Cardiomyoplasty in vitro: Host /Doner Cell Interactions

体外心肌成形术：宿主/供体细胞相互作用

• 批准号: 7145059
• 负责人: Nenad Bursac
• 金额: $ 22.43万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-06-15 至 2008-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7145059
• 关键词: age difference; autologous transplantation; bone marrow; calcium flux; cardiac myocytes; cell cell interaction; cell differentiation; cell transplantation; growth factor; heart electrical activity; immunocytochemistry; immunofluorescence technique; intercellular connection; laboratory rat; membrane potentials; mixed tissue /cell culture; myoblasts; tissue /cell culture

DESCRIPTION (provided by applicant): Implantation of exogenous donor cells into the heart (cellular cardiomyoplasty) is an emerging methodology for the treatment of post-ischemic ventricular remodeling. While initial clinical implantations of autologous skeletal myoblasts and bone marrow cells have been promising, the intrinsic potential of these cells to affect electro-mechanical function of surrounding heart tissue has still not been elucidated. Therefore, the main goal of this proposal is to systematically study the ability of different donor cells (i.e. skeletal myoblasts and mesenchymal stem cells, as compared to control cardiac myofibroblasts) to propagate electro-mechanical activity through a host cardiac network in vitro. We hypothesize that: 1) propagation of electrical activity through donor cells depends on their type and stage of differentiation, as well as the presence of not only electrical but also mechanical junctions between the donor-donor and host-donor cell pairs, and 2) electro- mechanical propagation in the donor cell implant can be improved through upregulation of the intercellular communication by specific growth factors. To test these hypotheses we propose to study electrical conduction through donor cells using a geometrically simplified, reproducible one-dimensional setting, i.e. the micropatterned cardiomyocyte strands with inserts made of donor cells. The propagation of cell membrane potentials and intracellular calcium transients in donor cells will be optically mapped in the presence of various differentiation agents and growth factors. Obtained results will be correlated with those from immunohistochemical and molecular analyses. The findings from this proposal are expected to elucidate potential of different donor cells to functionally integrate in the heart. Eventually, the proposed experimental framework will allow us to perform high throughput in vitro analysis of the factors that can improve electromechanical propagation of different donor cells. The final aim is to aid in the development of efficient and safe cell-based approaches for the treatment of regional heart injury due to ischemia, infarction, or congenital defects.

描述（由申请人提供）：将外源供体细胞植入心脏（细胞心肌成形术）是治疗缺血后心室重塑的新兴方法。虽然自体骨骼成肌细胞和骨髓细胞的初步临床植入前景广阔，但这些细胞影响周围心脏组织机电功能的内在潜力尚未阐明。因此，该提案的主要目标是系统地研究不同供体细胞（即骨骼肌母细胞和间充质干细胞，与对照心脏肌成纤维细胞相比）在体外通过宿主心脏网络传播机电活动的能力。我们假设：1）电活动通过供体细胞的传播取决于其类型和分化阶段，以及供体-供体和宿主-供体细胞对之间不仅存在电连接而且还存在机械连接，2）供体细胞植入物中的机电传播可以通过特定生长因子上调细胞间通讯来改善。为了测试这些假设，我们建议使用几何简化的、可重复的一维设置（即带有由供体细胞制成的插入物的微图案心肌细胞链）来研究通过供体细胞的电传导。在各种分化剂和生长因子的存在下，供体细胞中细胞膜电位和细胞内钙瞬变的传播将被光学绘制。获得的结果将与免疫组织化学和分子分析的结果相关联。该提案的研究结果有望阐明不同供体细胞在心脏中功能整合的潜力。最终，所提出的实验框架将使我们能够对可以改善不同供体细胞机电传播的因素进行高通量体外分析。最终目标是帮助开发有效且安全的基于细胞的方法来治疗由于缺血、梗塞或先天性缺陷引起的局部心脏损伤。