Quantification of Etoposide Hypersensitivity: A Sensitive, Functional Method for Assessing Pluripotent Stem Cell Quality

Frank J. Secreto, Xing Li, Alyson J. Smith, Elizabeth S. Bruinsma, Ester Perales-Clemente, Saji Oommen, Gresin Hawse, Sybil C.L. Hrstka, Bonnie K. Arendt, Emma B. Brandt, Dennis A Wigle, Timothy J Nelson

Research output: Contribution to journalArticle

2 Scopus citations

Abstract

Human induced pluripotent stem cells (hiPSC) hold great promise in diagnostic and therapeutic applications. However, translation of hiPSC technology depends upon a means of assessing hiPSC quality that is quantitative, high-throughput, and can decipher malignant teratocarcinoma clones from normal cell lines. These attributes are lacking in current approaches such as detection of cell surface makers, RNA profiling, and/or teratoma formation assays. The latter remains the gold standard for assessing clone quality in hiPSCs, but is expensive, time-consuming, and incompatible with high-throughput platforms. Herein, we describe a novel method for determining hiPSC quality that exploits pluripotent cells’ documented hypersensitivity to the topoisomerase inhibitor etoposide (CAS No. 33419-42-0). Based on a study of 115 unique hiPSC clones, we established that a half maximal effective concentration (EC50) value of <300 nM following 24 hours of exposure to etoposide demonstrated a positive correlation with RNA profiles and colony morphology metrics associated with high quality hiPSC clones. Moreover, our etoposide sensitivity assay (ESA) detected differences associated with culture maintenance, and successfully distinguished malignant from normal pluripotent clones independent of cellular morphology. Overall, the ESA provides a simple, straightforward method to establish hiPSC quality in a quantitative and functional assay capable of being incorporated into a generalized method for establishing a quality control standard for all types of pluripotent stem cells. Stem Cells Translational Medicine 2017;6:1829–1839.

Original languageEnglish (US)
Pages (from-to)1829-1839
Number of pages11
JournalStem cells translational medicine
Volume6
Issue number10
DOIs
StatePublished - Oct 1 2017

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Keywords

  • Etoposide
  • Functional
  • Hypersensitivity
  • Pluripotent stem cells
  • Quantification

ASJC Scopus subject areas

  • Developmental Biology
  • Cell Biology

Cite this

Secreto, F. J., Li, X., Smith, A. J., Bruinsma, E. S., Perales-Clemente, E., Oommen, S., Hawse, G., Hrstka, S. C. L., Arendt, B. K., Brandt, E. B., Wigle, D. A., & Nelson, T. J. (2017). Quantification of Etoposide Hypersensitivity: A Sensitive, Functional Method for Assessing Pluripotent Stem Cell Quality. Stem cells translational medicine, 6(10), 1829-1839. https://doi.org/10.1002/sctm.17-0116