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

University of Strathclyde, Department of Bioscience, Glasgow, UK

Chantevy Pou

University of Strathclyde, Department of Bioscience, Glasgow, UK

Honglin Niu

University of Strathclyde, Department of Bioscience, Glasgow, UK

Ji-Hu Zhang

University of Strathclyde, Department of Bioscience, Glasgow, UK

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

This version was published on September 1, 2005

Journal of Biomolecular Screening, Vol. 10, No. 6, 615-623 (2005)
DOI: 10.1177/1087057105276474


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