Birmingham J R, Jeska E L
Immunology. 1980 Dec;41(4):807-14.
Bovine peripheral blood monocytes were isolated from buffy coats of ACD-anticoagulated whole blood. Cells cultivated on plastic surfaces for 20 h were judged to be 95% monocytes based on nonspecific esterase-1 staining, uptake of latex particles and surface receptor characteristics. Bovine monocytes were maintained up to 80 days in vitro in Dulbecco's modified Eagle medium with 15% horse serum and 15% foetal bovine serum. Several morphological and physiological features of bovine monocytes were examined during the course of culture. Cell size and cytoplasmic spreading, granulation and vacuolization increased progressively. Multinucleated giant cells predominated during the latter stages of in vitro culture. A high percentage of bovine monocytes possessed C3 and IgG Fc receptors, whereas IgM Fc and sheep erythrocyte receptors were not detected. Phagocytosis was mediated by the IgG receptor, but not by the C3 receptor. Peroxidase activity declined in a linear fashion, with cells essentially negative after 8 days of culture. Total cell protein and acid phosphatase increased during cultivation. Lysozyme activity was undetectable in both lysates and supernatants of bovine monocyte. These findings are consistent with the concept of maturation of mononuclear phagocytes. The procedures for isolation and cultivation described in this paper will provide methodology for detailed study of bovine mononuclear phagocytes.
牛外周血单核细胞从ACD抗凝全血的血沉棕黄层中分离得到。基于非特异性酯酶-1染色、乳胶颗粒摄取和表面受体特征，在塑料表面培养20小时的细胞被判定为95%的单核细胞。牛单核细胞在含有15%马血清和15%胎牛血清的杜尔贝科改良伊格尔培养基中可在体外维持长达80天。在培养过程中检测了牛单核细胞的几种形态和生理特征。细胞大小、细胞质伸展、颗粒化和空泡化逐渐增加。在体外培养后期多核巨细胞占主导。高比例的牛单核细胞具有C3和IgG Fc受体，而未检测到IgM Fc和绵羊红细胞受体。吞噬作用由IgG受体介导，而非C3受体。过氧化物酶活性呈线性下降，培养8天后细胞基本呈阴性。培养过程中总细胞蛋白和酸性磷酸酶增加。在牛单核细胞的裂解物和上清液中均未检测到溶菌酶活性。这些发现与单核吞噬细胞成熟的概念一致。本文所述的分离和培养方法将为详细研究牛单核吞噬细胞提供方法。
