| RRC ID |
45496
|
| 著者 |
Yoshie S, Imaizumi M, Nakamura R, Otsuki K, Ikeda M, Nomoto Y, Wada I, Omori K.
|
| タイトル |
Generation of airway epithelial cells with native characteristics from mouse induced pluripotent stem cells.
|
| ジャーナル |
Cell Tissue Res
|
| 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.
|
| 巻・号 |
364(2)
|
| ページ |
319-30
|
| 公開日 |
2016-5-1
|
| DOI |
10.1007/s00441-015-2304-7
|
| PII |
10.1007/s00441-015-2304-7
|
| PMID |
26590823
|
| MeSH |
Activins / pharmacology
Animals
Cell Differentiation / physiology
Cells, Cultured
Embryoid Bodies / cytology*
Epithelial Cells / cytology*
Fibroblast Growth Factors / pharmacology
Hepatocyte Nuclear Factor 3-beta / metabolism
Induced Pluripotent Stem Cells / cytology*
Keratin-5 / metabolism
Mice
Mucin 5AC / metabolism
Nanog Homeobox Protein / metabolism
Receptors, CXCR4 / metabolism
Regeneration / physiology*
Respiratory Mucosa / cytology
Respiratory Mucosa / physiology*
Reverse Transcriptase Polymerase Chain Reaction
Tight Junctions / physiology
Tubulin / metabolism
|
| IF |
4.044
|
| 引用数 |
5
|
|
WOS 分野
|
CELL BIOLOGY
|
| リソース情報 |
| ヒト・動物細胞 |
iPS-MEF-Ng-20D-17(APS0001) |
