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MEFs cultured either with ambient or 3% O2. A. Viable cell count (VCC) per cm2 for MEFs cultured at 37°C with 5% CO2 and either ambient or 3% O2. B. Viability (%) for MEFs cultured at 37°C with 5% CO2 and either ambient or 3% O2. MEFs grown at 3% O2 maintained high viability and an average doubling rate of 45 h over 120 days of culture. The % viability and VCC per cm2 for MEFs grown at ambient O2 declined with each subsequent passage, and thus resulted in a negative doubling rate.
