Functional characterization of various channel-expressing central airway epithelial cells from mouse induced pluripotent stem cells
Functional characterization of various channel-expressing central airway epithelial cells from mouse induced pluripotent stem cells
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小鼠诱导多能干细胞表达多种通道的中央气道上皮细胞的功能表征
DOI：
10.1002/jcp.28254
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发表时间：
2019
影响因子：
5.6
通讯作者：
Hazama Akihiro
中科院分区：
文献类型：
--
作者：
Yoshie Susumu;Nakamura Ryosuke;Kobayashi Daisuke;Miyake Masao;Omori Koichi;Hazama Akihiro
诱导多能干细胞（iPSCs）的功能性中枢气道上皮细胞（CAECs）是一种有吸引力的中枢气道再生的潜在细胞来源。中央气道上皮，如气管上皮，由纤毛细胞、杯状细胞和基底细胞组成，具有重要的生理功能，如通过Cl -和水通道调节气道表面的水量，以及通过纤毛运动消除从外界环境吸入的颗粒。我们小组和其他研究小组之前的工作已经报道了从ipsc生成气道上皮细胞。然而，目前尚不清楚iPSC衍生的caec是否表达了调节气道表面水量所需的各种通道，以及这些通道是否正常运作。在本研究中，我们利用气液界面培养技术从iPSCs中添加激活素和bFGF生成caec。然后，我们通过使用卤化物离子敏感的黄色荧光蛋白和纤毛运动检测Cl -通道的基因表达和转运功能来评估iPSC衍生的caec的生理功能。逆转录聚合酶链反应和免疫组织化学表明，多种通道标志物，如囊性纤维化跨膜传导调节剂（CFTR）和水通道蛋白（AQP）存在于iPSC衍生的caec中。此外，还成功地证实了Cl−通道和CFTR的输运功能。最后，测量纤毛运动，观察到纤毛跳动频率（CBF）约为10 Hz。这些结果表明，我们的方法生成的caec具有与天然caec相似的生理功能。
Functional central airway epithelial cells (CAECs) from induced pluripotent stem cells (iPSCs) are an attractive potential cell source for central airway regeneration. The central airway epithelium, such as the tracheal epithelium, is composed of ciliated cells, goblet cells, and basal cells and has physiologically important functions such as the regulation of water volume on the airway surface by Cl−and water channels and the elimination of particles inhaled from the external environment by ciliary movement. Previous work from our group and from other research groups has reported the generation of airway epithelial cells from iPSCs. However, it remains unclear whether iPSC‐derived CAECs express the various channels that are required for the regulation of water volume on the airway surface and whether these channels function properly. In this study, we generated CAECs from iPSCs supplemented with activin and bFGF using air–liquid interface culture. We then evaluated the physiological functioning of the iPSC‐derived CAECs by examining the gene expression and transport functions of Cl−channels using a halide ion‐sensitive yellow fluorescent protein and ciliary movement. Reverse‐transcription polymerase chain reaction and immunohistochemistry indicated that various channel markers such as cystic fibrosis transmembrane conductance regulator (CFTR) and aquaporin (AQP) were present in iPSC‐derived CAECs. Furthermore, the transport functions of Cl−channels and CFTR were successfully confirmed. Finally, ciliary movement was measured, and a ciliary beating frequency (CBF) of approximately 10 Hz was observed. These results demonstrate that CAECs generated by our method have physiological functions similar to those of native CAECs.