Generation of airway epithelial cells with native characteristics from mouse induced pluripotent stem cells
Generation of airway epithelial cells with native characteristics from mouse induced pluripotent stem cells
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从小鼠诱导多能干细胞中产生具有天然特征的气道上皮细胞
DOI：
10.1007/s00441-015-2304-7
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发表时间：
2015
期刊：
影响因子：
3.6
通讯作者：
Omori K
中科院分区：
文献类型：
--
作者：
Yoshie S;Imaizumi M;Nakamura R;Otsuki K;Ikeda M;Nomoto Y;Wada I;Omori K
来源于诱导多能干细胞（iPS）的气道上皮细胞有望成为气道上皮再生的有用来源。我们对拟胚体（EB）形成和气液界面（ALI）方法的初步研究表明，小鼠iPS细胞可以分化为气道上皮细胞。然而，由小鼠iPS细胞产生的细胞是否具有天然气道上皮细胞的特征和表型仍未研究。在这项研究中，我们通过在无血清条件下培养EB并补充激活素和bFGF以及通过ALI方法产生气道上皮细胞，并根据其基因表达、免疫反应性、形态学和功能来表征iPS细胞衍生的气道上皮细胞。实时定量逆转录聚合酶链反应（RT-PCR）分析显示，诱导分化后，未分化细胞标志物Nanog的表达呈时间依赖性下降，而定形内胚层标志物Foxa 2和Cxcr 4则瞬时上调。然后采用RT-PCR和免疫组化方法检测气道上皮标志物Tubb 4a、Muc 5ac和Krt 5的表达。紧密连接的形成也证实了免疫染色和渗透性测定。通过苏木精和伊红染色和扫描电子显微镜分析表明，由小鼠iPS细胞产生的细胞形成气道上皮样组织并具有纤毛，纤毛的运动是可视化的并观察到是同步的。这些结果表明，由我们的方法产生的气道上皮细胞具有天然特性，并为损伤气道上皮的再生开辟了新的前景。
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 markerNanogdecreased time-dependently after the induction of differentiation, whereas definitive endoderm markersFoxa2andCxcr4were transiently up-regulated. Thereafter, the expression of airway epithelium markers such asTubb4a,Muc5ac, andKrt5was 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.
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影响因子：
4.6
作者：
Molly Weaver;J. Yingling;N. R. Dunn;S. Bellusci;Brigid L. M. Hogan
通讯作者：
Molly Weaver;J. Yingling;N. R. Dunn;S. Bellusci;Brigid L. M. Hogan
DOI：
10.1172/jci15217
发表时间：
2002-03
期刊：
The Journal of clinical investigation
影响因子：
--
作者：
M. Knowles;R. Boucher
通讯作者：
M. Knowles;R. Boucher
影响因子：
64.5
作者：
Takahashi, Kazutoshi;Yamanaka, Shinya
通讯作者：
Yamanaka, Shinya
单个动力蛋白分子的振荡运动模型。
影响因子：
2
作者：
D. Goedecke;T. Elston
通讯作者：
T. Elston
通过诱导多能干细胞衍生的器官芽移植生成人类肝脏组织。
DOI：
--
发表时间：
--
期刊：
影响因子：
--
作者：
武部貴則;小林眞司;矢吹雄一郎;鈴木啓;水野満;安村和正;広富浩一;鄭允文;前川二郎;谷口英樹;Takebe T;Enomura M;Zheng Y;Zhang R;Sekine K;Mizuno M;武部貴則;武部貴則;武部貴則;谷口英樹;Takanori Takebe;Takanori Takebe
通讯作者：
Takanori Takebe