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目的:建立兔不同部位单核/巨噬细胞(Mφ)的分离、培养和鉴定的方法,并在体外培养情况下研究其形态、生长特性及生理功能的差异。方法:6只健康新西兰兔以Fi-coll-Hypaque密度梯度离心法分离外周血单核细胞(Mo),以灌洗法分离肺泡巨噬细胞(AM)和腹腔巨噬细胞(PM),以酶消化法+Percoll密度梯度法分离肝脏枯否氏细胞(KC)。在光镜和透射电镜下观察形态学及微观结构的差异,采用墨汁吞噬试验及免疫细胞化学方法分别测定吞噬功能和表型的表达情况。结果:同一个体来源的4个部位贴壁细胞具有单核/巨噬细胞的形态学特征及免疫表型,有较强的吞噬能力,且表现出一定的差异性。墨汁吞噬功能:AM>PM、KC>Mo,MAC387表达:KC>AM>PM>Mo。结论:同一个体不同部位兔单核/巨噬细胞的功能及表型存在异质性。
OBJECTIVE: To establish a method for the isolation, culture and identification of mononuclear / macrophage (Mφ) in different parts of rabbit and study its morphological, growth characteristics and physiological functions in vitro. Methods: Peripheral blood mononuclear cells (Mo) were isolated from six healthy New Zealand rabbits by Fi-coll-Hypaque density gradient centrifugation. The alveolar macrophages (AM) and peritoneal macrophages (PM) Digestion method + Percoll density gradient method to isolate liver Kupffer cells (KC). Morphology and microstructure were observed under light and transmission electron microscopy. The phagocytosis and phenotypes were determined by ink phagocytosis and immunocytochemistry respectively. Results: The adherent cells derived from the same individual had morphological features and immunophenotypes of monocytes / macrophages with strong phagocytic ability and showed some differences. Ink phagocytosis: AM> PM, KC> Mo, MAC387 expression: KC> AM> PM> Mo. Conclusion: The function and phenotype of rabbit monocytes / macrophages in different parts of the same individual are heterogeneous.