Effects of Y-27632, a ROCK inhibitor on Human Corneal Endothelial Cells Cultured by Isolating Human Corneal Endothelial Progenitor cells

Purpose: Human corneal endothelial progenitor cells (HCEPs), which has been selectively isolated and differentiated into human corneal endothelial cells (HCECs), are crucial for repairing corneal endothelial damage. In this study, we evaluated the roles of a ROCK inhibitor, Y-27632, on the isolation and expansion of HCEPs, and assessed the in vitro effects of different concentrations of Y-27632 on the differentiated HCEPs (HCECs). Methods: HCEPs were isolated and expanded in a medium with and without 10 μM Y-27632, and then differentiated into HCECs in a medium with FBS. The characteristics of HCEP and differentiated HCEPs (HCECs) were confirmed by immunofluorescence staining. The proliferation, viability, morphology, and wound healing ability of HCECs were assessed in the presence of different concentrations of Y-27632. Results: Y-27632 enabled the isolation and expansion of HCEPs from the corneal endothelium. The differentiated HCEPs (HCECs) showed an optimal increase in proliferation and survival in the presence of 10 μM Y-27632. As the concentration of Y-27632 increased, differentiated HCEPs (HCECs) became elongated, and actin filaments were redistributed to the periphery of cells. Y-27632 also caused a concentration-dependent enhancement in the wound healing ability of differentiated HCEPs (HCECs). Conclusions: Y-27632 enabled the isolation and expansion of HCEPs. It also enhanced the proliferation, viability, and migration of differentiated HCEPs (HCECs).