Generation of a human iPSC-derived cardiomyocyte/fibroblast engineered heart tissue model.
Generation of a human iPSC-derived cardiomyocyte/fibroblast engineered heart tissue model.
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DOI：
10.12688/f1000research.139482.1
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发表时间：
2023
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动物模型已被证明是扩大我们对复杂心脏疾病的理解不可或缺的，但受到细胞生理学显著物种依赖性差异的阻碍。人诱导的多能干细胞衍生的心肌细胞（hiPSC-CM）在心脏疾病的建模中显示出巨大的前景，尽管在功能和结构成熟度方面存在限制。3D干细胞衍生的心脏模型代表了模拟心脏中存在的复杂微环境作为体外模型的一个步骤。将非肌细胞细胞类型（如心脏成纤维细胞）纳入工程化心脏组织模型（EHT）中可以帮助更好地重现人类心肌中存在的细胞与细胞和细胞与基质的相互作用。将人诱导的多能干细胞衍生的心脏成纤维细胞（hiPSC-CF）和hiPSC-CM整合到EHT模型中使得能够生成能够探索心脏病理生理学中存在的深奥的结构和电生理相互作用的遗传均质建模系统。此外，构建更多生理相关的3D心脏模型为心脏病研究中替代动物提供了巨大的潜力。在这里，我们描述了用于hiPSC-CM和hiPSC-CF的分化及其随后同化为EHT的有效且可重复的方案。所得EHT由纵向排列的iPSC-CM组成，与hiPSC-CF一起并入。与仅由hiPSC-CM组成的那些相比，具有hiPSC-CM和hiPSC-CF两者的EHT表现出较慢的搏动频率和增强的收缩力。修改后的协议可能有助于更好地阐明心肌中不同细胞类型之间的相互作用及其对结构重塑和心脏纤维化的贡献。
Animal models have proven integral to broadening our understanding of complex cardiac diseases but have been hampered by significant species-dependent differences in cellular physiology. Human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) have shown great promise in the modelling of cardiac diseases despite limitations in functional and structural maturity. 3D stem cell-derived cardiac models represent a step towards mimicking the intricate microenvironment present in the heart as an in vitro model. Incorporation of non-myocyte cell types, such as cardiac fibroblasts, into engineered heart tissue models (EHTs) can help better recapitulate the cell-to-cell and cell-to-matrix interactions present in the human myocardium. Integration of human-induced pluripotent stem cell-derived cardiac fibroblasts (hiPSC-CFs) and hiPSC-CM into EHT models enables the generation of a genetically homogeneous modelling system capable of exploring the abstruse structural and electrophysiological interplay present in cardiac pathophysiology. Furthermore, the construction of more physiologically relevant 3D cardiac models offers great potential in the replacement of animals in heart disease research. Here we describe efficient and reproducible protocols for the differentiation of hiPSC-CMs and hiPSC-CFs and their subsequent assimilation into EHTs. The resultant EHT consists of longitudinally arranged iPSC-CMs, incorporated alongside hiPSC-CFs. EHTs with both hiPSC-CMs and hiPSC-CFs exhibit slower beating frequencies and enhanced contractile force compared to those composed of hiPSC-CMs alone. The modified protocol may help better characterise the interplay between different cell types in the myocardium and their contribution to structural remodelling and cardiac fibrosis.