Growth Media for mammalian cell culture:

The media used for animal cell culture is a complex mixture of Amino acid, Monosaccharide, Vitamin, Inorganic ion, trace elements and broad spectrum antibiotics. The other key ingredients of cell culture is a natural medium which may be ani mal body fluids or medium of tissue extraction, including plasma, serum, lymph, chicken embryos leaching solution, etc. Serum, usually bovine or calf is the most commonly used natural medium. Serum provide a similar osmotic pressure and pH as of body environment. Serum enhances the cell attachment and provides extra nutrients, various hormones like growth factor that promotes healthy growth of the cell. In order to monitor the status of media, phenol red is added as a pH indicator. This will turn yellow if media becomes acidic otherwise media at pH 7.2-7.4 remains red.

Prepartion of cell culture medium

To explain the method of media preparation, we are taking the example of DMEM media (Figure 8.2). Measure 80 - 90% of the final volume of cell culture grade water. Add 13.4 gm dry powder medium to the water and mix to dissolve it completely. For each liter of DMEM, add 3.7g/L of sodium bicarbonate, mix completely and adjust the pH to 6.9 -7.1 using 1N NaOH or 1N HCl. Finally add cell culture grade water to the media to bring it to the final volume. Sterilize the solution using a sterilized membrane filter with a pore size of 0.22μm. Supplements, such as antibiotics and serum can be added to the sterilized solution using aseptic technique.

Figure 8.2: Sterilization of cell culture medium (a) filteration unit set up for the filteration of DMEM media through 0.22μm filter. (b) complete media containing 10% serum and 1% pencillin–streptomycin antibiotic.