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Functional and cytometric examination of different human lung epithelial cell types as drug transport barriers

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Abstract

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.

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Acknowledgments

This research was financially supported by grants from Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (NRF-2015R1C1A1A02036781 to M. C. Shin and 2015R1A6A3A01020598 to K. A. Min). Part of this work was funded by NIH Grant R01GM078200 to G. R. Rosania.

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Authors and Affiliations

  1. College of Pharmacy and Research Institute of Pharmaceutical Sciences, Gyeongsang National University, 501 Jinju Daero, Jinju, Gyeongnam, 52828, Republic of Korea

    Kyoung Ah Min & Meong Cheol Shin

  2. Department of Pharmaceutical Sciences, College of Pharmacy, University of Michigan, 428 Church St., Ann Arbor, MI, 48109, USA

    Kyoung Ah Min & Gus R. Rosania

  3. College of Pharmacy, Seoul National University, 1 Gwanak-ro, Gwanak-gu, Seoul, 08826, Republic of Korea

    Chong-Kook Kim

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  1. Kyoung Ah Min
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  2. Gus R. Rosania
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  4. Meong Cheol Shin
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Correspondence to Meong Cheol Shin.

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Min, K.A., Rosania, G.R., Kim, CK. et al. Functional and cytometric examination of different human lung epithelial cell types as drug transport barriers. Arch. Pharm. Res. 39, 359–369 (2016). https://doi.org/10.1007/s12272-015-0704-6

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  • DOI: https://doi.org/10.1007/s12272-015-0704-6

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