ROCK inhibitor Y-27632 increases the cloning efficiency of limbal stem/progenitor cells by improving their adherence and ROS-scavenging capacity.

Rho-associated coiled-coil kinase (ROCK) inhibitor Y-27632 has been shown to increase proliferative capacity and even immortalize primary keratinocytes. Here, we demonstrate that rabbit primary limbal epithelial cells (LECs) treated with Y-27632 also exhibited improved colony-forming efficiency by enhancing the expansion of the stem/progenitor cells. Moreover, Y-27632 treatment improved the rapid adherence of limbal stem/progenitor cells in the initial inoculation of primary cells. In addition, Y-27632 treatment elevated the intracellular glutathione level and decreased cellular reactive oxygen species (ROS) accumulation during the expansion of LECs. Therefore, ROCK inhibitor Y-27632 increased the cloning efficiency of rabbit limbal stem/progenitor cells by improving their adherence and ROS scavenging capacity.