In vitro circulation model driven by tissue-engineered dome-shaped cardiac tissue
In vitro circulation model driven by tissue-engineered dome-shaped cardiac tissue
复制标题
DOI：
10.1088/1758-5090/ac77c1
复制
发表时间：
2022-07-01
期刊：
影响因子：
9
通讯作者：
Shimizu, Tatsuya
中科院分区：
文献类型：
--
作者：
Kikuchi, Tetsutaro;Matsuura, Katsuhisa;Shimizu, Tatsuya
心脏是动物和人类的重要器官。随着多能干细胞可用性的增加，由培养的心肌细胞组成的三维心脏组织在体外药物评价中的应用已得到广泛研究。已经提出了几种模型来实现泵功能，这是心脏的原始功能。然而，还没有使用培养的心脏组织来模拟人体循环系统的模型。这项研究表明，利用细胞片堆叠技术制造的圆顶形心脏组织可以模仿人体循环系统，实现类似心脏的泵功能并循环培养基。首先，将人诱导多能干细胞分化为自主跳动的心肌细胞，并使用温度响应型培养皿创建心肌细胞细胞片。使用细胞片操纵器堆叠心肌细胞片和人真皮成纤维细胞片。然后将这种两层细胞片充气，形成基底直径为 8 毫米的圆顶形心脏组织。圆顶状心脏组织的体积随着自主跳动而变化。每搏输出量随着培养时间的延长而增加，并在第 21 天达到 21 +/- 8.9 μl (n = 6)。它还对 β 刺激剂和细胞外钙浓度有反应。还通过专用培养装置在等容条件下记录内部压力波动。第 19 天，在 0.5 mmHg 的基础压力下，脉动压力的峰值高度为 0.33 +/- 0.048 mmHg (n = 3)。当组织连接到应用了止回阀的流路时，它驱动平均流速约为 1 μl s(-1) 的定向流。此外，通过在三种流体阻力条件下同时测量压力和体积的变化来创建压力-体积（P-V）图。总之，该心脏模型可用于驱动多器官芯片的生物泵，并可使用 P-V 图进行更准确的体外药物评估。
The heart is an essential organ for animals and humans. With the increased availability of pluripotent stem cells, the use of three-dimensional cardiac tissues consisting of cultured cardiomyocytes in in vitro drug evaluation has been widely studied. Several models have been proposed for the realization of the pump function, which is the original function of the heart. However, there are no models that simulate the human circulatory system using cultured cardiac tissue. This study shows that a dome-shaped cardiac tissue fabricated using the cell sheet stacking technique can achieve a heart-like pump function and circulate culture medium, there by mimicking the human circulatory system. Firstly, human induced pluripotent stem cells were differentiated into autonomously beating cardiomyocytes, and cardiomyocyte cell sheets were created using temperature-responsive culture dishes. A cardiomyocyte sheet and a human dermal fibroblast sheet were stacked using a cell sheet manipulator. This two-layered cell sheet was then inflated to create a dome-shaped cardiac tissue with a base diameter of 8 mm. The volume of the dome-shaped cardiac tissue changed according to the autonomous beating. The stroke volume increased with the culture period and reached 21 +/- 8.9 mu l (n = 6) on day 21. It also responded to beta-stimulant and extracellular calcium concentrations. Internal pressure fluctuations were also recorded under isovolumetric conditions by dedicated culture devices. The peak heights of pulsatile pressure were 0.33 +/- 0.048 mmHg (n = 3) under a basal pressure of 0.5 mmHg on day 19. When the tissue was connected to a flow path that had check valves applied, it drove a directional flow with an average flow rate of approximately 1 mu l s(-1). Furthermore, pressure-volume (P-V) diagrams were created from the simultaneous measurement of changes in pressure and volume under three conditions of fluidic resistance. In conclusion, this cardiac model can potentially be used for biological pumps that drive multi-organ chips and for more accurate in vitro drug evaluation using P-V diagrams.