Differentiation of Bone-Marrow Stem Cells in a Laser Patterned Myocyte Coculture

激光图案化肌细胞共培养中骨髓干细胞的分化

• 批准号: 7414837
• 负责人: BRUCE Z GAO
• 金额: $ 14.31万
• 依托单位: CLEMSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-05-01 至 2012-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7414837
• 关键词: Accounting; Action Potentials; Address; Adult; Americas; Arts; Atomic Force Microscopy; Biomedical Engineering; Bone Marrow; Bone Marrow Stem Cell; Cardiac; Cardiac Myocytes; Cause of Death; Cell Communication; Cell Differentiation process; Cells; Cellular biology; Cessation of life; Clinical; Coculture Techniques; Connexin 43; Core Facility; Coupling; Cultured Cells; Data; Deposition; Development; Electrodes; Embryo; Engineering; Environment; Equilibrium; Equipment; Extracellular Matrix; Figs - dietary; Funding; Goals; Grant; Heart; Heart Diseases; In Vitro; Institution; Island; Knowledge; Lasers; Manuscripts; Measures; Mechanics; Microelectrodes; Microscopy; Modeling; Molecular and Cellular Biology; Monitor; Mus; Muscle Cells; Myocardial Infarction; Myocardium; N-Cadherin; Neonatal; Optics; Patch-Clamp Techniques; Patients; Pattern; Physics; Proliferating; Property; Regenerative Medicine; Regulation; Research; Research Personnel; Role; Staging; Statistically Significant; Stem cell transplant; Stem cells; Stress; Structure; System; Testing; Therapeutic; Time; Tissues; Traction; Training; United States National Institutes of Health; Woman; Writing; career; cell type; chemical property; electrical property; feeding; foot; functional restoration; genetically modified cells; improved; in vivo; injured; men; nano; patch clamp; programs; protein expression; repaired; skills training; vector

DESCRIPTION (provided by applicant): This proposal describes training in cardiac-cell biology for a PI whose background is in physics and engineering. The training includes 1) graduate courses in cell biology; 2) experimental-skill training in molecular and cellular biology; and 3) academic development, including developing a research network and writing manuscripts/proposals. The immediate goal is to improve the Pi's ability to obtain NIH funding, particularly an R01 grant. The long-term goal is to assist the PI in developing his academic career in biomedical engineering with a better balance between his biomedical and engineering proficiencies. To achieve these goals, the PI proposes to investigate the functional differentiation of bone-marrow stem cells in an engineered cardiac myocyte-coculture model. In this model, identical cell-culture microwells will be fabricated to allow creation of a defined stem cell-myocyte interface in each well through application of a laser beam. This model will provide statistically significant data on the regulation of stem-cell differentiation by various types of cocultured myocytes. To test the hypothesis that the microenvironments will regulate the functional differentiation of stem cells into myocytes, the following specific aims will be addressed: 1) Determine the microenvironment's role in regulating stem-cell differentiation into cardiac cells by changing the extracellular matrix, switching the coculture cell types, and genetically modifying the cell-cell juctional protein expressions; 2) Use atomic force microscopy to determine the stem cells' mechanical properties by measuring cell stiffness and traction force microscopy to evaluate the cells' mechanical coupling; 3) Determine the stem cells' electrical properties by studying action potentials using the patch clamp technique and evaluating electrical coupling using microelectrode arrays. Heart disease is America's leading cause of death for both men and women, accounting for nearly 40% of all annual deaths. By adding to our knowledge of the functional differentiation of bone-marrow stem cells into heart muscle cells, the Pi's research will contibute to the current effort in regenerative medicine to restore the the function of a damaged heart using the patient's own cells. The Pi's institution will provide more than 2,500 sq feet of lab space and unlimited access to all core facilities, which are staffed and provided with state-of-the-art equipment, in addition to 75% release time.

描述（由申请人提供）：该提案描述了对具有物理和工程学背景的 PI 的心脏细胞生物学培训。培训内容包括1）细胞生物学研究生课程； 2）分子和细胞生物学实验技能培训； 3) 学术发展，包括发展研究网络和撰写手稿/提案。近期目标是提高 Pi 获得 NIH 资助（特别是 R01 资助）的能力。长期目标是协助 PI 发展其在生物医学工程方面的学术生涯，并在其生物医学和工程能力之间取得更好的平衡。为了实现这些目标，PI 建议研究工程心肌细胞共培养模型中骨髓干细胞的功能分化。在该模型中，将制造相同的细胞培养微孔，以便通过应用激光束在每个孔中创建明确的干细胞-肌细胞界面。该模型将提供有关各种类型的共培养肌细胞对干细胞分化调节的统计显着数据。为了检验微环境将调节干细胞向心肌细胞功能分化的假设，将解决以下具体目标：1）通过改变细胞外基质、切换共培养细胞类型和基因修饰细胞-细胞连接蛋白表达来确定微环境在调节干细胞向心肌细胞分化中的作用； 2) 使用原子力显微镜通过测量细胞刚度来确定干细胞的机械特性，并使用牵引力显微镜来评估细胞的机械耦合； 3) 通过使用膜片钳技术研究动作电位并使用微电极阵列评估电耦合来确定干细胞的电特性。心脏病是美国男性和女性的首要死因，占年度死亡总数的近 40%。通过增加我们对骨髓干细胞向心肌细胞功能分化的了解，Pi 的研究将为当前再生医学领域利用患者自身细胞恢复受损心脏功能的努力做出贡献。 Pi 的机构将提供超过 2,500 平方英尺的实验室空间，并可无限制地使用所有核心设施，这些设施配备了工作人员并配备了最先进的设备，此外还有 75% 的发布时间。