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Serum-free media: standardizing cell culture systems

Michael Butler

The first attempts at culturing animal cells in vitro made use of biological fluids, such as serum or tissue extracts. It was in the 1950s that a scientific approach was adopted to determine the defined nutrients required for mammalian cell growth. The idea of a chemically defined media was pioneered by Eagle, who determined the minimum ingredients that were essential for the growth of a number of human cell lines. This led to the development of Eagle’s minimal essential media that consisted of 13 amino acids, 8 vitamins and 6 ionic species [1]. This formulation appeared to provide the requirements for the growth of a number of isolated cell lines if supplemented with animal-sourced serum. Higher cell densities were obtainable by increasing the component concentrations of Eagle’s minimal essential media and became established through basal formulations such as Dulbecco’s modification of Eagles medium (DMEM). Clonal cell growth of selected cell lines was obtained by enrichment with an enhanced range of nutritional components, largely through the early work of Ham to produce the well-known Ham’s F-12 medium, Sato had the ingenious idea of combining these two approaches to blend a basal media formulation – DMEM/F-12 – that has become widely used for the growth of multiple cell lines to high density [2]. However, despite the inclusion of up to 70 components in these well-defined basal media formulations, supplementation with dialyzed serum (~10%) is necessary to provide sustained growth of most cell lines.