Huang Song, Boda Bernadett, Vernaz Jimmy, Ferreira Emilie, Wiszniewski Ludovic, Constant Samuel
Epithelix, 14 Chemin des Aulx, Plan-Les-Ouates, CH-1228 Geneva, Switzerland.
Epithelix, 14 Chemin des Aulx, Plan-Les-Ouates, CH-1228 Geneva, Switzerland.
Eur J Pharm Biopharm. 2017 Sep;118:68-72. doi: 10.1016/j.ejpb.2016.12.006. Epub 2016 Dec 28.
We report here the establishment and characterization of an in vitro human small airway model (SmallAir™). The epithelial cells were isolated from the distal lungs by enzymatic digestion. After amplification, the cells were seeded on the microporous membrane of Transwell inserts. Once confluent, the cultures were switched to air-liquid interface. After 3weeks of culture, the epithelium became fully differentiated, with morphology of columnar epithelium, and a thickness of 10-15μm. Most significantly, CC-10, a specific marker of Club cells, was highly expressed in SmallAir™. CC-10 was detected by both immune-cytochemistry and Western Blot. As expected, SmallAir™ contained few Muc5-Ac positive cells (goblet cells). In contrast, CC-10 was not detected in MucilAir™, an in vitro model of the human nasal and bronchial epithelial model. Instead, Muc5-Ac was highly expressed in MucilAir™. However, both MucilAir™ and SmallAir™ contain basal cells and ciliated cells, showing cilia beating and mucociliary clearance. Clearly, MucilAir™ and SmallAir™ are two distinct airway epithelial models.
我们在此报告一种体外人小气道模型（SmallAir™）的建立与特性。上皮细胞通过酶消化从肺远端分离得到。扩增后，将细胞接种于Transwell小室的微孔膜上。细胞汇合后，培养转换为气液界面培养。培养3周后，上皮细胞完全分化，呈柱状上皮形态，厚度为10 - 15μm。最显著的是，Club细胞的特异性标志物CC - 10在SmallAir™中高表达。通过免疫细胞化学和蛋白质免疫印迹法均检测到CC - 10。正如预期，SmallAir™中Muc5 - Ac阳性细胞（杯状细胞）很少。相反，在人鼻和支气管上皮模型的体外模型MucilAir™中未检测到CC - 10。取而代之的是，Muc5 - Ac在MucilAir™中高表达。然而，MucilAir™和SmallAir™均含有基底细胞和纤毛细胞，表现出纤毛摆动和黏液纤毛清除功能。显然，MucilAir™和SmallAir™是两种不同的气道上皮模型。
