rhG-CSF对小鼠骨髓间充质干细胞的动员作用  

Effects of rhG-CSF on Mobilization of Mouse Mesenchymal Stem Cells

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,十堰442008 [2]郧阳医学院附属人民医院临床医学研究所,十堰442000 [3]武汉大学人民医院血液科,武汉430060

出　　处：《中国实验血液学杂志》2007年第4期790-794,共5页Journal of Experimental Hematology

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资助项目(编号2005CXG02)

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-CSF80μg/(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(p<0.01)。6小时取材较12小时,168小时取材形成的CFU-F数目多(p<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(p<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。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 × 10^6 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 （p 〈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 （p 〈0.01 ）. There was no obvious difference between CFU-F at 12 hours and at 168 hours （p 〉 0.05 ）. MSCs were positive for CD90 ,CD105 and negative for CD34 and CD133. MSCs were found to differentiate into adipocyte and osteocyte in vitro. The CFU-F of PBMNCs obtained and cultured in vitro in the same culture conditions could be observed after the rhG-CSF injection at 6 hours, but cloning efficiency was （0.50 ±0. 11 ） ×10^-6 MNCs and showed statistical difference as compared with control. It is concluded that rhG-CSF to mobilize hemopoietic stem cells can be used to induce mouse MSCs in vivo expansion, which showed the peak value within 6 hours after final injection of rhG-CSF, rhG-CSF have the mini- mobilization effect on murine MSCs derived from bone marrow.

关 键 词：重组人粒细胞集落刺激因子 骨髓间充质干细胞 CFU-F 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]