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Drug Delivery Characteristics of the Progenitor Bronchial Epithelial Cell Line VA10

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ABSTRACT

Purpose

To determine the integrity and permeability properties of the immortalized human VA10 bronchial epithelial cell line for its suitability as an in vitro drug permeation model.

Methods

Cells were grown under liquid-covered culture (LCC) or air-liquid interface (ALI) culture, characterized using electron microscopy and immunostaining. Integrity was measured using transepithelial electrical resistance (TER) and permeability of fluorescein sodium (Flu-Na). General permeability was established with dextrans and model drugs and P-glycoprotein (P-gp) function determined with bidirectional flux of rhodamine-123.

Results

ALI culture resulted in 2–3 cell layers with differentiation towards ciliated cells but LCC showed undifferentiated morphology. VA10 cells formed TJ, with higher TER in LCC than ALI (∼2500 vs. ∼1200 Ω*cm2) and Flu-Na permeability ∼1–2 × 10−7 cm/s. ALI cultured cells expressed P-gp and distinguished between compounds depending on lipophilicity and size, consistent with previous data from Calu-3 and 16HBE14o-cell lines.

Conclusions

ALI cultured cell layers capture the in vivo-like phenotype of bronchial epithelium and form functional cell barrier capable of discriminating between compounds depending on physiochemical properties. The VA10 cell line is an important alternative to previously published cell lines and a relevant model to study airway drug delivery in vitro.

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Abbreviations

3D:

three-dimensional

A:

surface area (cm2)

A-B:

apical-to-basolateral

ALI:

air-liquid interface

B-A:

basolateral to apical

BSA:

bovine serum albumin

CBF:

ciliary beating frequency

CFTR:

cystic fibrosis transmembrane conductance regulator

FD:

fluorescein isothiocyanate labeled dextran

Flu-Na:

fluorescein sodium

HBSS:

Hanks balanced salt solution

HPV-16:

human papilloma virus-16

LCC:

liquid-covered culture

NHBE:

normal human bronchial epithelial

Papp :

apparent permeability (cm/s)

PBS:

phosphate buffered saline

P-gp:

p-glycoprotein

Rb:

retinoblastoma tumor suppressor protein

Rh123:

rhodamine 123

SEM:

scanning electron microscopy

TER:

transepithelial electrical resistance (Ω*cm2)

TJ:

tight junction

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ACKNOWLEDGMENTS AND DISCLOSURES

Financial support from the Eimskip Fund of University of Iceland, the University of Iceland Research Fund, the Landspitali University Hospital Science Fund and the Bergthóru and Thorsteins Scheving Thorsteinssonar Fund is gratefully acknowledged. We thank Professor Magnus Karl Magnusson for critical discussion and good advice, Sigrún Kristjánsdóttir at the Pathology Department of Landspitali University Hospital for her contribution to the paraffin prepared samples and Bergthóra S. Snorradóttir at the University of Iceland for help with the HPLC.

Author information

Correspondence to Ólafur Baldursson.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Video of beating cilia of differentiated VA10 cells cultured at ALI for 14 days was taken with Leica DMI3000 inverted microscopy, 40x objective and DIC filter, focus directed at apical surface of the cell layer. Before imaging, the surface of the cells was immersed in PBS. The ciliary beating can clearly be seen on individual cells and patches of ciliated cells that cover 10-15% of the surface area. (WMV 1794 kb)

Supplementary Video

Video of beating cilia of differentiated VA10 cells cultured at ALI for 14 days was taken with Leica DMI3000 inverted microscopy, 40x objective and DIC filter, focus directed at apical surface of the cell layer. Before imaging, the surface of the cells was immersed in PBS. The ciliary beating can clearly be seen on individual cells and patches of ciliated cells that cover 10-15% of the surface area. (WMV 1794 kb)

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Benediktsdóttir, B.E., Arason, A.J., Halldórsson, S. et al. Drug Delivery Characteristics of the Progenitor Bronchial Epithelial Cell Line VA10. Pharm Res 30, 781–791 (2013) doi:10.1007/s11095-012-0919-x

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KEY WORDS

  • air-liquid interface culture
  • airway permeability
  • differentiation
  • drug delivery
  • human bronchial epithelial cells