Effect of the thawing process on cryopreserved arteries.
This study was designed to explore the changes that occur in cryopreserved pig arteries following different thawing procedures, before and after being placed in an in vitro flow circuit. Segments of minipig iliac artery were cryopreserved in complete minimal essential medium plus 10% dimethylsulphoxide and stored in liquid nitrogen at -196 degrees C for 30 days. Three study groups were established according to whether the arterial specimens were fresh (control, n = 20), cryopreserved and rapidly thawed (RT) at 37 degrees C (n = 22) or cryopreserved and subjected to controlled, automated slow thawing (ST) (n = 22). Half of the specimens of each group were subsequently placed in the flow circuit for 72 hr. Evaluation was made of morphological and ultrastructural changes. Cell damage was established using the TUNEL method. All cryopreserved specimens showed endothelial denudation that was most extensive in those subjected to rapid thawing. Slowly thawed specimens showed improved cell viability and organization of the vessel wall, compared to those thawed rapidly. Under conditions of flow, the damage induced by the freezing/thawing process was enhanced. These findings suggest that (a) slow thawing of cryopreserved arteries results in improved preservation of the structure and viability of vessels, and (b) the damage induced by freezing/thawing is enhanced when vessels are subjected to flow in an in vitro circuit.
Pascual
G
G
Department of Morphological Sciences and Surgery (Surgical Research Laboratory), Medical School, University of Alcala, Crta. Madrid-Barcelona Km 33,600, E-28871 Alcalá de Henares, Madrid, Spain.
García-Honduvilla
N
N
Rodríguez
M
M
Turégano
F
F
Bujan
J
J
Bellón
J M
JM
D000818:Animals; D002470:Cell Survival / Q000502:physiology; D015925:Cryopreservation / Q000379:methods*; D053938:DNA Fragmentation / Q000502:physiology; D004730:Endothelium, Vascular / Q000502:physiology / Q000503:physiopathology; D007083:Iliac Artery / Q000502:physiology*; D020287:In Situ Nick-End Labeling; D023421:Models, Animal; D008955:Models, Cardiovascular; D009131:Muscle, Smooth, Vascular / Q000502:physiology / Q000503:physiopathology; D013556:Swine, Miniature; D014654:Vascular Patency / Q000502:physiology