Department of Integrative Bioscience and Biomedical Engineering, Graduate School of Advanced Science and Engineering, TWIns, Waseda University, Tokyo, Japan.
Department of Modern Mechanical Engineering, School of Creative Science and Engineering, TWIns, Waseda University, Tokyo, Japan.
Tissue Eng Part C Methods. 2020 Sep;26(9):485-492. doi: 10.1089/ten.TEC.2020.0164.
Three-dimensional (3D) cardiac tissue reconstruction using tissue engineering technology is a rapidly growing area of regenerative medicine and drug screening development. However, there remains an urgent need for the development of a method capable of accurately measuring the contractile force of physiologically relevant 3D myocardial tissues to facilitate the prediction of human heart tissue drug sensitivity. To this end, our laboratory has developed a novel drug screening model that measures the contractile force of cardiac cell sheets prepared using temperature-responsive culture dishes. To circumvent the difficulties that commonly arise during the stacking of cardiomyocyte sheets, we established a stacking method using centrifugal force, making it possible to measure 3D myocardial tissue. Human induced pluripotent stem cell-derived cardiomyocytes were seeded in a temperature-responsive culture dish and processed into a sheet. The cardiac cell sheets were multilayered to construct 3D cardiac tissue. Measurement of the contractile force and cross-sectional area of the multilayered 3D cardiac tissue were then obtained and used to determine the relationship between the cross-sectional area of the cardiac tissue and its contractile force. The contractile force of the 1-, 3-, and 5-layer tissues increased linearly in proportion to the cross-sectional area. A result of 6.4 mN/mm, accounting for one-seventh of the contractile force found in adult tissue, was obtained. However, with 7-layer tissues, there was a sudden drop in the contractile force, possibly because of limited oxygen and nutrient supply. In conclusion, we established a method wherein the thickness of the cell sheets was controlled through layering, thus enabling accurate evaluation of the cardiac contractile function. This method may enable comparisons with living heart tissue while providing information applicable to regenerative medicine and drug screening models.
使用组织工程技术进行三维（3D）心脏组织重建是再生医学和药物筛选开发的一个快速发展领域。然而，仍然迫切需要开发一种能够准确测量生理相关 3D 心肌组织收缩力的方法，以促进对人体心脏组织药物敏感性的预测。为此，我们实验室开发了一种新的药物筛选模型，该模型可测量使用温度响应培养皿制备的心肌细胞片的收缩力。为了避免在堆积心肌细胞片时经常出现的困难，我们建立了一种使用离心力进行堆叠的方法，从而可以测量 3D 心肌组织。将人诱导多能干细胞衍生的心肌细胞播种到温度响应培养皿中，并加工成薄片。将心肌细胞片多层化以构建 3D 心肌组织。然后测量多层 3D 心肌组织的收缩力和横截面积，并用于确定心肌组织横截面积与其收缩力之间的关系。1、3 和 5 层组织的收缩力与横截面积成正比线性增加。获得了 6.4 mN/mm 的结果，占成人组织收缩力的七分之一。然而，对于 7 层组织，收缩力突然下降，可能是由于氧气和营养供应有限。总之，我们建立了一种通过分层控制细胞片厚度的方法，从而能够准确评估心脏收缩功能。这种方法可以与活体心脏组织进行比较，同时提供适用于再生医学和药物筛选模型的信息。
