Orthogonal experimental design has been applied to study the suitable cell culture medium and condition of conductive polypyrrole on rat keratinocyte cell cultures. The results showed that one thousand milliliters of complete minimum essential medium(CMEM) which contained 900 ml minimum essential medium(MEM), 10% fetal bovine serum(FBS), 1% NEAA, 6 ml insulin and 10 mg hydrocortisone was fit for keratinocyte cell culture. And the conductive polypyrrole obtained with 10 mA current density for 100 second switching times, given 100 mV electrical stimulation voltage for 2 hours, would stimulate the growth of rat keratinocyte cell in vitro cultures. The cell density under culture conditions described above was 1.30 times as high as that under routine culture conditions.