To begin, thaw one bottle of each feeder cell thawing media, supplement A, supplement B, and supplement C, in a 37 degrees Celsius water bath. Alternatively, refrigerate them at four degrees Celsius for 16 to 24 hours. Transfer the entire contents of each bottle of thawed medium into a 15 milliliter or 50 milliliter conical tube.
To prepare complete plating medium, take a bottle of the plating medium and add 4.5 milliliters of supplement B, followed by 11 milliliters of supplement A into the bottle. In a separate bottle, combine 100 milliliters of culture medium, one milliliter of supplement C, and 14 milliliters of supplement A to prepare the complete culture medium. Store the remaining supplement C and supplement A at minus 20 degrees Celsius until week two media preparation.
