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首页> 外文期刊>Physiological Reports >Epithelial monolayer culture system for real‐time single‐cell analyses
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Epithelial monolayer culture system for real‐time single‐cell analyses

机译：上皮单层培养系统，用于实时单细胞分析

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AbstractMany epithelial cells form polarized monolayers under in vivo and in vitro conditions. Typically, epithelial cells are cultured for differentiation on insert systems where cells are plated on a porous filter membrane. Although the cultured monolayers have been a standard system to study epithelial physiology, there are some limits: The epithelial cells growing inside the commercial inserts are not optimal to visualize directly through lenses on inverted microscopes. The cell images are optically distorted and background fluorescence is bright due to the filter membrane positioned between the cells and the lens. In addition, the cells are not easily accessible by electrodes due to the presence of tall side walls. Here, we present the design, fabrication, and practical applications of an improved system for analysis of polarized epithelial monolayers. This new system allows (1) direct imaging of cells without an interfering filter membrane, (2) electrophysiological measurements, and (3) detection of apical secretion with minimal dilution. Therefore, our culture method is optimized to study differentiated epithelial cells at the single-cell and subcellular levels, and can be extended to other cell types with minor modifications.
机译：摘要许多上皮细胞在体内和体外条件下会形成极化的单层膜。通常，在插入系统上培养上皮细胞以分化，在插入系统中将细胞铺板在多孔滤膜上。尽管培养的单层细胞已成为研究上皮生理的标准系统，但仍存在一些局限性：商业化插入物中生长的上皮细胞不是最佳的可通过倒置显微镜直接在镜片上可视化的方法。由于位于细胞和晶状体之间的滤膜，细胞图像发生了光学畸变，背景荧光明亮。另外，由于存在高侧壁，电池不容易被电极接近。在这里，我们介绍了一种用于极化上皮单层分析的改进系统的设计，制造和实际应用。这个新系统允许（1）在不干扰滤膜的情况下直接对细胞成像，（2）电生理学测量，以及（3）以最小的稀释度检测根尖分泌物。因此，我们的培养方法经过优化，可以在单细胞和亚细胞水平上研究分化的上皮细胞，并且可以稍加修改即可扩展到其他细胞类型。

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