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This version was published on March 1, 2007
Journal of Biomolecular Screening, Vol. 12, No. 2, 248-254 (2007)
DOI: 10.1177/1087057106297828
© 2007 Society for Biomolecular Sciences

Use of Cryopreserved Transiently Transfected Cells in High-Throughput Pregnane X Receptor Transactivation Assay

Zhengrong Zhu

Jaime Puglisi

David Connors

Jeremy Stewart

John Herbst

Bristol-Myers Squibb Company, Wallingford, Connecticut, New Jersey

Anthony Marino

Bristol-Myers Squibb Company, Lawrenceville, New Jersey

Michael Sinz

Jonathan O'Connell

Martyn Banks

Kenneth Dickinson

Angela Cacace

Bristol-Myers Squibb Company, Wallingford, Connecticut, New Jersey

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 (r2 > 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.

Key Words: cryopreservation • transient transfection • cell-based assay • high throughput • pregnane X receptor


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M. Agler, M. Prack, Yingjie Zhu, J. Kolb, K. Nowak, R. Ryseck, Ding Shen, M. E. Cvijic, J. Somerville, S. Nadler, et al.
A High-Content Glucocorticoid Receptor Translocation Assay for Compound Mechanism-of-Action Evaluation
J Biomol Screen, December 1, 2007; 12(8): 1029 - 1041.
[Abstract] [PDF]