Media and Additives

The medium is not only a food source but also represents the environment in which the cells live. Medium selection for the screen is dependent on the cell type and on the assay. The medium composition must support the desired cell function needed, and provide an optimal energy source as well as all important nutrients such as vitamins, growth factors, minerals, and essential lipids that the cells need [29, 30]. A media for cell production can differ from one used for plate generation, as the latter often contains antibiotics and HEPES. The optimal amount of fetal calf serum (FCS) and ingredients such as glucose and glutamine can be different for the production phase compared to the plating medium. For fluorescence read-outs, as well as some reporter assays, the presence of phenol red in the medium enhances the background. All medium components such as FCS, growth factors, hormones, vitamins, ions, and nucleotides should be evaluated for possible interference with the assay signal. FCS contains different amounts of stress-protecting factors such as vitamin E and ATP, as well as calcium, which modulates cell function and influences cell adhesion, proliferation [31], and differentiation [32].

Charges of media and additives must be checked for functionality for each assay, and should not be changed during HTS and development.

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