Structural coupling of cardiomyocytes and noncardiomyocytes: quantitative comparisons using a novel micropatterned cell pair assay
Structural coupling of cardiomyocytes and noncardiomyocytes: quantitative comparisons using a novel micropatterned cell pair assay
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DOI：
10.1152/ajpheart.91531.2007
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发表时间：
2008-07-01
影响因子：
4.8
通讯作者：
Bursac, Nenad
中科院分区：
文献类型：
--
作者：
Pedrotty, Dawn M.;Klinger, Rebecca Y.;Bursac, Nenad
心肌细胞和其他细胞之间的结构和功能相互作用的良好对照研究对于理解心脏病理生理学和进一步开发安全有效的细胞疗法至关重要。我们建立了一种新的体外测定方法，该方法由大量具有可重现形状、大小和细胞-细胞接触区域的单个微图案化细胞对组成。应用该测定来定量和比较由心肌细胞（CM）、心脏成纤维细胞（CF）、骨骼肌成肌细胞（SKM）和间充质干细胞（MSC）组成的5，000个细胞对中电（连接蛋白43）和机械（N-钙粘蛋白）偶联蛋白的表达和分布的频率。我们发现，对于所有细胞对类型，两个细胞之间的侧-侧接触形成的频率是端-端接触的4.5-14.3倍。连接蛋白43和N-钙粘蛋白均在所有同型CM对中表达，但仅在13.4-91.6%的含有非心肌细胞的对中表达，其中表达是连接性的（在细胞-细胞接触部位）或扩散性的（在细胞质内）。CM表达在同型对中仅为交界性，但在异型对中主要为弥漫性。非心肌细胞同型对表现出弥漫性表达的1.7-8.7倍，经常比交界性表达，这是增加2.6-4.4倍，在异型对。连接蛋白43和N-钙粘蛋白表达分别见于38.6 +/- 7.3和39.6 +/- 6.2%的CM-MSC对，21.9 +/- 5.0和13.6 +/- 1.9%的CM-SKM对，以及仅3.8-9.6%的CM-CF对。蛋白质表达和分布的测量频率稳定至少4天。在微图案化细胞对中描述的研究为与心脏功能和细胞疗法相关的细胞相互作用提供了新的线索。
Well-controlled studies of the structural and functional interactions between cardiomyocytes and other cells are essential for understanding heart pathophysiology and for the further development of safe and efficient cell therapies. We established a novel in vitro assay composed of a large number of individual micropatterned cell pairs with reproducible shape, size, and region of cell-cell contact. This assay was applied to quantify and compare the frequency of expression and distribution of electrical (connexin43) and mechanical (N-cadherin) coupling proteins in 5,000 cell pairs made of cardiomyocytes (CMs), cardiac fibroblasts (CFs), skeletal myoblasts (SKMs), and mesenchymal stem cells (MSCs). We found that for all cell pair types, side-side contacts between two cells formed 4.5-14.3 times more often than end-end contacts. Both connexin43 and N- cadherin were expressed in all homotypic CM pairs but in only 13.4-91.6% of pairs containing noncardiomyocytes, where expression was either junctional (at the site of cell-cell contact) or diffuse (inside the cytoplasm). CM expression was exclusively junctional in homotypic pairs but predominantly diffuse in heterotypic pairs. Noncardiomyocyte homotypic pairs exhibited diffuse expression 1.7-8.7 times more often than junctional expression, which was increased 2.6-4.4 times in heterotypic pairs. Junctional connexin43 and N- cadherin expression, respectively, were found in 38.6 +/- 7.3 and 39.6 +/- 6.2% of CM-MSC pairs, 21.9 +/- 5.0 and 13.6 +/- 1.9% of CM-SKM pairs, and in only 3.8-9.6% of CM-CF pairs. Measured frequencies of protein expression and distribution were stable for at least 4 days. Described studies in micropatterned cell pairs shed new light on cellular interactions relevant for cardiac function and cell therapies.