Bioengineering of whole hearts using human embryonic stem cells (hESCs)-derived cardiovascular progenitor cells (CPCs) and natural matrices is a promising approach to overcome organ donor shortage threatening millions of patients awaiting for heart transplantation. Here, we developed a novel strategy for generation of heart constructs by repopulating engineered decellularized rat hearts using hESCs-derived CPCs. Careful expansion of CPCs in a scalable stirred-suspension bioreactor combined with step-wise seeding (60 million cells in 3 steps of 20 million per 1.5 h) onto decellularized hearts containing immobilized basic fibroblast growth factor (bFGF) resulted in improved retention of CPCs and differentiation to cardiomyocytes, smooth muscle cells and endothelial cells as evaluated by immunohistochemistry and qRT-PCR. We observed spontaneous and synchronous contractions of humanized hearts after 12 days of perfusion as well as advanced alignment of myofilaments. Our study provides a robust platform for generation of artificial human hearts and resolves major bottlenecks hindering further development of this technology.

中文翻译：

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人类胚胎干细胞衍生的心血管祖细胞在bFGF束缚的天然基质中有效定居以构建收缩的人源化大鼠心脏

使用人类胚胎干细胞（hESCs）衍生的心血管祖细胞（CPC）和天然基质对整个心脏进行生物工程是一种有前途的方法，可以克服器官供体短缺的问题，这种短缺威胁着数百万等待心脏移植的患者。在这里，我们通过使用hESCs衍生的CPC重新填充工程化的脱细胞大鼠心脏，从而开发了一种生成心脏结构的新策略。在可扩展的搅拌悬浮生物反应器中小心地扩展CPCs，并在包含固定的碱性成纤维细胞生长因子（bFGF）的脱细胞心脏上分步播种（3个步骤，以6000万个细胞分3个步骤，每1.5h 2000万个），从而改善了CPCs和通过免疫组织化学和qRT-PCR评估可分化为心肌细胞，平滑肌细胞和内皮细胞。我们观察到人源化心脏的自发性和同步性收缩，经过12天的灌注以及肌丝的提前排列。我们的研究为生成人造人类心脏提供了一个强大的平台，并解决了阻碍该技术进一步发展的主要瓶颈。