Substrate Independent, Spatially Resolved, Stable Polymer Coatings for Studying Human Mesenchymal Stem Cells (hMSCs)

用于研究人间充质干细胞 (hMSC) 的基底独立、空间分辨、稳定的聚合物涂层

• 批准号: 1306482
• 负责人: Padma Gopalan
• 金额: $ 39万
• 依托单位: University of Wisconsin-Madison
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-01 至 2018-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1306482&HistoricalAwards=false
• 关键词: Substrate; Independent; Spatially; Resolved; Stable

This award by the Biomaterials program in the Division of Materials Research is a three-year plan of integrated research, education, and outreach on a polymeric coating for studying human mesenchymal stem cells (hMSCs). This award is cofunded by the Polymer program in the Division of Materials Research. Earlier studies strongly indicate that hMSC phenotype is strongly influenced by ligands derived from the natural cellular microenvironment, including cell adhesion peptides, heparin-binding peptides, and growth factor-mimetic peptides. This proposal is to develop polymer coatings based on copolymers of poly(ethylene glycol) with at most 10% of crosslinkable groups, which can be crosslinked into 30 nm mats for immobilization of peptides. These mats are chemically-defined for stable presentation of two putative pro-osteogenic peptides over an extended timeframe, and to explore in depth the effects on hMSC differentiation into osteoblasts. These polymer coatings with peptides present a stable cytophobic slate, and hence one can truly investigate the effect of the relevant peptides on hMSCs during the entire time course of osteogenic differentiation. Importantly, these coatings are thin films that can be applied to virtually any underlying substrate, which will lead to broad impact on cell culture studies and future design of biomedical devices. Because of the thin-film nature, the proposed use of X-ray photoelectron spectroscopy will quantify precisely the peptides presented on the polymer coatings. Further, this proposal will study the first systematic analysis of chemically defined, nanometer-scale presentation of a growth factor-receptor binding ligand, and it may lead to broad impact in view of the importance of receptor dimerization/clustering in virtually all growth factor signaling systems. The technological broader impact of this proposal would be in developing a new class of ultra-thin biomaterial coatings with unique set of properties for stem cell growth. In order to clearly understand the effects of micro environmental signals on stem cell behavior, there is a critical need for experimental systems that present specific signals, and complex signal combinations, to stem cells. The proposed platform may lead to widespread use of the proposed approaches by cell biologists in general. Ultimately, this line of research could result in identification of signals that promote lineage-specific stem cell differentiation, leading to development and optimization of substrates that promote stem cell differentiation in vitro and in vivo. More generally, the proposed adaptable and chemically tailorable thin films would be applicable to a broad range of fundamental questions in cell biology in which stable, controlled presentation of peptide ligands is desirable. As part of this grant, this project will: 1) develop Interdisciplinary Curriculum Development of two courses in two departments, one based in Materials Science & Engineering, and the second one based in Biomedical Engineering; 2) carry out dissemination and outreach activities by the development of hands-on exhibits on Stem Cells and Tissue Engineering that would provide case study materials for a Science and Social Media course; and 3) recruit and mentor female and underrepresented minority graduate and undergraduate students in research activities.

该奖项由材料研究部生物材料项目颁发，是一项为期三年的综合研究、教育和推广用于研究人类间充质干细胞 (hMSC) 的聚合物涂层的计划。该奖项由材料研究部聚合物项目共同资助。早期研究强烈表明，hMSC 表型受到天然细胞微环境中配体的强烈影响，包括细胞粘附肽、肝素结合肽和生长因子模拟肽。该提案旨在开发基于聚乙二醇共聚物的聚合物涂层，其可交联基团最多为 10%，可交联成 30 nm 垫用于固定肽。 这些垫经过化学定义，可在较长时间内稳定呈现两种推定的促成骨肽，并深入探索对 hMSC 分化为成骨细胞的影响。这些带有肽的聚合物涂层呈现出稳定的疏细胞板，因此可以真正研究相关肽在成骨分化的整个时间过程中对 hMSC 的影响。重要的是，这些涂层是薄膜，几乎可以应用于任何底层基材，这将对细胞培养研究和生物医学设备的未来设计产生广泛影响。由于薄膜的性质，建议使用 X 射线光电子能谱来精确量化聚合物涂层上存在的肽。此外，该提案将研究生长因子-受体结合配体的化学定义的纳米级呈现的首次系统分析，鉴于受体二聚化/聚类在几乎所有生长因子信号传导中的重要性，它可能会产生广泛的影响系统。该提案的技术更广泛的影响将是开发一种新型超薄生物材料涂层，具有独特的干细胞生长特性。为了清楚地了解微环境信号对干细胞行为的影响，迫切需要向干细胞呈现特定信号和复杂信号组合的实验系统。所提出的平台可能会导致细胞生物学家广泛使用所提出的方法。最终，这一系列研究可能会鉴定出促进谱系特异性干细胞分化的信号，从而开发和优化促进体外和体内干细胞分化的底物。更一般地，所提出的适应性和化学可定制薄膜将适用于细胞生物学中广泛的基本问题，其中需要稳定、受控的肽配体呈现。作为这笔赠款的一部分，该项目将：1）开发两个系的两门课程的跨学科课程开发，一门基于材料科学与工程，第二门基于生物医学工程； 2) 通过开发干细胞和组织工程实践展览来开展传播和外展活动，为科学和社交媒体课程提供案例研究材料； 3) 在研究活动中招募和指导女性和代表性不足的少数族裔研究生和本科生。