Tissue-engineered human embryonic stem cell-containing cardiac patches: evaluating recellularization of decellularized matrix
Tissue-engineered human embryonic stem cell-containing cardiac patches: evaluating recellularization of decellularized matrix
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DOI：
10.1177/2041731420921482
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发表时间：
2020-07-01
影响因子：
8.2
通讯作者：
de Carvalho, Antonio Carlos Campos
中科院分区：
文献类型：
--
作者：
Hochman-Mendez, Camila;Pereira de Campos, Dilza Balteiro;de Carvalho, Antonio Carlos Campos
脱细胞心脏细胞外基质支架具有保留的组成和结构，可用于组织工程中复制复杂的心脏细胞外基质。然而，评估再细胞化尝试后脱细胞心脏细胞外基质支架的心肌细胞再增殖的程度是具有挑战性的。在这里，我们描述了一个独特的组合的生化，生物力学，组织学和生理参数的量化再细胞化效率的组织工程心脏补丁相比，天然心脏组织。将人胚胎干细胞来源的心肌细胞接种于大鼠心房和心室去细胞的心肌细胞外基质补片中。共聚焦和原子力显微镜显示细胞外基质基底膜内的细胞整合，伴随着天然心脏组织被动力学性能的恢复。多电极阵列和免疫染色（连接蛋白43）被用来确定与电耦合的同步场电位。肌红蛋白含量（类似于60%）和肌节长度测量（>45% vs 2D培养）用于评价整合细胞的心肌细胞成熟。这些技术的组合使我们能够证明，随着细胞化效率的提高，心肌细胞成熟并同步电活动，组织的机械/生物化学性质向天然组织的那些改善。
Decellularized cardiac extracellular matrix scaffolds with preserved composition and architecture can be used in tissue engineering to reproduce the complex cardiac extracellular matrix. However, evaluating the extent of cardiomyocyte repopulation of decellularized cardiac extracellular matrix scaffolds after recellularization attempts is challenging. Here, we describe a unique combination of biochemical, biomechanical, histological, and physiological parameters for quantifying recellularization efficiency of tissue-engineered cardiac patches compared with native cardiac tissue. Human embryonic stem cell-derived cardiomyocytes were seeded into rat heart atrial and ventricular decellularized cardiac extracellular matrix patches. Confocal and atomic force microscopy showed cell integration within the extracellular matrix basement membrane that was accompanied by restoration of native cardiac tissue passive mechanical properties. Multi-electrode array and immunostaining (connexin 43) were used to determine synchronous field potentials with electrical coupling. Myoglobin content (similar to 60%) and sarcomere length measurement (>45% vs 2D culture) were used to evaluate cardiomyocyte maturation of integrated cells. The combination of these techniques allowed us to demonstrate that as cellularization efficiency improves, cardiomyocytes mature and synchronize electrical activity, and tissue mechanical/biochemical properties improve toward those of native tissue.