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Cell-Cycle Inhibitor Profiling by High-Content Analysis

  • Fabio Gasparri
  • Antonella Ciavolella
  • Arturo Galvani
Part of the Advances In Experimental Medicine And Biology book series (AEMB, volume 604)

The discovery of agents which disrupt cancer cell division by specifically targeting key components of the cell-cycle machinery represents a major focus of recent drug discovery efforts in Oncology. The drug discovery process can be greatly enhanced by multiparametric cellular analysis which can assist in confirmation, often in a few multiplexed assays, of the mechanism of action (MOA) of compounds identified through biochemical screening or similar in vitro methods. High-Content Analysis (HCA) is a technique based on automated microscopy which enables multiparametric analysis of fluorescent indicators to define cellular responses to compound treatment. Several distinct fluorescence channels can be acquired and analyzed within a single measure in the same cell population. Here we present a multiparametric HCA approach to characterize potential cell-cycle inhibitors in osteosarcoma U-2 OS adherent cell cultures. This approach allows monitoring of compound-induced cell-cycle perturbations by analyzing specific cellular markers such as nuclear morphology, DNA content or histone H3 phosphorylation. Moreover, the induction of DNA damage response or apoptosis can also be readily evaluated. By considering the profile of the investigated cellular markers at different compound concentrations, a fingerprint defines the cellular and molecular phenotype associated with each compound.

Keywords

BrdU Incorporation Nuclear Area Cellular Marker High Content Screening Automate Microscopy 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Springer 2007

Authors and Affiliations

  • Fabio Gasparri
    • 1
  • Antonella Ciavolella
    • 1
  • Arturo Galvani
    • 1
  1. 1.Department of Biology, Nerviano Medical SciencesItaly

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