PT  - JOURNAL ARTICLE
AU  - Naoki Kotani
AU  - Kazuya Maeda
AU  - Takao Watanabe
AU  - Mariko Hiramatsu
AU  - Li-kun Gong
AU  - Yi-an Bi
AU  - Toshiaki Takezawa
AU  - Hiroyuki Kusuhara
AU  - Yuichi Sugiyama
TI  - Culture-period-dependent changes in the uptake of transporter substrates in sandwich-cultured rat and human hepatocytes
AID  - 10.1124/dmd.111.038968
DP  - 2011 Jun 14
TA  - Drug Metabolism and Disposition
PG  - dmd.111.038968
4099  - http://dmd.aspetjournals.org/content/early/2011/06/14/dmd.111.038968.short
4100  - http://dmd.aspetjournals.org/content/early/2011/06/14/dmd.111.038968.full
AB  - 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 (CLbile,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 downregulated function of uptake transporters is a major cause 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 biliary clearance (CLbile,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 CLbile,vivo was achieved when based on CLbile,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 h or 96 h. Thus, the uptake activity in SCHH was maintained relatively well compared with that in SCRH.