title: Use of cryopreserved transiently transfected cells in high-throughput pregnane X receptor transactivation assay.
abstract: 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.affiliation: Bristol-Myers Squibb Company, Wallingford, CT 06492, USA. zhengrong.zhu@bms.com
authors.forename: Zhengrong
authors.initials: Z
authors.lastname: Zhu
authors.forename: Jaime
authors.initials: J
authors.lastname: Puglisi
authors.forename: David
authors.initials: D
authors.lastname: Connors
authors.forename: Jeremy
authors.initials: J
authors.lastname: Stewart
authors.forename: John
authors.initials: J
authors.lastname: Herbst
authors.forename: Anthony
authors.initials: A
authors.lastname: Marino
authors.forename: Michael
authors.initials: M
authors.lastname: Sinz
authors.forename: Jonathan
authors.initials: J
authors.lastname: O'Connell
authors.forename: Martyn
authors.initials: M
authors.lastname: Banks
authors.forename: Kenneth
authors.initials: K
authors.lastname: Dickinson
authors.forename: Angela
authors.initials: A
authors.lastname: Cacace
mesh_terms: D001681:Biological Assay / Q000379:methods*; D018938:Caco-2 Cells; D006528:Carcinoma, Hepatocellular / Q000473:pathology; D045744:Cell Line, Tumor; D002463:Cell Membrane Permeability / Q000187:drug effects; D015925:Cryopreservation / Q000379:methods*; D064420:Drug-Related Side Effects and Adverse Reactions; D006801:Humans; D008113:Liver Neoplasms / Q000473:pathology; D015735:Mifepristone / Q000378:metabolism / Q000494:pharmacology; D004364:Pharmaceutical Preparations / Q000378:metabolism*; D000077297:Pregnane X Receptor; D011987:Receptors, Steroid / Q000819:agonists / Q000037:antagonists & inhibitors / Q000378:metabolism*; D015203:Reproducibility of Results; D012293:Rifampin / Q000378:metabolism / Q000494:pharmacology; D013442:Sulfinpyrazone / Q000378:metabolism / Q000494:pharmacology; D015533:Transcriptional Activation; D014162:Transfection
chemical_list: D004364:Pharmaceutical Preparations; D000077297:Pregnane X Receptor; D011987:Receptors, Steroid; D015735:Mifepristone; D013442:Sulfinpyrazone; D012293:Rifampin