特定序列寡核苷酸对人骨髓间充质干细胞增殖能力的影响  

Proliferative Effect of ODN with Specific Sequences on Human Bone Marrow Mesenchymal Stem Cells

作　　者：李东文[1] 满大鹏[1] 杜晓岩[1] 马树明[1] 苗林[1] 康宁 董平 肖苒 王丽颖[3] 刘春秀[1] 

机构地区：[1]佳木斯大学附属第二医院口腔科,黑龙江佳木斯154007 [2]中国医学科学院整形外科医院研究中心 [3]吉林大学基础医学院分子生物学教研室

出　　处：《口腔医学研究》2014年第7期607-610,共4页Journal of Oral Science Research

基　　金：佳木斯大学重大项目培育基金项目(SZP2011-001)

摘　　要：目的:比较不同种类特定序列寡核苷酸(Oligonucleotide,ODN)对人骨髓间充质干细胞(human bone marrow mesenchymal stem cells,hBMSCs)增殖能力的影响。方法:密度梯度离心法分离培养hBMSCs,传至第3代进行成骨、成脂及成软骨诱导,鉴定其多向分化能力;将hBMSCs以不同密度接种(3.0×103/cm2、6.0×103/cm2、1.2×104/cm2、2.4×104/cm2),采用CCK-8连续检测7d,确立最佳铺板密度;依据ODN种类(MT01、FC003、Sa05f)和终浓度(0.5mg/L、1.0mg/L、2.0mg/L、4.0mg/L)分为12个实验组和1个对照组(加等量PBS),采用CCK-8连续检测3d,明确不同种类及浓度的ODN对hBMSCs增殖能力的影响。结果:分离培养的hBMSCs具有成骨、成脂和成软骨的多向分化能力;以6.0×103/cm2的接种密度铺板可获得良好的细胞生长曲线;hBMSCs在加入每种不同浓度的ODN后,与对照组相比,终浓度为0.5mg/L的ODN FC003在第1天检测到光密度值显著升高(P<0.01);终浓度为1.0mg/L和4.0mg/L的ODN Sa05f在第1天检测到光密度值均显著降低(分别为P<0.01,P<0.05);终浓度为2.0mg/L的ODN MT01在连续3d内的光密度值均明显升高,差异具有统计学意义(第1天P<0.01,第2,3天P<0.05)。结论:终浓度为2.0mg/L的ODN MT01可显著促进hBMSCs的增殖。Objective：To compare the effects of different kinds of ODN on the proliferation of human bone marrow mesenchymal stem cells.Methods：hBMSCs were isolated by density gradient centrifugation method.Firstly,the muti-differentiation capacity of cells at passage3was identified by osteogenic,adipogenic,and chondrogenic induction.Then,to achieve a standard growth curve,hBMSCs with different seeding densities was detected by CCK-8assay.Finally,the proliferation of hBMSCs cultured with 3kinds of ODN in a gradient of concentrations within first3days were detected by CCK-8to select the positive one and the optimal final concentration.Results：hBMSCs after committed induction stained postive with Alizarin red,oil red,and Toluidine blue.The standard growth curve was achieved when the seeding density was 6.0×103/cm2.Compared to the control group,the mean OD values of hBMSCs with ODN FC003at the concentration of 0.5mg/L was significantly higher on the first day.But with ODN Sa05f,the mean OD values were significantly lower with the concentrations of 1.0mg/L and 4.0mg/L on the first day.The mean OD values of hBMSCs with ODN MT01was significantly higher than control in the first 3days.（P〈0.05vs all of others）.Conclusion：ODN MT01with the concentration of 2.0mg/L can improve the proliferation of hBMSCs.

关 键 词：骨髓间充质干细胞 增殖 寡核苷酸 CCK-8 细胞周期 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]