兔骨髓间充质干细胞向心肌细胞分化的研究  

Study of Inducing Rabbit BMSC into Cadiocyte

作　　者：王新庄[1] 和小娥[1] 闫颖颖[1,2] 任卫青[1] 何俊丹[1] 吕婧玉[1] 

机构地区：[1]河南农业大学牧医工程学院,郑州450002 [2]西北农林科技大学动物医学学院,杨凌712100

出　　处：《畜牧兽医学报》2011年第10期1362-1367,共6页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：河南省基础与前沿技术研究计划(092300410081);国家肉牛;牦牛产业体系(30300324)

摘　　要：旨在研究5-氮杂胞苷(5-Aza)诱导兔骨髓间充质干细胞(Mesenchymal stem cells,MSCs)向心肌细胞分化的作用及适宜浓度。取兔股骨骨髓,分离并培养骨髓间充质干细胞,用5-Aza定向诱导向心肌样细胞分化。用倒置显微镜、免疫细胞化学法鉴定诱导后的细胞是否能表达心肌特异性蛋白α-横纹肌肌动蛋白(α-actin)和肌钙蛋白T(Cardiac troponin T,cTnT),计算各组细胞向心肌样细胞的诱导分化率。结果,5-Aza诱导MSCs后,细胞明显变宽,诱导1个月后的MSCs呈长杆状,走向趋于一致,相邻细胞间连接较紧密,相互融合。心肌细胞特异性蛋白α-actin和cTnT呈阳性表达,不同浓度的5-Aza诱导的MSCs心肌细胞特异性蛋白的表达率不一样,其中以10μmol.L-1组表达率最高。5-Aza作用兔MSCs体外培养可以诱导其向心肌样细胞分化,最佳浓度为10μmol.L-1。To investigate the method by which rabbit mesenchymal stem cells（MSCs）could be induced to cardiomyocytes and effect,MSCs isolated from rabbit femoral bone marrow were cultured in cell culture medium and in medium with 5-Azacytidine（5-Aza）.Whether the induced cells expressed cardiac-specific protein α-actin and cardiac troponin T（cTnT） was identified by immunocytochemistry and inverted microscope,and the differentiation rate in each group was calculated.The results showed that some MSCs gradually increased in size and formed a stick-like appearance after inducation in experimental group,one month after treatment,most of the long stick-like cells connected with adjoining cells and began to form myotube-like structures,the direction of the cells arraying was similar gradually.Cardiac cell-specific protein α-actin and cTnT expressed positively,the expression rate of cardiac cell-specific protein was different after MSCs was induced by different concentration 5-Aza,of which 10 μmol·L-1 5-Aza was the optimal concentration.These results indicate that MSCs can be induced to cardiomyocytes by 5-Aza in vitro,and which have the characteristics of cardiomyocytes in structure.The optimal concentration of 5-Aza inducing MSCs to cardiomyocytes is 10 μmol·L-1.

关 键 词：骨髓间充质干细胞 5-氮杂胞苷 心肌样细胞 兔 

分 类 号：S852.4[农业科学—基础兽医学]