Objective To improve the culture results of amniotic fluid by adding antioxidants.Methods Using Fractional factorial designs combined with the steepest ascent path approach,the concentration of antioxidants added in H medium was optimized to increase the growth speed of the passage fibroblast cells from amniotic fluid.And the performance of the optimized culture medium was tested with the growth index,the numbers of clones and metaphase of primary amniotic fluid cells culture.Results Vitamin E,L-ascorbic ac...