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

提案编号：1547693一种类型的生物制造涉及在二维(2D)和三维(3D)中将细胞放置在一起。为了更好地了解健康组织和患病组织在体内发生的情况，必须了解细胞的生长和死亡。就像在不杀死树枝的情况下修剪树枝一样，该项目的研究人员将使用一种名为细胞凋亡的受控自然过程来修剪2D和3D环境中的细胞组，以改善整体结构的功能。这些研究可以加深我们对如何控制正常细胞进入组织以及如何控制癌症等疾病过程的理解。这个项目中要研究的细胞的容器将包括使用3D打印机产生的生物反应器。该项目的技术部分寻求解决在2D和3D环境中生成和成形单元组件的挑战。该项目旨在了解正常细胞和癌细胞的生长过程，目标是实现更好的生物制造策略和对肿瘤生长的洞察。除了生长，这一奖项的研究人员还将使用两种类型的高纵横比结构(HARS)对细胞施加凋亡刺激，以控制的方式剪除细胞。在这个项目中，为了便于成像到更厚(0.5 mm)的3D细胞组件，将使用结合多光子显微镜的梯度折射率(GRIN)透镜。该项目使用的HARS材料包括中空的、不可降解的埃洛石，以及一种新型的、可生物降解的含铜生物复合材料。这两种HARS材料从直径的纳米尺度到长度的微尺度。这个项目中进行的实验将利用3D打印机产生的生物反应器，生物反应器的功能输出将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来检测谷氨酸和pH动态，以比较细胞产量和细胞生长在细胞凋亡修剪过程前后的变化。为了更好地模拟大脑中的动态过程，还将添加小胶质细胞来模拟细胞凋亡后的恢复。为该项目生成的3D打印生物反应器的代工厂将以生物反应器图像、.stl文件和动画的形式建立，并将进行测试，以与商业上可用的微流体设备集成，以检测例如随着时间的推移在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可用性越来越高，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.