Stress-free cell aggregation by using the CEPT cocktail enhances embryoid body and organoid fitness

Seungmi Ryu, Claire Weber, Pei Hsuan Chu, Ben Ernest, Vukasin M. Jovanovic, Tao Deng, Jaroslav Slamecka, Hyenjong Hong, Yogita Jethmalani, Hannah M. Baskir, Jason Inman, John Braisted, Marissa B. Hirst, Anton Simeonov, Ty C. Voss, Carlos A. Tristan, Ilyas Singeç

Research output: Contribution to journalArticlepeer-review


Embryoid bodies (EBs) and self-organizing organoids derived from human pluripotent stem cells (hPSCs) recapitulate tissue development in a dish and hold great promise for disease modeling and drug development. However, current protocols are hampered by cellular stress and apoptosis during cell aggregation, resulting in variability and impaired cell differentiation. Here, we demonstrate that EBs and various organoid models (e.g., brain, gut, kidney) can be optimized by using the small molecule cocktail named CEPT (chroman 1, emricasan, polyamines, trans-ISRIB), a polypharmacological approach that ensures cytoprotection and cell survival. Application of CEPT for just 24 h during cell aggregation has long-lasting consequences affecting morphogenesis, gene expression, cellular differentiation, and organoid function. Various qualification methods confirmed that CEPT treatment enhanced experimental reproducibility and consistently improved EB and organoid fitness as compared to the widely used ROCK inhibitor Y-27632. Collectively, we discovered that stress-free cell aggregation and superior cell survival in the presence of CEPT are critical quality control determinants that establish a robust foundation for bioengineering complex tissue and organ models.

Original languageEnglish
Article number015016
Issue number1
StatePublished - 11 Dec 2023
Externally publishedYes


  • CEPT cocktail
  • cytoprotection
  • embryoid body
  • iPS cells
  • organoid
  • pluripotent stem cell
  • polypharmacology

ASJC Scopus subject areas

  • Bioengineering
  • Biochemistry
  • Biotechnology
  • Biomedical Engineering
  • Biomaterials


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