Air-liquid interface culture of serially passaged human nasal epithelial cell monolayer for in vitro drug transport studies.
The objective of this study was to establish a drug transport study using human nasal epithelial (HNE) cell monolayers cultured by the air-liquid interface (ALI) method using serum-free medium (BEGM:DME/F12, 50:50). The cells were developed and characterized in comparison to those that have been previously cultured by the liquid-covered culture (LCC) method. The epithelial cell monolayer cultured by the ALI method resulted in a significantly higher transepithelial electrical resistance value (3,453 +/- 302 ohm x cm(2)) that was maintained (>1,000 ohm x cm(2)) for up to 20 days compared with that cultured by the LCC method. Observation by scanning electron microscopy revealed mature cilia after 2 weeks in the ALI culture, while flatten unhealthy ciliated cells were observed in the LCC method. After 21 days, higher level of MUC5AC and 8 mRNA were expressed in ALI culture which confirmed the secretory differentiation of HNE monolayers in vitro. No significant difference in the permeability coefficients of a model hydrophilic marker ((14)C-mannitol) and a lipophilic drug (budesonide) was observed between the two conditions on day 7. The passage 2-3 of the HNE monolayer using ALI condition retained the morphology and differentiated features of normal epithelium. Thus it would be a suitable model for in vitro nasal drug delivery studies.
Lee
Min-Ki
MK
College of Medicine, Pusan National University, Busan, South Korea.
Yoo
Jin-Wook
JW
Lin
Hongxia
H
Kim
You-Sun
YS
Kim
Dae-Duk
DD
Choi
Young-Min
YM
Park
Soon-Kew
SK
Lee
Chang-Hun
CH
Roh
Hwan-Jung
HJ
D001692:Biological Transport; D018929:Cell Culture Techniques; D017097:Electric Impedance; D006801:Humans; D008855:Microscopy, Electron, Scanning; D009297:Nasal Mucosa / Q000166:cytology* / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction