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

 描述(申请人提供)：心脏具有复杂的三维结构，由排列的、拉长的心肌细胞片组成，这些心肌细胞形成跨壁螺旋结构。心脏的三维微环境被证明有助于心肌细胞的发育，而心脏的三维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无法精确控制高密度细胞中的3D组织结构 体外培养的组织。因此，我们的目标是利用基于纳米加工的细胞片工程和人多能干细胞来源的心肌细胞(HPS-CMS)来设计能够概括各向异性螺旋心脏结构的新型3D心脏组织。为此，我们将利用开发的平台，包括温度响应性，纳米软化基质(TNFS)和凝胶浇注方法来制造3D，无支架的组织，逐层结构控制。我们将首先设计和表征多层心脏组织，作为一种改进的盘中之心模型。然后，我们将确定微环境对干细胞来源的心肌细胞成熟的影响，例如仿生纳米地形图线索，以及3D细胞密集的心脏微环境。最后，将构建具有不同结构的3D堆叠组织，并分析其收缩和电功能，作为心肌组织的生理模型。这些模型将被用来研究人类心脏组织的结构-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.