Grün Christoph, Pfeifer Jana, Liebsch Gregor, Gottwald Eric
Institute of Functional Interfaces, Karlsruhe Institute of Technology, Karlsruhe, Germany.
PreSens Precision Sensing GmbH, Regensburg, Germany.
Front Bioeng Biotechnol. 2023 Feb 20;11:1111316. doi: 10.3389/fbioe.2023.1111316. eCollection 2023.
Oxygen concentration plays a crucial role in (3D) cell culture. However, the oxygen content is usually not comparable to the situation, which is partly due to the fact that most experiments are performed under ambient atmosphere supplemented with 5% CO, which can lead to hyperoxia. Cultivation under physiological conditions is necessary, but also fails to have suitable measurement methods, especially in 3D cell culture. Current oxygen measurement methods rely on global oxygen measurements (dish or well) and can only be performed in 2D cultures. In this paper, we describe a system that allows the determination of oxygen in 3D cell culture, especially in the microenvironment of single spheroids/organoids. For this purpose, microthermoforming was used to generate microcavity arrays from oxygen-sensitive polymer films. In these oxygen-sensitive microcavity arrays (sensor arrays), spheroids cannot only be generated but also cultivated further. In initial experiments we could show that the system is able to perform mitochondrial stress tests in spheroid cultures to characterize mitochondrial respiration in 3D. Thus, with the help of sensor arrays, it is possible to determine oxygen label-free and in real-time in the immediate microenvironment of spheroid cultures for the first time.
氧气浓度在（3D）细胞培养中起着至关重要的作用。然而，氧气含量通常与实际情况不可比，部分原因是大多数实验是在补充了5%二氧化碳的环境气氛下进行的，这可能导致高氧。在生理条件下培养是必要的，但也缺乏合适的测量方法，尤其是在3D细胞培养中。目前的氧气测量方法依赖于整体氧气测量（培养皿或孔板），并且只能在2D培养中进行。在本文中，我们描述了一种系统，该系统能够测定3D细胞培养中的氧气含量，特别是在单个球体/类器官的微环境中。为此，采用微热成型技术从氧敏聚合物薄膜制备微腔阵列。在这些氧敏微腔阵列（传感器阵列）中，不仅可以生成球体，还可以进一步培养。在初步实验中，我们能够证明该系统能够在球体培养中进行线粒体应激测试，以表征3D中的线粒体呼吸。因此，借助传感器阵列，首次有可能在球体培养的直接微环境中无标记且实时地测定氧气。
