Functional characterization of various channel-expressing central airway epithelial cells from mouse induced pluripotent stem cells.

Abstract

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.