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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- Remove medium, rinse with 0.25% (w/v) trypsin - 0.53 mM EDTA solution
- Add 1 to 2 mL additional trypsin solution and allow flasks to remain at room temperature (or incubate at 37°C) until cells detach.
- Resuspend pellet in fresh medium and dispense into a culture flask.
Subcultivation Ratio: A ratio of 1:2 to 1:3 is recommended
Medium Renewal: Twice weekly
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Cryopreservation
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Freeze medium: Culture medium, 90%; DMSO, 5%; FBS ,5% Storage temperature: liquid nitrogen vapor phase
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Culture Conditions
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Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C
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