Functional assessment of cryopreserved human femoral arteries for pharmaceutical research.
An established method for cryopreservation that might preserve the vascular and endothelial responses of human femoral arteries (HFAs) to be transplanted as allografts was studied. HFAs were harvested from multiorgan donors and stored at 4 degrees C in saline solution before cryostorage. Thirty HFA rings were isolated and randomly assigned to one control group of unfrozen HFAs (eight rings) and one group of cryopreserved HFAs (22 rings). Cryopreservation was performed in RPMI solution containing dimethylsulfoxide (DMSO) and the rate of cooling was -1 degrees C/min until -40 degrees C and faster rates until -150 degrees C was reached. The contractile and relaxant responses of unfrozen and frozen/thawed arteries were assessed in organ bath by measurement of isometric force generated by the HFAs. After thawing, the maximal contractile responses to the contracting agonist tested (noradrenaline) were in the range of 43% of the responses in unfrozen HFAs. The endothelium-independent responses to sodium nitroprusside were not altered whereas the endothelium-dependent relaxant responses to acetylcholine were weakly altered. The cryopreservation method used provided a limited preservation of contractility of HFAs, a good preservation of the endothelium-independent relaxant responses, and a good preservation of endothelium-dependent relaxation. It is possible that further refinements of the cryopreservation protocol, such as a slower rate of cooling and a more controlled stepwise addition of DMSO, might allow better post-thaw functional recovery.
Rendal Vázquez
Maria Esther
ME
Cryobiology Unit, Complejo Hospitalario Juan Canalejo, La Coruña, Spain. Esther_Rendal@canalejo.cesga.es
Rodríguez Cabarcos
M
M
Martínez Santos
M V
MV
Fernández Mallo
R O
RO
Sánchez Ibáñez
J
J
Segura Iglesias
R
R
Matheu Capó
G
G
Filgueira Fernández
P
P
Pértega Díaz
S
S
Bermúdez González
T
T
Andión Núñez
C
C
D000109:Acetylcholine / Q000494:pharmacology; D015925:Cryopreservation; D003451:Cryoprotective Agents; D004121:Dimethyl Sulfoxide; D004730:Endothelium, Vascular / Q000187:drug effects / Q000502:physiology; D005260:Female; D005263:Femoral Artery / Q000187:drug effects / Q000502:physiology; D006801:Humans; D066298:In Vitro Techniques; D008297:Male; D008875:Middle Aged; D009638:Norepinephrine / Q000494:pharmacology; D014021:Tissue Preservation; D014661:Vasoconstriction / Q000187:drug effects; D014664:Vasodilation / Q000187:drug effects
D003451:Cryoprotective Agents; D000109:Acetylcholine; D009638:Norepinephrine; D004121:Dimethyl Sulfoxide