Influence of ischemic time and temperature on endothelial cell growth after transport

Background: The preparation of tissue-engineered material is a complex proc edure. The possibility to transport tissue between laboratories without loo sing endothelial cell (EC) function was examined. Methods: In 3 month old juvenile sheep (n = 6) a piece of vein (n = 14) was harvested and transported over 900 km to the tissue laboratory in Dulbecco 's Modified Eagle's Medium (=DMEM). Vein material of each animal was transp orted at 4 degreesC (Group I, n = 6) and 25 degreesC (Group II, n = 8). EC growth potential was evaluated in function of the medium temperature and th e ischemic time (between 8-24 hours). At the end of the first passage the E C of Group I and II were put together to save autologous serum of the sheep . After the 2(nd) passage the EC were cryopreserved at -80 degreesC to eval uate if EC viability would change. Results: The growth potential of hypothermic Group I was equal in 16.7% (n = 1), higher in 33.3% (n = 2) and lower in 50% (n = 3) than Group II which had the same ischemic time during transport. Increase in ischemic time up t o 24 hours showed no decrease of growth potential. Cryopreservation had no significant influence on EC viability. Viability at the end of the second p assage, after recultivation and at the end of the third passage was 97.4% /- 1.52, 95.5% +/- 1.34 and 94.5% +/- 1.08 respectively. Conclusions: In sheep there is no need to transport the EC at a temperature of 4 degreesC. Up to 24 hours growth potential and viability are maintaine d also at 25 degreesC.