东方田鼠肝内巨噬细胞的分离和鉴定  

Isolation and identification of macrophages from liver of Microtus fortis

作　　者：胡媛[1] 孙磊[1] 徐馀信[1] 曹建平[1] 

机构地区：[1]中国疾病预防控制中心寄生虫病预防控制所,卫生部寄生虫病原与媒介生物学重点实验室,世界卫生组织疟疾、血吸虫病和丝虫病合作中心,上海200025

出　　处：《中国血吸虫病防治杂志》2015年第3期282-284,共3页Chinese Journal of Schistosomiasis Control

基　　金：上海市自然科学基金面上项目(14ZR1444200)

摘　　要：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 separate and purify intrahepatic macrophages from Microtus fortis Mf and identify its phagocy-tosis. Methods The intrahepatic macrophages from Mf were separated and purified by perfusion collagenase digestion and density gradient centrifugation. The function of the cells was identified by FACS analysis and ink phagocytosis activity. Results The macrophage cells from the liver of Mf were obtained. These cells were bright and circular and grew adhering to the wall. The proportion of the living cells was 95%. The binding rate of these cells from Mf with anti-mouse CD14 antibody Clone Sa2-8 was about 50%of the rate of macrophage from C57BL/6 mice with this monoclonal antibody. The result of ink-phagocytosis ex-periment of macrophage cells from the liver of Mf was positive. Conclusion The method above mentioned is useful to separate and purify macrophage from the liver of Mf. The study builds the foundation for further research on macrophages of Mf against Schistosoma japonicum.

关 键 词：东方田鼠 日本血吸虫 巨噬细胞 分离 鉴定 天然免疫 

分 类 号：R383.24[医药卫生—医学寄生虫学]