Characterization of a Novel “Thaw and Go” Cell-based Transporter Model for Studying SLC Transporters
2015
Objective: Development and characterization of a novel cell-based transporter model to study regulatory agencies (USFDA/EMA) recommended and industry important SLC transporters. Methods: Cryopreserved HEK293 cells which transiently overexpress OATP1B1*1a (wild-type), OATP1B3, OAT1, OAT3, OCT1, OCT2, MATE1, MATE2-K, OATP1A2, OATP2B1, PEPT1, PEPT2, NTCP and empty vector, respectively, were thawed and plated on 24-well Poly-D-Lysine coated plates, and then refed with plating media in the presence or absence of 2mM sodium butyrate at 3-4 hours post plating. Uptake assays were performed at 24 hrs post plating. Following two washes, cells were pre-equilibrated with HBSS buffer for 10min, MATE1 and MATE2-K cells require extra incubation with HBSS buffer containing NH4Cl. The cells were then incubated with substrate or inhibitor for desired amount of time. After the assay was terminated, the cells were lysed and subjected to analysis. Results: The novel SLC transporter model, including OATP1B1*1a (wild-type), OAT...
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