Eppenberger H M, Zuppinger C
Department of Biology, Institute of Cell Biology, Swiss Federal Institute of Technology ETH, CH-8093 Zurich, Switzerland.
FASEB J. 1999;13 Suppl:S83-9. doi: 10.1096/fasebj.13.9001.s83.
Primary adult rat cardiomyocytes (ARC)in culture are shown to be a model system for cardiac cell hypertrophy in vitro. ARC undergo a process of morphological transformation and grow only by increase in cell size, however, without loss of the cardiac phenotype. The isolated cells spread and establish new cell-cell contacts, eventually forming a two-dimensional heart tissue-like synchronously beating cell sheet. The reformation of specific cell contacts (intercalated disks) is shown also between ventricular and atrial cardiomyocytes by using antibodies against the gap junction protein connexin-43 and after microinjection into ARC of N-cadherin cDNA fused to reporter green fluorescent protein (GFP) cDNA. The expressed fusion protein allowed the study of live cell cultures and of the dynamics of the adherens junction protein N-cadherin during the formation of new cell-cell contacts. The possible use of the formed ARC cell-sheet cells under microgravity conditions as a test system for the reformation of the cytoskeleton of heart muscle cells is proposed.
培养的原代成年大鼠心肌细胞(ARC)被证明是体外心脏细胞肥大的模型系统。ARC经历形态转化过程,仅通过细胞大小增加而生长,然而,不会丧失心脏表型。分离的细胞铺展并建立新的细胞间接触,最终形成二维心脏组织样同步跳动的细胞片。通过使用针对间隙连接蛋白连接蛋白-43的抗体以及将与报告绿色荧光蛋白(GFP)cDNA融合的N-钙黏蛋白cDNA显微注射到ARC中,还显示了心室和心房心肌细胞之间特定细胞接触(闰盘)的重建。表达的融合蛋白允许对活细胞培养物以及新细胞间接触形成过程中黏附连接蛋白N-钙黏蛋白的动力学进行研究。有人提出,在微重力条件下将形成的ARC细胞片细胞用作心肌细胞细胞骨架重建的测试系统。