کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
10162402 | 1114328 | 2014 | 7 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Establishment of Optimized MDCK Cell Lines for Reliable Efflux Transport Studies
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کلمات کلیدی
موضوعات مرتبط
علوم پزشکی و سلامت
داروسازی، سم شناسی و علوم دارویی
اکتشاف دارویی
پیش نمایش صفحه اول مقاله
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چکیده انگلیسی
MadinâDarby canine kidney (MDCK) cells transfected with human MDR1 gene (MDCKâMDR1) encoding for Pâglycoprotein (hPgp, ABCB1) are widely used for transport studies to identify drug candidates as substrates of this efflux protein. Therefore, it is necessary to rely on constant and comparable expression levels of Pgp to avoid false negative or positive results. We generated a cell line with homogenously high and stable expression of hPgp through sorting single clones from a MDCKâMDR1 cell pool using fluorescenceâactivated cell sorting (FACS). To obtain control cell lines for evaluation of crossâinteractions with endogenous canine Pgp (cPgp) wildâtype cells were sorted with a low expression pattern of cPgp in comparison with the MDCKâMDR1. Expression of other transporters was also characterized in both cell lines by quantitative realâtime PCR and Western blot. Pgp function was investigated applying the CalceinâAM assay as well as bidirectional transport assays using 3HâDigoxin, 3HâVinblastine, and 3HâQuinidine as substrates. Generated MDCKâMDR1 cell lines showed high expression of hPgp. Control MDCKâWT cells were optimized in showing a comparable expression level of cPgp in comparison with MDCKâMDR1 cell lines. Generated cell lines showed higher and more selective Pgp transport compared with parental cells. Therefore, they provide a significant improvement in the performance of efflux studies yielding more reliable results. © 2014 Wiley Periodicals, Inc. and the American Pharmacists Association J Pharm Sci 103:1298-1304, 2014
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Pharmaceutical Sciences - Volume 103, Issue 4, April 2014, Pages 1298-1304
Journal: Journal of Pharmaceutical Sciences - Volume 103, Issue 4, April 2014, Pages 1298-1304
نویسندگان
Dominik Gartzke, Gert Fricker,