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Cell and Tissue Research

, Volume 364, Issue 2, pp 319–330 | Cite as

Generation of airway epithelial cells with native characteristics from mouse induced pluripotent stem cells

  • Susumu Yoshie
  • Mitsuyoshi Imaizumi
  • Ryosuke Nakamura
  • Koshi Otsuki
  • Masakazu Ikeda
  • Yukio Nomoto
  • Ikuo Wada
  • Koichi OmoriEmail author
Regular Article

Abstract

Airway epithelial cells derived from induced pluripotent stem (iPS) cells are expected to be a useful source for the regeneration of airway epithelium. Our preliminary study of embryoid body (EB) formation and the air-liquid interface (ALI) method suggested that mouse iPS cells can differentiate into airway epithelial cells. However, whether the cells generated from mouse iPS cells had the character and phenotype of native airway epithelial cells remained uninvestigated. In this study, we generated airway epithelial cells from EBs by culturing them under serum-free conditions supplemented with Activin and bFGF and by the ALI method and characterized the iPS cell-derived airway epithelial cells in terms of their gene expression, immunoreactivity, morphology, and function. Analysis by quantitative real-time reverse transcription-polymerase chain reaction(RT-PCR) revealed that the expression of the undifferentiated cell marker Nanog decreased time-dependently after the induction of differentiation, whereas definitive endoderm markers Foxa2 and Cxcr4 were transiently up-regulated. Thereafter, the expression of airway epithelium markers such as Tubb4a, Muc5ac, and Krt5 was detected by RT-PCR and immunostaining. The formation of tight junctions was also confirmed by immunostaining and permeability assay. Analysis by hematoxylin and eosin staining and scanning electron microscopy indicated that the cells generated from mouse iPS cells formed airway-epithelium-like tissue and had cilia, the movement of which was visualized and observed to be synchronized. These results demonstrate that the airway epithelial cells generated by our method have native characteristics and open new perspectives for the regeneration of injured airway epithelium.

Keywords

iPS cells Airway epithelial cells Cilia Embryoid bodies Mouse 

Notes

Acknowledgment

We thank Ms. Etsuko Sato for excellent technical assistance.

Supplementary material

Fig. S1

Ciliary movement of mouse iPS-cell-derived airway epithelial cells. The bright-field images of the beating cilia were captured at 200 frames/s. The movie is shown at 30 frames/s. Bar 10 μm. (AVI 29523 kb)

Fig. S2

Ciliary movement of mouse trachea. The bright-field images of the beating cilia were captured at 200 frames/s. The movie is shown at 30 frames/s. Bar 10 μm. (MOV 10041 kb)

Fig.S3

Coordination of ciliary movement of mouse iPS-cell-derived airway epithelial cells. The movie is shown at 30 frames/s. Bar 200 μm. (MOV 10041 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Susumu Yoshie
    • 1
  • Mitsuyoshi Imaizumi
    • 1
  • Ryosuke Nakamura
    • 1
  • Koshi Otsuki
    • 1
  • Masakazu Ikeda
    • 1
  • Yukio Nomoto
    • 1
  • Ikuo Wada
    • 2
  • Koichi Omori
    • 1
    Email author
  1. 1.Department of OtolaryngologySchool of Medicine, Fukushima Medical University, Fukushima, JapanFukushima CityJapan
  2. 2.Departments of Cell ScienceSchool of Medicine, Fukushima Medical University, Fukushima, JapanFukushima CityJapan

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