The authors present their contribution to the improvement of methods suitable for the detection of the freezing and thawing damage of cells of cryopreserved venous grafts used for lower limb revascularization procedures. They studied the post-thaw viability of cells of the wall of cryopreserved venous grafts (CVG) immediately after thawing and after 24 and 48 h culture at +37 °C in two groups of six CVG selected randomly for slow thawing in the refrigerator and rapid thawing in a water bath at +37 °C. The grafts were collected from multi-organ and tissue brain-dead donors, cryopreserved, and stored in a liquid nitrogen vapor phase for five years. The viability was assessed from tissue slices obtained by perpendicular and longitudinal cuts of the thawed graft samples using in situ staining with fluorescence vital dyes. The mean and median immediate post-thaw viability values above 70% were found in using both thawing protocols and both types of cutting. The statistically significant decline in viability after the 48-h culture was observed only when using the slow thawing protocol and perpendicular cutting. The possible explanation might be the "solution effect damage" during slow thawing, which caused a gentle reduction in the graft cellularity. The possible influence of this phenomenon on the immunogenicity of CVG should be the subject of further investigations.

• Keywords
• Thawing method, cell viability, confocal microscopy, cryopreservation, fluorescence vital dyes, vascular allograft,
• MeSH
• Allografts diagnostic imaging   drug effects   MeSH
• Apoptosis drug effects   MeSH
• Tissue Donors MeSH
• Dimethyl Sulfoxide pharmacology   MeSH
• Fluorescent Dyes * MeSH
• Microscopy, Confocal methods   MeSH
• Cryopreservation methods   MeSH
• Cryoprotective Agents pharmacology   MeSH
• Humans MeSH
• Optical Imaging methods   MeSH
• Vascular Grafting methods   MeSH
• Femoral Vein diagnostic imaging   drug effects   MeSH
• Saphenous Vein diagnostic imaging   drug effects   MeSH
• Cell Survival drug effects   MeSH
• Freezing * MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Dimethyl Sulfoxide MeSH
• Fluorescent Dyes * MeSH
• Cryoprotective Agents MeSH