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应用胶原酶与链霉蛋白酶消化和分解破坏大鼠肝细胞的方法,获得非肝细胞;再经贴壁培养,成功地获得大量高纯度的枯否细胞。枯否细胞获得率为8.3×10~6个/每克肝组织,胞质内均含有在体吞噬的墨汁颗粒。培养成活的枯否细胞呈典型巨噬细胞形态,电镜观察下表而有发达的伪足和微绒毛等结构,胞质内含大量溶酶体。以绵羊红细胞(SRBC)检测枯否细胞免疫活性,90%以上的细胞在特异抗体介导下形成SRBC花环,并能活跃吞噬SRBC。电镜观察了枯否细胞花环和吞噬功能。本实验国内首次分离培养成功大鼠肝枯否细胞,并证明具有良好的免疫活性,这对进一步研究枯否细胞的功能,尤其是它对肿瘤的免疫监视作用有实际意义。
The method of collagenase and pronase digestion and decomposition to destroy rat hepatocytes was used to obtain non-hepatocytes. After culturing in vitro, a large number of high-purity Kupffer cells were successfully obtained. Kupffer cell acquisition rate of 8.3 × 10 ~ 6 / g liver tissue, the cytoplasm contains in vivo phagocytic ink particles. Kupffer cells were cultured to form a typical morphology of macrophages, electron microscopy under the watch with developed pseudopodia and microvilli and other structures, the cytoplasm contains a large number of lysosomes. The immune activity of Kupffer cell was detected by sheep erythrocytes (SRBC). More than 90% of the cells could form SRBC rosette under specific antibody and actively swallowed SRBC. Electron microscopy observed Kupffer cell rosette and phagocytosis. In this experiment, the first successful isolation and culture of Hepatic Fibroblasts in rats, and proved to have good immunocompetence, which is of practical significance for the further study of the function of Kupffer cells, especially its immune surveillance of tumor.