裸鼹鼠骨髓巨噬细胞分离培养方法的建立及其功能的初步研究  

Establishment of Method for Separation, Cultivation of Bone Marrow Macrophage in Naked Mole Rats and Preliminary Research on Function

作　　者：程继帅 李莉[2] 肖邦[1] 赵善民[1] 林丽芳[1] 丛薇[1] 汤球[1] 孙伟[1] 余琛琳[1] 杨文静[1] 崔淑芳[1] 

机构地区：[1]第二军医大学实验动物中心,上海200433 [2]第二军医大学教学保障处,上海200433

出　　处：《实验动物与比较医学》2016年第1期72-75,共4页Laboratory Animal and Comparative Medicine

摘　　要：目的 建立一种裸鼹鼠骨髓巨噬细胞分离培养的方法,并通过与小鼠骨髓巨噬细胞进行比较研究其吞噬功能.方法 分离裸鼹鼠骨髓细胞,在含60 ng/mL巨噬细胞集落刺激因子（M-CSF）完全培养基的诱导下,体外贴壁培养6～7 d,采用倒置显微镜观察细胞的生长状态.裸鼹鼠骨髓细胞诱导结束后,采用流式细胞术检测贴壁细胞表面抗原CD11b和F4/80的阳性率,鉴定骨髓巨噬细胞的纯度;采用异硫氰酸荧光素（FITC）-dextran根据荧光强度比较裸鼹鼠和ICR小鼠骨髓巨噬细胞的吞噬功能.结果 通过本方法获得的贴壁细胞具有典型的巨噬细胞的形态特征,且细胞表面抗原CD1 1b和F4/80的阳性率均高于80％;裸鼹鼠与ICR小鼠骨髓巨噬细胞的吞噬率分别为99.87％士0.13％和88.79％＆#177;0.90％.结论 本文建立的方法是一种简单实用的体外分离培养裸鼹鼠骨髓巨噬细胞的方法,且能够获得高纯度、高活性的巨噬细胞.裸鼹鼠骨髓巨噬细胞的吞噬功能高于小鼠骨髓巨噬细胞.Objective To establish the method for the primary culture of bone marrow macrophage in naked mole rats and study the phagocytosis compared with mice. Method The bone marrow cells from the hint leg of naked mole rats were detached. The cells were induced by 60 ng/mL Macrophage colony-stimulating factor （M-CSF） and cultured for 6-7 days in vitro. During the culture, the cell was observed in microscope. After induced, the flow cytometry was conducted to detect the purity of macrophage and the phagocytosis of macrophage with mouse as control. Result Through this method, the adherent cells with typical morphological characteristics were obtained and the positive rate of the cell surface antigen F4/80 and CD 1 1b were higher than 80%. The phagocytosis rate of naked mole rats bone marrow macrophage was 99.87% ± 0.13%, but the ICR mice was 88.79% ± 0.90%. Conclusion This method is simple and practical for separation and culture of highly purified and functional naked mole rats bone marrow macrophage in vitro. It indicates that phagocytosis of naked mole rats bone marrow macrophage is higher than that of mice.

关 键 词：裸鼹鼠 骨髓巨噬细胞 分离培养 功能 

分 类 号：Q95-33[生物学—动物学]