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

• Published in: PloS one Vol. 15; no. 5; p. e0233057
• Main Authors: So, Seongjun, Lee, Yeonmi, Choi, Jiwan, Kang, Seoon, Lee, Ji-Yoon, Hwang, Julie, Shin, Joosung, Dutton, James R, Seo, Eul-Ju, Lee, Beom Hee, Kim, Chong Jai, Mitalipov, Shoukhrat, Oh, Soo Jin, Kang, Eunju
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 12.05.2020; Public Library of Science (PLoS)
• Subjects: Apoptosis; Biology and Life Sciences; Cell differentiation; Cell survival; Chromosomes; Differentiation; EDTA; Enzyme inhibitors; Epithelium; Freezing; Kinase inhibitors; Kinases; Life sciences; Medical research; Medical schools; Medicine; Medicine and Health Sciences; Melting; Methods; Neural crest; Physical Sciences; Pluripotency; Researchers; Retinal pigment epithelium; Rho-associated kinase; Rocks; Stem cell research; Stem cells; Survival; Thawing; South Korea; Oregon; United States > US
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0233057

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.