Susceptibility of human pluripotent stem cells (hPSCs), such as human iPS and embryonic stem (ES) cells, to single-cell dissociation has been a large obstacle to develop the efficient manipulation techniques required for stem cell research. When hPSCs are completely dissociated into single cells, programmed cell death (apoptosis) is immediately induced. A specific inhibitor of Rho-associated protein ldnase (ROCK inhibitor), Y-27632, is of particular interest as a useful reagent that allows hPSCs to escape the dissociation-induced apoptosis. ROCK inhibitor has been used in a variety of applications associated with cell dissociation in the process of stem cell research, such as passaging, expansion, cryopreservation, gene transfer, differentiation induction, and cell sorting, suggesting that it may be a crucial reagent for the handling of hPSCs. This article reviews the current applications of ROCK inhibitors to stem cell research from the viewpoint of quality control of hPSCs. (C) 2012, The Society for Biotechnology, Japan. All rights reserved.