Through isolation and pure culture of Shaneng milk goat fibroblasts with skin tissue, fibroblast line was established. On conditions of the same cryoprotectant and FCS concentration Pure Shaneng milk goat fibroblasts cultured were cryopreservated in five cryopreservation methods with different pre-cool and balance ways and pre-cool time, through the contrast and analyses of cell vigor after five months cryopreservation, method 3 (70% cell suspension -f 20%FCS + 10% DMSO; cryopreservation procedure: first fibroblasts were pre-cooled and balanced for half hour at 4 C , and second hanged in gaseous nitrogen in close to the mouth of nitrogen canister for 4 hours, and finally sunk into liquid nitrogen. ) was the best cryopreservation method, it could maintain 84. 8% of living cell rate through cell vigor test after cells were revived and stained by Trypan Blue and 85. 4% of cell adherence rate after cells were revived and cultured for 48 hours, living cell rate and cell adherence rate of method 3 superior to other four methods.