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Complete Growth Medium
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HITES medium supplemented with 5% fetal bovine serum
The base medium for this cell line is ATCC-formulated DMEM:F12 Medium Catalog No.30-2006. To make the complete growth medium,add the following components to the base medium- 0.005 mg/ml Insulin
- 0.01 mg/ml Transferrin
- 30nM Sodium selenite (final conc.)
- 10 nM Hydrocortisone (final conc.)
- 10 nM beta-estradiol (final conc.)
- extra 2mM L-glutamine (for final conc. of 4.5 mM)
- 5% fetal bovine serum (final conc.)
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Subculturing
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Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:6 is recommended Medium Renewal: Twice per week 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 37C) 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 with an additional 5% fetal bovine serum, 95%; DMSO, 5%
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