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Use of Reduced Temperature Cell Pausing to Enhance Flexibility of Cell-Based AssaysPfizer Global Research & Development, Sandwich, Kent, UK
Pfizer Global Research & Development, Sandwich, Kent, UK
Pfizer Global Research & Development, Sandwich, Kent, UK, darren.cawkill{at}pfizer.com Construction and supply of cell-based reagents for in vitro plate-based screens are often highlighted as a bottleneck within drug discovery. Recent years have seen the successful application of both cryopreservation and automation to increase the capacity and flexibility of cell provision. However, routine cell culture remains a fixed experimental process that requires cells to be prepared and used at specific times. We have investigated the potential of reduced temperature incubation to be used as a simple methodology for stopping and starting cell growth and introduce further flexibility into cell provision. Our results show that incubation of CHOK1, HEK293, and 1321N1 cells at 23 °C arrested growth while maintaining cell viability. Recovery of these paused cells at 37 °C resulted in resumption of normal cell growth and target protein function. Experiments demonstrated that paused cells, expressing either a recombinant G-protein-coupled receptor or an ion channel, performed comparably with the equivalent continuously cultured cells in a 384-well cell-based assay. This simple technique offers the potential to introduce flexibility into cell culture experiments and processes that were previously considered to be fixed. (Journal of Biomolecular Screening 2009:716-722)
Key Words: cell culture • cell pausing • cell-based assays • reagent provision • cell supply
This version was published on July
1, 2009 Journal of Biomolecular Screening, Vol. 14, No. 6,
716-722 (2009) |
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