粒细胞集落刺激因子对小鼠骨髓间充质干细胞的作用  

Effects of Recombinant Human Granulocyte Colony Stimulating Factor on Mesenchymal Stem Cells in Mice

作　　者：刘岐焕[1] 程范军[1] 陈龙[1] 唐俊明[2] 王家宁[2] 高清平[3] 

机构地区：[1]郧阳医学院附属东风医院血液科,湖北十堰442008 [2]郧阳医学院附属人民医院临床医学研究所,湖北十堰442000 [3]武汉大学人民医院血液科,湖北武汉430060

出　　处：《武汉大学学报（医学版）》2007年第3期336-340,F0003,共6页Medical Journal of Wuhan University

基　　金：湖北省自然科学基金资助项目(编号:2005ABA081);郧阳医学院科研基金(编号:2005CXG02)

摘　　要：目的:研究不同剂量下重组人粒细胞集落刺激因子(rhG-CSF)对小鼠骨髓间充质干细胞的动员作用。方法:32只小鼠随机分成4组,分别给予0,20,40,80μg/kg rhG-CSF皮下注射,每日1次,连续5 d,最后一次注射后6 h分别取骨髓和外周血,以1×106分别接种于含10%胎牛血清的DMEM培养基的12孔板中培养10 d,计数成纤维样集落(CFU-F)的个数,对形成的CFU-F消化后收集细胞,流式细胞术检测表面抗原表达并进行成脂、成骨诱导。结果:骨髓所形成的CFU-F与rhG-CSF剂量成正比,外周血也有类似骨髓的成纤维样集落,但出现的频率极低。骨髓CFU-F流式检测呈CD34-、CD133-、CD90+、CD105+,具有成脂、成骨分化的能力。结论:rhG-CSF可促进骨髓间充质干细胞增殖,且随rhG-CSF剂量增大,促增殖作用越强。rhG-CSF也有动员小鼠间充质干细胞的能力,但作用较弱。Objective. To evaluate the effects of rhG-CSF on the mobilization of mesenchymal stem cells （MSCs） at different dose in mice. Methods： Thirty-two mice were randomized divided into four groups as control group treated with normal saline, and 20, 40 and 80 g/L rhG-CSF per day treatment groups. The bone marrow and peripheral blood cells were obtained at 6 h after the final administration. The bone marrow mononuclear cells （BMMNCs） and peripheral blood mononuclear cells （PBMNCs） were seeded into a 12-well DMEM plate containing 10% FBS at a concentration of 1 × 10^8 per well. The colony-forming unit-fibroblast （CFU-F） was evaluated after 10 days. The cells were collected by trypsinization. The surface antigens CD34, CD133, CD90 and CD105 were analyzed through flow cytometry. Results： The number of CFU-F was gradually increasing after culturing of BMMNCs in rhG-CSF groups according with the increase of rhG-CSF dose, and was higher than in control group（all P〈0. 01）. PBMNCs were obtained and cultured in vitro in the same culture conditions, and the CFU-F of PBMNCs could be observed after the rhG-CSF administration. But it appeared at a very low frequency. MSCs were positive for CD90 and CD105, and negative for CD34 and CD133. MSCs had the ability to differentiate into adipocytes and osteocytes in vitro. Conclusion： The number of CFU-F after BMMNCs in vitro culture was increased significantly after rhG-CSF administration, with the increase of rhG-CSF dose, the number of CFU-F in the BMMNCs gradually increased; rhG-CSF had the mini-mobilization effect on mice.

关 键 词：间充质干细胞 粒细胞集落刺激因子 增殖 动员 

分 类 号：R457.7[医药卫生—治疗学]