Method for selective ablation of undifferentiated human pluripotent stem cell populations for cell-based therapies

JCI Insight. 2021 Apr 8;6(7):e142000. doi: 10.1172/jci.insight.142000.


Human pluripotent stem cells (PSCs), which are composed of embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), provide an opportunity to advance cardiac cell therapy-based clinical trials. However, an important hurdle that must be overcome is the risk of teratoma formation after cell transplantation due to the proliferative capacity of residual undifferentiated PSCs in differentiation batches. To tackle this problem, we propose the use of a minimal noncardiotoxic doxorubicin dose as a purifying agent to selectively target rapidly proliferating stem cells for cell death, which will provide a purer population of terminally differentiated cardiomyocytes before cell transplantation. In this study, we determined an appropriate in vitro doxorubicin dose that (a) eliminates residual undifferentiated stem cells before cell injection to prevent teratoma formation after cell transplantation and (b) does not cause cardiotoxicity in ESC-derived cardiomyocytes (CMs) as demonstrated through contractility analysis, electrophysiology, topoisomerase activity assay, and quantification of reactive oxygen species generation. This study establishes a potentially novel method for tumorigenic-free cell therapy studies aimed at clinical applications of cardiac cell transplantation.

Keywords: Cardiology; Cardiovascular disease; Stem cell transplantation; Stem cells.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Apoptosis / drug effects
  • Cardiotoxicity / etiology
  • Cardiotoxicity / prevention & control
  • Cell Death / drug effects
  • Cell Differentiation / drug effects
  • Cell Proliferation / drug effects
  • Cell- and Tissue-Based Therapy / adverse effects
  • Cell- and Tissue-Based Therapy / methods*
  • Dose-Response Relationship, Drug
  • Doxorubicin / administration & dosage*
  • Doxorubicin / pharmacology
  • Embryonic Stem Cells / drug effects*
  • Embryonic Stem Cells / transplantation
  • Gene Expression Regulation / drug effects
  • Human Embryonic Stem Cells / cytology
  • Human Embryonic Stem Cells / drug effects
  • Humans
  • Mice
  • Mice, SCID
  • Myocytes, Cardiac / drug effects*
  • Pluripotent Stem Cells / cytology*
  • Reactive Oxygen Species / metabolism
  • Teratoma / prevention & control


  • Reactive Oxygen Species
  • Doxorubicin