PLoS ONE (Jan 2020)

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

  • Seongjun So,
  • Yeonmi Lee,
  • Jiwan Choi,
  • Seoon Kang,
  • Ji-Yoon Lee,
  • Julie Hwang,
  • Joosung Shin,
  • James R Dutton,
  • Eul-Ju Seo,
  • Beom Hee Lee,
  • Chong Jai Kim,
  • Shoukhrat Mitalipov,
  • Soo Jin Oh,
  • Eunju Kang

DOI
https://doi.org/10.1371/journal.pone.0233057
Journal volume & issue
Vol. 15, no. 5
p. e0233057

Abstract

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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.