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Gene-modified embryonic stem cell test to characterize chemical risks.

Kohei Kitada1, Akane Kizu2, Takeshi Teramura3

  • 1Department of Obstetrics and Gynecology, Osaka City University, Graduate School of Medicine, 1-4-3 Asahimachi, Abeno-ku, Osaka, 545-8585, Japan.

Environmental Science and Pollution Research International
|July 25, 2015
PubMed
Summary

This study developed a high-throughput cell growth inhibition test using mouse embryonic stem (ES) cells to assess chemical toxicity. Gene-modified ES cells help categorize chemicals by their effects on apoptosis and genotoxicity.

Keywords:
Chemical risksCytotoxicityGene modificationHigh-throughput analysisMouse embryonic stem cellsp53

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Area of Science:

  • Toxicology and Pharmacology
  • Genetics and Molecular Biology
  • Cell Biology

Background:

  • Chemical toxicity assessment is crucial for risk evaluation.
  • The p53 tumor suppressor gene plays a key role in apoptosis and senescence.
  • Gene-modified cell lines offer advanced models for toxicological studies.

Purpose of the Study:

  • To establish a high-throughput cell growth inhibition assay for chemical toxicity screening.
  • To utilize p53 gene-modified mouse embryonic stem (ES) cells for chemical categorization.
  • To investigate the role of p53 in cellular responses to chemical exposure.

Main Methods:

  • A 96-well plate-based MTT assay was employed for high-throughput screening.
  • Wild-type and p53-deficient mouse ES cells were used to assess differential cytotoxicity.
  • Chemicals tested included diethylnitrosamine (DEN), methyl methanesulfonate (MMS), and N-methyl-N-nitrosourea (NMU).

Main Results:

  • p53-deficient ES cells showed increased resistance to DEN, suggesting impaired apoptosis or senescence.
  • Genomic instability, indicated by chromosome aberrations, was higher in p53-deficient ES cells.
  • No significant cytotoxicity differences were observed for MMS and NMU between cell types.

Conclusions:

  • The ES cell-based assay effectively prioritizes chemicals based on cytotoxic properties.
  • Utilizing gene-modified ES cells allows for categorization based on specific cellular responses.
  • This system aids in evaluating chemical effects on apoptosis, genotoxicity, and general cytotoxicity.