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Open Access Research

ATP-binding cassette transporters in immortalised human brain microvascular endothelial cells in normal and hypoxic conditions

Christian Lindner12, Alexander Sigrüner3, Franziska Walther1, Ulrich Bogdahn1, Pierre O Couraud4, Gert Schmitz3 and Felix Schlachetzki1*

Author Affiliations

1 Department of Neurology, University of Regensburg, Bezirksklinikum Regensburg, Regensburg, Germany

2 Department of Psychiatry, University of Regensburg, Bezirksklinikum Regensburg, Regensburg, Germany

3 Institute of Clinical Chemistry and Laboratory Medicine, Regensburg University Medical Center, Regensburg, Germany

4 Departement de biologie cellulaire (CNRS UMR 8104), Institut Cochin, Université Paris Descartes, Paris, France and INSERM U567, Departement de biologie cellulaire, Institut Cochin, Paris, France

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Experimental & Translational Stroke Medicine 2012, 4:9  doi:10.1186/2040-7378-4-9

Published: 3 May 2012

Abstract

Background

Rapid reperfusion following ischemia is the most effective therapy in stroke therapy. However, the success may be compromised by ischemia & reperfusion (I/R) injury and at the human blood–brain barrier (BBB), therefore the effects on transendothelial transport are of special interest. Current studies suggest the ATP-binding cassette (ABC) transporters to be regulated upon ischemic stroke in a way that impedes the effects of drug therapy. The immortalised human brain microvascular endothelial cell line hCMEC/D3 provides most of the unique properties of the BBB with respect to transport and might be a reliable in vitro model to study transendothelial transport after I/R.

Methods

We exposed hCMEC/D3 cells to 24 hours of hypoxia alone and to hypoxia followed by 60 min of reoxygenisation as an in vitro model for I/R. Western blot showed mild upregulation of hypoxia inducible factor (HIF-1α) after hypoxia alone and RNA lysates were analysed with a well-established real-time RT-PCR-based TaqMan low-density array detecting 47 of 48 known human ABC transporters.

Results

No significant increases of ABC mRNA expression levels were detected neither in hypoxic nor in I/R samples. However, slight decrease of ABCC1 in hypoxic and I/R samples and of ABCA10 and ABCD3 in I/R samples was observed.

Conclusion

Our data suggests that hCMEC/D3 cell line and – at the moment – in vitro models in general are a poor basis for stroke research but may be enhanced by co-culturing more cells of the neurovascular unit inducing an overall ischemic response at the BBB.

Keywords:
Blood–brain barrier; Ischemia/reperfusion injury; Hypoxia-inducible factor; Multidrug resistance; ABC transporters; Stroke