From March 1st to April 27th, 2008 and from March 3rd to May 1st, 2009, the semen samples from four breeding Giant pandas which distributed in Qinling Mountains (Ailuropoda melanoleuca qinlingensis) were used for semen cryopreservation experiments in Shaanxi Rare Wildlife Rescue and Breeding Research Center. The sperm motility and acrosome integrity were tested after straw cryopreservation of semen with 4 diluents, glucose-fructose-trisodium citrate-yolk-glycerin-antibiotics, glucose-sucrose-trisodium citrate-yolk-glycerin-antibiotics, glucose-trisodium citrate-yolk-glycerin-antibiotics, and TEST imported form the USA(added with 3.5% glycerin), and with 2 methods, direct cooling balance method, and gradual cooling balance method. The results showed that the percentages of sperm motility and intact acrosome were 46.25%±11.67% and 80.75%±7.89% respectively in diluent 1; 48.75%±8.54% and 84.50%±7.59% in diluent TEST. There was no significant difference between these two diluents (P>0.05), but both were better than diluent 2 and diluent 3 (P<0.01). After freezing and thawing, the percentages of sperm motility and intact acrosome were 45.67%±10.54% and 81.37%±8.42% respectively in cryopreservation method 1, significantly higher than those of method 2 (P<0.01). The percentage of sperm abnormality was 23.50%±3.51%,in method 1, significantly lower than that of method 2 (P<0.01). Thus, we conclude that method 1 (with diluent 1) is good for semen straw cryopreservation in Giant Panda.