Smedsrød B, Pertoft H, Eggertsen G, Sundström C
Cell Tissue Res. 1985;241(3):639-49. doi: 10.1007/BF00214586.
This paper presents a study on the structure and function of Kupffer cells (KC) and liver endothelial cells (LEC) isolated by a simple and rapid technique involving 1) perfusion of the liver with collagenase; 2) cell separation by means of density centrifugation in Percoll; and 3) cell culture, taking advantage of the fact that KC and LEC differ in their preferences for growth substrate. The KC, which attach and spread under serum-free conditions on surfaces of glass or plastic during the first 15 min in culture exhibit a typical macrophage-like morphology including membrane ruffling and a heterogenous content of vacuoles. Moreover, these cells express (a) Fc receptors (FcR) for binding and phagocytosis of erythrocytes covered with immune globulin G (E-IgG), and (b) complement receptors (CR) for binding and serum dependent phagocytosis of erythrocytes covered with either human C3b or mouse inactivated C3b (iC3b). The cells also bind fluid phase fluoresceinated C3b. Approximately 30% of the KC express immune response-associated (Ia)-antigens. The LEC attach and spread on fibronectin coated surfaces, but not on glass or plastic surfaces, during the first two hours in culture with or without serum, and are morphologically distinct from KC. Cultured LEC are well spread out with no membrane ruffling and with numerous large vesicles surrounding the regularly shaped nucleus. These cells bind, but do not ingest E-IgG via the FcR, but no binding of fluid phase C3b or particle fixed C3b or iC3b can be observed. Incubation of LEC with fluorescein amine conjugates of ovalbumin or formaldehyde treated serum albumin, but not with fluoresceinated native serum albumin, results in accumulation of fluorescence specifically localized in the large perinuclear vesicles. Neither KC nor any other cell types tested have the ability to accumulate fluorescence upon incubation with these compounds. Ia-antigens are not present on the LEC. Cytochemical demonstration of unspecific esterase, acid phosphatase, and peroxidase reveals different patterns and intensities of staining in KC as compared to LEC.
本文介绍了一项关于枯否细胞（KC）和肝内皮细胞（LEC）结构与功能的研究。该研究采用了一种简单快速的技术，包括：1）用胶原酶灌注肝脏；2）通过在 Percoll 中进行密度离心分离细胞；3）利用 KC 和 LEC 对生长底物偏好不同的特点进行细胞培养。在无血清条件下，培养最初 15 分钟内，KC 附着并铺展在玻璃或塑料表面，呈现典型的巨噬细胞样形态，包括膜皱褶和含有不同内容物的液泡。此外，这些细胞表达：（a）用于结合和吞噬覆盖有免疫球蛋白 G 的红细胞（E-IgG）的 Fc 受体（FcR）；（b）用于结合和血清依赖性吞噬覆盖有人 C3b 或小鼠灭活 C3b（iC3b）的红细胞的补体受体（CR）。这些细胞还结合液相荧光素标记的 C3b。约 30%的 KC 表达免疫反应相关（Ia）抗原。在有或无血清的培养最初两小时内，LEC 附着并铺展在纤连蛋白包被的表面，但不附着在玻璃或塑料表面，且在形态上与 KC 不同。培养的 LEC 铺展良好，无膜皱褶，围绕规则形状的细胞核有许多大泡。这些细胞通过 FcR 结合但不摄取 E-IgG，未观察到对液相 C3b 或颗粒固定 C3b 或 iC3b 的结合。用卵清蛋白或甲醛处理的血清白蛋白的荧光胺缀合物孵育 LEC，但不用荧光素标记的天然血清白蛋白孵育，会导致荧光积聚，特异性定位于大的核周泡中。KC 或其他任何测试的细胞类型在与这些化合物孵育时都没有积累荧光的能力。LEC 上不存在 Ia 抗原。非特异性酯酶、酸性磷酸酶和过氧化物酶的细胞化学显示，与 LEC 相比，KC 具有不同的染色模式和强度。
