Assessment of viability of long-term stored cryopreserved allogeneic venous grafts using vital fluorescence dyes and confocal microscopy
Abstract
Eight grafts of vena sahpena magna collected from deceased donors were examined.Controlled freezing in the presence of 10% DMSO was performed within 24 hours of collection. This was followed by storage in liquid nitrogen vapors for 3 years (2 grafts) or 5 years (6 grafts).
Thawing was followed by viability testing using fluorescent dyesaccording to Johnson and Rabinovitch at the Imaging Methods Service Laboratory, BIOCEV. After dividing into 3 segments, viability was determined immediately, after 24 and 48 hours of culturing in Dulbecco's medium in a CO 2 incubator at + 37 ° C.
Immediatelyafter thawing, the mean viability was in vertical sections (KR): 81%, SD 6.5, median 87.5%, in longitudinal sections (PR): 73.5%, SD 4.8, median 76.5%. After 48 hours it decreased in KR to 60.6%, SD 6.8, median 66.5%, in PR it was 74.8%, SD 5.9, median 79%.
The results showed very good preservation of cell viability and good stabilityof results after short-term cultivation.