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目的对东方田鼠(Microtus fortis,Mf)肝内巨噬细胞进行分离纯化和功能鉴定。方法采用在体灌注、胶原酶消化及梯度密度离心相结合的方法分离Mf肝内巨噬细胞,并采用流式和墨汁吞噬功能实验进行巨噬细胞的功能鉴定。结果通过此方法获得Mf肝脏来源的巨噬细胞,分离的细胞呈透亮圆形,贴壁生长,活细胞比例为95%。Mf来源的巨噬细胞与抗鼠CD14流式单抗(Clone:Sa2-8)阳性结合率是小鼠来源巨噬细胞与该抗体结合率的50%左右。Mf肝脏来源的巨噬细胞墨汁吞噬实验阳性。结论该方法能有效地分离纯化Mf肝内巨噬细胞,可为进一步研究东方田鼠肝脏巨噬细胞抗血吸虫机制奠定基础。
Objective To isolate, purify and characterize intrahepatic macrophages of Microtus fortis (Mf). Methods Mf intrahepatic macrophages were isolated by in vivo perfusion, collagenase digestion and gradient density centrifugation, and the function of macrophages was evaluated by flow cytometry and ink phagocytosis assay. Results Mf liver-derived macrophages were obtained by this method. The isolated cells were translucent and circular with adherent growth. The percentage of viable cells was 95%. The positive binding rate of Mf-derived macrophages to anti-murine CD14 monoclonal antibody (Clone: Sa2-8) was about 50% of the binding rate of mouse-derived macrophages to this antibody. Mf liver-derived macrophage ink phagocytosis test was positive. Conclusion The method can effectively separate and purify Mf intrahepatic macrophages and lay a foundation for further study on the mechanism of anti-schistosoma infection in liver voles.