Excitation propagation in three-dimensional engineered hearts using decellularized extracellular matrix
Excitation propagation in three-dimensional engineered hearts using decellularized extracellular matrix
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DOI：
10.1016/j.biomaterials.2014.05.080
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发表时间：
2014-09-01
期刊：
影响因子：
14
通讯作者：
Komuro, Issei
中科院分区：
文献类型：
--
作者：
Yasui, Haruyo;Lee, Jong-Kook;Komuro, Issei
利用脱细胞细胞细胞外基质构建三维心脏组织可能是一种新的技术，可以创造一个“器官样”的心脏结构。然而，要设计出功能上与天然心脏相当的人工心脏，还有许多问题有待解决，包括稳定的兴奋传播。为了阐明这一点，我们研究了工程组织的传导特性。我们用新生大鼠心肌细胞重新填充脱细胞心脏，然后，我们使用高分辨率相机观察自发搏动的兴奋传播。我们还进行了免疫荧光染色，以检查形态学方面。活组织成像显示GFP标记的分离的心肌细胞通过冠状动脉外渗迁移到间质空间。用表达Ca2+指示蛋白（GCaMP2）的心脏细胞重新填充的工程心脏进行光学成像实验。尽管工程心脏通常显示出良好组织的稳定兴奋传播，但心脏也表现出致心律失常的倾向，如无序传播。免疫荧光研究显示心肌细胞、内皮细胞和平滑肌细胞随机混合排列。再细胞化心肌细胞排列紊乱，连接蛋白43表达明显减少。总之，我们成功地证明了再细胞化的心脏作为一个“完整的器官”表现出动态的兴奋传播。我们的策略可以为构建3D工程心脏提供必要的信息，在功能上与天然心脏相当。(C)2014爱思唯尔有限公司版权所有。
Engineering of three-dimensional (3D) cardiac tissues using decellularized extracellular matrix could be a new technique to create an "organ-like" structure of the heart. To engineer artificial hearts functionally comparable to native hearts, however, much remain to be solved including stable excitation-propagation. To elucidate the points, we examined conduction properties of engineered tissues. We repopulated the decellularized hearts with neonatal rat cardiac cells and then, we observed excitation-propagation of spontaneous beatings using high resolution cameras. We also conducted immunofluorescence staining to examine morphological aspects. Live tissue imaging revealed that GFP-labeled-isolated cardiac cells were migrated into interstitial spaces through extravasation from coronary arteries. Engineered hearts repopulated with Ca2+-indicating protein (GCaMP2)-expressing cardiac cells were subjected to optical imaging experiments. Although the engineered hearts generally showed well-organized stable excitation-propagation, the hearts also demonstrated arrhythmogenic propensity such as disorganized propagation. Immunofluorescence study revealed randomly-mixed alignment of cardiomyocytes, endothelial cells and smooth muscle cells. The recellularized hearts also showed disarray of cardiomyocytes and markedly decreased expression of connexin43. In conclusion, we successfully demonstrated that the recellularized hearts showed dynamic excitation-propagation as a "whole organ". Our strategy could provide prerequisite information to construct a 3D-engineered heart, functionally comparable to the native heart. (C) 2014 Elsevier Ltd. All rights reserved.