Defined Engineered Human Myocardium With Advanced Maturation for Applications in Heart Failure Modeling and Repair.
Defined Engineered Human Myocardium With Advanced Maturation for Applications in Heart Failure Modeling and Repair.
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DOI：
10.1161/circulationaha.116.024145
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发表时间：
2017-05-09
期刊：
影响因子：
37.8
通讯作者：
Zimmermann WH
中科院分区：
文献类型：
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作者：
Tiburcy M;Hudson JE;Balfanz P;Schlick S;Meyer T;Chang Liao ML;Levent E;Raad F;Zeidler S;Wingender E;Riegler J;Wang M;Gold JD;Kehat I;Wettwer E;Ravens U;Dierickx P;van Laake LW;Goumans MJ;Khadjeh S;Toischer K;Hasenfuss G;Couture LA;Unger A;Linke WA;Araki T;Neel B;Keller G;Gepstein L;Wu JC;Zimmermann WH
推进干细胞衍生的心肌细胞的结构和功能成熟仍然是疾病建模、药物筛选和心脏修复应用的关键挑战。在这里，我们试图在特定条件下，将工程化人心肌（EHM）中的心肌细胞成熟推向成人表型。我们系统地研究了在无血清条件下从胚胎和诱导多能干细胞衍生的心肌细胞和具有器官型功能的成纤维细胞产生EHM的细胞组成、基质和培养基条件。我们采用形态学，功能和转录组分析基准成熟EHM。EHM显示了出生后心肌的重要结构和功能特征，包括：（1）具有M带的杆状心肌细胞组装成功能性合胞体;（2）收缩力与真实出生后心肌相似;（3）正性力-频率-反应;（4）对β-肾上腺素能刺激的正性肌力反应通过经典的β1-和β2-肾上腺素能受体信号通路介导;（5）心肌细胞的收缩力和收缩力与正常出生后心肌相似。和（5）通过转录组谱分析的先进分子成熟的证据。EHM对慢性儿茶酚胺毒性的反应是收缩功能障碍、心肌细胞肥大、心肌细胞死亡和NT-proBNP释放;所有这些都是心力衰竭的经典标志。此外，我们证明了EHM的可扩展性，根据预期的心脏修复的临床需求。我们提供了一种普遍适用的技术的概念验证，用于在限定的无血清条件下从胚胎和诱导多能干细胞衍生的心肌细胞工程化大规模人类心肌，用于疾病建模和心脏修复。
Advancing structural and functional maturation of stem cell-derived cardiomyocytes remains a key challenge for applications in disease modelling, drug screening, and heart repair. Here, we sought to advance cardiomyocyte maturation in engineered human myocardium (EHM) towards an adult phenotype under defined conditions. We systematically investigated cell composition, matrix and media conditions to generate EHM from embryonic and induced pluripotent stem cell-derived cardiomyocytes and fibroblasts with organotypic functionality under serum-free conditions. We employed morphological, functional, and transcriptome analyses to benchmark maturation of EHM. EHM demonstrated important structural and functional properties of postnatal myocardium, including: (1) rod-shaped cardiomyocytes with M-bands assembled as a functional syncytium; (2) systolic twitch forces at a similar level as observed in bona fide postnatal myocardium; (3) a positive force-frequency-response; (4) inotropic responses to β-adrenergic stimulation mediated via canonical β1- and β2-adrenoceptor signaling pathways; and (5) evidence for advanced molecular maturation by transcriptome profiling. EHM responded to chronic catecholamine toxicity with contractile dysfunction, cardiomyocyte hypertrophy, cardiomyocyte death, and NT-proBNP release; all are classical hallmarks of heart failure. Additionally, we demonstrate scalability of EHM according to anticipated clinical demands for cardiac repair. We provide proof-of-concept for a universally applicable technology for the engineering of macro-scale human myocardium for disease modelling and heart repair from embryonic and induced pluripotent stem cell-derived cardiomyocytes under defined, serum-free conditions.