Potential Use of ES Cells for Cardiac Tissue Engineering

ES 细胞在心脏组织工程中的潜在用途

• 批准号: 7339853
• 负责人: WILLIAM C CLAYCOMB
• 金额: $ 17.75万
• 依托单位: LSU HEALTH SCIENCES CENTER
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-01-15 至 2009-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7339853
• 关键词: Adult; Autologous; Biomedical Engineering; Cardiac; Cardiac Myocytes; Cardiovascular Diseases; Cause of Death; Cells; Cellular Morphology; Characteristics; Collagen; Condition; Congenital Abnormality; Custom; Data; Developed Countries; Developing Countries; Development; Device Designs; Devices; ES Cell Line; Electric Stimulation; Engineering; Fibronectins; Gene Expression; Genes; Goals; Heart; Histology; Immunohistochemistry; In Vitro; Mechanics; Methodology; Morbidity - disease rate; Morphology; Mus; Muscle; Myocardium; NIH Program Announcements; Natural regeneration; Numbers; Operative Surgical Procedures; Polymerase Chain Reaction; Population; Proliferating; Property; Reconstructive Surgical Procedures; Research; Research Personnel; Research Project Grants; Reverse Transcription; Source; Stretching; Support of Research; Techniques; Testing; Therapeutic; Tissue Engineering; Tissues; Transmission Electron Microscopy; Undifferentiated; United States; Ventricular; Work; concept; design; embryonic stem cell; human embryonic stem cell; injured; innovation; mortality; novel strategies; repaired; response

DESCRIPTION (provided by applicant): This revised application is being submitted in response to the program announcement (PA) "Exploratory/ Developmental (R21) Bioengineering Research Grants (EBRG PA-03-058). Cardiovascular disease is the number one cause of death in the United States. Adult cardiomyocytes do not proliferate and therefore cardiac muscle cannot regenerate when the cells are permanently injured. We have recently isolated several pure populations of cardiomyocytes from murine ES cells which express cardiac specific genes. We propose to use these cells as a renewable source in tissue engineered constructs. To create these constructs, the cells will be mixed with collagen and fibronectin. The constructs will be exposed to multiple mechanical or electrical stimulation regiments using a custom designed device. As the heart develops, gene expression of ventricular cardiomyocytes changes to that of terminally differentiated cardiomyocytes. We are hypothesizing that mechanical load and electrical stimulation will induce our genetically selected cardiomyocytes to become better organized and more highly differentiated. Preliminary data has provided proof of concept for the creation of a construct using either HL-1 cardiomyocytes or the genetically selected ES cells. This data also shows that a mechanical load can be applied to the constructs using the device and that the mechanical load has a direct effect on the morphology of the construct and its cellular components. The proposed research defines a plan that will enable us to optimize the conditions for creating a cardiac tissue engineered construct and to investigate the effect that mechanical load and electrical stimulation has on the differentiation of ES cardiomyocytes. Reverse transcription PCR will be utilized to evaluate the effect that mechanical and electrical stimulation has on cardiac gene expression. The cellular morphology of the construct will be assessed with histology, immunohistochemistry, and transmission electron microscopy. In addition, the mechanical properties of the construct will be tested and compared to normal heart muscle. The significance of this work is that if we are successful in our goals then this methodology could be applied to the use of human ES cells for the therapeutic repair of injured cardiac muscle or the creation of artificial cardiac tissue for surgical repair or as a source of tissue for reconstructive surgery of congenital birth defects.

描述（由申请人提供）：本修订申请是为了响应项目公告（PA）“探索/开发（R21）生物工程研究赠款（EBRG PA-03-058）”而提交的。心血管疾病是美国的头号死因。成年心肌细胞不增殖，因此当细胞永久性损伤时，心肌不能再生。我们最近从表达心脏特异性基因的小鼠ES细胞中分离出了几个纯的心肌细胞群。我们建议使用这些细胞作为组织工程构建的可再生来源。为了创建这些构建体，细胞将与胶原蛋白和纤连蛋白混合。使用定制设计的器械将结构暴露于多种机械或电刺激方案。 随着心脏的发育，心室心肌细胞的基因表达改变为终末分化的心肌细胞的基因表达。我们假设机械负荷和电刺激将诱导我们的基因选择的心肌细胞变得更好地组织和更高度分化。初步数据提供了使用HL-1心肌细胞或遗传选择的ES细胞创建构建体的概念证明。该数据还表明，可以使用该器械对结构施加机械载荷，并且机械载荷对结构及其细胞组分的形态有直接影响。拟议的研究定义了一个计划，使我们能够优化创建心脏组织工程构建体的条件，并研究机械负荷和电刺激对ES心肌细胞分化的影响。逆转录PCR将用于评估机械和电刺激对心脏基因表达的影响。将采用组织学、免疫组织化学和透射电子显微镜评估构建体的细胞形态。此外，将测试结构的机械性能，并与正常心肌进行比较。这项工作的意义在于，如果我们成功地实现了我们的目标，那么这种方法可以应用于人类ES细胞的使用，用于治疗性修复受伤的心肌或创建用于手术修复的人工心脏组织，或作为先天性出生缺陷重建手术的组织来源。

项目成果

Embryonic stem cell-derived cardiomyocytes harbor a subpopulation of niche-forming Sca-1+ progenitor cells.

• DOI: 10.1007/s11010-010-0661-9
• 发表时间: 2011-03
• 期刊: MOLECULAR AND CELLULAR BIOCHEMISTRY
• 影响因子: 4.3
• 作者: Lam, May L.;Hashem, Sherin I.;Claycomb, William C.
• 通讯作者: Claycomb, William C.

Store-operated calcium entry is present in HL-1 cardiomyocytes and contributes to resting calcium.

• DOI: 10.1016/j.bbrc.2011.10.133
• 发表时间: 2011-12-09
• 期刊: BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
• 影响因子: 3.1
• 作者: Touchberry, Chad D.;Elmore, Chris J.;Nguyen, Tien M.;Andresen, Jon J.;Zhao, Xiaoli;Orange, Matthew;Weisleder, Noah;Brotto, Marco;Claycomb, William C.;Wacker, Michael J.
• 通讯作者: Wacker, Michael J.