
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culture medium 95%; DMSO, 5%
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