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First published on January 26, 2007, doi:10.1177/1087057106297828

Journal of Biomolecular Screening 2007;12:248.

A more recent version of this article appeared on March 1, 2007

Article

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

Zhengrong Zhu1*, Jaime Puglisi1, David Connors1, Jeremy Stewart1, John Herbst1, Anthony Marino2, Michael Sinz1, Jonathan O'Connell1, Martyn Banks1, Kenneth Dickinson1, Angela Cacace1

1 Bristol-Myers Squibb Company, Wallingford, Connecticut
2 Bristol-Myers Squibb Company, Lawrenceville, New Jersey.

* To whom correspondence should be addressed. E-mail: zhengrong.zhu{at}bms.com.


  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 (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, PXR


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[Abstract] [PDF]