Improved survival of fulminant liver failure by transplantation of microencapsulated cryopreserved porcine hepatocytes in mice.
The aim of this study was to establish hepatocyte isolation in pigs, and to evaluate function of isolated hepatocytes after encapsulation, cryopreservation, and transplantation (Tx) in a mouse model of fulminant liver failure (FLF). After isolation, porcine hepatocytes were microencapsulated with alginate-poly-L-Lysine-alginate membranes and cryopreserved. In vitro, albumin production of free and encapsulated hepatocytes were measured by enzyme linked-immunoadsorbent assay. In vivo, encapsulated hepatocytes were transplanted into different groups of mice with FLF and the following experimental groups were performed: group 1, Tx of empty capsules; group 2, Tx of free primary porcine hepatocytes; group 3, Tx of fresh encapsulated porcine hepatocytes; group 4, Tx of cryopreserved encapsulated porcine hepatocytes. In vitro, fresh or cryopreserved encapsulated porcine hepatocytes showed a continuous decreasing metabolic function over 1 week (albumin and urea synthesis, drug catabolism). In vivo, groups 1 and 2 showed similar survival (18% and 25%, respectively, p > 0.05). In groups 3 and 4, Tx of fresh or cryopreserved encapsulated porcine hepatocytes significantly increased survival rate to 75% and 68%, respectively (p < 0.05). Primary porcine hepatocytes maintained metabolic functions after encapsulation and cryopreservation. In mice with FLF, Tx of encapsulated xenogeneic hepatocytes significantly improved survival. These results indicate that porcine hepatocytes can successfully be isolated, encapsulated, stored using cryopreservation, and transplanted into xenogeneic recipients with liver failure and sustain liver metabolic functions.
Mei
Jie
J
Surgical Research Unit, Department of Surgery, University Hospital Geneva, Geneva, Switzerland.
Sgroi
Antonino
A
Mai
Gang
G
Baertschiger
Reto
R
Gonelle-Gispert
Carmen
C
Serre-Beinier
Véronique
V
Morel
Philippe
P
Bühler
Leo H
LH
D000818:Animals; D002478:Cells, Cultured; D015925:Cryopreservation / Q000379:methods*; D004339:Drug Compounding; D006085:Graft Survival; D022781:Hepatocytes / Q000378:metabolism / Q000637:transplantation*; D017114:Liver Failure, Acute / Q000378:metabolism / Q000628:therapy*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D046529:Microscopy, Electron, Transmission; D013552:Swine; D014183:Transplantation, Heterologous / Q000379:methods*