![[Feedback]](/icons/banner/feedbackACT.gif){kind=icon}
![[For Subscribers]](/icons/banner/subscriberACT.gif){kind=icon}
![[Archive]](/icons/banner/archiveACT.gif){kind=icon}
![[Search]](/icons/banner/searchACT.gif){kind=icon}
![[Contents]](/icons/banner/tocACT.gif){kind=icon}
|
|
|
|
|
||
Vol. 27, Issue 3, 327-335, March 1999
XenoTech, Limited Liability Company,
Kansas City, Kansas (A.M., D.M., K.C., A.P.);
Department of
Pharmacology, Toxicology and Therapeutics, Center for Environmental and
Occupational Health, University of Kansas Medical Center, Kansas City,
Kansas (R.D., A.P.); and
School of Pharmacy, Division of Pharmaceutics,
University of North Carolina, Chapel Hill, North Carolina
(E.L.)
In the present study, we evaluated the inducibility of cytochrome
P-450 (CYP) CYP1A, CYP2B, CYP3A, and CYP4A by 
-naphthoflavone, phenobarbital, dexamethasone, and clofibric acid, respectively, in
primary hepatocyte cultures prepared from both fresh and cryopreserved rat hepatocytes. Rat hepatocytes were successfully thawed and cultured
after cryopreservation in liquid nitrogen for up to 1 month. Percentage
of total recovery, viable cell recovery, and final viability of the
cells were 68%, 72%, and 85%, respectively. Regardless of whether
they were cryopreserved or not, cultured hepatocytes exhibited
near-normal morphology. Treatment of cryopreserved hepatocytes with
-naphthoflavone caused an 8-fold increase in 7-ethoxyresorufin
O-dealkylase (CYP1A1/2) activity, with an
EC50 of 1.5 µM; treatment with phenobarbital caused a
26-fold increase in 7-pentoxyresorufin O-dealkylase
(CYP2B1/2) activity, with an EC50 of 10 µM; treatment
with dexamethasone caused a 10-fold increase in testosterone
6-hydroxylase (CYP3A1/2) activity, with an EC50 of 1.3 µM, whereas treatment with clofibric acid caused a 3-fold increase in
lauric acid 12-hydroxylase (CYP4A1-3) activity, with an
EC50 of 170 µM. The induction of CYP1A, CYP2B, CYP3A, and
CYP4A enzymes by these inducers was confirmed by Western
immunoblotting. The patterns of P-450 induction in
cryopreserved rat hepatocytes, in terms of concentration response,
reproducibility, magnitude, and specificity of response, were similar
to those observed in freshly isolated hepatocytes. Additionally, the
magnitude and specificity of induction was similar to that observed in
vivo in rats. In conclusion, under the conditions examined,
cryopreserved rat hepatocytes appear to be a suitable in vitro system
for evaluating xenobiotics as inducers of P-450 enzymes.
This article has been cited by other articles:
|
|
 |
|
  C. Lu, C. Berg, S. R. Prakash, F. W. Lee, and S. K. Balani Prediction of Pharmacokinetic Drug-Drug Interactions Using Human Hepatocyte Suspension in Plasma and Cytochrome P450 Phenotypic Data. III. In Vitro-in Vivo Correlation with Fluconazole Drug Metab. Dispos., July 1, 2008; 36(7): 1261 - 1266. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Wortham, M. Czerwinski, L. He, A. Parkinson, and Y.-J. Y. Wan Expression of Constitutive Androstane Receptor, Hepatic Nuclear Factor 4{alpha}, and P450 Oxidoreductase Genes Determines Interindividual Variability in Basal Expression and Activity of a Broad Scope of Xenobiotic Metabolism Genes in the Human Liver Drug Metab. Dispos., September 1, 2007; 35(9): 1700 - 1710. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Lu, G. T. Miwa, S. R. Prakash, L.-S. Gan, and S. K. Balani A Novel Model for the Prediction of Drug-Drug Interactions in Humans Based on in Vitro Cytochrome P450 Phenotypic Data Drug Metab. Dispos., January 1, 2007; 35(1): 79 - 85. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Lu, P. Li, R. Gallegos, V. Uttamsingh, C. Q. Xia, G. T. Miwa, S. K. Balani, and L.-S. Gan Comparison of Intrinsic Clearance in Liver Microsomes and Hepatocytes from Rats and Humans: Evaluation of Free Fraction and Uptake in Hepatocytes Drug Metab. Dispos., September 1, 2006; 34(9): 1600 - 1605. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. A. Graham, B. Goodwin, R. V. Merrihew, W. L. Krol, and E. L. LeCluyse Cloning, Tissue Expression, and Regulation of Beagle Dog CYP4A Genes Toxicol. Sci., August 1, 2006; 92(2): 356 - 367. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M.-Y. Lee, C. B. Park, J. S. Dordick, and D. S. Clark From the Cover: Metabolizing enzyme toxicology assay chip (MetaChip) for high-throughput microscale toxicity analyses PNAS, January 25, 2005; 102(4): 983 - 987. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. D. R. Lloyd, S. Orr, D. P. Berry, and A. R. Dennison Development of a Protocol for Cryopreservation of Hepatocytes for Use in Bioartificial Liver Systems Ann. Clin. Lab. Sci., April 1, 2004; 34(2): 165 - 174. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Madan, R. A. Graham, K. M. Carroll, D. R. Mudra, L. A. Burton, L. A. Krueger, A. D. Downey, M. Czerwinski, J. Forster, M. D. Ribadeneira, et al. Effects of Prototypical Microsomal Enzyme Inducers on Cytochrome P450 Expression in Cultured Human Hepatocytes Drug Metab. Dispos., April 1, 2003; 31(4): 421 - 431. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Houle, J. Raoul, J.-F. Levesque, K. S. Pang, D. A. Nicoll-Griffith, and J. M. Silva Retention of Transporter Activities in Cryopreserved, Isolated Rat Hepatocytes Drug Metab. Dispos., April 1, 2003; 31(4): 447 - 451. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. A. Graham, A. Downey, D. Mudra, L. Krueger, K. Carroll, C. Chengelis, A. Madan, and A. Parkinson In Vivo and in Vitro Induction of Cytochrome P450 Enzymes in Beagle Dogs Drug Metab. Dispos., November 1, 2002; 30(11): 1206 - 1213. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. E. Burczynski, M. McMillian, J. B. Parker, S. Bryant, A. Leone, E. R. Grant, J. M. Thorne, Z. Zhong, R. A. Zivin, and M. D. Johnson Cytochrome P450 Induction in Rat Hepatocytes Assessed by Quantitative Real-Time Reverse-Transcription Polymerase Chain Reaction and the RNA Invasive Cleavage Assay Drug Metab. Dispos., September 1, 2001; 29(9): 1243 - 1250. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. Robertson, H. H. DeCory, A. Madan, and A. Parkinson In Vitro Inhibition and Induction of Human Hepatic Cytochrome P450 Enzymes by Modafinil Drug Metab. Dispos., June 1, 2000; 28(6): 664 - 671. [Abstract] [Full Text]
|
|
|
|
|
|
E. LeCluyse, P. Bullock, A. Madan, K. Carroll, and A. Parkinson Influence of Extracellular Matrix Overlay and Medium Formulation on the Induction of Cytochrome P-450 2B Enzymes in Primary Cultures of Rat Hepatocytes Drug Metab. Dispos., August 1, 1999; 27(8): 909 - 915. [Abstract] [Full Text]
|
|
 |
 |
 |
|
 |
 |
 |
