Different Culture Conditions Affect Drug Transporter Gene Expression, Ultrastructure, and Permeability of Primary Human
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RESEARCH PAPER
Different Culture Conditions Affect Drug Transporter Gene Expression, Ultrastructure, and Permeability of Primary Human Nasal Epithelial Cells Mateja Erdani Kreft 1 & Larisa Tratnjek 1 & Eva Lasič 1 & Neli Hevir 2 & Tea Lanišnik Rižner 3 & Katja Kristan 3,4 Received: 14 April 2020 / Accepted: 6 August 2020 # Springer Science+Business Media, LLC, part of Springer Nature 2020
ABSTRACT Purpose This study aimed to characterize a commercially available primary human nasal epithelial cell culture and its gene expression of a wide range of drug transporters under different culture conditions. Methods Human nasal cells were cultured in three different types of culture media at the air-liquid (A-L) or liquid-liquid (L-L) interfaces for 1 or 3 wks. The effects of the different cell culture conditions were evaluated using light and electron microscopy, transepithelial electrical resistance (TEER) measurements, permeation studies with dextran, and gene expression profiling of 84 drug transporters. Results The type of culture medium affected cell ultrastructure, TEER, and dextran permeation across epithelia. The expression of 20 drug transporter genes depended on the culture interface and/or time in culture; the A-L interface and longer time in culture favored higher expression levels of five ABC and seven SLC transporters. Conclusions Culture conditions influence the morphology, barrier formation, permeation properties, and drug transporter expression of human nasal epithelial cells, and this must be taken into consideration during the establishment and Electronic supplementary material The online version of this article (https://doi.org/10.1007/s11095-020-02905-w) contains supplementary material, which is available to authorized users. * Katja Kristan [email protected] 1
Institute of Cell Biology, Faculty of Medicine, University of Ljubljana, Ljubljana, Slovenia
2
Lek Pharmaceuticals d.d, Biopharma Process & Product Development, Mengeš, Slovenia
3
Institute of Biochemistry, Faculty of Medicine, University of Ljubljana, Ljubljana, Slovenia
4
Lek Pharmaceuticals, d.d, Sandoz Development Center Slovenia, Ljubljana, Slovenia
validation of in vitro models. A thorough characterization of a nasal epithelial model and its permeability properties is necessary to obtain an appropriate standardized model for the design of aerosol therapeutics and drug transport studies.
KEY WORDS Primary nasal epithelial cell culture . TEER . drug transporters . intranasal administration . gene expression
ABBREVIATIONS ABC AQP A-L DIV hAECN L-L SEM SLC TEER TEM
ATP-binding cassette Aquaporin Air-liquid interface Days in vitro Human airway epithelial cells of nasal origin Liquid-liquid interface Scanning electron microscopy Solute carrier Transepithelial electrical resistance Transmission electron microscopy
INTRODUCTION The administration of aerosol therapeutics via the nasal route is becoming an appealing alternative for drug delivery. This non-invasive method relies on a leaky epithelium with an extensively vascular
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