Engineered Early Embryonic Cardiac Tissue (EEECT)

描述（由申请人提供）：本R21提案的目的是开发和验证一种组织工程范例，该范例使用从早期鸡胚心脏中分离的细胞有效地产生三维（3D）功能心肌，称为“工程化早期胚胎心脏组织或EEECT”。胚胎发育（苍蝇，鱼，青蛙，鸡，小鼠等）作为独特的实验模型系统，用于心血管细胞命运作图，用于与形态发生和畸形相关的基因发现，用于定义形态发生的生理和生物力学。注定形成心脏和血管的细胞来自几个来源，包括内脏中胚层、神经嵴、心前区和心外膜器官。这些细胞迁移，克隆增殖，分化，并诱导其他细胞沿着心血管谱系的时空定义的形态学模式，响应于生物力学和代谢应力。心血管组织工程已成为一个领域，这是提供新的治疗选择的管理范围广泛的疾病，包括结构性心脏病和心力衰竭。近年来，一些研究小组成功地构建了工程化心脏组织（ECTs）。然而，目前成功临床植入ECTs的技术障碍包括ECTs有限的增殖能力，将ECTs整合到成熟心肌的高度组织化的多细胞和各向异性收缩机制中的后勤挑战，以及移植的组织工程化细胞和组织的不确定的体内“自然史”。我们的实验室一直致力于开发新的方法来研究在体内和体外的生物力学调控的发展心肌。我们现在正在应用这一专业知识开发一种独特的工程化早期胚胎心脏组织（EEECT），这将提供一个强大的体外模型，以继续我们对心肌分化和适应生物力学负荷的研究，并为心血管修复提供一个潜在的体外工程化心肌来源。我们的初步数据与3D培养的胚胎鸡心肌细胞提供了一个证明的原则，胚胎心肌细胞增殖和分化的文化，响应机械负荷，并开发收缩特性类似于天然发展心肌。我们一直专注于使用胚胎心脏细胞，因为它们比新生儿和成熟细胞具有更大的增殖能力，并且具有内在的分化和适应能力。 具体目标1。开发和功能表征3D工程化早期胚胎心脏组织（EEECT）。 具体目标2。定义机械应力对EEECT结构和收缩功能的影响。我们建议从早期胚胎心肌中开发EEECT的重要性是在受控的生物力学环境中研究心肌细胞分化和适应的独特机会。我们的长期目标是评估EEECT作为一种新型的生物材料来修复畸形或损伤的心肌。

DESCRIPTION (provided by applicant): The purpose of this R21 proposal is to develop and validate a tissue engineering paradigm that uses cells isolated from the early staged embryonic chick heart to efficiently generate a 3-dimensional (3D) functioning myocardium termed "Engineered Early Embryonic Cardiac Tissue or EEECT". Developing embryos (fly, fish, frog, chicken, mouse, etc..) serve as unique experimental model systems for cardiovascular cell fate mapping, for gene discovery related to morphogenesis and malformations, for defining the physiology and biomechanics of morphogenesis. The cells destined to form the heart and blood vessels arise from several sources including the lateral splanchnic mesoderm, neural crest, anterior heart field, and proepicardial organ. These cells migrate, clonally proliferate, differentiate, and induce other cells along cardiovascular lineages in spatio-temporally defined morphometric patterns that are responsive to biomechanical and metabolic stresses. Cardiovascular tissue engineering has emerged as a field which is providing novel therapeutic options for the management of a wide range of diseases including structural heart disease and heart failure. Recently several research groups have succeeded in constructing Engineered Cardiac Tissues (ECTs). However, current technical barriers to the successful clinical implantation of ECTs include the limited proliferative capacity of ECTs, the logistical challenges of integrating ECTs into the highly organized multicellular and anisotropic contractile machinery of the mature myocardium, and the uncertain in-vivo "natural-history" of transplanted tissue engineered cells and tissues. Our laboratory has focused on developing novel approaches to investigate the in vivo and in vitro biomechanical regulation of the developing myocardium. We are now applying that expertise to develop a unique Engineered Early Embryonic Cardiac Tissue (EEECT) which will provide a robust in vitro model to continue our investigation of myocardial differentiation and adaptation to biomechanical load and also provide a potential in vitro source of engineered myocardium for cardiovascular repair. Our preliminary data with 3D culture of embryonic chick cardiac cells have provided a proof of principle that embryonic cardiomyocytes proliferate and differentiate in culture, respond to mechanical load, and develop contractile properties similar to native developing myocardium. We have focused on the use of embryonic cardiac cells due to their greater proliferative capacity versus neonatal and mature cells and their intrinsic ability to differentiate and adapt. Specific Aim 1. Develop and functionally characterize 3D Engineered Early Embryonic Cardiac Tissue (EEECT). Specific Aim 2. Define the impact of mechanical stress on the architecture and contractile function of EEECT. The significance of our proposal to develop EEECT from early embryonic myocardium is the unique opportunity to investigate cardiomyocyte differentiation and adaptation in a controlled biomechanical environment. Our long term goal is to evaluate EEECT as a novel biomaterial to repair the malformed or injured myocardium.