A versatile polypharmacology platform promotes cytoprotection and viability of human pluripotent and differentiated cells

Nat Methods. 2021 May;18(5):528-541. doi: 10.1038/s41592-021-01126-2. Epub 2021 May 3.


Human pluripotent stem cells (hPSCs) are capable of extensive self-renewal yet remain highly sensitive to environmental perturbations in vitro, posing challenges to their therapeutic use. There is an urgent need to advance strategies that ensure safe and robust long-term growth and functional differentiation of these cells. Here, we deployed high-throughput screening strategies to identify a small-molecule cocktail that improves viability of hPSCs and their differentiated progeny. The combination of chroman 1, emricasan, polyamines, and trans-ISRIB (CEPT) enhanced cell survival of genetically stable hPSCs by simultaneously blocking several stress mechanisms that otherwise compromise cell structure and function. CEPT provided strong improvements for several key applications in stem-cell research, including routine cell passaging, cryopreservation of pluripotent and differentiated cells, embryoid body (EB) and organoid formation, single-cell cloning, and genome editing. Thus, CEPT represents a unique poly-pharmacological strategy for comprehensive cytoprotection, providing a rationale for efficient and safe utilization of hPSCs.

Publication types

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

MeSH terms

  • Cell Culture Techniques
  • Cell Differentiation / drug effects*
  • Cell Survival / drug effects*
  • Cryopreservation / methods
  • Cryoprotective Agents / chemistry
  • Cryoprotective Agents / pharmacology*
  • Gene Expression Regulation / drug effects
  • High-Throughput Screening Assays
  • Humans
  • Pluripotent Stem Cells / drug effects*
  • Pluripotent Stem Cells / physiology
  • Polypharmacology*
  • rho-Associated Kinases / antagonists & inhibitors
  • rho-Associated Kinases / genetics
  • rho-Associated Kinases / metabolism


  • Cryoprotective Agents
  • rho-Associated Kinases