Functional recovery of human mesenteric and coronary arteries after cryopreservation at -196 degrees C in a serum-free medium.
OBJECTIVE
Long-term patency of cryopreserved vascular grafts is determined by maintained cellular and tissue viability, which implies preservation of various biochemical, smooth muscle, and endothelial functions. Therefore, it was investigated whether the presence of fetal calf serum (FCS) in the cryomedium improves the postthaw contractile and endothelial function of human arteries.


METHODS
Rings from human mesenteric (HMA) and left circumflex coronary arteries (HCA) obtained from organ donors were randomized into three groups and studied either unfrozen or after storage for 3 to 6 weeks at -196 degrees C while suspended in Krebs-Henseleit solution without or with 20% FCS as the vehicles and 1.8 mol/L dimethyl sulfoxide and 0.1 mol/L sucrose as cryoprotecting agents. The samples were slowly frozen to -70 degrees C and then stored in liquid nitrogen. Before use, the tissues were thawed within 3 minutes in a 40 degrees C water bath.


RESULTS
After thawing the sensitivity to various agonists and maximal responses to the endothelium-independent relaxing agent sodium nitroprusside were unchanged. However, after cryopreservation of HMA was performed without and with FCS, maximal contractile responses to noradrenaline were significantly reduced to 10.1 +/- 0.7 gm and 9.9 +/- 0.9 gm compared with 13.3 +/- 0.6 gm in unfrozen HMA (mean +/- SEM, n = 15). After cryopreservation of HCA was performed without and with FCS, maximal contractile responses to prostaglandin F2 alpha (6.9 +/- 0.4 gm in unfrozen HCA) were significantly reduced to 4.3 +/- 0.3 gm and 3.8 +/- 0.2 gm (mean +/- SEM, n = 6). In both types of arteries cryopreservation also attenuated significantly the endothelium-dependent relaxant responses to bradykinin during U46619 (10 nmol/L)-induced tone. In HMA the maximal bradykinin-induced relaxation (85% +/- 4%) was significantly diminished to 29% +/- 7% and 38% +/- 9% after cryopreservation without and with FCS (mean +/- SEM, n = 6). In HCA maximal bradykinin-induced relaxation (88% +/- 4%) was significantly diminished to 26% +/- 10% and 36% +/- 11% after cryopreservation without and with FCS (mean +/- SEM, n = 6). This result was reflected by a marked endothelial denudation in all groups of cryopreserved arteries. Neither functional nor morphologic preservation of the endothelial cell lining was significantly improved by FCS supplementation of the cryomedium.


CONCLUSIONS
Cryopreservation diminished contractile and endothelium-dependent relaxant responses of human arteries. The presence of FCS in the cryomedium did not modify these changes.
Müller-Schweinitzer
E
E
Department of Internal Medicine, Clinical Pharmacology, University Hospital, Basel, Switzerland.
Mihatsch
M J
MJ
Schilling
M
M
Haefeli
W E
WE
D000818:Animals; D001920:Bradykinin / Q000494:pharmacology; D002417:Cattle; D002470:Cell Survival; D003331:Coronary Vessels / Q000033:anatomy & histology / Q000187:drug effects / Q000502:physiology* / Q000637:transplantation; D015925:Cryopreservation; D003451:Cryoprotective Agents / Q000008:administration & dosage; D016895:Culture Media, Serum-Free; D004121:Dimethyl Sulfoxide / Q000008:administration & dosage; D015237:Dinoprost / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000378:metabolism / Q000502:physiology; D005312:Fetal Blood; D005615:Freezing; D005947:Glucose / Q000008:administration & dosage; D006801:Humans; D008638:Mesenteric Arteries / Q000033:anatomy & histology / Q000187:drug effects / Q000502:physiology* / Q000637:transplantation; D009131:Muscle, Smooth, Vascular / Q000378:metabolism / Q000502:physiology; D009584:Nitrogen; D009599:Nitroprusside / Q000494:pharmacology; D009638:Norepinephrine / Q000494:pharmacology; D019188:Organ Preservation Solutions / Q000008:administration & dosage; D013395:Sucrose / Q000008:administration & dosage; D014022:Tissue Survival; D014325:Tromethamine / Q000008:administration & dosage; D014654:Vascular Patency; D014661:Vasoconstriction; D014662:Vasoconstrictor Agents / Q000494:pharmacology; D014665:Vasodilator Agents / Q000494:pharmacology
D003451:Cryoprotective Agents; D016895:Culture Media, Serum-Free; C074097:Krebs-Henseleit solution; D019188:Organ Preservation Solutions; D014662:Vasoconstrictor Agents; D014665:Vasodilator Agents; D014325:Tromethamine; D009599:Nitroprusside; D013395:Sucrose; D015237:Dinoprost; D005947:Glucose; D009584:Nitrogen; D001920:Bradykinin; D009638:Norepinephrine; D004121:Dimethyl Sulfoxide