重组人红细胞生成素促进体外培养的人骨髓间充质干细胞增殖  

Erythropoietin Promotes Proliferation of Human Bone Marrow Mesenchymal Stem Cells In Vitro

作　　者：曾琴兵[1] 程范军[1] 张卫国[2] 唐俊明[3] 陈龙[1] 刘歧焕[1] 高清平[4] 王家宁[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]郧阳医学院附属东风医院骨科,湖北十堰442008 [3]郧阳医学院临床医学研究所,湖北十堰442000 [4]武汉大学人民医院血液科,湖北武汉430060

出　　处：《中国实验血液学杂志》2008年第6期1392-1397,共6页Journal of Experimental Hematology

基　　金：湖北省自然科学基金(编号2005ABA081);湖北省教育厅重点项目(编号D200724004);湖北省科技攻关项目(编号2007AA301C14-1);湖北省卫生厅科研基金项目(编号QJX2005-18);郧阳医学院优秀中青年科技创新团队资助(编号2005CXG02)

摘　　要：为了观察重组人红细胞生成素(rhEPO)对人骨髓间充质干细胞(MSCs)增殖活性的影响,抽取健康志愿者的骨髓液,贴壁培养获取第3代细胞,通过细胞形态特征、细胞表面抗原及诱导分化的方法进行MSCs鉴定;取经过鉴定的第3代MSC(P3-MSC)加入不同浓度rhEPO(0.5、1、5、10、50U/ml)后培养,应用细胞计数法及MTT法检测细胞增殖,用流式细胞术(FCM)分析细胞周期。结果发现:骨髓贴壁培养获取的第3代细胞同时表达CD105和CD90,不表达CD34和CD45,并可被诱导分化为脂肪、骨及软骨细胞,被鉴定为MSC。MTT结果显示EPO处理组的光密度值(OD)均高于对照组(p<0.05);50U/ml浓度EPO组作用最显著,细胞计数法获得的结果与其一致。FCM细胞周期分析表明,rhEPO能明显降低G0/G1期细胞比例,并提高S+G2/M期细胞比例,与对照组相比,差异均具有显著统计学意义(p<0.01)。结论:EPO能促进体外培养的人骨髓MSCs增殖,该效应与EPO调节其进入细胞增殖周期有关。This study was aimed to investigate the effects of recombinant human erythropoietin （rhEPO） on proliferation of human bone marrow-derived mesenchymal stem cells（MSCs） in vitro. The aspirates of the bone marrow from bealty volunteers were seeded in culture medium. Then MSCs were isolated according to characteristics adhering to the plastics. After three passages in culture, bone marrow-derived adherent cells were identified by growing morphological features, cell surface antigens and differentiation into multi-lineages. Then P3-MSCs which had been identified were incubated with different concentrations of rhEPO （0. 5, 1, 5, 10 and 50 U/ml ）. Subsequently, proliferation of MSCs was measured by MTT assay, as well as cell counts. At the same time, cell cycle was detected by flow cytometry （FCM）. The results indicated that the expressions of CD90 and CD105 in P3 bone morrow-derived adherent cells were positive, while the expressions of CD34 and CD45 were negative, and these cells could differentiate into adipocytes, osteocytes and chondrocytes in induction media. MTT assay showed that the optical density （OD） of group treated with EPO was significantly higher than that in the control group（p 〈 0.05 ）, and the group treated with 50 U/ml EPO achieved the most predominant effects. The results of cell count were coincident with that of MTT assay. Furthermore, the cell cycle analysis by FCM revealed that rhEPO could relatively decrease the cell ratio in G0/G1 phase, and increase the cell ratio in S and G2/M phases. As compared with the control group, all those differences were statistically significant （p 〈 0.01 ）. It is concluded that erythropoietin can promote proliferation of human bone marrowmesenchymal stem cells in vitro, which may be correlated with the increasable entry into S and M phases of cell cycle of MSCs adjusted by EPO.

关 键 词：红细胞生成素 间充质干细胞 细胞周期 细胞增殖 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]