Complete Growth Medium
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Alpha minimum essential medium with 0.001 mM methotrexate, 90%; dialyzed heat-inactivated fetal bovine serum, 10%
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Subculturing
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Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:8 is recommended Medium Renewal: Add fresh medium at the time of subculture Rinse cell sheet 2 times with fresh 0.25% trypsin, remove trypsin and allow the culture to stand for 5 to 10 minutes at 37C. Add fresh medium, aspirate and dispense into new flasks.
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