The Rho-associated kinase inhibitor fasudil can replace Y-27632 for use in human pluripotent stem cell research

PLoS One. 2020 May 12;15(5):e0233057. doi: 10.1371/journal.pone.0233057. eCollection 2020.

Abstract

Poor survival of human pluripotent stem cells (hPSCs) following freezing, thawing, or passaging hinders the maintenance and differentiation of stem cells. Rho-associated kinases (ROCKs) play a crucial role in hPSC survival. To date, a typical ROCK inhibitor, Y-27632, has been the primary agent used in hPSC research. Here, we report that another ROCK inhibitor, fasudil, can be used as an alternative and is cheaper than Y-27632. It increased hPSC growth following thawing and passaging, like Y-27632, and did not affect pluripotency, differentiation ability, and chromosome integrity. Furthermore, fasudil promoted retinal pigment epithelium (RPE) differentiation and the survival of neural crest cells (NCCs) during differentiation. It was also useful for single-cell passaging of hPSCs and during aggregation. These findings suggest that fasudil can replace Y-27632 for use in stem research.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine / analogs & derivatives*
  • 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine / pharmacology
  • Amides / pharmacology
  • Cell Differentiation / drug effects
  • Cell Line
  • Cell Proliferation / drug effects
  • Cell Survival / drug effects
  • Embryonic Stem Cells / cytology
  • Embryonic Stem Cells / drug effects
  • Humans
  • Induced Pluripotent Stem Cells / cytology
  • Induced Pluripotent Stem Cells / drug effects
  • Neural Crest / cytology
  • Neural Crest / drug effects
  • Pluripotent Stem Cells / cytology*
  • Pluripotent Stem Cells / drug effects*
  • Protein Kinase Inhibitors / pharmacology
  • Pyridines / pharmacology
  • Retinal Pigment Epithelium / cytology
  • Retinal Pigment Epithelium / drug effects
  • Stem Cell Research
  • rho-Associated Kinases / antagonists & inhibitors*

Substances

  • Amides
  • Protein Kinase Inhibitors
  • Pyridines
  • Y 27632
  • 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine
  • rho-Associated Kinases
  • fasudil

Grants and funding

The study was supported by grants from the National Research Foundation of Korea (NRF-2015K1A4A3046807 and 2017M3A9F8031039) and intramural grants (2018-796) from the Asan Institute for Life Sciences, Asan Medical Center. There was no additional external funding received for this study.