Long-term maintenance of drug-metabolizing enzyme activities in rat hepatocytes after cryopreservation

被引:32
作者
Fautrel, A
Joly, B
Guyomard, C
Guillouzo, A
机构
[1] BIOPRED RENNES ATALANTE,RENNES,FRANCE
[2] INSERM U49,UNITE RECH HEPATOL,RENNES,FRANCE
关键词
D O I
10.1006/taap.1997.8253
中图分类号
R9 [药学];
学科分类号
1007 ;
摘要
Recent studies have demonstrated that freshly isolated adult hepatocytes from various species can be hypothermically preserved for a short period or cryopreserved for a prolonged period before seeding in primary culture. This study was designed to determine whether rat hepatocytes could be maintained functional for a prolonged period after either hypothermic preservation or cryopreservation. Cold storage was carried out in University of Wisconsin solution (UW) and freezing in Leibovitz medium added with 10% fetal calf serum and 16% dimethyl sulfoxide. Rat hepatocytes were then set up either in pure conventional culture or in coculture with rat liver epithelial cells. Various functions were measured over 4- and 15-day periods, i.e., albumin secretion rate, deethylation of ethoxyresorufin and phenacetin, dealkylation of pentoxyresorufin, glucuronidation and sulfoconjugation of paracetamol, and N-acetylation of procainamide. No major differences were observed between unfrozen, frozen, and UW-preserved cells. While in pure culture all the functions tested were markedly decreased after 3 or 4 days, they remained high over the 15-day period in coculture, being either maintained or increased after 7-12 days compared to initial values. These results clearly demonstrate that when maintained under suitable culture conditions, rat hepatocytes can fully recover after hypothermic preservation or cryopreservation and therefore represent a suitable in vitro model system for pharmacotoxicological studies. (C) 1997 Academic Press.
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页码:110 / 114
页数:5
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