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将1～5日龄的大白鼠(乳鼠)肝组织剪成小块,用低浓度的胶原酶(0.05％)与胰蛋白酶(0.06％)等量混合液直接消化,离心分离获得较多量的肝MΦ,并在含20％小牛血清的1640培养基中接种,在37℃恒温培养箱内静止培养。最初分离的MΦ为圆形,比肝细胞大,折光性强,6h后已贴壁生长。MΦ生长3～5d时,通过相差显微镜观察到形态多样,大小不一,具有变形运动及吞噬功能,胞质内充满折光性强的粗大颗粒。0.1％苔盼蓝试验后,HE、瑞氏染色可见,胞质内有被吞噬的苔盼蓝颗粒及死亡的白细胞。经组织化学观察,ACP与ANAE活性强阳性,SDH活性弱阳性,PAP法显示Con-A受体染色,可见细胞膜周边呈强阳性,其余阳性反应。同时细微突起也很清晰,糖衣很厚,
The liver tissue of 1 to 5 days old rats (suckling mice) was cut into small pieces, and directly digested with a low concentration of collagenase (0.05%) and trypsin (0.06%) in equal volumes and centrifuged to obtain a large amount of Liver MΦ was inoculated in 1640 medium containing 20% fetal bovine serum and cultured at 37°C in a static incubator. The initially isolated MΦ was round, larger than hepatocytes, and had a strong refractive index, and had adherent growth after 6 hours. When MΦ grew for 3 to 5 days, it was observed by phase-contrast microscopy that its morphology was varied, its sizes varied, and it had deformed movement and phagocytosis. The cytoplasm was filled with coarse particles with high refractive index. After 0.1% trypan blue test, HE and Wright’s staining showed phagocytized trypan blue particles and dead white blood cells in the cytoplasm. The histochemical observation showed that ACP and ANAE activity were strongly positive and SDH activity was weakly positive. PAP staining showed that the Con-A receptor was stained, showing strong positive staining around the cell membrane and positive reaction in the rest. At the same time, the tiny protrusions are also very clear and the sugar coating is very thick.