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(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，该范例使用从早期胚胎小鸡心脏分离的细胞来高效地生成被称为工程早期胚胎心脏组织或EEECT的三维(3D)功能心肌。发育中的胚胎(苍蝇、鱼、青蛙、鸡、老鼠等)作为心血管细胞命运图谱的独特实验模型系统，用于发现与形态发生和畸形相关的基因，定义形态发生的生理学和生物力学。预定形成心脏和血管的细胞来自几个来源，包括内脏外侧中胚层、神经脊、心前区和心前器官。这些细胞沿着心血管谱系迁移、克隆性增殖、分化和诱导其他细胞，这些细胞以时空定义的形态计量学模式对生物力学和代谢应激做出反应。心血管组织工程学已经成为一个为治疗包括结构性心脏病和心力衰竭在内的多种疾病提供新的治疗选择的领域。最近，几个研究小组成功构建了工程心脏组织(ECTs)。然而，目前ECTs临床移植成功的技术障碍包括ECTs的增殖能力有限，将ECTs整合到成熟心肌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细胞和组织在体内的“自然历史”不确定。我们的实验室致力于开发新的方法来研究发育中的心肌的体内和体外生物力学调节。我们正在应用这些专业知识开发一种独特的工程化早期胚胎心脏组织(EEECT)，它将为我们继续研究心肌分化和对生物力学负荷的适应提供一个强大的体外模型，并为心血管修复提供潜在的体外工程化心肌来源。我们的鸡胚胎心肌细胞3D培养的初步数据证明了胚胎心肌细胞在培养中增殖和分化，对机械负荷做出反应，并发展出与自然发育的心肌相似的收缩特性。我们将重点放在胚胎心肌细胞的使用上，因为它们比新生儿和成熟细胞具有更强的增殖能力，以及它们固有的分化和适应能力。 具体目的1.三维工程化早期胚胎心脏组织(EEECT)的研制及功能鉴定。 明确机械应力对EEECT结构和收缩功能的影响。我们提出的从早期胚胎心肌发展EEECT的意义在于，这是在受控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价EEECT作为一种修复畸形或受损心肌的新型生物材料。