title: Culture period-dependent changes in the uptake of transporter substrates in sandwich-cultured rat and human hepatocytes.
abstract: Sandwich-cultured hepatocytes (SCH) are a useful tool for evaluating hepatobiliary drug transport in vitro. Some studies have investigated the in vitro-in vivo correlations of the biliary clearance of drugs using SCH. In most cases, the biliary clearance observed in vivo correlated well with the predicted clearance, but the predicted absolute values were underestimated when based on in vitro experiments with SCH. We hypothesized that the down-regulated function of uptake transporters is one of the causes of this underestimation. Therefore, the uptake of taurocholate, digoxin, pravastatin, and rosuvastatin was investigated in sandwich-cultured rat hepatocytes (SCRH) cultured for 5, 24, 48, and 96 h, and the predicted hepatic clearance from in vitro uptake clearance (CL(H, vitro)) was calculated with a dispersion model. In SCRH cultured for 96 h, the saturable uptake of taurocholate, digoxin, pravastatin, and rosuvastatin decreased to 7.5, 3.3, 64, and 23%, respectively, of their uptake in hepatocytes cultured for 5 h, and a better prediction of in vivo hepatic clearance (CL(H, vivo)) was achieved when based on CL(H, vitro) of 5-h-cultured hepatocytes. These results suggest that the uptake activity is considerably reduced in cell culture, even in a sandwich-culture format. In a similar study, we also examined taurocholate and rosuvastatin in sandwich-cultured human hepatocytes (SCHH). Unlike in SCRH, the saturable uptake of these compounds did not differ markedly in SCHH cultured for 5 or 96 h. Thus, the uptake activity in SCHH was maintained relatively well compared with that in SCRH.
authors.affiliation: Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-0033 Japan.
authors.forename: Naoki
authors.initials: N
authors.lastname: Kotani
authors.forename: Kazuya
authors.initials: K
authors.lastname: Maeda
authors.forename: Takao
authors.initials: T
authors.lastname: Watanabe
authors.forename: Mariko
authors.initials: M
authors.lastname: Hiramatsu
authors.forename: Li-kun
authors.initials: LK
authors.lastname: Gong
authors.forename: Yi-an
authors.initials: YA
authors.lastname: Bi
authors.forename: Toshiaki
authors.initials: T
authors.lastname: Takezawa
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Kusuhara
authors.forename: Yuichi
authors.initials: Y
authors.lastname: Sugiyama
mesh_terms: D000818:Animals; D001646:Bile / Q000378:metabolism; D001692:Biological Transport; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D004077:Digoxin / Q000493:pharmacokinetics*; D005464:Fluorobenzenes / Q000493:pharmacokinetics*; D022781:Hepatocytes / Q000378:metabolism*; D006801:Humans; D008658:Inactivation, Metabolic; D008099:Liver / Q000378:metabolism; D008297:Male; D026901:Membrane Transport Proteins / Q000378:metabolism*; D017035:Pravastatin / Q000493:pharmacokinetics*; D011743:Pyrimidines / Q000493:pharmacokinetics*; D051381:Rats; D017207:Rats, Sprague-Dawley; D000068718:Rosuvastatin Calcium; D013449:Sulfonamides / Q000493:pharmacokinetics*; D013656:Taurocholic Acid / Q000493:pharmacokinetics*
chemical_list: D005464:Fluorobenzenes; D026901:Membrane Transport Proteins; D011743:Pyrimidines; D013449:Sulfonamides; D013656:Taurocholic Acid; D004077:Digoxin; D000068718:Rosuvastatin Calcium; D017035:Pravastatin