Document Detail


Use of cryopreserved transiently transfected cells in high-throughput pregnane X receptor transactivation assay.
MedLine Citation:
PMID:  17259590     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Cryopreserved, transiently transfected HepG2 cells were compared to freshly transfected HepG2 cells for use in a pregnane X receptor (PXR) transactivation assay. Assay performance was similar for both cell preparations; however, cryopreserved cells demonstrated less interassay variation. Validation with drugs of different PXR activation potencies and efficacies demonstrated an excellent correlation (r(2) > 0.95) between cryopreserved and fresh cells. Cryopreservation did not change the effect of known CYP3A4 inducers that have poor cell permeability, indicating that cryopreservation had little effect on membrane permeability. In addition, cryopreserved HepG2 cells did not exhibit enhanced susceptibility to cytotoxic compounds compared to transiently transfected control cells. The use of cryopreserved cells enables this assay to run with enhanced efficiency.
Authors:
Zhengrong Zhu; Jaime Puglisi; David Connors; Jeremy Stewart; John Herbst; Anthony Marino; Michael Sinz; Jonathan O'Connell; Martyn Banks; Kenneth Dickinson; Angela Cacace
Publication Detail:
Type:  Comparative Study; Journal Article     Date:  2007-01-26
Journal Detail:
Title:  Journal of biomolecular screening     Volume:  12     ISSN:  1087-0571     ISO Abbreviation:  J Biomol Screen     Publication Date:  2007 Mar 
Date Detail:
Created Date:  2007-03-12     Completed Date:  2007-04-20     Revised Date:  2011-05-23    
Medline Journal Info:
Nlm Unique ID:  9612112     Medline TA:  J Biomol Screen     Country:  United States    
Other Details:
Languages:  eng     Pagination:  248-54     Citation Subset:  IM    
Affiliation:
Bristol-Myers Squibb Company, Wallingford, CT 06492, USA. zhengrong.zhu@bms.com
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MeSH Terms
Descriptor/Qualifier:
Biological Assay / methods*
Caco-2 Cells
Carcinoma, Hepatocellular / pathology
Cell Line, Tumor
Cell Membrane Permeability / drug effects
Cryopreservation / methods*
Drug Toxicity
Humans
Liver Neoplasms / pathology
Mifepristone / metabolism,  pharmacology
Pharmaceutical Preparations / metabolism*
Receptors, Steroid / agonists,  antagonists & inhibitors,  metabolism*
Reproducibility of Results
Rifampin / metabolism,  pharmacology
Sulfinpyrazone / metabolism,  pharmacology
Transcriptional Activation*
Transfection
Chemical
Reg. No./Substance:
0/Pharmaceutical Preparations; 0/Receptors, Steroid; 0/pregnane X receptor; 13292-46-1/Rifampin; 57-96-5/Sulfinpyrazone; 84371-65-3/Mifepristone

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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