巨噬细胞,动物造模 z-bio 2021-11-25 216

　　Objective: To establish a high-purity method for the isolation and culture of alveolar macrophages to provide materials for the study of the biological characteristics of macrophages and the study of the survival mechanism of intracellular parasites.

　　Method: Lavage goat lungs with PBS several times in a sterile environment. After collecting the cells, use peripheral blood mononuclear cell separation solution combined with density gradient centrifugation to separate goat alveolar macrophages from the mixed cells; cultured at 10% In the cell culture medium of fetal bovine serum, observe the cell morphology under an inverted microscope, and use its phagocytic function on chicken red blood cells to detect cell viability; use flow cytometry to detect CD14, a characteristic marker on the surface of mononuclear macrophages.

　　Results: The obtained adherent cells had typical morphological characteristics of macrophages, with pseudopods and protrusions, showing round and irregular shapes, rich cytoplasm, and large cell bodies; 54.5% of cultured macrophages were phagocytosed Chicken erythrocytes have strong phagocytic activity. After being continuously cultured in vitro for nearly a month, 93.7% of the cells can express CD14 antigen specifically 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.