The Effects of Wnt Protein on Proliferation and Stemness Maintenance of Corneal Limbal Stem Cells (CLSCs)

ABSTRACT

PURPOSE: To evaluate the effects of the Wnt protein on proliferation and stemness maintenance of cultured corneal limbal stem cells. METHODS: We examined the expression of Wnt proteins by Western blot analysis. We then evaluated the effects of Wnt on cell proliferation by colony forming efficiency. beta-catenin activation using Wnt proteins was examined by immunocytochemistry. We also examined the effects of Wnt on proliferation and stemness maintenance by reverse transcriptase polymerase chain reaction of p63 and connexin43. RESULTS: Wnt has a different effect on corneal epithelial stem cells. Colony forming efficiency was also significantly higher in treated Wnt2 and Wnt4 cells compared with controls. The Wnt2 and Wnt4 treated cells showed nuclear accumulation of beta-catenin. In addition, the limbal stem cell marker p63 was strongly expressed in Wnt2, Wnt4 Wnt5a, and Wnt5b. Connexin43 mRNA was also strongly expressed in Wnt5a, Wnt5b and Wnt7b cells. CONCLUSIONS: We suggest that Wnt2 and Wnt4 could lead to more effective proliferation and stemness maintenance for human corneal epithelial stem cells.