| Budget Amount *help |
¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1997: ¥1,000,000 (Direct Cost: ¥1,000,000)
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| Research Abstract |
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.
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