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

Research output: Contribution to journalArticlepeer-review

11 Scopus citations


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.

Original languageEnglish (US)
Article numbere0233057
JournalPloS one
Issue number5
StatePublished - May 2020

Bibliographical note

Funding Information:
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. We thank Jumi Park, Doin Kim, Hyunjeong Kim, Yuri Han, Mustafa Zafer Karagozlu, the Confocal Imaging Core, FACS Core Laboratory, and the Medical Genetics Center at Asan Medical Center for their technical support. We are grateful to the Center for Embryonic Cell and Gene Therapy, Oregon Health & Science University, for providing hESCs. We also would like to thank the DMPK core facility at the ConveRgence mEDIcine research cenTer (CREDIT), Asan Medical Center, for conducting molecular stability analysis and Patrick Walsh, Vincent Truong, and Caitlin Hill at the Stem Cell Institute, University of Minnesota, for transferring expertise in hPSC culture and differentiation.

Publisher Copyright:
© 2020 So et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.


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