Engineered Early Embryonic Cardiac Tissue (EEECT)

工程化早期胚胎心肌组织 (EEECT)

• 批准号: 7061616
• 负责人: Bradley Barth Keller
• 金额: $ 17.52万
• 依托单位: CHILDREN'S HOSP PITTSBURGH/UPMC HLTH SYS
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-05-15 至 2007-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7061616
• 关键词: bioengineering /biomedical engineering; biomaterial interface interaction; biomechanics; biotechnology; cardiac myocytes; cell differentiation; cell proliferation; chick embryo; collagen; confocal scanning microscopy; cytoskeletal proteins; embryo /fetus tissue /cell culture; histogenesis; mechanical stress; mechanoreceptors; myocardium; myosins; three dimensional imaging /topography; tissue /cell culture; tissue engineering; tissue support frame; troponin

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.

描述（由申请人提供）：该R21提案的目的是开发和验证一种组织工程范式，该范式使用了从早期阶段的胚胎小鸡心脏隔离的细胞来有效产生以“工程性早期胚胎性心脏组织或EEECT”的3维（3D）发挥功能的心肌”。开发胚胎（蝇，鱼，青蛙，鸡，小鼠等）是用于心血管细胞命运映射的独特实验模型系统，用于与形态发生和畸形有关的基因发现，以定义形态发生的生理和生物力学。注定要形成心脏和血管的细胞由多种来源引起，包括外侧斜膜中胚层，神经冠，心脏前部场和前心脏器官。这些细胞在时空定义的形态计量学模式中沿着心血管谱系沿着心血管谱系迁移，分化和诱导其他细胞，这些模式对生物力学和代谢应激有反应。心血管组织工程已成为一个领域，该领域正在为治疗多种疾病提供新的治疗选择，包括结构性心脏病和心力衰竭。最近，一些研究小组成功地构建了工程性心脏组织（ECTS）。然而，当前对ECT成功临床植入的技术障碍包括ECT的有限增殖能力，将ECT纳入成熟心肌的高度有组织的多细胞和各向异性收缩机械的后勤挑战，以及不确定的“自然疗程”细胞和组织中的“自然”和“自然”。我们的实验室专注于开发新的方法来研究发育心肌的体内和体外生物力学调节。现在，我们正在应用该专业知识来开发独特的早期胚胎心脏组织（EEECT），该组织将提供强大的体外模型，以继续我们研究心肌分化和适应生物力学负荷，并为工程性心肌的体外来源提供心血管修复的潜在来源。我们具有胚胎鸡心脏细胞3D培养的初步数据提供了一个原理证明，胚胎心肌细胞在培养中增殖和分化，对机械负荷反应并发展类似于与天然发育心肌相似的收缩特性。由于胚胎心脏细胞的增殖能力更大，新生儿和成熟细胞及其分化和适应的内在能力，我们专注于胚胎心脏细胞的使用。 特定目标1。开发并在功能上表征3D工程的早期胚胎心脏组织（EEECT）。 具体目的2。定义机械应力对EEECT结构和收缩功能的影响。我们提出从早期胚胎心肌发展EEECT的提议的意义是研究在受控生物力学环境中进行心肌细胞分化和适应性的独特机会。我们的长期目标是评估EEECT作为一种新型生物材料来修复畸形或受伤的心肌。