Cardiomyocytes can be generated from marrow stromal cells in vitro
Cardiomyocytes can be generated from marrow stromal cells in vitro
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DOI：
10.1172/jci5298
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发表时间：
1999-03-01
影响因子：
15.9
通讯作者：
Ogawa, S
中科院分区：
文献类型：
--
作者：
Makino, S;Fukuda, K;Ogawa, S
我们从小鼠骨髓基质细胞中分离出一个心肌细胞系（CMG）。将基质细胞永生化，用5-氮杂胞苷处理，并反复筛选自发跳动的细胞。细胞表现出成纤维细胞样的形态，但在5-氮胞苷处理后，类似于30%的细胞的形态发生了变化; 1周后它们与相邻细胞连接，形成肌管样结构，2周后开始自发搏动，3周后同步搏动。它们表达心房利钠肽和脑利钠肽，并用抗肌球蛋白、抗结蛋白和抗辅肌动蛋白抗体染色。电镜下可见心肌样超微结构，包括典型的肌节、位于中央的细胞核和心房颗粒。这些细胞具有几种类型的动作电位，如窦房结样和心室细胞样动作电位。所有细胞都有一个长的动作电位持续时间或平台，一个相对浅的静息膜电位，和起搏器样舒张晚期缓慢去极化。对肌球蛋白重链、肌球蛋白轻链和α-肌动蛋白等收缩蛋白基因亚型的分析表明，它们的肌肉表型与胎儿心室肌细胞的表型相似。这些细胞在5-氮胞苷处理前表达Nkx 2.5/Csx、GATA 4、TEF-1和MEF-2C mRNA，在处理后表达MEF-2A和MEF-2D。这种新的细胞系为研究心肌细胞分化提供了一个强有力的模型。
We have isolated a cardiomyogenic cell line (CMG) from murine bone marrow stromal cells. Stromal cells were immortalized, treated with 5-azacytidine, and spontaneously beating cells were repeatedly screened. The cells showed a fibroblast-like morphology, but the morphology changed after 5-azacytidine treatment in similar to 30% of the cells; they connected with adjoining cells after one week, formed myotube-like structures, began spontaneously beating after two weeks, and beat synchronously after three weeks. They expressed atrial natriuretic peptide and brain natriuretic peptide and were stained with anti-myosin, anti-desmin, and anti-actinin antibodies. :Electron microscopy revealed a cardiomyocyte-like ultrastructure, including typical sarcomeres, a centrally positioned nucleus, and atrial granules. These cells bad several types of action potentials, such as sinus node-like and ventricular cell-like action potentials. All cells had a long action potential duration or plateau, a relatively shallow resting membrane potential, and a pacemaker-like late diastolic slow depolarization. Analysis of the isoform of contractile protein genes, such as myosin heavy chain, myosin light chain, and a-actin, indicated that their muscle phenotype was similar to that of fetal ventricular cardiomyocytes. These cells expressed Nkx2.5/Csx, GATA4, TEF-1, and MEF-2C mRNA before 5-azacytidine treatment and expressed MEF-2A and MEF-2D after treatment. This new cell line provides a powerful model for the study of cardiomyocyte differentiation.