Fully automatic flow-based device for monitoring of drug permeation across a cell monolayer
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A novel flow-programming setup based on the sequential injection principle is herein proposed for on-line monitoring of temporal events in cell permeation studies. The permeation unit consists of a Franz cell with its basolateral compartment mixed under mechanical agitation and thermostated at 37 °C. The apical compartment is replaced by commercially available Transwell inserts with a precultivated cell monolayer. The transport of drug substances across epithelial cells genetically modified with the P-glycoprotein membrane transporter (MDCKII–MDR1) is monitored on-line using rhodamine 123 as a fluorescent marker. The permeation kinetics of the marker is obtained in a fully automated mode by sampling minute volumes of solution from the basolateral compartment in short intervals (10 min) up to 4 h. The effect of a P-glycoprotein transporter inhibitor, verapamil as a model drug, on the efficiency of the marker transport across the cell monolayer is thoroughly investigated. The analytical features of the proposed flow method for cell permeation studies in real time are critically compared against conventional batch-wise procedures and microfluidic devices.
KeywordsFully automated system Permeation study P-glycoprotein transporter Rhodamine 123 Sequential injection analysis
The authors gratefully acknowledge the financial support of the Ministry of Education of the Czech Republic, project no. SVV 260 184, Grant Agency of the Charles University, project no. 159415, and the Grant Agency of the Czech Republic, project no. 15-10781S/P206. This work received financial support from the European Union (FEDER funds through COMPETE) and Portuguese National Funds (FCT, Fundação para a Ciência e Tecnologia) through project UID/Multi/04378/2013. M.M. is grateful to the Spanish Ministry of Science and Competitiveness for financial support through projects CTM2014-56628-C3-3-R and CTM2014-61553-EXP. Special thanks to Mgr. Lucie Hyršová for cell cultivation and kinetic profiles evaluation.
Compliance with ethical standard
Conflict of interest
The authors declare no conflict of interest.
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