title: [Evaluation of induction of drug-metabolizing enzyme mRNAs in primary cultures of cryopreserved human and cynomolgus monkey hepatocytes].
abstract: We introduce a method for measuring drug-metabolizing enzymes and transporters using real-time one-step RT-PCR with TaqMan probes. This method has the advantages of high sensitivity, simplicity, and linearity of quantification over a wide range of mRNA concentrations, making it particularly suitable for evaluating large numbers of samples, as required in expression profile determinations. We also introduce the use of primary cultures of cryopreserved human and cynomolgus monkey hepatocytes as an enzyme induction model. Furthermore, we introduce the combination of real-time one-step RT-PCR and hepatocytes for evaluating the potency of various drugs in inducing drug-metabolizing enzymes and transporters. We discuss the usefulness of evaluating the gene induction of drug-metabolizing enzymes and transporters following exposure to drugs in human and cynomolgus monkey hepatocytes. The combination of real-time one-step RT-PCR and primary cultures of cryopreserved hepatocytes is useful for preclinical drug evaluation and for screening to evaluate the induction potency of new drug candidates.
authors: New Product and Business Development, Research and Development Center, Otsuka Pharmaceutical Factory, Inc., Tokushima, Japan. nisimums@otsukakj.co.jp
authors: Masuhiro
authors: M
authors: Nishimura
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D015925:Cryopreservation; D051544:Cytochrome P-450 CYP3A / Q000096:biosynthesis* / Q000235:genetics*; D004121:Dimethyl Sulfoxide; D004790:Enzyme Induction; D015971:Gene Expression Regulation, Enzymologic; D022781:Hepatocytes / Q000201:enzymology*; D006801:Humans; D008252:Macaca fascicularis; D012333:RNA, Messenger / Q000032:analysis*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods
chemical_list: D012333:RNA, Messenger; D051544:Cytochrome P-450 CYP3A; D004121:Dimethyl Sulfoxide