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Complete Growth Medium
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Minimum essential medium (Eagle) in Earle's BSS with non-essential amino acids, sodium pyruvate (1 mM) and 0.1% lactalbumin hydrolysate, 90%; fetal bovine serum, 10%. Maintain pH at 7.3 to 7.4
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Subculturing
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Protocol: Remove medium, and rinse with 0.25% trypsin, 0.53 mM 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. Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:4 is recommended Medium Renewal: Every 2 to 3 days
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Cryopreservation
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Freeze medium: Complete medium 95%; DMSO 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: 37.0°C
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