The aim at optimizing the procedure for preparing an immune-deficient hindlimb ischemia mouse, a model of hindlimb in immunodeficient mouse is made by combining the most effective method for. each model. Hindlimb ischemia mouse model is made by burning the femoral artery and examined the effect on 3 to 5-month-old mouse and 6 to 12 month-old mouse. Mice with the best effective for hindlimb ischemia are used in the immunosuppression protocol. There were 3 groups set up in the study to create model of hindlimb ischemia, in which the dosages were as follows (injected for 4 days): (1) cyclophosphomide 50mg/kg, (2) busufan 20mg/kg combined with cyclophosphomide 50mglkg, (3) busulfan 40mg/kg combined with cyclophosphomide 50mg/kg, from which to select the most effective regimen for stability. The results showed (1) 100 percent of the mices in 6 to 12-month old mouse group showed more advantages when being used to make the hindlimb ischemia model
(2) busulfan 20 mg/kg combined cyclophosphomide 50 mg/kg for 4 days, then inject cyclophosphomide 50mg/kg 3 daysltimes for the highest efficiency immunodeficiency and effective immunosuppression was maintained stable for 15 days. (3) The combination of the two regimens have created a mouse model both hindlimb ischemic, immunodeficiency stable for at least 15 days.