Objective To get limbal stem cells(LSCs)by primary culture and explore the influence of insulin on LSCs’proliferation. Methods The cells from the rabbit corneal limbal tissue were cultured and identified by immunohistochemical staining and transmission electron microscopy. The LSCs were treated with 5mg/L, 10mg/L, 20mg/L, 40mg/L, 80mg/L and 100mg/L insulin respectively. Then methyl thiazolyl tetrazolium(MTT)assay was used to explore the influence of insulin on LSCs’ proliferation. Results The experiment groups at the concentration of 5mg / L and 10mg / L had significantly higher cell viability than that of the control group（EM>P/EM>0.05）, and so were the 5mg/L group and 10mg/L group（EM>P/EM>0.05）. But there were no significant difference between the experimental groups at th