Animal model for liver cell banking from non-heart beating donors after prolonged ischaemia time.
OBJECTIVE
Although there is a growing interest on the use of non-heart beating donors to enlarge the liver donor pool, livers with prolonged warm ischaemia time are not currently considered for organ transplantation. We hypothesised that these organs may represent a source of hepatocytes for cell transplantation and/or use in bioartificial liver devices. Thus, we investigated if prolonged ischaemia could influence the recovery and viability of functional hepatocytes dissociated from rat livers.


METHODS
Hepatocytes were isolated from the liver within 15 min after death (t=15 min) and after 4, 8 and 12h of ischaemia. Cells were either maintained in culture or cryopreserved. In all products, we evaluated cell recovery and viability, hepatocyte markers and cellular functions, including albumin and urea production.


RESULTS
The number of cells per gram of tissue was similar at 15 min, 4 and 8h, while it was significantly decreased at 12h. About 0.2 x 10(6) viable cells expressing hepatocyte markers and producing albumin and urea were isolated up to 8h of ischaemia per gram of tissue.


CONCLUSIONS
Recovery of viable and functional hepatocytes seems possible after prolonged ischaemia time. These data warrant the evaluation of hepatocyte isolation from human livers of non-heart beating donors.
Porretti
L
L
Center of Transfusion Medicine, Cell Therapy and Cryobiology, IRCCS Fondazione Ospedale Policlinico, Mangiagalli e Regina Elena, Via Francesco Sforza 35, 20122 Milan, Italy. l.porretti@policlinico.mi.it
Gatti
S
S
Gramignoli
R
R
Colombo
F
F
Lopa
R
R
Cattaneo
A
A
Scalamogna
M
M
Colombo
G
G
Rossi
G
G
Bonino
F
F
Rebulla
P
P
Prati
D
D
D000818:Animals; D002469:Cell Separation / Q000379:methods; D002470:Cell Survival; D015925:Cryopreservation; D006323:Heart Arrest; D022781:Hepatocytes / Q000637:transplantation*; D007511:Ischemia; D008099:Liver / Q000098:blood supply* / Q000473:pathology; D023421:Models, Animal; D051381:Rats; D017207:Rats, Sprague-Dawley; D013997:Time Factors; D014015:Tissue Banks; D014019:Tissue Donors