High-content imaging (HCI) is a powerful method for quantifying biological effects in vitro. Historically, HCI has been applied to adherent cells growing in monolayers. With the advent of confocal versions of HCI devices, researchers now have the option of performing analyses on 3D cell cultures. However, some obstacles remain in integrating the third dimension, such as limited light penetration and less sophisticated image analysis. Here, we report the development of an HCI technique for imaging human bronchial 3D organotypic air-liquid interface (ALI) cultures (hBR-ALI). In this method, we monitored differentiation status through HCI evaluation markers representative of ciliated epithelial cells and goblet cells (Muc5AC [mucin 5AC]). As a second use case for demonstrating the utility of this technique, we induced goblet cell hyperplasia in hBR-ALI by using interleukin (IL)-13. Our results demonstrate the utility of the HCI technique for imaging hBR-ALI grown on Transwell inserts. This technique may be expanded to other cell culture systems, such as skin epithelia and 3D intestinal systems.

中文翻译：

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体外基于高内涵成像的支气管3D器官型气液界面培养的表型分析。

高内涵成像（HCI）是在体外定量生物学效应的强大方法。从历史上看，HCl已应用于单层生长的贴壁细胞。随着共聚焦版本的HCI设备的出现，研究人员现在可以选择对3D细胞培养物进行分析。但是，在集成三维空间时仍存在一些障碍，例如光穿透力有限和图像分析不够复杂。在这里，我们报告了用于成像人支气管3D有机型气液界面（ALI）文化（hBR-ALI）的HCI技术的发展。在这种方法中，我们通过代表纤毛上皮细胞和杯状细胞（Muc5AC [粘蛋白5AC]）的HCI评估标记监测分化状态。作为展示该技术实用性的第二个用例，我们通过使用白介素（IL）-13诱导hBR-ALI中的杯状细胞增生。我们的结果证明了HCI技术可用于在Transwell插入片段上生长的hBR-ALI成像。该技术可以扩展到其他细胞培养系统，例如皮肤上皮细胞和3D肠道系统。