The invention discloses application of regulatory T cells in preparing a drug for treating autoimmune diabetes and an expanding cultivation solution and method thereof. The expanding cultivation solution of the regulatory T cells is prepared from, by volume, 35.274-42.69 parts of serum-free cultivation solutions, 1.25-2.5 parts of 4-(2-hydroxyerhyl)piperazine-1-erhanesulfonic acid buffer solution, 0.5-1 part of penicillin-streptomycin solution, 0.5-1 part of L-glutamine, 0.05-0.1 part of 2-mercaptoethanol, 0.00769-0.01154 part of recombinant interleukin-2, 0.001-0.01 part of rapamycin and 5-10 parts of AB serum. According to the application of the regulatory T cells in preparing the drug for treating the autoimmune diabetes, through targeted research on the regulatory T cells and the pathogenesis of the autoimmune diabetes, applicants find that the regulatory T cells has relatively good therapeutic effects at the aspect of treating the autoimmune diabetes.