In-vitro cell culture models of the nasal epithelium: A comparative histochemical investigation of their suitability for drug transport studies
In-vitro cell culture models of the nasal epithelium: A comparative histochemical investigation of their suitability for drug transport studies
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DOI：
10.1023/a:1016038119909
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发表时间：
1996-07-01
影响因子：
3.7
通讯作者：
Kissel, T
中科院分区：
文献类型：
--
作者：
Werner, U;Kissel, T
目的。评价不同的体外细胞培养模型对研究药物通过细胞单层转运的适用性。用凝集素结合特性对牛鼻甲细胞(BT；ATCC CRL 1390)、人鼻中隔肿瘤细胞(RPMI，2650；ATCC CCL 30)和原代培养的人鼻上皮细胞进行了形态和组织化学特征的研究。通过肌动蛋白染色和跨上皮电阻测量来监测培养过程中紧密连接的形成。用荧光标记的凝集素对人鼻腔呼吸道上皮薄片的结合模式进行了表征。杯状细胞粘液经PNA、WGA和SEA染色，证实鼻腔粘液中分别含有N-乙酰氨基葡萄糖、N-乙酰氨基半乳糖和半乳糖残基。纤毛细胞显示N-乙酰氨基葡萄糖的结合部位，WGA染色，而ConA，甘露糖部分，标记上皮细胞的顶端细胞质。该组织中不存在DBA的结合部位。比较BT、RPMI2650和三种凝集素(PNA、WGA、ConA)的原代培养模型以及细胞内肌动蛋白染色和跨上皮电阻测量，发现只有原代培养的人鼻上皮细胞出现分化的上皮细胞、纤毛化的鼻细胞和粘液产生杯状细胞，形成融合的单层紧密连接的细胞。在所研究的体外细胞培养模型中，只有原代培养的人鼻细胞似乎适合于药物转运研究。
Purpose. To evaluate different in-vitro cell culture models for their suitability to study drug transport through cell monolayers.Methods. Bovine turbinate cells (BT; ATCC CRL 1390), human nasal septum tumor cells (RPMI, 2650; ATCC CCL 30), and primary cell cultures of human nasal epithelium were characterized morphologically and histochemically by their lectin binding properties. The development of tight junctions in culture was monitored by actin staining and transepithelial electrical resistance measurements.Results. The binding pattern of thin-sections of excised human nasal respiratory epithelium was characterized using a pannel of fluorescently-labelled lectins. Mucus in goblet cells was stained by PNA, WGA and SEA, demonstrating the presence of terminal N-acetylglucosaminne N-acetylgalactosamine and galactose residues respectively in the mucus of human nasal cells. Ciliated cells revealed binding sites for N-acetylglucosamine, stained by WGA, whereas Con A, characteristic for mannose moieties, labelled the apical cytoplasm of epithelial cells. Binding sites for DBA were not present in this tissue. Comparing three different cell culture models: BT, RPMI 2650, and human nasal cells in primary culture using three lectins (PNA, WGA, Con A) as well as intracellular actin staining and transepithelial electrical resistance measurements we found, that only human nasal epithelial cells in primary culture showed differentiated epithelial cells, ciliated nasal cells and mucus producing goblet cells, which developed confluent cell monolayers with tight junctions.Conclusions. Of the in-vitro cell culture models studied, only human nasal cells in primary culture appears to be suitable for drug transport studies.