[Viability and differentiation of human hepatocytes immunoprotected by macroencapsulation and transplanted in rats].
OBJECTIVE
To determine the viability and differentiation of human hepatocytes immunoprotected by encapsulation and transplanted in rats without immunosuppression.


METHODS
Freshly isolated human hepatocytes were encapsulated in hollow fibers and transplanted in the peritoneal cavity of immunocompetent rats. The fibers were explanted for analysis at D3, D7 and D14 following transplantation. Morphological features under light and electron microscopy and gene expression were compared to those of non-transplanted encapsulated hepatocytes (D0). Human cytochrome P450 3A and albumin mRNAs were quantified by Northern blot. Cytochrome P450 3A proteins were detected by Western blot and cytochrome P450 3A enzyme activity was assessed by measuring the formation of 6beta-hydroxytestosterone by high performance liquid chromatography.


RESULTS
Transplanted hepatocytes were more than 60 % viable and exhibited morphological criteria of hepatocytic differentiation up to D7. Albumin and cytochrome P450 3A transcripts were also detected up to D14. At D3 and D7, albumin mRNA levels were of 30 %, compared to control D0 hepatocytes, while cytochrome P450 3A5 and cytochrome P450 3A4 mRNA levels were 65 % and 0 %, respectively. Cytochrome P450 3A immunoreactivity was detected by Western blot up to D14 and 6beta-hydroxylase activity was 17 % at D3 compared to D0, supporting with disappearance of cytochrome P450 3A4 mRNA.


CONCLUSIONS
Human hepatocytes remain viable for a short period, following encapsulation and intraperitoneal transplantation in rat. Other experimental conditions need to be tested to prevent or delay a decrease in hepatocyte specific gene expression.
Nicoluzzi
J E
JE
INSERM U402, Paris, France.
Barbu
V
V
Baudrimont
M
M
Lakehal
F
F
Becquemont
L
L
Chafaï
N
N
Delelo
R
R
Sarkis
R
R
Honiger
J
J
Housset
C
C
Balladur
P
P
D000818:Animals; D001189:Aryl Hydrocarbon Hydroxylases; D015152:Blotting, Northern; D015153:Blotting, Western; D002454:Cell Differentiation / Q000502:physiology*; D002470:Cell Survival; D017690:Cell Transplantation / Q000379:methods*; D051544:Cytochrome P-450 CYP3A; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics; D005786:Gene Expression Regulation / Q000502:physiology; D006801:Humans; D008099:Liver / Q000166:cytology* / Q000648:ultrastructure; D008297:Male; D010089:Oxidoreductases, N-Demethylating / Q000235:genetics; D051381:Rats; D011917:Rats, Inbred Lew; D012709:Serum Albumin / Q000235:genetics; D014021:Tissue Preservation / Q000379:methods*; D014183:Transplantation, Heterologous / Q000379:methods*
D012709:Serum Albumin; D003577:Cytochrome P-450 Enzyme System; D001189:Aryl Hydrocarbon Hydroxylases; D051544:Cytochrome P-450 CYP3A; D010089:Oxidoreductases, N-Demethylating