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目的：研究鼠肝Ｋｕｐｆｅｒ细胞的分离、培养和鉴定。方法：用链酶蛋白酶和胶原酶原位灌流，ｎｙｃｏｄｅｎｚ密度梯度离心分离大鼠Ｋｕｐｐｆｅｒ细胞，再经贴壁培养，并应用免疫组织化学、吞噬功能试验，电镜等方法进行鉴定。结果：本法能成功地获得高纯度的Ｋｕｐｆｅｒ细胞，Ｋｕｐｆｅｒ细胞得率为３～５×１０６／肝，贴壁后呈典型的星形及多角形，免疫组化染色示ｌｙｓｏｚｙｍｅ阳性，胸浆内见吞噬的ｉｎｄｉａｉｎｋ及ｌａｔｅｘｂｅａｄｓ颗粒，电镜观察细胞表面有发达的伪足、微绒毛，胞浆内含大量溶酶体及吞噬ｌａｔｅｘｂｅａｄｓ颗粒。结论：本实验所用的Ｋｕｐｆｅｒ细胞分离培养方法，简单易行，可靠，细胞纯度高，可用于进一步研究Ｋｕｐｆｅｒ细胞的生物学功能
Objective: To study the isolation, culture and identification of murine Kupfer cells. Methods: Rat Kuppfer cells were isolated by in situ perfusion with streptavidin and collagenase and centrifuged by nycodenz gradient centrifugation. The cells were cultured in adherent culture and identified by immunohistochemistry, phagocytosis test and electron microscopy. Results: Kupfer cells with high purity were successfully obtained in this method. The yield of Kupfer cells was 3 ~ 5 × 106 / liver, typical astrocytes and polygons were adherent, immunohistochemical staining showed lysozyme positive, intrathoracic See devoured indiaink and latexbeads particles, electron microscopy of cell surface developed pseudopodia, microvilli, cytoplasm containing a large number of lysosomes and phagocytic latexbeads particles. Conclusion: The method of Kupfer cell isolation and culture used in this study is simple, reliable, high purity and can be used to further study the biological function of Kupfer cells