IDF: Controlled Network Disruption and Spatiotemporal Sampling of Microperfused 3-D Neural Cultures

培养和操纵细胞的能力对于理解正常和疾病过程的基本机制至关重要。尤其是三维粗大的神经培养物，可以模拟脑组织，但由于缺乏输送营养和清除废物的血液供应而受到限制。迫切需要开发新技术来满足这一需求，并产生有效的脑组织模型。这项跨学科研究的智力价值在于一种新的培养模型的工程设计，该模型包括大脑中的主要细胞类型：神经元、星形胶质细胞和小胶质细胞，以及一个高度可控的微流体系统，该系统在整个培养过程中灌注营养物质，并允许在正常条件下和细胞损伤期间对细胞培养基进行废物清除和取样。一些新的创新元素将被纳入：1)包括炎症细胞（小胶质细胞），以创建一个更真实的细胞模型；2)引入一种独特的基于超声的损伤模型，在培养物中产生局部损伤；3)加入微流体进行灌注和取样。因此，总体目标是创建一个强大而复杂的神经组织等同物，将忠实地代表大脑，并研究炎症触发后小胶质细胞的作用。在任务1中，选择最合适的构建块来创建一个用于研究炎症的新颖复杂的3-D神经系统。此外，微流体将集成到包括灌注和采样能力。此外，将建立一个新的创伤损伤模型，为使用高度可控和可调的方法详细研究损伤机制提供手段。在任务2中，将测试小胶质细胞在损伤反应中的作用，因为假设损伤小胶质细胞释放的细胞因子会增加细胞死亡。这项研究意义重大，因为强大的培养工具结合了多种细胞类型和微流体灌注和采样，为确定正常和损伤细胞的机制提供了前所未有的空间和时间控制水平。这一研究方向的更广泛影响将是开发可用于众多应用的极其新颖的神经组织等效物。预计通过这种方法实现的下一代培养系统将彻底改变神经细胞培养的方式，因为考虑了细胞类型之间的复杂相互作用，并通过微灌注模拟微循环。具有这些功能的三维组织模型将推动基础科学发现转化为工业、政府和医学突破。这些技术发现将与大学、政府和行业研究人员分享，重点是合作，并最终对那些受创伤性损伤或其他神经系统疾病影响的人产生影响。

0933506LaPlacaThe ability to grow and manipulate cells in culture is crucial to understanding basic mechanisms of both normal and disease processes. Three-dimensional thick neural cultures, in particular, mimic brain tissue, but are limited by the lack of a blood supply to deliver nutrients and remove waste. There is a critical need to develop new technology to address this need and produce valid brain tissue models. The intellectual merit of this interdisciplinary research lies in the engineering of a new culture model that includes the major cell types in the brain: neurons, astrocytes, and microglia, together with a highly controllable microfluidic system that perfuses nutrients throughout the culture and permits waste removal and sampling of the cell culture media during periods of both normal conditions and cell injury. Several new innovative elements will be incorporated: 1) include inflammatory cells (microglia) to create a more realistic cell model; 2) introduce a unique, ultrasound-based injury model to produce local injury within the culture; and 3) incorporate microfluidics for perfusion and sampling. Thus, the overall objective is to create a robust and complex neural tissue equivalent that will faithfully represent brain and to investigate the role of microglia following inflammatory triggers. In Task 1, the most appropriate building blocks are chosen to create a novel, complex 3-D neural system for studying inflammation. In addition, microfluidics will be integrated to include perfusion and sampling capabilities. Furthermore, a new traumatic injury model will be developed, providing a means for detailed study of injury mechanisms using highly controllable and tunable methodology. In Task 2, the role of the microglia in the injury response will be tested, as cytokines released from injured microglia are hypothesized to increase cell death. This research is highly significant, as robust culture tools that incorporate multiple cell types and microfluidic perfusion and sampling offer unprecedented levels of spatial and temporal control for determining mechanisms of both normal and injured cells. The broader impact of this research direction will be the development of extremely novel neural tissue equivalents that can be used for numerous applications. It is expected that the next generation of culture systems realized by this approach will revolutionize the way neural cell culturing is done, as the complex interactions among cell types are considered and microcirculation is mimicked through microperfusion. Three-dimensional tissue models with these capabilities will push forward the translation of basic science discoveries for industry, government, and medical breakthroughs. The technical findings will be shared with university, government, and industry researchers with emphasis on collaboration and ultimately having an impact on those affected with traumatic injury or other neurological disorders.