Research on activation and differentiation of airway epithelial cells by using of 3-dimensional culture system.

我们成功地将豚鼠气管上皮细胞培养在人羊膜上。在空气界面饲养条件下，3周内观察到纤毛细胞。然而，在该培养系统中未发现任何杯状细胞或基底细胞。将豚鼠气管上皮细胞与人羊膜下生长的豚鼠肺成纤维细胞共培养。这种共培养促进上皮细胞分化为纤毛细胞、杯状细胞和基底细胞。此外，上皮层形成假多层，上皮细胞看起来更立方比没有成纤维细胞。此外，我们还寻找在上皮细胞和成纤维细胞共培养条件下促进假多层膜形成的因素。我们在单独的上皮细胞培养基上添加了几种生长因子，如EGF-1、IGF-1、PDGF、TGF-β、HGF或FGF-2。支气管哮喘是一种慢性气道炎症性疾病，气道壁重塑是其发展过程中的重要现象。慢性哮喘患者气道内可见炎性细胞浸润，基底膜增厚，杯状细胞增生。气道上皮细胞可能在这一过程中起重要作用。因此，阐明气道重塑的机制对支气管哮喘的研究具有不可忽视的意义。

We succeeded in the culture of guinea-pig tracheal epithelial cells on the human amnion membrane. Ciliated cells were observed during 3 weeks in the condition of air-interface feeding. However, any goblet or basal cells were not found in this culture system. The tracheal epithelial cells of the guinea pig were co-cultured with fibroblasts of the guineapig lung, which grew under the human amnion membrane. This co-culture facilitated the differentiation of epithelial cells into ciliated cells, goblet cells and basal cells. Moreover, epithelial layr formed a pseudomultilayr and epithelial cells looked more cuboidal than that without fibroblasts. Also, we looked for the factor that facilitated pseudomultilayr in the co-culture-condition of epithelial cells and fibroblasts. We added several growth factors such as EGF-1, IGF-1, PDGF,TGF-beta, HGF or FGF-2 on the culture medium of epithelial cells alone. Only FGF-2 facilitated pseudomultilayr in the epithelial cell culture.Bronchial asthma has been recognized as chronic airway inflammatory disease, in which developing process remodeling of the airway wall is a critical phenomenon. We can observe inflammatory cell infiltration into the airway, thickening of the basement membrane and hyperplasia of goblet cells in the airway of chronic asthmatics. Airway epithelial cell may play an important role in those process. Then, it is not negligible to elucidate the mechanism of remodeling of the airway on the research of bronchial asthma.