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.
Bibliographical noteFunding 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.
© 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.