Kinetics of cellular viability in warm versus cold ischemia conditions of kidney preservation. A biometric study.
We have determined the kinetics of the cellular viability ratio (CVR), defined as the number of living cells over the total cell count, in pig kidneys using propidium iodide and fluorescein diacetate staining, as a function of time and preservation conditions. The kidneys were preserved in warm or cold ischemia in order to mimic the conditions of transplantation from non-heart-beating donors or multiple removal with optimal preservation of the graft, respectively. To determine the CVR, the cells were obtained by a fine-needle aspiration biopsy, which minimizes the damage to the graft. A biometric analysis by regression enabled the determination of the time dependence for warm ischemia (CVR(t) = 80.0 x e(-0.733-t)(+2.7/-0.36)) and for cold ischemia (CVR(t) = 80.0 x e(-0.022-t)(+1.57/-0.64)) with a confidence interval of 95%. These master curves allow us to predict, under the described conditions, the CVR after a given ischemia time. The half-life of the cells can be deduced from the time-dependent CVR(t), and is 0.64 hr (38 min) for warm ischemia and 21.4 hr for cold ischemia. Further, the CVR for a given kidney can be used to assess its condition at removal: if the CVR is below 48% at 2 hr after removal, one can conclude that the organ has suffered a period of warm ischemia.
Savioz
D
D
Unité de Transplantation et Service de Chirurgie Digestive, Hôpital Universitaire Cantonal de Gèneve, Switzerland.
Bolle
J F
JF
Graf
J D
JD
Jeanjacquot
A
A
Savioz
M
M
Dietler
G
G
Favre
H
H
Leski
M
M
Morel
D
D
Morel
P
P
D000818:Animals; D001699:Biometry; D001707:Biopsy, Needle; D002452:Cell Count; D002470:Cell Survival; D015925:Cryopreservation; D005452:Fluoresceins; D006207:Half-Life; D006358:Hot Temperature; D007511:Ischemia / Q000473:pathology; D007668:Kidney / Q000166:cytology*; D007700:Kinetics; D011309:Preservation, Biological; D011419:Propidium; D012044:Regression Analysis; D012079:Renal Circulation; D013194:Staining and Labeling; D013552:Swine
D005452:Fluoresceins; D011419:Propidium; C018506:diacetylfluorescein