Introduction

The definitive treatment for severe heart failure is transplantation. However, only a small number of heart transplants are performed each year due to donor shortages. Therefore, novel treatment approaches based on artificial organs or regenerative therapy are being developed as alternatives. We have developed a technology known as cell sheet-based tissue engineering that enables the fabrication of functional three-dimensional (3D) tissue. Here, we report a new technique for engineering human cardiac tissue with perfusable blood vessels. Our method involved the layering of cardiac cell sheets derived from human induced pluripotent stem cells (hiPSCs) on a vascular bed derived from porcine small intestinal tissue.

Methods

For the vascular bed, a segment of porcine small intestine was harvested together with a branch of the superior mesenteric artery and a branch of the superior mesenteric vein. The small intestinal tissue was incised longitudinally, and the mucosa was resected. Human cardiomyocytes derived from hiPSCs were co-cultured with endothelial cells and fibroblasts on a temperature-responsive dish and harvested as a cardiac cell sheet. A triple-layer of cardiac cell sheets was placed onto the vascular bed, and the resulting construct was subjected to perfusion culture in a bioreactor system.

Results

The cardiac tissue on the vascular bed pulsated spontaneously and synchronously after one day of perfusion culture. Electrophysiological recordings revealed regular action potentials and a beating rate of 105 ± 13/min (n = 8). Furthermore, immunostaining experiments detected partial connection of the blood vessels between the vascular bed and cardiac cell sheets.

Conclusions

We succeeded in engineering spontaneously beating 3D cardiac tissue in vitro using human cardiac cell sheets and a vascular bed derived from porcine small intestine. Further development of this method might allow the fabrication of functional cardiac tissue that could be used in the treatment of severe heart failure.

中文翻译：

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生成用于体外创建三维心脏组织的大规模血管床。

介绍

严重心力衰竭的最终治疗是移植。然而，由于供体短缺，每年只进行少量心脏移植。因此，正在开发基于人工器官或再生疗法的新型治疗方法作为替代方案。我们开发了一种称为基于细胞片的组织工程技术，可以制造功能性三维 (3D) 组织。在这里，我们报告了一种用可灌注血管设计人体心脏组织的新技术。我们的方法涉及在源自猪小肠组织的血管床上对源自人类诱导多能干细胞 (hiPSC) 的心脏细胞片层进行分层。

方法

对于血管床，取一段猪小肠以及肠系膜上动脉的一个分支和肠系膜上静脉的一个分支。纵向切开小肠组织，切除黏膜。源自 hiPSC 的人心肌细胞与内皮细胞和成纤维细胞在温度响应培养皿上共培养，并作为心脏细胞片收获。将三层心脏细胞片放置在血管床上，并将所得构建体在生物反应器系统中进行灌注培养。

结果

灌注培养一天后，血管床上的心脏组织自发、同步地搏动。电生理记录显示有规律的动作电位和 105 ± 13/min (n = 8) 的跳动率。此外，免疫染色实验检测到血管床和心脏细胞片之间的血管部分连接。

结论

我们使用人类心脏细胞片和源自猪小肠的血管床成功地在体外设计了自发跳动的 3D 心脏组织。这种方法的进一步发展可能允许制造可用于治疗严重心力衰竭的功能性心脏组织。