Nanoscale 3D Cell Sheet Engineering: A Platform for Studying the Cardiac Microenvironment and Tissue-Level Heart Structure-Function Relationships

 描述（由申请人提供）：心脏具有复杂的3D结构，由排列的细长心肌细胞片组成，形成透壁螺旋结构。心脏的3D微环境已被证明有助于心肌细胞的发育，心脏的3D螺旋结构对适当的功能至关重要。然而，目前的组织工程策略不能精确地控制细胞密集的3D组织结构。 体外培养组织。因此，我们的目标是通过利用基于纳米制造的细胞片工程和人多能干细胞衍生的心肌细胞（hPS-CM）来设计新的3D心脏组织，其可以重现各向异性的螺旋状心脏结构。要做到这一点，我们将利用一个开发的平台，包括一个温敏，nanofabricated substratum（TNFS）和凝胶铸造方法来制造3D，无支架组织与逐层结构控制。我们将首先设计和表征多层心脏组织作为改进的心脏在一个盘子模型。然后，我们将确定微环境对干细胞衍生的心肌细胞成熟的影响，例如仿生纳米地形学线索，以及3D细胞密集的心脏微环境。最后，将制造具有不同结构的3D堆叠组织，并作为心肌组织的生理模型分析其收缩和电功能。这些模型将用于研究人体心脏组织的结构-功能关系。

 DESCRIPTION (provided by applicant): The heart has a complex, 3D structure comprised of sheets of aligned, elongated cardiomyocytes which form a transmural helical structure. The 3D microenvironment of the heart has been shown to assist in the development of cardiomyocytes and the 3D helical structure of the heart is crucial to appropriate function. However, current tissue engineering strategies are unable to precisely control 3D tissue architecture in cell- dense tissues in vitro. Thus, we aim to engineer novel 3D cardiac tissue which can recapitulate the anisotropic, helical heart structure by utilizing nanofabrication-based cell sheet engineering and human pluripotent stem cell-derived cardiomyocytes (hPS-CMs). To do so, we will utilize a developed platform consisting of a thermoresponsive, nanofabricated substratum (TNFS) and a gel casting method to fabricate 3D, scaffold-free tissues with layer-by-layer structural control. We will first engineer and characterize multilayered cardiac tissues as an improved heart-in-a-dish model. We will then determine the microenvironmental effects on stem cell-derived cardiomyocyte maturity, such as biomimetic nanotopographical cues, as well as the 3D cell-dense cardiac microenvironment. Finally, the 3D stacked tissues will be fabricated with varying structures and analyzed for contractile and electrical function as physiological models of myocardial tissue. These models will be used to investigate the structure-function relationship of human cardiac tissue.