Struber Andreas, Auer Georg, Fischlechner Martin, Wickstrom Cody, Reiter Lisa, Lutsch Eric, Simon-Nobbe Birgit, Marozin Sabrina, Lepperdinger Günter
Department of Biosciences and Medical Biology, University Salzburg, A-5020 Salzburg, Austria.
Sarcura GmbH, A-3400 Klosterneuburg, Austria.
Bioengineering (Basel). 2022 Feb 3;9(2):60. doi: 10.3390/bioengineering9020060.
The wide use of 3D-organotypic cell models is imperative for advancing our understanding of basic cell biological mechanisms. For this purpose, easy-to-use enabling technology is required, which should optimally link standardized assessment methods to those used for the formation, cultivation, and evaluation of cell aggregates or primordial tissue. We thus conceived, manufactured, and tested devices which provide the means for cell aggregation and online monitoring within a hanging drop. We then established a workflow for spheroid manipulation and immune phenotyping. This described workflow conserves media and reagent, facilitates the uninterrupted tracking of spheroid formation under various conditions, and enables 3D-marker analysis by means of 3D epifluorescence deconvolution microscopy. We provide a full description of the low-cost manufacturing process for the fluidic devices and microscopic assessment tools, and the detailed blueprints and building instructions are disclosed. Conclusively, the presented compilation of methods and techniques promotes a quick and barrier-free entry into 3D cell biology.
广泛使用3D器官型细胞模型对于增进我们对基本细胞生物学机制的理解至关重要。为此，需要易于使用的赋能技术，该技术应将标准化评估方法与用于细胞聚集体或原始组织形成、培养和评估的方法进行最佳链接。因此，我们构思、制造并测试了能够在悬滴内实现细胞聚集和在线监测的装置。然后，我们建立了球体操作和免疫表型分析的工作流程。所描述的工作流程节省了培养基和试剂，便于在各种条件下不间断地跟踪球体形成，并通过3D落射荧光反卷积显微镜进行3D标记分析。我们全面描述了流体装置和微观评估工具的低成本制造过程，并公开了详细的蓝图和构建说明。总之，所展示的方法和技术汇编促进了对3D细胞生物学的快速且无障碍的入门。
