FRESH™ 3D bioprinted cardiac tissue, a bioengineered platform for in vitro pharmacology.
FRESH™ 3D bioprinted cardiac tissue, a bioengineered platform for in vitro pharmacology.
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作者：
在药物开发中迫切需要人类心脏生理学的预测模型来评估化合物对人体组织的影响。心肌细胞的体外二维单层培养物提供了生化和细胞读数，体内动物模型提供了全身心血管反应的信息。然而，由于这些模型对成人心血管生理学的不完全再现，因此在这些模型中仍然存在显著的差距。最近在从工程心脏组织开发体外模型方面的努力已经证明了在三维组织结构中使用人诱导多能干细胞衍生的心肌细胞（hiPSC-CM）来弥合这一差距的潜力。在这里，我们通过实施FRESH™ 3D生物打印来推进这一范例，以中等通量、孔板格式构建人类心脏组织，该组织具有受控的组织结构、定制的细胞组成和类似天然的生理功能，特别是在其药物反应中。我们将hiPSC-CM、内皮细胞和成纤维细胞组合在细胞生物墨水中，并且FRESH™ 3D以稳定在组织夹具上的薄组织条的形式生物打印该混合物。我们表明，心脏组织可以直接在24孔板格式中制造，由密度和高度对齐的hiPSC-CM组成，> 6亿个细胞/mL，并且在14天内，表现出可再现的钙瞬变和16 cm/s的快速传导速度。用β-肾上腺素能受体激动剂异丙肾上腺素询问这些心脏组织，显示与正变时性和变力性一致的反应。用钙通道阻滞剂维拉帕米治疗证明了hiPSC-CM衍生的心脏组织的预期反应。这些结果证实，FRESH™ 3D生物打印心脏组织代表了一个提供人体生理反应数据的体外平台。
There is critical need for a predictive model of human cardiac physiology in drug development to assess compound effects on human tissues. In vitro two-dimensional monolayer cultures of cardiomyocytes provide biochemical and cellular readouts, and in vivo animal models provide information on systemic cardiovascular response. However, there remains a significant gap in these models due to their incomplete recapitulation of adult human cardiovascular physiology. Recent efforts in developing in vitro models from engineered heart tissues have demonstrated potential for bridging this gap using human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) in three-dimensional tissue structure. Here, we advance this paradigm by implementing FRESH™ 3D bioprinting to build human cardiac tissues in a medium throughput, well-plate format with controlled tissue architecture, tailored cellular composition, and native-like physiological function, specifically in its drug response. We combined hiPSC-CMs, endothelial cells, and fibroblasts in a cellular bioink and FRESH™ 3D bioprinted this mixture in the format of a thin tissue strip stabilized on a tissue fixture. We show that cardiac tissues could be fabricated directly in a 24-well plate format were composed of dense and highly aligned hiPSC-CMs at >600 million cells/mL and, within 14 days, demonstrated reproducible calcium transients and a fast conduction velocity of ∼16 cm/s. Interrogation of these cardiac tissues with the β-adrenergic receptor agonist isoproterenol showed responses consistent with positive chronotropy and inotropy. Treatment with calcium channel blocker verapamil demonstrated responses expected of hiPSC-CM derived cardiac tissues. These results confirm that FRESH™ 3D bioprinted cardiac tissues represent an in vitro platform that provides data on human physiological response.
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影响因子：
4.3
作者：
Yoshida, Azumi;Sekine, Waki;Homma, Jun;Sekine, Hidekazu;Itoyama, Yu Yamasaki;Sasaki, Daisuke;Matsuura, Katsuhisa;Kobayashi, Eiji;Shimizu, Tatsuya
通讯作者：
Shimizu, Tatsuya
影响因子：
48
作者：
Cho, Sangkyun;Discher, Dennis E.;Wu, Joseph C.
通讯作者：
Wu, Joseph C.
影响因子：
13.6
作者：
Hinton TJ;Jallerat Q;Palchesko RN;Park JH;Grodzicki MS;Shue HJ;Ramadan MH;Hudson AR;Feinberg AW
通讯作者：
Feinberg AW
影响因子：
16.1
作者：
Feric NT;Radisic M
通讯作者：
Radisic M
影响因子：
4.3
作者：
Burnett SD;Blanchette AD;Chiu WA;Rusyn I
通讯作者：
Rusyn I