摘要
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。
Aims:Kupffer cells are hepatic macrophages that reside in the lumen of hepatic sinusoids.Isolation and culture of kupffer cells are important to study the pathogenesis of liver diseases. Methods:Kuppffer cells were isolated from liver of Wistar rats by in situ perfusion with pronase and collagenase and density gradient centrifugation with Nycodenz,and established maintenance cultures of purified kupffer cells according to the attached time and cultured conditiones; Kuppffer cells were identificated by immunohistochemistry,endocytosis,and ultrastructure,et al. Results:Kupffer cells were isolated sucessfully with high purity ,the yield was 3～5×10 6/liver. kupffer cells showed particles of india ink and latex beads in cytoplasm,and lysozyme positive by immunohistochemistry staining;kupffer cells had appearance with numerous lamellipodia, microfilamentous and lysosomal structures,and latex particles of endocytosis under the transmission electron microsopy. Conclusions: The technique for isolation of kupffer cells described here is simple,reliable,used to futher study biologic functions of kupffer cells.
出处
《上海医学》
CAS
CSCD
北大核心
1998年第11期630-632,共3页
Shanghai Medical Journal
基金
国家自然科学基金
