摘要
目的:探讨5-氮胞苷和心肌内环境对骨髓基质干细胞(mesenchymalstemcells,MSCs)向心肌细胞转化的影响。方法:体外分离培养纯化MSCs,用Brdu标记MSCs,用不同浓度的5-氮胞苷诱导MSCs3周后,计算心肌细胞转化率。利用免疫组化方法测定诱导后MSCs中心肌特异性抗体(肌钙蛋白Ⅰ,肌凝蛋白重链)的存在。在体实验中,将体外标记的MSCs分为对照组,未诱导组,诱导组,移植给正常鼠,于移植后第1,2和4周取鼠心肌组织的HE病理切片镜下观察,使用免疫组化法检测Brdu阳性细胞中心肌特异性抗体的存在并观察MSCs向心肌细胞转化情况。结果:5-氮胞苷可以在体外诱导MSCs分化为心肌样细胞,而且在5μmol/L浓度的情况下,心肌样细胞转化率最高。同时,心肌内环境也可以诱导MSCs分化为心肌样细胞,而且避免了5-氮胞苷的副作用。结论:MSCs在5-氮胞苷和心肌内环境的作用下均可以分化为心肌样细胞,并具有心肌特异性蛋白结构,可以作为心肌细胞移植的理想种子来源。
AIM:To investigate the influence of 5-azacytidine(5-Aza) and myocardial internal environment on the differentiation of bone marrow mesenchymal stem cells(MSCs) into cardiomyocytes. METHODS:Bone marrow MSCs were isolated,cultured,purified and then labeled with Brdu in vitro.The ratio of MSCs differentiated into cardiomyocytes was calculated after induced with 5-Aza of different density for three weeks in vitro.The specific cardiomyocyte antibody(Troponin Ⅰand myosin heavy chain) in induced MSCs was testified by immunohistochemical method.In the experiment in vivo,the labeled MSCs were divided into three groups:control group,non-induction group and induction group.The labeled MSCs were transplanted into normal mice respectively,and then the HE pathological sections were observed under the microscope at 1,2 and 4 weeks after transplantation.The specific cardiomyocyte antibody in Brdu positive cells objectively was tested with immunohistochemical method,and the differentiation of MSCs into cardiomyocytes was observed. RESULTS:MSCs could be differentiated into cardiomyocytes by 5-Aza in vitro,and reached the highest differentiation ratio when 5-Aza at 5 μmol/L. Moreover,the myocardial internal environment could induce MSCs to differentiate into cardiomyocytes and avoid the side effects caused by 5-Aza. CONCLUSION:MSCs can differentiate into cardiomyocytes under the condition of 5-Aza and in the myocardial internal environment.The induced MSCs express the specific cardiomyocyte antibody,which can be as the better seed cells for the transplantation of cardiomyocytes.
出处
《中国临床康复》
CAS
CSCD
北大核心
2005年第3期140-142,i002,共4页
Chinese Journal of Clinical Rehabilitation
基金
国家自然科学基金资助项目(30170369)
黑龙江省自然科学基金资助项目(9826)~~
