Survival and differentiation of porcine hepatocytes encapsulated by semiautomatic device and allotransplanted in large number without immunosuppression.
OBJECTIVE
The aim of this study was to evaluate the survival and functions of porcine hepatocytes transplanted in large quantities in the peritoneal cavity of allogeneic animals following semiautomatic encapsulation.


METHODS
Isolated porcine hepatocytes and a polymer solution composed of AN69 were coextruded through a double lumen spinneret. Minitubes containing hepatocytes were transplanted in the peritoneal cavity of 12 pigs (4 x 10(9) cells/animal) in the absence of immunosuppressive therapy. Seven, 15, and 21 days after transplantation, minitubes was collected and processed for analyses. The morphology was examined under light and electron microscopy. Albumin synthesis was assessed by semi-quantitative reverse transcription-polymerase chain reaction. Cytochrome P450 3A (CYP3A) gene expression was analyzed by Western blot and by testosterone 6-beta-hydroxylation assay.


RESULTS
The device allowed to encapsulate 55 x 10(6) hepatocytes/min. Hepatocytes exhibited normal structural and ultrastructural features up to day 21. Albumin gene expression decreased progressively between days 0 and 21. The amount of CYP3A protein and 6-beta-hydroxylase activity were approximately 2-fold lower at days 7 and 15 than in freshly encapsulated hepatocytes, and further decreased thereafter.


CONCLUSIONS
The preservation of hepatocyte functions during 1-2 weeks is encouraging for potential short-term use of such bioartificial liver in future clinical application.
Benoist
S
S
Research Unit 402, INSERM, Hospital Saint-Antoine, Paris, France.
Sarkis
R
R
Chafaï
N
N
Barbu
V
V
Honiger
J
J
Lakehal
F
F
Becquemont
L
L
Baudrimont
M
M
Capeau
J
J
Housset
C
C
Nordlinger
B
B
D000418:Albumins / Q000235:genetics; D000818:Animals; D001189:Aryl Hydrocarbon Hydroxylases; D002454:Cell Differentiation; D002470:Cell Survival; D017690:Cell Transplantation / Q000295:instrumentation / Q000379:methods*; D051544:Cytochrome P-450 CYP3A; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D015194:Diffusion Chambers, Culture; D004867:Equipment Design; D015870:Gene Expression; D022781:Hepatocytes / Q000166:cytology* / Q000378:metabolism / Q000637:transplantation*; D007165:Immunosuppression Therapy; D008854:Microscopy, Electron; D010089:Oxidoreductases, N-Demethylating / Q000378:metabolism; D013552:Swine; D013997:Time Factors; D014184:Transplantation, Homologous
D000418:Albumins; D003577:Cytochrome P-450 Enzyme System; D001189:Aryl Hydrocarbon Hydroxylases; D051544:Cytochrome P-450 CYP3A; D010089:Oxidoreductases, N-Demethylating