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J Thorac Cardiovasc Surg 1996;111:218-230
© 1996 Mosby, Inc.

SURGERY FOR ACQUIRED HEART DISEASE

EFFECTS OF STORAGE TEMPERATURE AND FETAL CALF SERUM ON THE ENDOTHELIUM OF PORCINE AORTIC VALVES

Antwerp, Belgium

Received for publication Nov. 21, 1994. Accepted for publication Feb. 22, 1995. Address for reprints: Xiao J. Feng, MD, Department of Pharmacology, University of Antwerp (UIA), Universiteitsplein 1, B-2610 Wilrijk, Belgium.

Abstract

Endothelial integrity and function may be an important determinant for long-term success of allograft heart valves. To determine the optimal storage temperatures for preservation of long-term endothelial function in porcine aortic valves, different storage temperatures and times were investigated. Fresh valves were either (1) stored at 4° C, with or without 10% fetal calf serum supplement, for 1, 2, 4, 7, 14, 21, or 28 days; (2) cryopreserved for 2, 4, or 8 weeks at -80º C or -170° C; (3) cryopreserved in long-term storage (as long as 1 year), with or without fetal calf serum, at -170° C. Viability of endothelial cells was assessed through measurement of the production of prostacyclin in basal and bradykinin-stimulated conditions, during in vitro incubation of the valve cusps at 37° C. Endothelial morphologic variations in valves stored at 4° C were evaluated by scanning electron microscopy. With storage at 4° C, after 4 days the valves already produced significantly less (p < 0.05) prostacyclin than fresh preparations in both basal (0.21 ± 0.04 versus 3.56 ± 0.03 ng · ml^-1 · cm^-2) and stimulated conditions (4.17 ± 0.36 vs 24.23 ± 1.83). Morphologic changes could not yet be distinguished with scanning electron microscopy at that time. When the storage period was extended, the levels of prostacyclin further diminished; after 14 days, prostacyclin release could no longer be detected. In cryopreserved valves, prostacyclin production was similar for as long as 2 weeks of storage either at -80° C or at -170° C in basal (2.69 ± 0.63 vs 2.93 ± 0.51) and stimulated (16.43 ± 3.19 vs 16.50 ± 2.57, = 6) conditions. After 8 weeks, no prostacyclin release could be detected in valves stored at -80° C. After 6 months storage at -170° C, the prostacyclin production was significantly (p < 0.05) reduced compared with fresh valves; it then remained constant for as long as 1 year. The valves stored with fetal calf serum produced significantly (p < 0.05) less prostacyclin than did those without fetal calf serum. For longer cryopreserved banking, we recommend storing heart valves at -170° C instead of at -80° C to maintain viability of endothelial cells. Fetal calf serum would harm endothelial viability during long-term cryopreservation. (J THORAC CARDIOVASC SURG 1996;111:218-30)

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