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微环境对体外间充质干细胞向心肌样细胞分化的作用
Role of microenvironment on in vitro differentiation of mesenchymal stem cells into cardiomyogenic cells
【摘要】 目的研究心肌组织中何种细胞对骨髓间充质干细胞(MSCs)向心肌样细胞定向分化起决定性作用。方法分离培养骨髓MSCs、心肌细胞(CM)和内皮细胞(EC)。MSCs经BrdU标记后与CM、EC分别共培养和单独培养,通过形态学、免疫细胞化学和双重免疫细胞化学鉴定。结果分离培养的MSCs、CM和EC纯度高于95%。标记MSCs与CM共培养后,细胞具有心肌细胞样形态。4周时,有44%的细胞BrdU与肌动蛋白(sarcomeric actin)或连接蛋白-43(connexin-43)双重免疫细胞化学染色阳性。与EC共培养以及单独培养时,MSCs形态变化不明显,无双染阳性细胞出现。结论CM对骨髓MSCs向心肌样细胞的定向分化具有重要作用。
【Abstract】 Objective To find the cells in myocardium controlling in cardiomyogenic differentiation of marrow derived-mesenchymal stem cells(MSCs).Methods Rat marrow derived-MSCs,cardiomyocytes(CM)and endothelial cells(EC)were isolated and cultured,then MSCs were incubated with the CM or EC or cultured alone after labeled by 5-Bromo-2-deoxyuridine(BrdU)for 72 hours.The cells were identified by morphology,immunocytochemistry and double immunocytochemistry.Results More than 95% of the cells isolated and cultured belonged to MSCs,CM or EC.Incubated with CM,most MSCs labeled by BrdU showed cardiomyocyte-like morphology.After 4 weeks of cultivation,positive staining of double immunocytochemistry of BrdU and Sarcomeric Actin or Connexin-43(CX-43)were observed in 44% cells.However,there was no obvious change in morphology and no positive staining of double immunocytochemistry was recorded in MSCs incubated with EC or cultured alone after 1、2、3 and 4 weeks of cultivation.Conclusion MSCs can differentiate into cardiomyogenic cells in cardiomyocyte microenvironment and cardiomyocytes may play a crucial role in the differentiation.
【Key words】 bone marrow; mesenchymal stem cells; cardiomyocyte; endothelial cell;
- 【文献出处】 基础医学与临床 ,Basic & Clinical Medicine , 编辑部邮箱 ,2008年01期
- 【分类号】R329
- 【被引频次】1
- 【下载频次】135