Complete Growth Medium
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A 4:1 mixture of Dulbecco's modified Eagle's medium and Medium 199, 89%; chicken embryo extract, 1%; horse serum, 10%
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Subculturing
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Subcultivation Ratio: An inoculation density of 4000 viable cells per sq. cm. of flask or dish surface area is recommended Medium Renewal: Twice per week Remove medium, rinse the cell sheet once with fresh trypsin (0.25%), EDTA (0.03%) solution, remove trypsin and allow the flask to sit at room temperature (or at 37C) until the cells detach (about 10 minutes). Add fresh medium, aspirate and dispense into new flasks at a density of 2000 cells/sq. cm.
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Cryopreservation
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Culture medium, 95%; DMSO, 5%
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