Min Kyoung Ah, Rosania Gus R, Shin Meong Cheol
College of Pharmacy and Research Institute of Pharmaceutical Sciences, Gyeongsang National University, 501 Jinju Daero, Jinju, Gyeongnam, 52828, Republic of Korea.
Department of Pharmaceutical Sciences, College of Pharmacy, University of Michigan, 428 Church St., Ann Arbor, MI, 48109, USA.
Cell Biochem Biophys. 2016 Jun;74(2):191-203. doi: 10.1007/s12013-016-0719-8. Epub 2016 Jan 27.
To facilitate drug development for lung delivery, it is highly demanding to establish appropriate airway epithelial cell models as transport barriers to evaluate pharmacokinetic profiles of drug molecules. Besides the cancer-derived cell lines, as the primary cell model, normal human bronchial epithelial (NHBE) cells have been used for drug screenings because of physiological relevance to in vivo. Therefore, to accurately interpret drug transport data in NHBE measured by different laboratories, it is important to know biophysical characteristics of NHBE grown on membranes in different culture conditions. In this study, NHBE was grown on the polyester membrane in a different medium and its transport barrier properties as well as cell architectures were fully characterized by functional assays and confocal imaging throughout the days of cultures. Moreover, NHBE cells on inserts in a different medium were subject to either of air-interfaced culture (AIC) or liquid-covered culture (LCC) condition. Cells in the AIC condition were cultivated on the membrane with medium in the basolateral side only, whereas cells with medium in apical and basolateral sides under the LCC condition. Quantitative microscopic imaging with biophysical examination revealed distinct multilayered architectures of differentiated NHBE cells, suggesting NHBE as functional cell barriers for the lung-targeting drug transport.
为促进肺部给药的药物开发,建立合适的气道上皮细胞模型作为转运屏障以评估药物分子的药代动力学特征至关重要。除了癌症衍生的细胞系外,作为原代细胞模型,正常人支气管上皮(NHBE)细胞因其与体内生理相关性而被用于药物筛选。因此,为准确解读不同实验室测量的NHBE中的药物转运数据,了解在不同培养条件下在膜上生长的NHBE的生物物理特性很重要。在本研究中,NHBE在不同培养基中的聚酯膜上生长,其转运屏障特性以及细胞结构在整个培养过程中通过功能测定和共聚焦成像进行了全面表征。此外,在不同培养基中培养在插入物上的NHBE细胞分别置于气液界面培养(AIC)或液封培养(LCC)条件下。AIC条件下的细胞仅在基底侧有培养基的情况下在膜上培养,而LCC条件下的细胞在顶端和基底侧都有培养基。通过生物物理检查进行的定量显微镜成像揭示了分化的NHBE细胞独特的多层结构,表明NHBE作为肺部靶向药物转运的功能性细胞屏障。