Min Kyoung Ah, Rosania Gus R, Kim Chong-Kook, 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.
Arch Pharm Res. 2016 Mar;39(3):359-69. doi: 10.1007/s12272-015-0704-6. Epub 2016 Jan 8.
To develop inhaled medications, various cell culture models have been used to examine the transcellular transport or cellular uptake properties of small molecules. For the reproducible high throughput screening of the inhaled drug candidates, a further verification of cell architectures as drug transport barriers can contribute to establishing appropriate in vitro cell models. In the present study, side-by-side experiments were performed to compare the structure and transport function of three lung epithelial cells (Calu-3, normal human bronchial primary cells (NHBE), and NL-20). The cells were cultured on the nucleopore membranes in the air-liquid interface (ALI) culture conditions, with cell culture medium in the basolateral side only, starting from day 1. In transport assays, paracellular transport across all three types of cells appeared to be markedly different with the NHBE or Calu-3 cells, showing low paracellular permeability and high TEER values, while the NL-20 cells showed high paracellular permeability and low TEER. Quantitative image analysis of the confocal microscope sections further confirmed that the Calu-3 cells formed intact cell monolayers in contrast to the NHBE and NL-20 cells with multilayers. Among three lung epithelial cell types, the Calu-3 cell cultures under the ALI condition showed optimal cytometric features for mimicking the biophysical characteristics of in vivo airway epithelium. Therefore, the Calu-3 cell monolayers could be used as functional cell barriers for the lung-targeted drug transport studies.
为了开发吸入药物，人们使用了各种细胞培养模型来研究小分子的跨细胞转运或细胞摄取特性。对于吸入候选药物的可重复高通量筛选，进一步验证细胞结构作为药物转运屏障有助于建立合适的体外细胞模型。在本研究中，进行了平行实验以比较三种肺上皮细胞（Calu-3细胞、正常人支气管原代细胞（NHBE）和NL-20细胞）的结构和转运功能。从第1天开始，将细胞在气液界面（ALI）培养条件下培养在核孔膜上，仅在基底外侧加入细胞培养基。在转运实验中，所有三种类型细胞的旁细胞转运与NHBE或Calu-3细胞明显不同，显示出低旁细胞通透性和高跨上皮电阻值，而NL-20细胞显示出高旁细胞通透性和低跨上皮电阻。共聚焦显微镜切片的定量图像分析进一步证实，与多层的NHBE和NL-20细胞相比，Calu-3细胞形成了完整的细胞单层。在三种肺上皮细胞类型中，ALI条件下的Calu-3细胞培养物在模拟体内气道上皮的生物物理特性方面表现出最佳的细胞计量学特征。因此，Calu-3细胞单层可作为肺靶向药物转运研究的功能性细胞屏障。
