Improving high-content-screening assay performance by using division-arrested cells

Assay Drug Dev Technol. 2005 Oct;3(5):515-23. doi: 10.1089/adt.2005.3.515.

Abstract

As cell-based assays are used more commonly in robotic high-throughput compound screening, cells themselves have become critical reagents. Thus, it has become essential to produce cell reagents with high consistency and quality. We experimented with cells division-arrested with low-level mitomycin C treatment and demonstrate that they perform with better consistency than non-division-arrested counterparts in high-content screening imaging assays. We propose that for cell-based screening, it is possible to uncouple the cell production process from the screening process. Cells can be produced en masse, treated to become irreversibly division-arrested, and cryopreserved. These "ready-to-use" reagents can be thawed, plated, and used in screening with improved consistency and convenience.

Publication types

  • Comparative Study
  • Evaluation Study

MeSH terms

  • Antibiotics, Antineoplastic / administration & dosage
  • Apoptosis / drug effects
  • Biological Assay / methods*
  • Cell Culture Techniques / methods*
  • Cell Cycle / drug effects*
  • Cell Cycle / physiology
  • Cell Line
  • Cell Proliferation / drug effects
  • Cell Survival / drug effects
  • Dose-Response Relationship, Drug
  • Drug Evaluation, Preclinical / methods*
  • HeLa Cells
  • Humans
  • Kidney / cytology*
  • Kidney / drug effects*
  • Kidney / physiology
  • Lethal Dose 50
  • Mitomycin / administration & dosage*

Substances

  • Antibiotics, Antineoplastic
  • Mitomycin