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用链霉蛋白酶E直接灌流小鼠肝脏,选择性消化肝脏实质性细胞而得到非实质性细胞(non-parenchymal cells,NPC),收获量为(34.1±5.7)×10~6细胞/g组织。将NPC培养过夜,Kupffer细胞(KC)贴附玻璃而把它与内皮细胞分离。在细胞悬液中KC与内皮细胞的相对分配为25%和75%。在光学显微镜和扫描电子显微镜下观察贴附玻璃的KC具有典型巨噬细胞的形态学特征,胞膜上的Fc和C3b受体,以及吞噬功能都保存高活性,完整无损。本法对研究肝脏抗肿瘤和抗感染免疫功能是一种十分有用的技术。
The liver of mice was directly perfused with Pronase E, and the substantial cells of the liver were selectively digested to obtain non-parenchymal cells (NPC), and the harvested amount was (34.1±5.7)×10-6 cells/g. NPCs were cultured overnight and Kupffer cells (KC) were attached to glass to separate them from endothelial cells. The relative distribution of KC to endothelial cells in cell suspensions was 25% and 75%. Observing the glass-attached KC under the light microscope and scanning electron microscope has the morphological characteristics of typical macrophages. The Fc and C3b receptors on the cell membrane and the phagocytosis function are all kept highly active and intact. This method is a very useful technique for studying liver anti-tumor and anti-infection immune functions.