动物造模,巨噬细胞 z-bio 2022-11-08 544

　　Objective: To establish a high-purity method for separating and culturing alveolar macrophages, and to provide materials for studying the biological characteristics of macrophages and the survival mechanism of intracellular parasites.

　　Methods: Goat lungs were lavaged with PBS for several times in a sterile environment, and after cells were collected, goat alveolar macrophages were isolated from mixed cells with peripheral blood mononuclear cell separation medium combined with density gradient centrifugation; cultured in 10% In the cell culture medium of fetal bovine serum, the morphology of cells was observed under an inverted microscope, and the phagocytosis of chicken erythrocytes was used to detect cell activity; flow cytometry was used to detect CD14, a characteristic marker on the surface of mononuclear macrophages.

　　Results: The obtained adherent cells had typical morphological characteristics of macrophages, with pseudopodia and protrusions, round and irregular shapes, abundant cytoplasm, and large cell bodies; 54.5% of the cultured macrophages phagocytosed Chicken erythrocytes have strong phagocytic activity; 93.7% of the cells can still express CD14 antigen specifically after being continuously cultured in vitro for nearly a month, and have a macrophage-specific immunophenotype.

　　Conclusion: The goat alveolar macrophages obtained in this study have high purity and good biological activity, and provide a cell model for the study of the disease mechanism of intracellular parasites.