Abstract: The primary cultured cells of horse skin tissue were obtained by explant tissue culture and were purified by enzymes digest. Firstly, Horse skin fibroblasts were frozen in 12 different freezing protective media, which contained different concentration of DMSO, EG, GC and NCS. Secondly, Horse skin fibroblasts were frozen in two different cryo-preservation methods. Efficiency of attachment at 24 h was used to evaluate the effects of cryopreservation. The results of experiment showed that DMEM supplemented with 10% DMSO and 20% NCS was the most efficient cryo-preserving media (efficiency of attachment at 24 h after thawing, 84.98%). The effects of cryopreservation in programming-cryo-preservation were obviously better than those in manual cryo-preservation (86.32%, 79.98%)(P<0.05).

Key words: horse; skin fibroblast; culture in vitro; freezing