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DMEM/F12 Medium is a 1:1 mixture of Dulbecco’s Modified Essential Medium (DMEM) and Ham’s F-12 Medium, commonly used in growing a variety of mammalian cultures. DMEM/F12 is rich and complex, containing all 21 amino acids, 10 vitamins, glucose, zinc, and iron among other components. Together, it combines the high amount of glucose, amino acids and vitamins of DMEM, and the diversified components of F-12. It is a synthetic medium containing no serum or proteins and thus may require supplementation depending on the type of cell culture. Some commercial formulations may or may not include glutamine or phenol red.

In recent years, researchers have reported the culture of a variety of cell lines in serum-free medium that contained, instead of serum, a supplement of nutrients, growth factors and hormones. For example, Mather and Sato reported the successful cultivation of Leydig cells and Sertoli cells in serum-free medium that contained insulin, transferrin, epidermal growth factor, leutinizing hormone or follicle stimulating hormone, somatomedin and growth hormone. Although the hormones and their concentrations are specific for the type of cell under study, the medium found to be most satisfactory for studies of this type was a 1:1 mixture of Dulbecco’s Modified Eagle’s Medium (DMEM) and Ham’s F-12 Nutrient Mixture. HEPES buffer is included in the formulation at a final concentration of 15 mM to compensate for the loss of buffering capacity incurred by eliminating serum.