rhGCSF对小鼠骨髓间充质干细胞动员作用.docVIP

rhGCSF对小鼠骨髓间充质干细胞动员作用 　 作者：刘岐焕，程范军，陈龙，唐俊明，王家宁，高清平 【摘要】 为了研究rhGCSF体内应用对小鼠骨髓间充质干细胞（MSC）的动员作用，将30只小鼠随机分为2组：rhGCSF组和对照组，分别给予皮下注射rhGCSF 80 μg /（kg·d）和等量生理盐水,连续5天，在最后1次注射后分别于6，12和168小时取小鼠骨髓和外周血，以1×106 单个核细胞（MNC）接种于12孔培养板14天，对形成的成纤维细胞集落（CFUF）进行计数，用胰蛋白酶消化后应用流式细胞术测定表面抗原并进行成脂，成骨诱导。结果表明： rhGCSF组骨髓形成的CFUF数均较对照组明显增加(plt;0.01)。6小时取材较12小时,168小时取材形成的CFUF数目多(plt;0.01)，12小时与168小时CFUF差异无统计学意义。流式细胞术检测骨髓CFUF显示CD34－、CD133－、CD90＋、CD105＋，骨髓CFUF具有成脂、成骨分化的能力。rhGCSF组6小时取材外周血具有类似骨髓的CFUF，出现的频率为（0.50±0.11）×10-6，与对照组比较差异有显著性（plt;0.05）。结论： rhGCSF可使小鼠骨髓间充质干细胞增殖加速，但时间短暂，其高峰在rhGCSF应用5天后的6小时内。rhGCSF对小鼠间充质干细胞也有动员能力，但作用较弱。 【关键词】 骨髓间充质干细胞 Effects of rhGCSF on Mobilization of Mouse Mesenchymal Stem Cells Abstract To evaluate the effects of rhGCSF on mobilization of mesenchymal stem cells (MSCs) of mouse bone marrow at different time point, thirty mice were randomly divided into rhGCSF treatment group and control group. The mice were subcutaneously injected with rhGCSF in a dose of 80 μg/kg or saline for 5 days. The bone marrow and peripheral blood were obtained at time points of 6, 12, 168 hours after final injection of rhGCSF or saline. Bone marrow mononuclear cells (BMMNCs) were seeded at density of 1×106 MNCs onto 12well plate for culture expansion in DMEM supplemented with 10% FBS,and the number of colony forming unit  fibroblast (CFUF) was counted after 14 days. The cells were collected by trypsinization and the surface antigens CD34, CD133, CD90 and CD105 were analyzed by flow cytometry. The multidifferentiation of MSCs were done in the culture condition of inducedadipocyte and osteocyte. Peripheral blood MNCs examination was same as the bone marrow. The results indicated that the number of CFUF of bone marrow in rhGCSF group was more than that in control group (plt;0.01)，the number of CFUF in rhGCSF group at 6 hours was more than that at 12 hours and 168 hours, respectively (plt;0.01). There was no obvious difference between CFUF at 12 hours and at 168 hours (pgt;0