Maintenance of total cytochrome P-450 content in rat hepatocyte culture and the abundance of CYP1A2 and CYP2B1/2 mRNAs.
mRNAs encoding cytochrome P-450s CYP1A2 and CYP2B1/2 have been quantified in rat hepatocytes cultured for periods up to 72 h under several different culture conditions that maintain total cytochrome P-450 content. When hepatocytes were cultured at either 37 or 30 degrees C in Williams E media, both CYP1A2 and CYP2B1/2 mRNAs declined dramatically. However, when cultured at 30 degrees C for 24 h, the decline in these mRNAs was not as great as that observed in cells grown at 37 degrees C. The addition of dimethyl sulphoxide to cells grown at 37 degrees C did not affect the rate of disappearance of the CYP1A2 or CYP2B1/2 mRNAs. These mRNAs also declined rapidly in cells grown in 'P-450 medium' i.e. RPMI 1640 medium without cyst(e)ine but supplemented with 0.1 mM-delta-aminolaevulinic acid. However, the levels of CYP2B1/2 mRNAs were maintained when hepatocytes were cultured in Williams E medium supplemented with 0.5 mM-metyrapone. These conditions did not, however, maintain the levels of CYP1A2 mRNA.
Padgham
C R
CR
DH Department of Toxicology, St. Bartholomew's Hospital Medical College, London, U.K.
Paine
A J
AJ
Phillips
I R
IR
Shephard
E A
EA
D000818:Animals; D015152:Blotting, Northern; D002478:Cells, Cultured; D003470:Culture Media; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics* / Q000378:metabolism; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D008099:Liver / Q000201:enzymology*; D008297:Male; D008797:Metyrapone / Q000494:pharmacology; D009693:Nucleic Acid Hybridization; D012333:RNA, Messenger / Q000378:metabolism*; D051381:Rats; D013696:Temperature
D003470:Culture Media; D012333:RNA, Messenger; D003577:Cytochrome P-450 Enzyme System; D004121:Dimethyl Sulfoxide; D008797:Metyrapone