剪切应力与心肌细胞裂解液联合诱导骨髓间充质干细胞向心肌分化的研究  

Research on cardiomyogenic differentiation of bone marrow mesenchymal stem cells induced by shear stress combined with lysate of myocardial cells

作　　者：黄艳[1] 樊瑜波[1] 

机构地区：[1]北京航空航天大学生物与医学工程学院生物力学与力生物学教育部重点实验室,北京100191

出　　处：《医用生物力学》2011年第3期211-216,共6页Journal of Medical Biomechanics

基　　金：国家自然科学基金项目资助(10925208);高等学校博士学科点专项科研基金资助(20091102110031)

摘　　要：目的研究剪切应力与心肌细胞裂解液联合应用对骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)心肌向分化的影响。方法分离新生乳鼠的心肌细胞并制成心肌细胞裂解液,自成年大鼠骨髓中分离培养BMSCs,分为对照组(A组)、剪切应力组(B组)、心肌细胞裂解液组(C组)和剪切应力+心肌细胞裂解液组(D组)。用RT-RCR、免疫细胞化学染色和Western blotting检测诱导后BMSCs心肌细胞相关因子的表达。结果 B、C、D组均表达心肌细胞相关因子的mRNA和蛋白,D组表达明显高于B组或C组。结论剪切应力与心肌细胞裂解液联合应用可以更好地诱导BMSCs分化为心肌样细胞,为科研和临床应用提供种子细胞,有助于生物反应器的合理设计。Objective To investigate the effect of shear stress combined with lysate of myocardial cells on cardiomyogenic differentiation of bone marrow mesenchymal stem cells（BMSCs）.Methods Myocardial cells isolated from neonatal rats were lysed by repeated freezing and defrosting.BMSCs isolated from the marrow of adult rat tibia and femur were randomly assigned to four groups： Group A was BMSCs cultured in complete medium under static state as the control;Group B was BMSCs subjected to laminar shear stress with a parallel plate-type device;Group C was BMSCs induced by lysate of myocardial cells;Group D was BMSCs subjected to shear stress combined with lysate of myocardial cells.The cardiomyogenic differentiation of BMSCs was analyzed by RT-PCR,immunocytochemistry and Western blotting.Results Shear stress or/and lysate of myocardial cells treatment induced the expression of cardiomyocyte-related markers at both mRNA and protein levels.A combination of shear stress and lysate of myocardial cells had a stronger effect on the expression of cardiomyocyte-related markers than either treatment alone.Conclusions Shear stress combined with lysate of myocardial cells treatment could induce BMSC differentiation into cardiomyocytes,which provided an approach to obtain more committed differentiated cells for research or for clinic,and as reference for the design of appropriate biological reactor.

关 键 词：剪切应力 心肌细胞裂解液 骨髓间充质干细胞 心肌细胞 分化 

分 类 号：Q66[生物学—生物物理学]