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BioMed Research International
Volume 2013 (2013), Article ID 676845, 11 pages
http://dx.doi.org/10.1155/2013/676845
Research Article

P-Glycoprotein-Activity Measurements in Multidrug Resistant Cell Lines: Single-Cell versus Single-Well Population Fluorescence Methods

1Laboratory of Ecotoxicology UPRES EA 3222, IFRMP 23, University of Le Havre, 76058 Le Havre Cedex, France
2Department of Genetic Medicine, Weill Cornell Medical College, New York, NY 10022, USA
3Stem Cell and Microenvironment Laboratory, Weill Cornell Medical College in Qatar, Doha 24144, Qatar

Received 17 June 2013; Accepted 29 September 2013

Academic Editor: Beric Henderson

Copyright © 2013 Jennifer Pasquier et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Background. P-gp expression has been linked to the efflux of chemotherapeutic drugs in human cancers leading to multidrug resistance. Fluorescence techniques have been widely applied to measure the P-gp activity. In this paper, there is a comparison between the advantages of two fluorescence approaches of commonly available and affordable instruments: the microplate reader (MPR) and the flow cytometer to detect the P-gp efflux activity using calcein-AM. Results. The selectivity, sensibility, and reproducibility of the two methods have been defined. Our results showed that the MPR is more powerful for the detection of small inhibition, whereas the flow cytometry method is more reliable at higher concentrations of the inhibitors. We showed that to determine precisely the inhibition efficacy the flow cytometry is better; hence, to get the correct Emax and EC50 values, we cannot only rely on the MPR. Conclusion. Both techniques can potentially be used extensively in the pharmaceutical industry for high-throughput drug screening and in biology laboratories for academic research, monitoring the P-gp efflux in specific assays.