Design and formulation of functional pluripotent stem cell-derived cardiac microtissues
Design and formulation of functional pluripotent stem cell-derived cardiac microtissues
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DOI：
10.1073/pnas.1311120110
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发表时间：
2013-12-03
影响因子：
11.1
通讯作者：
Zandstra, Peter W.
中科院分区：
文献类型：
--
作者：
Thavandiran, Nimalan;Dubois, Nicole;Zandstra, Peter W.
获得强大和信息丰富的人类心脏组织模型将加速基于药物的心脏病治疗策略。尽管付出了巨大的努力，但高保真成人样人类心脏组织类似物的生成仍然具有挑战性。我们使用组织收缩和组装力学的计算模型，并结合微制造的约束来指导设计排列的和功能性的3D人类多能干细胞(HPSC)来源的心脏微组织，我们称之为心脏微丝(CMWS)。该平台的微型化避免了组织血管形成的需要，并使基于图像的更高吞吐量的CMW药物反应性分析成为可能。可以使用机电刺激和细胞成分来调整CMW的组织属性。具体地说，控制3D组织在排列的胶原中的自组装，以及使用点刺激电极起搏，被发现可以促进心脏成熟相关基因的表达和体内类似电信号的传播。此外，筛选一系列hPSC来源的心肌细胞比率发现，75%的NKX2 Homeobox 5(NKX2-5)+心肌细胞和25%的分化簇90 OR(CD90)+非心肌细胞优化了组织重塑动力学，并产生了增强的结构和功能特性。最后，我们展示了优化的平台在心律失常的心动过速模型中的实用性，这是心脏电生理学的一个方面，以前没有在3D体外hPSC衍生的心脏微组织模型中概括。我们的CMW平台确定的设计标准应该会加速人体心脏组织功能体外预测性分析的发展。
Access to robust and information-rich human cardiac tissue models would accelerate drug-based strategies for treating heart disease. Despite significant effort, the generation of high-fidelity adult-like human cardiac tissue analogs remains challenging. We used computational modeling of tissue contraction and assembly mechanics in conjunction with microfabricated constraints to guide the design of aligned and functional 3D human pluripotent stem cell (hPSC)-derived cardiac microtissues that we term cardiac microwires (CMWs). Miniaturization of the platform circumvented the need for tissue vascularization and enabled higher-throughput image-based analysis of CMW drug responsiveness. CMW tissue properties could be tuned using electromechanical stimuli and cell composition. Specifically, controlling self-assembly of 3D tissues in aligned collagen, and pacing with point stimulation electrodes, were found to promote cardiac maturation-associated gene expression and in vivo-like electrical signal propagation. Furthermore, screening a range of hPSC-derived cardiac cell ratios identified that 75% NKX2 Homeobox 5 (NKX2-5)+ cardiomyocytes and 25% Cluster of Differentiation 90 OR (CD90)+ nonmyocytes optimized tissue remodeling dynamics and yielded enhanced structural and functional properties. Finally, we demonstrate the utility of the optimized platform in a tachycardic model of arrhythmogenesis, an aspect of cardiac electrophysiology not previously recapitulated in 3D in vitro hPSC-derived cardiac microtissue models. The design criteria identified with our CMW platform should accelerate the development of predictive in vitro assays of human heart tissue function.