Humna ES Cells for Cardiac Tissue Engineering

描述（由申请人提供）：本补充申请旨在扩展R24 HL64387“工程化心脏形态发生：干细胞和支架”资助的工作，该工作专注于工程化心肌组织的发展。该奖项的一个主要目的是研究干细胞和干细胞衍生的实质细胞在已建立的和新的组织工程支架上的相互作用。根据提议和资助，父母资助将这些研究限制在小鼠干细胞上。然而，最近我们对人类胚胎干细胞（hES）细胞的理解取得了进展，以及对小鼠和人类胚胎干细胞之间细胞行为的关键差异的认识表明，现在我们对胚胎干细胞-支架相互作用的研究扩展到包括人类胚胎干细胞实验是至关重要的。人类胚胎干细胞（NIH代码：WA07）的培养将被扩大、分化和分离，以产生Hes细胞衍生的心肌细胞。人类胚胎干细胞或胚胎干细胞衍生的心肌细胞将被植入已建立的和新的支架上，包括：胶原凝胶；微纤维阵列和纤维多孔结构；在本项目的资助下，开发并投入使用了几种多孔聚合物支架。我们将评估细胞毒性和细胞增殖。我们将评估支架内的增殖和细胞组织，例如心脏标记物的表达和嵌入盘的形成。拟议的工作是我们资助的目标的直接和合乎逻辑的延伸，即使用植入支架的小鼠胚胎干细胞来评估这些相同的参数。我们期望拟建的人类胚胎干细胞和hES细胞衍生的心肌细胞的研究结果将极大地帮助寻找适合心脏组织工程的细胞来源，并将阐明人类胚胎干细胞在组织工程中的使用参数。人类胚胎干细胞的多能性与组织工程策略的成功相结合，具有巨大的潜力，可以解决用于治疗的组织和器官供应极为有限的问题，因此成功将有望对再生医学的实践产生巨大影响。

DESCRIPTION (provided by applicant): This supplemental application seeks to extend the work funded under R24 HL64387 "Engineered Cardiac Morphogenesis: Stem Cells and Scaffolds" which is focused on the development of engineered cardiac muscle tissue. A key aim of this parent award (alluded to in the title) is to study the interactions of stem cells and stem cell-derived parenchymal cells on both established and novel tissue engineering scaffolds. As proposed and funded, the parent grant limited these investigations to murine stem cells. However, recent advancements in our understanding of human embryonic stem (hES) cells, as well as recognition of key differences in cell behavior between ES cells from murine and human origin, suggest that it is crucial to extend now our investigations of ES cell-scaffold interactions to include human ES cell experiments. Cultures of human ES ceils (NIH code: WA07) will be expanded, differentiated, and fractionated to yield Hes cell-derived cardiomyocytes. Either human ES cells or the hES cell-derived cardiomyoctyes will be seeded on established and novel scaffolds including: collagen gels; microfiber arrays and fiberporous structures; and several porous polymeric scaffolds developed and in use under the parent award. We will evaluate cell toxicity and cell proliferation. We will assess proliferation and cellular organization within the scaffolds, e.g. expression of cardiac markers and the formation of intercalated disks. The proposed work is a direct and logical extension of our funded aims to evaluate these same parameters using murine ES cells seeded on scaffolds. We expect the results of the proposed work with human ES cells and hES cell-derived cardiomyocytes will greatly aid the search for cell sources suitable for cardiac tissue engineering, and will clarify parameters for the use of human ES cells in tissue engineering in general. The demonstrated pluripotentiality of human ES cells combined with success in tissue engineering strategies has tremendous potential to address the vastly limited supply of tissues and organs for therapeutic use, and thus success will be expected to impart enormous impact to the practice of regenerative medicine.