Since Harry Eagle's first reports, studies have been underway to determine the media requirements for individual cell types. The medium found to be most satisfactory for these studies as a 1:1 mixture of Dulbecco's Modified Eagle's Medium (DMEM) and Ham's F-12 Nutrient Mixture. To help compensate for the buffering capacity lost with the removal of serum, HEPES can be added in some modifications at a final concentration of 15 mM
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