Secretory cell differentiation and mucus secretion in cultures of human nasal epithelial cells.

我们已经开发了一种空气-液体界面培养系统，用于人鼻上皮细胞，其在限定的无血清培养基中分化为粘膜纤毛表型。将从鼻息肉获得的分离的细胞培养在胶原凝胶基质上。在汇合时，细胞失去分化细胞的特征，并且通过去除顶部培养基并且仅从底部隔室进料来产生气-液界面。在气-液界面中7天后，观察到分泌细胞和纤毛细胞的分化，并且分泌细胞的数量仅增加。在21天时，约一半的上皮细胞用阿新蓝-高碘酸-希夫（AB-PAS）或单克隆抗体HCS 18染色，该抗体针对上皮分泌（杯状）细胞特异性的人鼻粘蛋白。狭缝印迹分析显示，抗体反应性鼻粘蛋白分泌的文化，只有顶端侧，分泌的粘蛋白的量增加，与分泌细胞分化的程度相一致。这些结果表明，人鼻上皮细胞的分泌细胞分化可以通过气-液界面培养物诱导，并且这些细胞分泌人鼻粘蛋白，其对鼻上皮分泌（杯状）细胞是特异性的。本研究中开发的抗粘蛋白单克隆抗体的培养系统应可用于研究各种药理学和生理学操作后人鼻上皮细胞的极化粘液分泌。

We have developed an air-liquid interface culture system for human nasal epithelial cells that differentiate into mucociliary phenotypes in a defined serum-free medium. Dissociated cells obtained from nasal polyp were cultured on a collagen gel substrate. At confluence, the cells lost characteristics of differentiated cells, and an air-liquid interface was produced by removing the apical medium and feeding only from the bottom compartment. After 7 days in an air-liquid interface, secretory cell and ciliated cell differentiation was observed, and the number of secretory cells only increased. At 21 days, about half of the epithelial cells were stained with alcian blue-periodic acid-Schiff (AB-PAS) or an monoclonal antibody HCS18, that was directed against human nasal mucins specific for epithelial secretory (goblet) cells. Slot blot analysis revealed that the antibody-reactive nasal mucins were secreted only the apical side of the cultures, and the amount of secreted mucins increased, coincident with the degree of secretory cell differentiation. These results shows that secretory cell differentiation of human nasal epithelial cells can be induced by air-liquid interface cultures, and these cells secrete human nasal mucins, that are specific for nasal epithelial secretory (goblet) cells. This culture system with an anti-mucin monoclonal antibody developed n this study should be useful for studying polarized mucus secretion from human nasal epithelial cells following various pharmacological and physiological manipulations.