Abstract: To provide materials for further study on the biological characteristics and immunological functions of monocyte macrophage,bovine peripheral blood mononuclear cells were isolated by using bovine lymphocyte separation medium and incubated in RPMI 1640 medium supplemented with 10% fetal bovine serum for 5 days. The cell morphology was observed by inverted microscope and surface characteristic marker CD14 of monocyte macrophages was detected by using flow cytometry. The results showed that the obtained adherent cells had the morphological characteristics and immunophenotype of macrophage. Through the morphological observation and identification by flow cytometry, bovine peripheral blood monocyte macrophage was successfully obtained.