Stem cells are vital for tissue regeneration, and their dysfunction can lead to diseases like aplastic anemia and Alzheimer's disease. Limbal epithelial stem cells (LESCs) maintain corneal transparency as the sole source of corneal epithelium. Dysfunction or deficiency of LESCs causes corneal opacity, neovascularization or corneal blindness, known as limbal stem cell deficiency (LSCD). Current LSCD treatment mainly relies on stem cell transplantation, which faces challenges like limited cell sources and immune rejection, making therapy challenging and often ineffective. This study introduces a cell‐free, stemness‐enhanced eye drop (SEED@) to stimulate autologous LESCs regeneration. SEED@, a thermosensitive Pluronic F‐127 hydrogel loaded with versican, can promote LESCs proliferation, chemotaxis, and migration while maintaining their stemness via the Wnt/β‐catenin signaling pathway. In vivo, Pluronic F‐127 alone can lubricate the cornea and promote corneal epithelial repair in mouse corneal alkali burn model. In addition, SEED@ forms a thin, transparent layer on the ocular surface, enabling sustained versican release for up to 24 h. Notably, twice‐daily application of SEED@ significantly accelerated corneal regeneration in a rabbit LSCD model, achieving approximately 95% corneal epithelial repair by day 14. Collectively, SEED@ shows great potential for treating LSCD and offers a promising drug delivery platform for ocular diseases.