Experimental transplantation of hepatocytes in cases of toxic acute liver failure. An allograft model.
The aim of this experimental study was to modify a rat liver-cell harvesting technique and to evaluate the efficacy of allogeneic liver cell transplantation (Tx) using cyclosporin A immunosuppression in rats with N-dimethylonitrosamine (N-DMNA)-induced acute liver failure (ALF). Twenty male Wistar rats, weighing 190-320 g, were used as donors. Hepatocytes were harvested by the use of a modification of the Seglen portal vein collagenase perfusion technique (type V/1.3 mg/ml), which resulted in the isolation of a mean of 8000 viable clusters of hepatocytes per donor (viability was measured using the trypan blue exclusion test). The male Lewis recipients and controls received 20 mg/kg N-DMNA i.v., and were then divided in three groups. Group 1 (n = 5) received no treatment, group 2 (n = 10) received 5000 clusters of freshly isolated hepatocytes (FIH) in the spleen 24 h after the administration of N-DMNA- and group 3 (n = 10) received 5000 clusters of FIH beneath the renal capsule, 24 h after the administration of N-DMNA. All groups were treated with cyclosporin A 20 mg/kg per day i.p.. SGOT and bilirubin values were measured and all surviving rats were sacrificed on day 14. All rats in group 1 died of histologically confirmed liver necrosis within 72 h. The 14-day survival was 60% in group 2 and 50% in group 3. The post-Tx SGOT values reached their maximum on days 3-4 (group 2, mean 754; group 3, mean 529) and were only slightly elevated on day 14 (group 2 = 75, group 3 = 48). The post-Tx bilirubin values reached their maximum on days 3-5 (group 2 = 1.1, group 3 = 1) but failed to return to normal until day 14. Autopsy and histological examination of the surviving animals showed well-preserved hepatocellular spherical aggregates in the spleen and hepatocellular "cords" in the kidney accompanied by signs of regeneration of the native liver. We concluded that the hepatocyte Tx in a rat experimental allo-Tx model improved the survival rate and the SGOT values in cases of toxic ALF. Survival rates between the two different sites of Tx were similar.
Second Department of Surgery, University of Athens Medical School, Areteion Hospital, Greece.
N
N
Arkadopoulos
A
A
Papalois
T H
TH
Pataryas
B
B
Golematis
J
J
Papadimitriou
D000818:Animals; D001219:Aspartate Aminotransferases / Q000097:blood; D001663:Bilirubin / Q000097:blood; D017690:Cell Transplantation / Q000379:methods*; D016572:Cyclosporine / Q000627:therapeutic use; D004128:Dimethylnitrosamine / Q000633:toxicity; D006085:Graft Survival; D022781:Hepatocytes / Q000166:cytology*; D017114:Liver Failure, Acute / Q000139:chemically induced / Q000628:therapy*; D008297:Male; D051381:Rats; D011917:Rats, Inbred Lew; D017208:Rats, Wistar; D013997:Time Factors; D014184:Transplantation, Homologous / Q000276:immunology / Q000379:methods* / Q000502:physiology
D016572:Cyclosporine; D001219:Aspartate Aminotransferases; D004128:Dimethylnitrosamine; D001663:Bilirubin