血管内皮细胞对联合培养体系中骨髓间充质干细胞侏儒症相关转录因子表达及成骨分化的影响  

Effects of vascular endothelial cells on runt-related transcription factor 2 expression and osteogenic differentiation of bone marrow mesenchymal stem cells in a co-cultivation system

作　　者：杨瑞年[1] 刘流[1] 王福科[1] 赵德萍[2] 

机构地区：[1]昆明医学院第一附属医院头颈外科,云南省昆明市650032 [2]昆明医学院第一附属医院实验中心,云南省昆明市650032

出　　处：《中国组织工程研究》2012年第19期3447-3452,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金资助项目(30960388;81160239)~~

摘　　要：背景:血管内皮细胞能够促进骨髓间充质干细胞向成骨方向转变,为干细胞的成骨分化提供营养支持。目的:观察人脐静脉血管内皮细胞在联合培养体系中对人骨髓间充质干细胞形态、生长、细胞分化及其侏儒相关转录因子基因表达的影响。方法:在联合培养装置中,取第3代人骨髓间充质干细胞与第3代人脐静脉血管内皮细胞联合培养(实验组),设立单纯骨髓间充质干细胞培养组(对照组),于培养第4,6,10天观察骨髓间充质干细胞形态变化,细胞碱性磷酸酶活性及侏儒相关转录因子基因表达情况。结果与结论:与对照组比较,实验组中细胞形态呈多样化,后期部分细胞之间出现了连接及融合,成骨分化明显。对照组各个时间点细胞碱性磷酸酶活性无明显变化,实验组各时间点碱性磷酸酶活性高于对照组(P<0.05),且实验组第6,8天细胞侏儒相关转录因子基因表达量为对照组4倍左右(P<0.01)。表明在联合培养体系中,静脉内皮细胞能促进骨髓间充质干细胞侏儒相关转录因子基因的表达,并诱导其向成骨细胞方向分化。BACKGROUND： Vascular endothelial cells can promote osteogenic differentiation of bone marrow mesenchymal stem cells （BMSCs）, which provide nutrition supports for osteogenic differentiation of stem cells. OBJECTIVE： To investigate the effects of human umbilical vein endothelial cells （HUVECs） on the morphology, growth, differentiation of BMSCs as well as the runt-related transcription factor 2 （Runx2） gene expression in a co-cultivation system. METHODS： In a co-cultivation system, passage 3 BMSCs and passage 3 HUVECs were cultured together and served as experimental group. Passage 3 BMSCs were simply cultured and served as control group. At 4, 6, 10 days of culture, BMSCs morphology, alkaline phosphatase activity and Runx2 gene expression were determined. RESULTS AND CONCLUSION： Compared with control group, BMSCs morphology was diverse, and cell conjunction and fusion appeared, and osteogenic differentiation was obvious during anaphase in the experimental group. At each time point, there was no obvious change in alkaline phosphatase activity in the control group, and alkaline phosphatase activity was significantly higher in the experimental group than in the control group （P 〈 0.05）. At 6 and 8 days, Runx2 gene expression in the experimental group was about 4 times that in the control group （P 〈 0.01）. This suggests that in the co-cultivation system, intravenous endothelial cells can promote Runx2 expression in BMSCs and induce osteogenic differentiation of BMSCs.

关 键 词：血管内皮细胞 骨髓间充质干细胞 侏儒相关转录因子(Runx-2) 联合培养 实时定量PCR 

分 类 号：R394.2[医药卫生—医学遗传学]