Abstract: This experiment contain two parts.The purpose of the first part is to test the effect of different concentration and combination of digestive enzymes on the separation of bovine mammary tissue.The purpose of the second part is to test the effect of different FBS concentrations on the culture of bovine mammary epithelial cells.The result indicate that accumulation of bovine mammary epithelial cells is highest for treatment 0.25% Trypsin＋0.20% collagenase Ⅱ as compared with other treatments. The bovine mammary epithelial cells purification for 5% and 10% FBS treatments are better than other treatments,and there are no significant difference for the bovine mammary epithelial cells number between 5% and 10% FBS treatment （P0.05）,but these two treatments are much higher than other treatments （P0.05）.