QY1骨髓多能间质干细胞系向心肌细胞和血管内皮细胞分化  

Differentiation of QY1 bone marrow pluripotential mesenchymal stem cell line cells into cardiomyocytes and vascular endothelial cells in vitro

作　　者：杨晶[1] 谢祁阳[1] 向红霞[1] 

机构地区：[1]中南大学湘雅医学院生理系

出　　处：《中南大学学报（医学版）》2007年第1期93-98,共6页Journal of Central South University ：Medical Science

基　　金：教育部留学归国人员科研启动基金;湖南省杰出中青年专家科技专项计划(01JZY2100);海南省教育厅高校科研重点资助(Hjkj200228);海南省卫生厅科研重点资助(200201);海南省自然科学基金资助(30216)~~

摘　　要：目的:探索QY1骨髓多能间质干细胞系在体外向心肌细胞、血管内皮细胞分化的能力;优化骨髓多能间质干细胞向心肌细胞分化的体外适宜诱导条件;并初步探讨骨髓间质干细胞向成心肌成血管细胞分化的能力。方法:分别用5-氮胞苷、血管内皮生长因子对QY1骨髓多能间质干细胞进行定向诱导分化。利用免疫组化、RT-PCR鉴定分化后的细胞。结果:在传代培养至72h时,加入10μmol/L5-氮胞苷诱导24h后换液培养,以后每7d换液1次,培养14d进行再次诱导的条件下,细胞能出现自发性搏动的心肌细胞和肌管样结构,诱导率达(39.47±0.56)%。心肌特异性的表面抗体α-横纹肌肌动蛋白、心肌连接蛋白-43表达阳性,RT-PCR检测有心肌特异性因子α-肌球蛋白重链表达。在用血管内皮生长因子诱导48h后,出现呈直线排列的血管内皮细胞;7d后,有部分区域细胞相互连接呈血管内皮细胞网状;14d时免疫组化检测,发现诱导后的细胞血管内皮特异性表面抗体CD31和VIII因子表达阳性。结论:QY1骨髓多能间质干细胞系细胞在体外特定条件下具有分化为心肌细胞和血管内皮细胞的能力。Objective To explore the differentiation potential of QY1 bone marrow mesenchymal stem cell （MSCs）line cells into cardiacmyocytes and vascular endothelial cells in vitro, to optimize the suitable conditions of MSCs differentiating into cardiomyocytes in vitro, and to examine the potentials of MSCs differentiating into cardiomyogenesis and vasculogenesis. Methods Specifically committed differentiation inductive medium was employed, including 5-azacytidine for cardiomyogenesis and vascular endothelial growth factor for vasculogenesis in culture respectively in vitro. The differentiated cells were identified by immunohistochemistry and molecular biology. Results MSCs line cells had been cultured in the normal culture medium for 72 hours, then the differentiation inductive medium including 10 μmol/L 5-azacytidine was added into the normal culture dishes for 24 hours only. After that the culture medium was changed back to the normal culture medium. Normal culture medium was changed every 7 days. The second induction was performed after 14 days. The differentiated cells treated with 5-azacytidine could beat spontaneously and formed myotube structures in the optimal induction conditions, and the differentiation rate was （ 39.47 ± 0.56 ） %. The differentiated cells expressed specific cardiomyocytic proteins identified by the positive immunohistochemistry staining with anti-α-sarcomeric antibody and anti-Cx-43 antibody, and also expressed the α-myosin heavy chain examined by RT-PCR. The differentiated cells began to appear as the lined up vascular endothelial cells after 48 hour treatment with vascular endothelial growth factor. Some of the differentiated cells connected each other to form vascular endothelial web-like structure after 7 day treatment with vascular endothelial growth factor. On 14 d after treating with vascular endothelial growth factor, the differentiated cells were identified by immunohistochemistry staining. The expressions of both specific surface antibody CD31 and factor Ⅷ for vascular e

关 键 词：QY1骨髓间质干细胞 分化 心肌细胞 血管内皮细胞 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]