| Background and Objective: Cryopreservation has been extensively used in cryomedicine. Vitrification is considered as the best method for the cryopreservation of different organisms. However, how to improve the viability of the preserved skin is still unsatisfactory. It has reported that the reaction of tissues to hypothermy involves alterations in cell-cell and cell-matrix interactions mediated by cellular cytoskeleton suggesting it would take a important role in the cryopreservation. But the mechanism is not entirely cleared. As we know, the normal function of skin is highly dependent on the epidermal cytoskeleton system. Disruption and dysfunction of cytoskeleton may result in impairment of dermal function. The aim of this study was to investigate the changes of epidermal cytoskeleton treated by different temperatures and to look for a better cryoprotectant(CPA) for skin..Methods: In the first part of this thesis, the skins were stored in four different temperature: 4℃, —20℃, —80℃, —196℃, comparing with the fresh skin. After we observed the changes of cutaneous cells by microscopy, four associated proteins (α-tubulin, β- tubulin, β-actin and cytoskeratin ) have been studied after 7days, 14days and 21 days. Then we analyzed the expression of these proteins by H.E. staining and immunohistochemical staining after different treatment. We also observed the ultrastructure of microfilament, microtuble and intermediate filament by electron microscope. Finally, we used RT-PCR and Western blot to investigate the changes of the content of p-actin treated by different temperature.In the second part of this thesis, the cryopreserved skins were treated with DMSO/ Propyleneglycol, DMSO, DMEM and trehalose/DMSO, comparing with...
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