EAGER: Biomanufacturing: Multi-scale high-aspect ratio structures (HARS) for constructing dynamic 2D and 3D cellular bioreactors

PI: DeCoster, Mark A.提案号：1547693一种生物制造涉及将细胞在二维（2D）和三维（3D）中放置在一起。为了更好地了解身体健康和病变组织中发生的情况，必须了解细胞的生长和细胞死亡。就像修剪一棵树的枝干而不杀死它一样，该项目的研究人员将使用一种被称为细胞凋亡的受控自然过程，在2D和3D环境中修剪细胞群，以改善整体结构的功能。这些研究可以增强我们对如何控制正常细胞形成组织以及如何控制癌症等疾病过程的理解。在这个项目中要研究的细胞容器将包括使用3D打印机生成的生物反应器。该项目的技术组件旨在解决在2D和3D环境中生成和塑造细胞组件的挑战。该项目旨在了解正常细胞和癌细胞的生长过程，以实现更好的生物制造策略和洞察肿瘤生长。除了生长之外，EAGER奖的研究人员还将使用两种类型的高纵横比结构（HARS）对细胞进行凋亡刺激，以一种可控的方式修剪细胞。为了便于在本项目中成像成更厚（0.5 mm）的3D细胞组件，将使用梯度指数（GRIN）透镜结合多光子显微镜。该项目中使用的HARS材料包括一种中空的、不可降解的高岭土和一种新型的、可生物降解的含铜生物复合材料。两种HARS材料从直径的纳米尺度扩展到长度的微米尺度。本项目中进行的实验将利用3D打印机生成的生物反应器和生物反应器的功能输出，包括使用快速生长的胶质瘤细胞系和生长较慢的（正常）星形胶质细胞原代培养模型检测谷氨酸和pH动态，以比较细胞输出与凋亡修整过程前后的生长情况。为了更好地模拟大脑的动态过程，还将添加小胶质细胞来模拟细胞凋亡后的恢复。将以生物反应器图像的形式建立为该项目生成的3D打印生物反应器的铸造厂。STL文件和动画，并将测试与市售的微流体设备集成，以检测，例如，随着时间的推移，生物反应器中发生的化学变化。考虑到3d打印技术和设计软件的日益普及，预计该项目的结果将影响未来的生物制造战略和教育材料。

PI: DeCoster, Mark A. Proposal Number: 1547693One type of Biomanufacturing involves placing cells together in both two dimensions (2D) and three dimensions (3D). To better approximate what happens in the body in both healthy and diseased tissues, the growth of cells, as well as cell death must be understood. Much like pruning the limbs of a tree without killing it, the investigators of this project will use a controlled, natural process called apoptosis to prune groups of cells in both 2D and 3D environments to improve the function of the overall construct. These studies could enhance our understanding of how to control normal cell formations into tissues and how to control disease processes such as cancer. The containers for the cells to be studied in this project will include bioreactors generated using 3D printers. The technological components of this project seek to address the challenges of generating and shaping assemblies of cells in both 2D and 3D environments. The project aims to understand the growth processes of both normal and cancer cells, with the goal of achieving better biomanufacturing strategies and insight into tumor growth. Beyond just growth, the investigators of this EAGER award will also apply apoptotic stimuli to cells using two types of high-aspect ratio structures (HARS), to prune away cells in a controlled manner. To facilitate imaging into thicker (0.5 mm) 3D cell assemblies in this project, gradient index (GRIN) lenses combined with multi-photon microscopy will be used. The HARS materials used in this project include a hollow, non-degradable halloysite, and a novel, biodegradable biocomposite containing copper. Both HARS materials scale from the nano-dimension in diameter to the micro-dimension in length. The experiments carried out in this project will utilize bioreactors generated using 3D printers and functional outputs from the bioreactors will include detection of glutamate and pH dynamics using a fast-growing glioma cell line and slower growing (normal) astrocyte primary culture model to compare cellular outputs with growth before and after the pruning process of apoptosis. To better approximate dynamic processes in the brain, microglia will also be added to model recovery after apoptosis. A foundry of 3D printed bioreactors generated for the project will be established in the form of bioreactor images, .stl files, and animations, and will be tested for integration with commercially available millifluidic devices to detect, for example, chemical changes occurring in the bioreactors over time. Results from this project are anticipated to impact future biomanufacturing strategies and educational materials considering the increasing availability of 3D-printing technology and design software.