Abstract: Objective To study the changes in cryo-preserved high molecular weight epidermal keratin and analyze the effect of cryobiology on epidermal cytoskeleton. Methods Adult human skin samples were divided into fresh group,4℃ group,-20℃ group,-80℃ group and-196℃ group.Changes in epidermal keratin were observed under an electron microscope with immunohistochemical staining 14 days after cryopreservation.Keratin level in skin tissue was analyzed with an image analyzer. Results The expression level of keratin decreased with the decreasing preservation temperature in 4℃ group,-20℃ group,and-80℃ group,and was similar in-196℃ group and fresh group. Conclusion Hypothermia injury of skin is closely correlated with cytoskeleton,and keratin plays an important role in the mechanism of cryobiology.