Nanopatterning of advanced research tools to harness the mechanobiology of cell-matrix interaction for stem cell expansion

先进研究工具的纳米图案利用细胞-基质相互作用的机械生物学来进行干细胞扩增

• 批准号: RGPIN-2016-04043
• 负责人: Yim, Evelyn
• 金额: $ 2.04万
• 依托单位: University of Waterloo
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2019
• 资助国家: 加拿大
• 起止时间: 2019-01-01 至 2020-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=682619
• 关键词: Nanopatterning; advanced; research; tools; harness

Pluripotent stem cells (PSC), which can give rise to different cell types of all three germ layers, are excellent models to study tissue development and lineage commitment in vitro. While conventional culture conditions for PSCs focus on biochemical definition, biophysical cues that are present in the stem cell niche are not incorporated in vitro. Our previous studies have demonstrated the significant influence of topography on stem cell fate determination through the involvement and restructuring of focal adhesion. Yet the biophysical regulation of PSC pluripotency maintenance and lineage commitment in the in vitro niche remains a mystery. We hypothesize that 1) biophysical cues such as nanotopography regulate pluripotency maintenance via a focal adhesion-signaling pathway; 2) a suitable biophysical environment and biochemical factor can work synergistically to enhance human PSC maintenance; and 3) advanced nanofabrication techniques can fabricate nanopatterned tools and 3D bioreactors for stem cell expansion.***Nanofabrication technologies enable breakthroughs in microelectronics, optics, and in vitro biological systems to study cell-matrix interaction. However, 2 key factors limit the application of nanotopography for stem cell-biology studies and industrial scale-up of cell expansion: the properties of the patterning materials, and conventional 2D patterned culture surfaces, respectively. Therefore, in the proposed research program, we aim to ***1) develop nanopatterned research tools with tunable mechanical properties and refractive index to enable the study of cell-matrix interactions through super-resolution microscopy; ***2) investigate the cell-matrix interaction during pluripotency maintenance to harness the optimal biophysical conditions that enhance PSC expansion; ***3) develop a 3D nanopatterned bioreactor for PSC expansion; and ***4) apply bioreactor and nanotopography tools in scaling up the production of PSC for research.***Our innovative approaches and research program will develop new research tools to study stem cells. With the new tools, the program will lead to new insights in stem cell biology research by providing a deeper understanding of the biophysical regulation of PSC lineage commitment. This will be a significant contribution to the stem cell and mechanobiology research communities. A minimalistic, biophysically- and biochemically-defined niche for stem cell expansion will also provide an effective means to meet the rigorous demand for consistent and reproducible cells needed for different applications such as biological studies or drug development. ***Ultimately, this program will yield new, potentially patentable devices and concepts that are applicable to many research areas and will help grow Canada's biotechnology industries. It will also produce the next generation of bioengineers for these vital academic and industrial communities.

多能干细胞（PSC）可以产生所有三个胚层的不同细胞类型，是体外研究组织发育和谱系定型的优秀模型。虽然 PSC 的传统培养条件侧重于生化定义，但干细胞生态位中存在的生物物理线索并未纳入体外。我们之前的研究表明，地形通过粘着斑的参与和重组对干细胞命运决定产生显着影响。然而，体外小生境中 PSC 多能性维持和谱系定型的生物物理调节仍然是一个谜。我们假设 1）生物物理线索（例如纳米形貌）通过粘着斑信号通路调节多能性维持； 2）适宜的生物物理环境和生化因子可以协同作用，增强人体PSC的维持； 3) 先进的纳米加工技术可以制造用于干细胞扩增的纳米图案工具和 3D 生物反应器。***纳米加工技术实现了微电子、光学和体外生物系统的突破，以研究细胞与基质的相互作用。然而，有两个关键因素限制了纳米形貌在干细胞生物学研究和细胞扩增的工业放大中的应用：图案化材料的特性和传统的二维图案化培养表面。 因此，在拟议的研究计划中，我们的目标是***1）开发具有可调机械性能和折射率的纳米图案研究工具，以便能够通过超分辨率显微镜研究细胞-基质相互作用； ***2) 研究多能性维持过程中的细胞-基质相互作用，以利用增强 PSC 扩增的最佳生物物理条件； ***3) 开发用于 PSC 扩展的 3D 纳米图案生物反应器； ***4) 应用生物反应器和纳米形貌工具扩大 PSC 的生产以用于研究。***我们的创新方法和研究计划将开发新的研究工具来研究干细胞。 借助新工具，该项目将通过提供对 PSC 谱系承诺的生物物理调节的更深入了解，为干细胞生物学研究带​​来新的见解。 这将是对干细胞和机械生物学研究界的重大贡献。一个简约的、生物物理和生物化学定义的干细胞扩增利基也将提供一种有效的方法来满足不同应用（如生物学研究或药物开发）对一致和可重复细胞的严格需求。 ***最终，该计划将产生新的、可能获得专利的设备和概念，适用于许多研究领域，并将有助于发展加拿大的生物技术产业。它还将为这些重要的学术和工业界培养下一代生物工程师。