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 (CLH, 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 (CLH, vivo) was achieved when based on CLH, 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.
Footnotes
This study was supported in part by the Ministry of Health, Labor and Welfare of Japan [Grants-in-Aid for Research on Publicly Essential Drugs and Medical Devices]; and the Japan Society for the Promotion of Science [Grant-in-Aid for Scientific Research 20249008].
Article, publication date, and citation information can be found at http://dmd.aspetjournals.org.
doi:10.1124/dmd.111.038968.
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ABBREVIATIONS:
- SCH
- sandwich-cultured hepatocyte(s)
- SCRH
- sandwich-cultured rat hepatocytes
- SCHH
- sandwich-cultured human hepatocytes
- BSA
- bovine serum albumin
- FBS
- fetal bovine serum
- HBSS
- Hanks' balanced salt solution
- OATP/Oatp
- organic anion transporting polypeptide
- NTCP/Ntcp
- Na+-taurocholate cotransporting polypeptide.
- Received February 18, 2011.
- Accepted June 13, 2011.
- Copyright © 2011 by The American Society for Pharmacology and Experimental Therapeutics
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