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Journal of Biomolecular Screening
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Article

Evaluation of Division-Arrested Cells for Cell-Based High-Throughput Screening and Profiling

Mary Ellen Digan*, Chantevy Pou, Honglin Niu, Ji-Hu Zhang

Lead Discovery Center/Discovery Technologies, Novartis Institutes for Biomedical Research, Cambridge, MA.

* To whom correspondence should be addressed. E-mail: mary.digan{at}novartis.com.


  Abstract

Just-in-time cell supply for cell-based high-throughput screening (HTS) is frequently problematic. In addition to scheduling and logistical issues, quality issues and variability due to passage effect, cell cycle, or confluency contribute to day-to-day signal variability in the course of cell-based HTS campaigns. Cell division-arrest and cryopreservation technologies permit the use of cells as assay-ready reagents for HTS and other cell-based profiling and structure-activity studies. In this report, the authors compare division-arrested and dividing cells in 2 assay types that are dependent on movement of proteins within or through cell membranes: a receptor tyrosine kinase assay involving A431 cells responsive to epidermal growth factor, and a secretion reporter assay, which measures secretion of a reporter gene, secreted alkaline phosphatase. In both assays, dividing and division-arrested cells yielded similar basal and maximal signals at a given cell density. Similar IC50s were obtained for reference inhibitors in each assay, type in both dividing and division-arrested cells. In addition, for the secretion reporter assay, when comparing IC50s obtained from 44 compounds randomly chosen from a primary screening hit list, the rank order of potency obtained from dividing cells and division-arrested cells was essentially identical. Furthermore, the results show that, under certain assay conditions, data generated using division-arrested cells are less variable than those generated using dividing cells. In summary, the results suggest that, in many cases, division-arrested cells can substitute for dividing cells and offer certain advantages for cell-based assays.

Key Words: high-throughput screening, cell-based assay, receptor tyrosine kinase, division-arrest, cryopreservation, secretion assay, assay comparison

First published on August 15, 2005, doi:10.1177/1087057105276474

Journal of Biomolecular Screening 2005;10:615.

A more recent version of this article appeared on September 1, 2005

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