Immortalization of rat hepatocytes by fusion with hepatoma cells. I. Cloning of a hepatocytoma cell line with bile canaliculi.
Hepatocytoma (HPCT) hybrid cells were obtained by fusion of cultured rat hepatocytes with Fao Reuber hepatoma cells H35 by polyethylene glycol treatment. Surviving cells were cloned in HAT (hypoxanthine-aminopterine-thymidine)/ouabain medium and propagated in cell lines over 80 passages. Morphological criteria were chosen to allow differentiation of the clones into two types of cells: 1) Type I cells which formed irregular cell layers, lacked contact inhibition and resembled the parental Fao hepatoma cells and 2) type II cells, which proliferated in monolayer cultures, exhibited contact inhibition during growth in culture plates and formed bile canaliculi thereby resembling cultured hepatocytes by phenotype. Bile canaliculi were absent in type I clones and Fao cells. One particular type II clone 1E3 was studied in detail. These cells formed bile canaliculi sealed by tight junctions and were comparably polarized as cultured hepatocytes. They expressed canalicular antigen B10, canalicular aminopeptidase N, and even secreted the fluorescent bile acid derivative NBD-cholate into the canalicular lumen. This type of HPCT cells lacked malignancy by tests in vivo and in vitro, and contained 110 +/- 5 chromosomes. The cells were considered to represent an immortalized hepatocyte-like cell line.
Petzinger
E
E
Institut für Pharmakologie und Toxikologie, Justus-Liebig-Universität, Giessen, Germany.
Föllmann
W
W
Blumrich
M
M
Walther
P
P
Hentschel
J
J
Bette
P
P
Maurice
M
M
Feldmann
G
G
D000818:Animals; D001648:Bile Canaliculi / Q000166:cytology*; D002459:Cell Fusion / Q000502:physiology; D002462:Cell Membrane / Q000201:enzymology; D002470:Cell Survival / Q000502:physiology; D002999:Clone Cells / Q000502:physiology; D006822:Hybrid Cells / Q000166:cytology*; D007621:Karyotyping; D008114:Liver Neoplasms, Experimental / Q000473:pathology*; D008297:Male; D051381:Rats; D017208:Rats, Wistar