Surface-Directed Differentiation of Human Mesenchymal Stem Cells on Orthogonal Peptide Concentration Gradient Surfaces

人间充质干细胞在正交肽浓度梯度表面上的表面定向分化

• 批准号: 1105329
• 负责人: Matthew Becker
• 金额: $ 42万
• 依托单位: University of Akron
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-15 至 2014-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1105329&HistoricalAwards=false
• 关键词: Surface; Directed; Differentiation; Human; Mesenchymal

This award by the Biomaterials program in the Division of Materials Research to University of Akron is in support of research that seeks to quantify the concentration dependence and spatial nature of ligand-receptor interactions, which influence cell differentiation pathways in human mesenchymal stem cells. Future advances in regenerative medicine will require the use of more than one peptide or bioactive molecule to drive stem cells into well-defined specifically differentiated populations. While many investigations have focused on individual molecular interactions at discrete concentrations, a complete interactome of the concentration dependence of extra cellular matrix-derived, osteogenic growth peptide and short peptide fragments of bone morphogenic proteins singly and in combination has remained elusive. The gradient approach also affords a method to decouple local signaling that occurs due to cell-integrin interactions from soluble paracrine effects. When identified, this information will drive the design of higher-order bioactive and biomimetic materials for use in several applications where synthetic materials contact biological systems. The aims of the proposal outline an approach to fabricate and characterize both unidirectional and orthogonal peptide concentration (10-200 pm/cm2 for unidirectional and 10-100 pm/cm2 for orthogonal) gradients. Using a multidimensional characterization approach with surface spectroscopy, immunohistochemistry, automated fluorescence microscopy and real-time polymerase chain reaction, the project will capture the concentration thresholds and synergistic signaling events that influence cell proliferation, lineage commitment and population heterogeneities. The multidisciplinary research ecosystem will offer training opportunities to researchers at all levels (high school, undergraduate, graduate and postdoctoral) at the cutting edge of materials chemistry, chemical biology and regenerative medicine. In partnership with a local high school, real world research activities will be incorporated into age-appropriate modules into upper level biology courses and will be used to teach fundamental concepts and relationships between both physics and chemistry.One of the major remaining barriers to advancing regenerative medicine into mainstream applications is the issue of reliable, safe cell sources. In the near term, stem cells for surgical implantation must come from the person needing the replacement part (autologous sourcing). This limitation will require a pre-surgical isolation of stem cells followed by an expansion protocol ex vivo (outside the body) to generate enough stem cells for the scaffold seeding. The existing synthetic materials are insufficient for these protocols. This proposal outlines an approach to identify the optimal combinations and concentrations of small biomimetic peptides to yield large quantities of well-defined stem cell populations. The autologous approach is free of most safety and ethical concerns that have been raised by various populations. If successful, the identified concentrations will have far reaching impact on our understanding of how individual and combinations of peptides influence cell differentiation and furthermore improve the quantity and quality of well-defined stem cell populations available for research and clinical investigations. The multidisciplinary research environment offers opportunities for training at all levels (high school, undergraduate, graduate and postdoctoral). The project involves training students, undergraduate, and high school students from a local school. The age appropriate educational modules will be designed to teach fundamental principles to high school biology courses at various levels using advanced research topics. It is critical to encourage students at this age to understand the importance of engineering principles and how they relate to advancing human health. This project is expected to support the nation's efforts to increase the numbers and diversity of the engineering student pipeline that require one to reach down and providing unique research opportunities to underrepresented/minority students populations which encourage the pursuit of science and engineering careers.

生物材料计划对阿克伦大学的材料研究划分的奖项支持研究，该研究旨在量化配体 - 受体相互作用的浓度依赖性和空间性质，这会影响人间质干细胞中的细胞分化途径。 再生医学的未来进步将需要使用多个肽或生物活性分子将干细胞驱动到定义明确的分化种群中。尽管许多研究集中在离散浓度下的单个分子相互作用上，但单独的细胞基质衍生，成骨生长肽和骨形态蛋白的短肽片段的浓度依赖性的完全相互作用依赖于骨形形态蛋白的短肽片段，并且组合在一起仍然难以捉摸。梯度方法还提供了一种将由于可溶性旁分泌作用引起的细胞紧密相互作用而导致的局部信号传导的方法。 确定后，此信息将推动高阶生物活性和仿生材料的设计，以在合成材料接触生物系统的几种应用中使用。该提案的目的概述了一种制造和表征单向和正交肽浓度（单向10-200 pm/cm2的方法），正交梯度为10-100 pm/cm2。 该项目使用表面光谱，免疫组织化学，自动荧光显微镜和实时聚合酶链反应的多维表征方法，该项目将捕获浓度阈值和协同信号事件，从而影响细胞增殖，谱系承诺和人群异质性。 多学科研究生态系统将为材料化学，化学生物学和再生医学的最前沿的各个级别的研究人员（高中，本科，研究生和博士后）提供培训机会。 与当地一所高中合作，现实世界的研究活动将被纳入适合年龄的模块中，并将用于教授物理和化学之间的基本概念和关系。 在短期内，需要进行手术植入的干细胞必须来自需要替换部分的人（自体源）。 该限制将需要在手术前分离干细胞，然后进行膨胀方案离体（体外），以产生足够的干细胞以进行支架播种。 现有的合成材料不足以适用于这些协议。 该提案概述了一种鉴定小型仿生肽的最佳组合和浓度以产生大量定义明确的干细胞群体的方法。 自体态方法不含各种人群提出的大多数安全和道德问题。 如果成功的话，确定的浓度将对我们对肽的个体和组合如何影响细胞分化以及改善可用于研究和临床研究的明确定义明确的干细胞种群的数量和质量的理解产生很大的影响。 多学科研究环境为各级培训提供了机会（高中，本科，研究生和博士后）。 该项目涉及培训来自当地学校的学生，本科生和高中生。 适合年龄的教育模块将旨在使用高级研究主题向高中生物学课程教授基本原则。 鼓励这个年龄的学生了解工程原则的重要性以及他们如何与人类健康的联系。预计该项目将支持美国为增加工程学生管道的数量和多样性的努力，这些努力需要一个人来降低并提供独特的研究机会，以使代表性不足/少数民族学生的人群鼓励人们追求科学和工程职业。