Noguchi Y, Uchida Y, Endo T, Ninomiya H, Nomura A, Sakamoto T, Goto Y, Haraoka S, Shimokama T, Watanabe T
Department of Pulmonary Medicine, University of Tsukuba, Ibaraki, Japan.
Biochem Biophys Res Commun. 1995 May 16;210(2):302-9. doi: 10.1006/bbrc.1995.1661.
We have developed a culture system of guinea pig tracheal epithelial cells using the epithelium-denuded human amnion as a source of basement membrane. Culture medium fluid over the epithelial cells was replaced by air after 7-day immersion culture and thereafter maintained for 2 weeks. Electron microscopical observations revealed that the height of epithelial cells and the ratio of ciliated epithelial cells looked like that of normal guinea pigs epithelium growth in 2 weeks after the air interface, but that no goblet cells could be found. In order to study cell polarity, we measured endothelin-1 levels in the media of apical and basal sides of the epithelial cell monolayer by means of the enzyme-linked immunosorbent assay. The endothelin-1 content of the submucosal side was over 30 times higher than that of the apical side. These findings suggest that ET-1 would be mainly released from airway epithelial cells toward the submucosal side.
我们利用去上皮的人羊膜作为基底膜来源，开发了一种豚鼠气管上皮细胞培养系统。在进行7天的浸没培养后，将上皮细胞上的培养基液替换为空气，然后维持2周。电子显微镜观察显示，在空气界面培养2周后，上皮细胞的高度和纤毛上皮细胞的比例看起来与正常豚鼠上皮的生长情况相似，但未发现杯状细胞。为了研究细胞极性，我们通过酶联免疫吸附测定法测量了上皮细胞单层顶端和基底侧培养基中的内皮素-1水平。黏膜下层侧的内皮素-1含量比顶端侧高30倍以上。这些发现表明，ET-1主要从气道上皮细胞释放到黏膜下层侧。
