Confocal Microscopy and Calcium Images of Keratinocytes Exposed to Sulfur Mustard

C. During the first week in culture, their medium was a 3:1 mixture of Dulbecco’s Modified Eagle’s Medium (DMEM) and Ham’s F-12 supplemented with 5% FCS, 0.1 nM cholera toxin, 0.4 |xg/ml hydrocortisone, 10 ng/ml epidermal growth factor, and 50 p-g/ml gentamicin sulfate. On d 7, the feeder layer was removed by vigorous aspiration. HEK cultures were subsequently maintained on serum-free keratinocyte growth medium (Clonetics, San Diego, CA) containing 0.15 mM calcium. All media and additives, unless otherwise stated, were obtained from the Sigma Chemical Co., St. Louis, MO.