Air-liquid interface culture of serially passaged human nasal epithelial cell monolayer for in vitro drug transport studies
Air-liquid interface culture of serially passaged human nasal epithelial cell monolayer for in vitro drug transport studies
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DOI：
10.1080/10717540500177009
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发表时间：
2005-09-01
期刊：
影响因子：
6
通讯作者：
Roh, HJ
中科院分区：
文献类型：
--
作者：
Lee, MK;Yoo, JW;Roh, HJ
本研究的目的是利用无血清培养基（BEGM:DME/F12,50:50）通过气液界面（ALI）方法培养的人鼻上皮（HNE）单层细胞建立药物转运研究。与之前通过液体覆盖培养 (LCC) 方法培养的细胞进行比较，对这些细胞进行了开发和表征。与LCC方法培养的上皮细胞单层相比，ALI方法培养的上皮细胞单层产生显着更高的跨上皮电阻值（3,453 +/- 302 ohm X cm(2)），并维持（>1,000 ohm X cm(2)）长达20天。扫描电镜观察发现，ALI培养2周后纤毛成熟，而LCC方法则观察到扁平的不健康纤毛细胞。 21天后，ALI培养物中MUC5AC和8 mRNA表达较高水平，这证实了HNE单层的体外分泌分化。第 7 天，两种条件下的模型亲水标记物（C-14-甘露醇）和亲脂性药物（布地奈德）的渗透系数没有观察到显着差异。使用 ALI 条件的 HNE 单层第 2-3 代保留了正常上皮的形态和分化特征。因此，它是体外鼻腔药物递送研究的合适模型。
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 (C-14-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.