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Related Experiment Video

Updated: Feb 22, 2026

Profiling Individual Human Embryonic Stem Cells by Quantitative RT-PCR
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Quantification of Etoposide Hypersensitivity: A Sensitive, Functional Method for Assessing Pluripotent Stem Cell

Frank J Secreto1,2, Xing Li1,3, Alyson J Smith1,2

  • 1Program for Hypoplastic Left Heart Syndrome-Center for Regenerative Medicine, Mayo Clinic, Rochester, Minnesota, USA.

Stem Cells Translational Medicine
|September 20, 2017
PubMed
Summary

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Human induced pluripotent stem cells (hiPSC) quality is crucial for diagnostics and therapeutics. A novel etoposide sensitivity assay (ESA) offers a quantitative, high-throughput method to assess hiPSC quality and distinguish malignant from normal clones.

Area of Science:

  • Stem Cell Biology
  • Cellular Quality Control
  • Drug Sensitivity Assays

Background:

  • Human induced pluripotent stem cells (hiPSC) have significant therapeutic and diagnostic potential.
  • Current methods for assessing hiPSC quality are often slow, expensive, and not high-throughput.
  • Distinguishing malignant teratocarcinoma clones from normal hiPSC lines is a critical challenge.

Purpose of the Study:

  • To develop a novel, quantitative, and high-throughput method for assessing hiPSC quality.
  • To establish a functional assay that correlates with established hiPSC quality metrics.
  • To differentiate malignant from normal pluripotent stem cell clones.

Main Methods:

  • Exploited the hypersensitivity of pluripotent cells to the topoisomerase inhibitor etoposide.
Keywords:
EtoposideFunctionalHypersensitivityPluripotent stem cellsQuantification

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  • Developed an etoposide sensitivity assay (ESA) to determine the half maximal effective concentration (EC50).
  • Studied 115 unique hiPSC clones, analyzing EC50 values after 24-hour etoposide exposure.

    • Main Results:

    • An EC50 value <300 nM of etoposide correlated positively with high-quality hiPSC markers (RNA profiles, colony morphology).
    • The ESA detected quality differences related to culture maintenance.
    • The assay successfully distinguished malignant from normal pluripotent clones, irrespective of morphology.

    Conclusions:

    • The etoposide sensitivity assay (ESA) provides a simple, quantitative, and functional method for hiPSC quality control.
    • ESA is compatible with high-throughput platforms, addressing limitations of current assays.
    • This assay can be integrated into a generalized quality control standard for all pluripotent stem cells.