Isolation of MDCK cells with low expression of mdr1 gene and their use in membrane permeability screening
The Madin-Darby canine kidney (MDCK) cell line is frequently used for permeability screening in drug discovery. It contains endogenous transporters, most prominently canine multidrug resistance P-glycoprotein (Mdr1), which can interfere with studies of P-glycoprotein substrate assessment and permeab...
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| Format: | Article |
| Language: | English |
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Sciendo
2022-06-01
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| Series: | Acta Pharmaceutica |
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| Online Access: | https://doi.org/10.2478/acph-2022-0003 |
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| author | BOKULIĆ ANA PADOVAN JASNA STUPIN-POLANČEC DARIJA MILIĆ ASTRID |
| author_facet | BOKULIĆ ANA PADOVAN JASNA STUPIN-POLANČEC DARIJA MILIĆ ASTRID |
| author_sort | BOKULIĆ ANA |
| collection | DOAJ |
| description | The Madin-Darby canine kidney (MDCK) cell line is frequently used for permeability screening in drug discovery. It contains endogenous transporters, most prominently canine multidrug resistance P-glycoprotein (Mdr1), which can interfere with studies of P-glycoprotein substrate assessment and permeability measurements. Because MDCK wild type (WT) is genetically heterogeneous, an isolation procedure was investigated in this study to obtain the subclonal line with low P-glycoprotein expression. The best clone obtained had up to 3-fold lower amprenavir efflux and P-glycoprotein expression in comparison to WT. Of 12 standard compounds tested that exhibited active efflux in WT cells, 11 showed a decrease in efflux in the isolated clone. However, the decrease was not below the cut-off value of 2, indicating residual P--glycoprotein activity. Clone isolation via the limiting dilution method, combined with bidirectional amprenavir permeability for clone selection, successfully identified MDCK clones with substantially lower P-glycoprotein efflux and has been demonstrated as a useful tool for assessing passive permeability in early drug discovery. |
| format | Article |
| id | doaj-art-d04d9511221d4d84b64c1374aeb5fd7f |
| institution | Kabale University |
| issn | 1846-9558 |
| language | English |
| publishDate | 2022-06-01 |
| publisher | Sciendo |
| record_format | Article |
| series | Acta Pharmaceutica |
| spelling | doaj-art-d04d9511221d4d84b64c1374aeb5fd7f2025-02-02T02:10:54ZengSciendoActa Pharmaceutica1846-95582022-06-0172227528810.2478/acph-2022-0003Isolation of MDCK cells with low expression of mdr1 gene and their use in membrane permeability screeningBOKULIĆ ANA0PADOVAN JASNA1STUPIN-POLANČEC DARIJA2MILIĆ ASTRID3Fidelta Ltd., 10000Zagreb, CroatiaFidelta Ltd., 10000Zagreb, CroatiaFidelta Ltd., 10000Zagreb, CroatiaFidelta Ltd., 10000Zagreb, CroatiaThe Madin-Darby canine kidney (MDCK) cell line is frequently used for permeability screening in drug discovery. It contains endogenous transporters, most prominently canine multidrug resistance P-glycoprotein (Mdr1), which can interfere with studies of P-glycoprotein substrate assessment and permeability measurements. Because MDCK wild type (WT) is genetically heterogeneous, an isolation procedure was investigated in this study to obtain the subclonal line with low P-glycoprotein expression. The best clone obtained had up to 3-fold lower amprenavir efflux and P-glycoprotein expression in comparison to WT. Of 12 standard compounds tested that exhibited active efflux in WT cells, 11 showed a decrease in efflux in the isolated clone. However, the decrease was not below the cut-off value of 2, indicating residual P--glycoprotein activity. Clone isolation via the limiting dilution method, combined with bidirectional amprenavir permeability for clone selection, successfully identified MDCK clones with substantially lower P-glycoprotein efflux and has been demonstrated as a useful tool for assessing passive permeability in early drug discovery.https://doi.org/10.2478/acph-2022-0003permeabilitymdckefflux ratioclone isolationp-glycoproteinqrt-pcrlc-ms/ms |
| spellingShingle | BOKULIĆ ANA PADOVAN JASNA STUPIN-POLANČEC DARIJA MILIĆ ASTRID Isolation of MDCK cells with low expression of mdr1 gene and their use in membrane permeability screening Acta Pharmaceutica permeability mdck efflux ratio clone isolation p-glycoprotein qrt-pcr lc-ms/ms |
| title | Isolation of MDCK cells with low expression of mdr1 gene and their use in membrane permeability screening |
| title_full | Isolation of MDCK cells with low expression of mdr1 gene and their use in membrane permeability screening |
| title_fullStr | Isolation of MDCK cells with low expression of mdr1 gene and their use in membrane permeability screening |
| title_full_unstemmed | Isolation of MDCK cells with low expression of mdr1 gene and their use in membrane permeability screening |
| title_short | Isolation of MDCK cells with low expression of mdr1 gene and their use in membrane permeability screening |
| title_sort | isolation of mdck cells with low expression of mdr1 gene and their use in membrane permeability screening |
| topic | permeability mdck efflux ratio clone isolation p-glycoprotein qrt-pcr lc-ms/ms |
| url | https://doi.org/10.2478/acph-2022-0003 |
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