In recent years, researchers have reported the culture of a variety of cell lines in serum-free media that contains a supplement of nutrients, growth factors, and hormones in the place of serum. Mather and Sato (BBRC, 1985) reported the successful cultivation of Leydig 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 optimal for these studies was a 1:1 mixture of Dulbecco's Modified Eagle's Medium (DMEM) and Ham's F-12 Nutrient Mixture. This mixture known as DMEM/F12 medium contains 21 amino acids, 10 vitamins, glucose, iron, and zinc, among other components.
DMEM/F12 is a richer and more complex media than DMEM. HEPES buffer can be also included in the formulation at a final concentration of 15 mM to compensate for the loss of buffering capacity that is incurred by eliminating serum. This synthetic medium does not contain any serum, proteins, lipids, or growth factors and thus may require supplementation with fetal bovine serum (FBS) depending on the type of cell culture. Some commercial formulations may or may not include glutamine or phenol red.
The advanced basal media DMEM/F12 is often used in applications that require low-serum or serum-free media, such as 3D organoid culture or stem cell culture. DMEM/f12 media can also be used to help optimize cellular transfections that require low-serum or serum-free conditions.
In additional to stem cell culture, which often require a reduced serum or serum-free environment, primary cells can be cultured in DMEM/F12 media. This includes fibroblasts, MDCK cells, neuronal cell lines, glial cells, and smooth muscle cells, which exhibit equivalent or superior growth and function to primary cells that were supplemented with high concentrations of serum. Cells cultured with DMEM/F12 media also display the same cell morphology as cells cultured in high serum conditions.
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