Effects of Y-27632, a Rho-associated Kinase Inhibitor, on Human Corneal Endothelial Cells Cultured by Isolating Human Corneal Endothelial Progenitor Cells
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Purpose: Human corneal endothelial progenitor cells (HCEPs), which has been selectively isolated and differentiated intohuman corneal endothelial cells (HCECs), are crucial for repairing corneal endothelial damage. In this study, we evaluated theroles of a Rho-assisted kinase (ROCK) inhibitor, Y-27632, on the isolation and expansion of HCEPs, and assessed the in vitroeffects of different concentrations of Y-27632 on the differentiated HCEPs. Methods: HCEPs were isolated and expanded in a medium with and without 10μM Y-27632, and then differentiated intoHCECs in a medium with fetal bovine serum. The characteristics of HCEPs and differentiated HCEPs were confirmed by immunofluorescencestaining. The proliferation, viability, morphology, and wound-healing ability of differentiated HCEPs wereassessed 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 HCEPsshowed an optimal increase in proliferation and survival in the presence of 10μM Y-27632. As the concentration of Y-27632increased, differentiated HCEPs became elongated, and actin filaments were redistributed to the periphery of cells. Y-27632also caused a concentration-dependent enhancement in the wound-healing ability of differentiated HCEPs. Conclusions: Y-27632 enabled the isolation and expansion of HCEPs. It also enhanced the proliferation, viability, and migrationof differentiated HCEPs.
Abstract
Purpose: Human corneal endothelial progenitor cells (HCEPs), which has been selectively isolated and differentiated intohuman corneal endothelial cells (HCECs), are crucial for repairing corneal endothelial damage. In this study, we evaluated theroles of a Rho-assisted kinase (ROCK) inhibitor, Y-27632, on the isolation and expansion of HCEPs, and assessed the in vitroeffects of different concentrations of Y-27632 on the differentiated HCEPs. Methods: HCEPs were isolated and expanded in a medium with and without 10μM Y-27632, and then differentiated intoHCECs in a medium with fetal bovine serum. The characteristics of HCEPs and differentiated HCEPs were confirmed by immunofluorescencestaining. The proliferation, viability, morphology, and wound-healing ability of differentiated HCEPs wereassessed 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 HCEPsshowed an optimal increase in proliferation and survival in the presence of 10μM Y-27632. As the concentration of Y-27632increased, differentiated HCEPs became elongated, and actin filaments were redistributed to the periphery of cells. Y-27632also caused a concentration-dependent enhancement in the wound-healing ability of differentiated HCEPs. Conclusions: Y-27632 enabled the isolation and expansion of HCEPs. It also enhanced the proliferation, viability, and migrationof differentiated HCEPs.
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