Increased communication among nasal epithelial cells in air-liquid interface culture.
OBJECTIVE
The retinoid acid (RA) sufficient air-liquid interface (ALI) cell culture model, but not the classical submerged single layer (SSL) cell culture model, can achieve ciliary differentiation of nasal epithelial cells. Because gap junction mediated intercellular communication (GJIC) may contribute to differentiation in numerous cell types, this study compared the extent of GJIC and the expression of Connexin 43 (Cx43) in nasal epithelial cells in both SSL and ALI cultures.


METHODS
Cell morphology was examined via optical and scanning electron microscope, and the number of cells with ciliary beating were counted. Lucifer Yellow dye transfer test using the scrape loading method was performed to assess the GJIC. Cx43 expression was measured with reverse-transcription polymerase chain reaction (RT-PCR) and quantitative (Q)-PCR.


RESULTS
Nasal epithelial cells in ALI culture exhibited increased numbers of ciliated cells compared with SSL culture during the 3-week culture period. On day 20, GJIC was increased in ALI culture (ALI % - SSL % = 9.6 +/- 1.2%, n = 5). Accordingly, Cx43 expression was increased via RT-PCR (4.22-fold) and Q-PCR (5.3 +/- 1.1-fold, n = 5) examination.


CONCLUSIONS
RA sufficient ALI culture manifested more differentiated nasal epithelial cell status with ciliogenesis. Cx43, being the responsible molecule for GJIC, increased in parallel. Consequently, as in primary cultured limbal epithelial cells, Cx43 expression and extent of GJIC may serve as markers for the differentiation status of nasal epithelial cells.
Yeh
Te-Huei
TH
Department of Otolaryngology, National Taiwan University Hospital, College of Medicine, National Taiwan University, Taipei, Taiwan. yth@ha.mc.ntu.edu.tw
Tsai
Ching-Hwa
CH
Chen
Yuh-Shyang
YS
Hsu
Wei-Chung
WC
Cheng
Chiung-Hsiang
CH
Hsu
Chuan-Jen
CJ
Lee
Shiann-Yann
SY
D002450:Cell Communication / Q000187:drug effects / Q000502:physiology*; D002452:Cell Count; D002478:Cells, Cultured; D018031:Connexin 43 / Q000235:genetics / Q000378:metabolism; D003470:Culture Media / Q000494:pharmacology*; D004848:Epithelium / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D015870:Gene Expression; D006801:Humans; D008855:Microscopy, Electron, Scanning; D009297:Nasal Mucosa / Q000187:drug effects / Q000502:physiology / Q000648:ultrastructure*; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014212:Tretinoin
D018031:Connexin 43; D003470:Culture Media; D012333:RNA, Messenger; D014212:Tretinoin