Objective To establish an efficient and reliable method of isolating，purifying and identifying macrophages from mouse uterus. Methods Five female mice were killed and their uteri were obtained. Mechanical dissection and digestive enzymes were used to prepare cell suspension of mouse uterus. Uterine macrophages were separated by density gradient centrifugation. And these cells were purified by anchoring cultivation. Then the cell viability was assessed by trypan blue exclusion staining. The phagocytosis of cell was evaluated by phagocytosis of chicken erythrocytes. The purity of cell was determined by acid phosphatase staining，αacetic acid naphthol esterase staining and surface specific antigen CD14 staining. Results By using this new method，the cell viability was（85.35±0.43）％. And the purity of uterine macrophages was （86.12±1.19）%,（