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The Characterization of the Human Nasal Epithelial Cell Line RPMI 2650 Under Different Culture Conditions and Their Optimization for an Appropriate in vitro Nasal Model

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ABSTRACT

Purpose

The further characterization of the cell line RPMI 2650 and the evaluation of different culture conditions for an in vitro model for nasal mucosa.

Methods

Cells were cultured in media MEM or A-MEM at air-liquid (A-L) or liquid-liquid (L-L) interfaces for 1 or 3 weeks. Different cryopreservation methods and cell culture techniques were evaluated with immunolabelling of junctional proteins, ultrastructural analysis using electron microscopy, transepithelial electrical resistance (TEER) measurements, permeation studies with dextran and jacalin, and gene expression profiling of 84 drug transporters.

Results

Cell proliferation and differentiation depended on the used medium. The established epithelia expressed occludin, claudin-1, and E-cadherin under all conditions. Cells grown at the A-L interface formed more layers and exhibited a higher TEER and lower dextran and jacalin permeability than at the L-L interface, where cells morphologically exhibited a more differentiated phenotype. The expression of ABC and SLC transporters depended on culture duration and interface.

Conclusions

The RPMI 2650 cells form a polarized epithelium resembling nasal mucosa. However, different culture conditions have a significant effect on cell ultrastructure, barrier integrity, and gene expression, and should be considered when using this cell line as an in vitro model for drug permeability studies and screening of nasal drug candidates.

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Abbreviations

A :

Surface area

ABC:

ATP-binding cassette

A-L:

Air-liquid interface

c 0 :

Initial donor concentration

dQ/dt :

Flux of dextran-FITC or jacalin-fluorescein across the RPMI 2650 cell barrier

L-L:

Liquid-liquid interface

Papp :

Apparent permeability coefficient

SEM:

Scanning electron microscopy

SLC:

Solute carrier

TEER:

Transepithelial electrical resistance

TEM:

Transmission electron microscopy

TJ:

Tight junction

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ACKNOWLEDGMENTS AND DISCLOSURES

The authors thank Sanja Čabraja, Sabina Železnik, Nada Pavlica and Linda Štrus for technical assistance. The study was supported by Lek Pharmaceuticals, d.d., Sandoz Development Center Slovenia and by Slovenian Research Agency (Grants No P3-0108, P1-170).

Author information

Authors and Affiliations

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

    Mateja Erdani Kreft, Urška Dragin Jerman & Eva Lasič

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

    Luka Peternel & Katja Kristan

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

    Tea Lanišnik Rižner, Neli Hevir-Kene & Katja Kristan

Authors
  1. Mateja Erdani Kreft
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  2. Urška Dragin Jerman
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  3. Eva Lasič
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  4. Tea Lanišnik Rižner
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  5. Neli Hevir-Kene
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  6. Luka Peternel
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Correspondence to Katja Kristan.

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Kreft, M.E., Jerman, U.D., Lasič, E. et al. The Characterization of the Human Nasal Epithelial Cell Line RPMI 2650 Under Different Culture Conditions and Their Optimization for an Appropriate in vitro Nasal Model. Pharm Res 32, 665–679 (2015). https://doi.org/10.1007/s11095-014-1494-0

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