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

Research Project

Project/Area Number	08671960
Research Category	Grant-in-Aid for Scientific Research (C)
Allocation Type	Single-year Grants
Section	一般
Research Field	Otorhinolaryngology
Research Institution	Mie University
Principal Investigator	SHIMIZU Takeshi Mie University Hospital, Lecture, 医学部・附属病院, 講師 (00206202)
Project Period (FY)	1996 – 1997
Project Status	Completed (Fiscal Year 1997)
Budget Amount *help	¥1,000,000 (Direct Cost: ¥1,000,000) Fiscal Year 1997: ¥1,000,000 (Direct Cost: ¥1,000,000)
Keywords	mucus / secretion / goblet cell / secretory cell / cell culture / nasal epithelium / monoclonal antibody / MUC gene / ムチン / マクロライド / サイトカイン / 細胞分化 / 線毛細胞
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.