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Culturing of Human Nasal Epithelial Cells at the Air Liquid Interface
Published on: October 8, 2013
Development of rhinovirus study model using organ culture of turbinate mucosa
Yong Ju Jang1, Si Hyeong Lee, Hyon-Ja Kwon
1Department of Otolaryngolgy, Asan Medical Center, University of Ulsan College of Medicine, 388-1 Pungnap-2dong, Songpa-gu, Seoul 138-736, South Korea. jangyj@amc.seoul.kr
Abstract:
To better understand the pathophysiology of rhinovirus (RV) infection, a development of a study model using organ culture of turbinate mucosa was sought. Inferior turbinate mucosal tissues were cultured using air-liquid interface methods, on a support of gelfoam soaked in culture media. RV-16 was applied to the mucosal surface and washed off, and histological changes were evaluated. The success of RV infection was assayed by semi-nested RT-PCR of the mucosal surface fluid taken 48h after incubation. Intracellular RVs were visualized by in situ hybridization (ISH). Secretion of the cytokines, IL-6 and IL-8, into the culture media was quantitated by ELISA. After 7 days of culture, the turbinate mucosae did not show significant damage. A PCR product indicating successful RV infection was detected in 5 out of 10 mucosal tissues. ISH showed a very small number of positively stained cells focally located in the epithelial layer. In the beginning 24h after infection, secretion of IL-6 and IL-8 into the culture media of infected mucosae was significantly greater than into the media of control mucosae. Our results indicate that the air-liquid interface organ culture of turbinate mucosa could serve as an acceptable in vitro model for studying RV infection.
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