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Different Culture Conditions Affect Drug Transporter Gene Expression, Ultrastructure, and Permeability of Primary Human Nasal Epithelial Cells

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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 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.

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Abbreviations

ABC:

ATP-binding cassette

AQP:

Aquaporin

A-L:

Air-liquid interface

DIV:

Days in vitro

hAECN:

Human airway epithelial cells of nasal origin

L-L:

Liquid-liquid interface

SEM:

Scanning electron microscopy

SLC:

Solute carrier

TEER:

Transepithelial electrical resistance

TEM:

Transmission electron microscopy

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Author information

Authors and Affiliations

  1. Institute of Cell Biology, Faculty of Medicine, University of Ljubljana, Ljubljana, Slovenia

    Mateja Erdani Kreft, Larisa Tratnjek & Eva Lasič

  2. Lek Pharmaceuticals d.d, Biopharma Process & Product Development, Mengeš, Slovenia

    Neli Hevir

  3. Institute of Biochemistry, Faculty of Medicine, University of Ljubljana, Ljubljana, Slovenia

    Tea Lanišnik Rižner & Katja Kristan

  4. Lek Pharmaceuticals, d.d, Sandoz Development Center Slovenia, Ljubljana, Slovenia

    Katja Kristan

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  1. Mateja Erdani Kreft
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Correspondence to Katja Kristan.

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Supplemental Table 1

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Supplemental Table 2

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Supplemental Fig. 1

The main ultrastructural characteristics of hAECN. These characteristics were observed irrespective of the media used. Rough endoplasmic reticulum (ER) is scarce in the cytoplasm, the Golgi apparatus (GA) is simple, normal mitochondria (M) and endocytotic compartments (E) are present, and bundles of cytoskeletal filaments (arrows) are abundant. Scale bar: 500 nm. (PNG 1048 kb)

High Resolution (TIF 15447 kb)

Supplemental Fig. 2

Immunofluorescence labelling of tight junctional protein occludin (A–C), adherent junction protein E-cadherin (D–F), and cytokeratin 7 (CK7, G–I) in hAECN of passage P3 maintained in Epithelix, PromoCell, and A-MEM for 1 wk. Nuclei are blue. Scale bar: 10 μm. (PNG 4399 kb)

High Resolution (TIF 73936 kb)

Supplemental Fig. 3

Transepithelial permeation of dextran-FITC across hAECN, RPMI2560, and Calu-3 epithelial models at 30 min (left) and 3 h (right) (n = 3–4, mean ± SE). The cells were maintained in A-MEM at the A-L or L-L interface. (PNG 170 kb)

High Resolution (TIF 556 kb)

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Kreft, M.E., Tratnjek, L., Lasič, E. et al. Different Culture Conditions Affect Drug Transporter Gene Expression, Ultrastructure, and Permeability of Primary Human Nasal Epithelial Cells. Pharm Res 37, 170 (2020). https://doi.org/10.1007/s11095-020-02905-w

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  • DOI: https://doi.org/10.1007/s11095-020-02905-w

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