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.

2nd Department of Surgery Department of Cardiovascular Surgery 1st Medical Faculty Charles University and General University Hospital 128 08 Prague Czech Republic

Department of Anatomy Histology and Embryology Medical Faculty in Hradec Králové Charles University 500 03 Hradec Králové Czech Republic

Department of Cell Biology Charles University Viničná 7 128 00 Prague Czech Republic

Department of Transplantation Surgery Institute for Clinical and Experimental Medicine 140 21 Prague Czech Republic

Imaging Methods Core Facility at BIOCEV Operated by Faculty of Science Charles University 252 50 Prague Czech Republic

Technology Centre of the Czech Academy of Sciences 160 00 Prague Czech Republic

Tissue Bank University Hospital 500 05 Hradec Králové Czech Republic

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