Toward Automated Additive Manufacturing of Living Bio-Tubes Using Ring-Shaped Building Units
Toward Automated Additive Manufacturing of Living Bio-Tubes Using Ring-Shaped Building Units
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DOI：
10.1177/2472630320920896
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发表时间：
2020-05-26
期刊：
影响因子：
2.7
通讯作者：
Morgan, Jeffrey R.
中科院分区：
文献类型：
--
作者：
Manning, Kali L.;Feder, Jacob;Morgan, Jeffrey R.
组织工程在很大程度上局限于学术研究机构，在商业环境中取得的成功有限。为了帮助解决这个问题，需要做更多的工作来开发新的组织相关技术的自动化制造工艺。在这篇文章中，我们描述了漏斗导向器的自动化，这是一种增材制造方法，使用活组织环作为构建单元来形成生物管。我们开发了一种基于96孔板和改进的现成液体处理机器人的方法，以检索、执行实时质量控制并将组织环转移到漏斗导向器。将细胞接种到含有特殊设计的琼脂糖微模具的96孔板中，细胞自组装并形成环形微组织，可以使用液体处理机器人取回。我们表征了时间、细胞类型和模具几何形状对环形微组织形态的影响，以告知建筑部件的最佳使用。我们编程并修改了现成的液体处理机器人，以从96孔板中检索环形微组织，并且我们制造了定制的照明移液管，以在漏斗引导器中存款之前可视化每个环形微组织。在液体-空气界面的成像提出了通过控制照明条件和液体曲率来克服的挑战。根据这些图像，我们在工作流程中纳入了基于目视检查和形态标准的实时质量控制步骤，以在使用前评估每个环。我们用这个系统来制造生物管的内皮细胞与管腔对齐。
Tissue engineering has been largely confined to academic research institutions with limited success in commercial settings. To help address this issue, more work is needed to develop new automated manufacturing processes for tissue-related technologies. In this article, we describe the automation of the funnel-guide, an additive manufacturing method that uses living tissue rings as building units to form bio-tubes. We developed a method based on 96-well plates and a modified off-the-shelf liquid-handling robot to retrieve, perform real-time quality control, and transfer tissue rings to the funnel-guide. Cells seeded into 96-well plates containing specially designed agarose micromolds self-assembled and formed ring-shaped microtissues that could be retrieved using a liquid-handling robot. We characterized the effects of time, cell type, and mold geometry on the morphology of the ring-shaped microtissues to inform optimal use of the building parts. We programmed and modified an off-the-shelf liquid-handling robot to retrieve ring-shaped microtissues from the 96-well plates, and we fabricated a custom illuminated pipette to visualize each ring-shaped microtissue prior to deposit in the funnel guide. Imaging at the liquid-air interface presented challenges that were overcome by controlling lighting conditions and liquid curvature. Based on these images, we incorporated into our workflow a real-time quality control step based on visual inspection and morphological criteria to assess each ring prior to use. We used this system to fabricate bio-tubes of endothelial cells with luminal alignment.