Improving consistency of cell-based assays by using division-arrested cells.
In this article we describe the use of division-arrested cells for cell-based assays designed for high-throughput screening. Cells are the most critical and variable reagent for cell-based high-throughput screening. The robustness of robotic screening depends on the quality and consistency of cell reagents. We demonstrate that for most cell types commonly used for high-throughput screening, cells can be irreversibly division-arrested by mitomycin C treatment at doses that cause no apparent toxicity or obvious change to the cell signaling properties we measured. Our data also suggest that division-arrested cells perform favorably compared to regular growing cells in reporter and calcium flux assays, two platforms most commonly used in robotic screening. Division arrest technology effectively uncouples the process of cell production from robotic screening and brings the convenience of having quality-approved cell reagent on demand for cell-based high-throughput screening.
Fursov
Natalie
N
Cell & Molecular Technologies, Inc., North Wales, PA 08865, USA.
Cong
Mei
M
Federici
Mark
M
Platchek
Michael
M
Haytko
Peter
P
Tacke
Roland
R
Livelli
Thomas
T
Zhong
Zhong
Z
D000818:Animals; D001681:Biological Assay / Q000379:methods*; D018929:Cell Culture Techniques / Q000379:methods*; D002453:Cell Cycle / Q000187:drug effects / Q000502:physiology*; D002460:Cell Line; D015925:Cryopreservation; D004305:Dose-Response Relationship, Drug; D015195:Drug Design; D006801:Humans; D007668:Kidney / Q000166:cytology* / Q000187:drug effects*; D051379:Mice; D016685:Mitomycin / Q000494:pharmacology*; D041681:NIH 3T3 Cells; D011933:Reagent Kits, Diagnostic; D015203:Reproducibility of Results; D012371:Robotics / Q000379:methods*; D012680:Sensitivity and Specificity
D011933:Reagent Kits, Diagnostic; D016685:Mitomycin