Fractionation of rat hepatocyte subpopulations with varying metabolic potential, proliferative capacity, and retroviral gene transfer efficiency.
The liver contains hepatocytes with varying ploidy and gene expression. To isolate cells on the basis of ploidy for analyzing mechanisms concerning cell proliferation and differentiation, we used Percoll gradients to separate F344 rat hepatocyte subpopulations. Specific fractions were enriched in polyploid (H2 fraction) or diploid (H3 and H4 fractions) hepatocytes containing glycogen and glucose-6-phosphatase. H4 cells were relatively smaller with greater nuclear/cytoplasmic ratios, less complex cytoplasm, and higher serum albumin or ceruloplasmin biosynthetic rates. H2 fraction cells were larger with lesser nuclear/cytoplasmic ratio, more complex cytoplasm, and more cytochrome P450 activity. Phenotypic marking showed that H4 cells originated in zone one and H2 cells in zones two or three of the liver lobule. H4 cells showed much greater mitogenic responsiveness to human hepatocyte growth factor. Retroviral gene transfer, which requires both viral receptors and cellular DNA synthesis, was significantly more efficient in H4 cells. The findings indicated that small diploid and large polyploid hepatocytes show unique biological differences. The ability to isolate hepatocytes of varying maturity is relevant for mechanisms concerning liver growth control and hepatic gene expression.
Rajvanshi
P
P
Division of Gastroenterology, Marion Bessin Liver Research Center, Bronx, New York, 10461, USA.
Liu
D
D
Ott
M
M
Gagandeep
S
S
Schilsky
M L
ML
Gupta
S
S
D000818:Animals; D002455:Cell Division / Q000187:drug effects / Q000235:genetics; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured; D018014:Gene Transfer Techniques; D017228:Hepatocyte Growth Factor / Q000494:pharmacology; D008099:Liver / Q000166:cytology* / Q000378:metabolism* / Q000821:virology; D008297:Male; D011003:Ploidies; D011205:Povidone; D051381:Rats; D011916:Rats, Inbred F344; D012190:Retroviridae / Q000235:genetics*; D012822:Silicon Dioxide
C016039:Percoll; D017228:Hepatocyte Growth Factor; D012822:Silicon Dioxide; D011205:Povidone