High conservation of both phase I and II drug-metabolizing activities in cryopreserved rat liver slices.
1. Xenobiotic-metabolizing enzymes, including both cytochrome P450 and phase II-conjugating systems, have been characterized in rat liver slices cryopreserved in 12 or 18% dimethylsulphoxide (DMSO). 2. Several cytochrome P450 isoforms in rat liver slices metabolized testosterone to a variety of hydroxylated products. The rates of formation of these same products were well maintained during cryopreservation of the slices in both 12 or 18% DMSO. 3. After cryopreservation of rat liver slices in 18% DMSO, the rates of metabolism of ropivacaine to 3-hydroxyropivacaine, 4-hydroxyropivacaine and PPX (all catalysed by different cytochrome P450 isoforms) were approximately 94, 79 and 82% respectively of the corresponding rates observed with fresh slices. 4. The rates of conjugation of 7-hydroxycoumarin and 1-naphthol by rat liver slices were significantly decreased after cryopreservation in 12% DMSO, but they were maintained when the concentration of this cryopreservant was increased to 18% 5. After cryopreservation in 12% DMSO, the mitochondrial reduction of the tetrazolium salt MTT by rat liver slices was significantly lowered. In contrast, slices cryopreserved in 18% DMSO demonstrated no significant decrease in their capacity to reduce MTT. 6. Thus, in agreement with previous studies, it was found that cytochrome P450-dependent activities are retained after cryopreservation of liver slices. Although phase II-conjugating enzyme activities are more sensitive to cryopreservation, it was shown that increasing the concentration of DMSO present during cryopreservation could circumvent the problem. This modification improves the usefulness of cryopreserved rat liver slices as a tool in drug metabolism studies.
Sohlenius-Sternbeck
A K
AK
Preclinical Development, AstraZeneca R&D Södertälje, Sweden.
Floby
E
E
Svedling
M
M
Orzechowski
A
A
D000577:Amides / Q000378:metabolism; D000818:Animals; D015925:Cryopreservation; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D004121:Dimethyl Sulfoxide; D006900:Hydroxylation; D007527:Isoenzymes / Q000378:metabolism; D008099:Liver / Q000201:enzymology*; D008297:Male; D008930:Mitochondria, Liver / Q000201:enzymology; D009284:Naphthols / Q000378:metabolism; D010084:Oxidation-Reduction; D051381:Rats; D017207:Rats, Sprague-Dawley; D000077212:Ropivacaine; D013739:Testosterone / Q000378:metabolism; D013778:Tetrazolium Salts / Q000378:metabolism; D013844:Thiazoles / Q000378:metabolism
D000577:Amides; D007527:Isoenzymes; D009284:Naphthols; D013778:Tetrazolium Salts; D013844:Thiazoles; C029350:1-naphthol; D013739:Testosterone; D000077212:Ropivacaine; D003577:Cytochrome P-450 Enzyme System; C022616:thiazolyl blue; D004121:Dimethyl Sulfoxide