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骨髓基质细胞向心肌样细胞分化的体外研究。

In vitro study of the differentiation of bone marrow stromal cells into cardiomyocyte-like cells.

机构信息

Department of Clinical Biochemistry, Faculty of Medicine, Ilam University of Medical Sciences, Ilam, Iran.

出版信息

Mol Cell Biochem. 2012 Feb;361(1-2):315-20. doi: 10.1007/s11010-011-1117-6. Epub 2011 Oct 19.

Abstract

Bone marrow multipotent stromal cells (BMSCs) have the ability to transdifferentiate into various cell types, including: osteoblasts, chondrocytes, adipocytes, neurons, and cardiomyocytes. This study aimed to differentiate the BMSCs into cardiomyocyte. BMSCs were exposed to 5-azacytidine for 24 h. Seven days after the induction of cell differentiation by 5-azacytidine, the cardiomyogenic cells were stained by fushin and binucleated cells were counted and compared with the neonate cardiomyocyte as positive control. In addition, immunofluorescence analysis and western blot were performed using the antibodies against α-actinin, desmin, troponin T, and β-myosin heavy chain. Our results showed that there was no significant difference between the number of binucleated cells within the cardiomyogenic cell group and positive control group; however, a statistically significant difference was observed between both of these groups and undifferentiated cell group (P < 0.005). In addition, after 5-azacytidine treatment, BMSCs had a higher expression of cardiac-specific markers such as desmin, α-actinin, troponin T and β-myosin heavy chain compared with the untreated groups (P < 0.005). We concluded that 5-azacytidine is an effective inducer for the differentiation of BMSCs into cardiomyocytes and could produce a population of binucleated cells, which express α-actinin, desmin, troponin T, and β-myosin heavy chain, four markers of cardiomyocytes.

摘要

骨髓间充质干细胞(BMSCs)具有向多种细胞类型转分化的能力,包括成骨细胞、软骨细胞、脂肪细胞、神经元和心肌细胞。本研究旨在将 BMSCs 分化为心肌细胞。BMSCs 用 5-氮杂胞苷处理 24 小时。在 5-氮杂胞苷诱导细胞分化 7 天后,用 fushin 对心肌细胞进行染色,并计数双核细胞,并与新生心肌细胞作为阳性对照进行比较。此外,使用抗α-肌动蛋白、结蛋白、肌钙蛋白 T 和β-肌球蛋白重链的抗体进行免疫荧光分析和 Western blot。我们的结果表明,心肌细胞组和阳性对照组的双核细胞数量之间没有显著差异;然而,这两个组与未分化细胞组之间存在统计学差异(P < 0.005)。此外,在 5-氮杂胞苷处理后,BMSCs 中与心脏特异性标志物如结蛋白、α-肌动蛋白、肌钙蛋白 T 和β-肌球蛋白重链的表达水平高于未经处理的对照组(P < 0.005)。我们得出结论,5-氮杂胞苷是诱导 BMSCs 分化为心肌细胞的有效诱导剂,并可产生一群表达α-肌动蛋白、结蛋白、肌钙蛋白 T 和β-肌球蛋白重链的双核细胞,这四种标志物均为心肌细胞的标志物。

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