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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

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【作者】 张本斯李庄杨新文洪虹洪永华

【Author】 ZHANG Ben-si1,LI Zhuang2,YANG Xin-wen1,HONG Hong1,HONG Yong-hua1(1.Department of Anatomy,Faculty of Basic Medicine;2.Department of Thoracic and Cardiovascular Surgery,the Affiliated Hospital,Dali University,Dali 671000,China)

【机构】 大理学院基础医学院人体解剖学教研室大理学院附属医院胸心外科大理学院基础医学院人体解剖学教研室 云南大理671000云南大理671000

【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.

【基金】 云南省教育厅科研基金(04Y142C)
  • 【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
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