骨髓间充质干细胞的体外趋化与单核细胞趋化蛋白1的作用  

Chemotaxis in vitro of bone marrow mesenchymal stem cells and the role of monocyte chemotactic protein-1

作　　者：孟玲[1] 刘国树[1] 高艳红[2] 李艳华[2] 裴雪涛[2] 

机构地区：[1]解放军总医院老年心肾科,北京市100853 [2]解放军军事医学科学院野战输血研究所干细胞研究室,北京市100850

出　　处：《中国临床康复》2005年第19期68-70,i001,共4页Chinese Journal of Clinical Rehabilitation

基　　金：国家"九七三"重点基础研究发展规划资助项目(2001CB509906;G1999053903);国家"八六三"高技术研究发展计划资助项目(2003AA205160;2002AA205051);北京市科委资助项目(H020220010190)~~

摘　　要：目的:观察单核细胞趋化蛋白1对骨髓间充质干细胞体外增殖和迁移的作用,及单核细胞趋化蛋白1抗体对该作用的影响。方法:实验于2003-12/2004-02在军事医学科学院野战输血研究所干细胞生物实验室完成。取10只健康雄性Wistar大鼠骨髓,体外分离、培养骨髓间充质干细胞,选择第2代作为实验细胞。观察0,5,25,50,75,150μg/L单核细胞趋化蛋白1对大鼠骨髓间充质干细胞的生物学作用;通过细胞迁移实验,观察0,5,25,50,75,150μg/L单核细胞趋化蛋白1对骨髓间充质干细胞的体外趋化作用,以及加入单核细胞趋化蛋白1抗体中和后其对骨髓间充质干细胞趋化作用的变化;应用反转录多聚酶链反应方法,检测大鼠骨髓间充质干细胞有无单核细胞趋化蛋白1相关受体细胞趋化因子受体2的表达。结果:①二甲基偶氮唑盐法测定细胞增殖实验结果:在体外,单核细胞趋化蛋白1以浓度梯度(0,5,25,50,75,150μg/L)依赖方式促进大鼠骨髓间充质干细胞的增殖,但作用较温和。②细胞迁移实验结果:单核细胞趋化蛋白1以浓度梯度(0,5,25,50,75,150μg/L)依赖方式促进大鼠骨髓骨髓间充质干细胞的定向迁移,该作用显著(P<0.05)。③抗体中和后的迁移实验:75μg/L质量浓度单核细胞趋化蛋白1对骨髓间充质干细胞的促迁移作用可被单核细胞趋化蛋白1中和抗体明显减弱(P<0.05)。④反转录聚合酶链反应方法证实骨髓间充质干细胞表达单核细胞趋化蛋白1的相关受体细胞趋化因子受体2。结论:在体外,单核细胞趋化蛋白1以浓度梯度依赖方式引起骨髓间充质干细胞的定向迁移,而且该作用可被单核细胞趋化蛋白1中和抗体减弱。单核细胞趋化蛋白1可能通过与细胞趋化因子受体2结合后发挥作用。本实验为了将影响因素相对简单化,便于观察其中的相互关系,并非将体外实验结果完全等同于在体情况。AIM:To observe the proliferation and migration of bone marrow mesenchymal stem cells(BMMSCs) enhanced by monocyte chemotactic protein-1(MCP-1) and investigate the influence of MCP 1 antibody on it.METHODS:This study was completed in the Biological Laboratory for Stem Cells,Beijing Institute of Transfusion Medicine Affiliated to Chinese Academy of Military Medical Sciences from December 2003 to February 2004.BMMSCs were separated from the bone marrow of 10 healthy male Wistar rats and cultured in vitro,and cells of the second generation were used as experimental cells.The biological effect of MCP 1(0,5,25,50,75 and 150 μ g/L) on BMMSCs was observed.The chemotaxis in vitro of MSCs by MCP 1 of the above concentration was observed as well through cell migration test.After its neutral antibody was added,the effect of MCP 1 on BMMSC chemotaxis was observed.It was examined if rat BMMSCs expressed chemokine receptor type 2(CKR 2) related to MCP 1 by using reverse transcription polymerase chain reaction(RT PCR) method.RESULTS:① It was shown that MCP 1 enhanced the proliferation of rat BMMSCs slightly in vitro in the manner of concentration dependence with the concentration gradient of 0,5,25,50,75 and 150 μ g/L.② In the cell migration test,MCP 1 improved the directional migration of rat BMMSCs significantly in the manner of concentration dependence with the concentration gradient of 0,5,25,50,75 and 150 μ g/L(P< 0.05).③ The migration of BMMSCs induced by MCP 1 with the concentration of 75 μ g/L could be depressed significantly by MCP 1 neutral antibody(P< 0.05).④ BMMSCs expressed the mRNA of CKR 2 related to MCP 1,which was confirmed by RT PCR method.CONCLUSION:MCP 1 enhances directional migration of rat BMMSCs significantly in vitro in a concentration dependent manner,which then can be suppressed by MCP 1 neutral antibody.After combined with CKR 2 of MSCs,MCP 1 may play its role.In our study,the results from in vitro experiment were not equal to those from the in vivo one completely in order to relatively sim

关 键 词：干细胞 骨髓细胞 单核细胞化学吸引蛋白质1 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]