Selective bipotential differentiation of mouse embryonic hepatoblasts in vitro.
A line of hepatic endoderm cells, hepatoblast cell line 3 (HBC-3), was derived from the liver diverticulum of the mouse on day 9.5 of gestation by culture on a mitomycin C treated STON+ feeder layer in a hepatoblast culture medium consisting of Dulbecco's modified Eagle's medium, nonessential amino acids, fetal calf serum, and beta-mercaptoethanol. This line, HBC-3, stains positively for alpha-fetoprotein, albumin, and cytokeratin 14 (CK-14), protein markers expressed by the embryonic liver diverticulum, indicating that HBC-3 cells retain an undifferentiated hepatoblast phenotype. HBC-3 cells acquire hepatocyte-like ultrastructural characteristics, including bile canaliculi, peroxisomes, and glycogen granules, when maintained in culture for 3 weeks without passage. Treatment with dimethylsulfoxide or sodium butyrate induces a rapid hepatocytic differentiation. The cells cease to express alpha-fetoprotein and CK-14, maintain albumin expression, and become positive for glucose-6-phosphatase activity (a profile consistent with differentiation along the hepatocyte lineage). On Matrigel, HBC-3 cells form elaborate ductular structures, which are positive for gamma-glutamyl transpeptidase and CK-14 and CK-19 and do not express detectable amounts of albumin, a phenotypic change consistent with differentiation along the bile ductular lineage. Thus, HBC-3 cells behave in culture as bipotential hepatoblasts and provide a model system to identify factors that regulate bipotential differentiation in the liver.
Rogler
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Marion Bession Liver Center, Department of Medicine, Albert Einstein College of Medicine, Bronx, NY 10461, USA.
D000818:Animals; D015415:Biomarkers; D002087:Butyrates / Q000494:pharmacology; D020148:Butyric Acid; D002454:Cell Differentiation; D002460:Cell Line; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D004622:Embryo, Mammalian / Q000166:cytology; D019084:Fluorescent Antibody Technique, Indirect; D015870:Gene Expression; D006651:Histocytochemistry; D007150:Immunohistochemistry; D007633:Keratins / Q000378:metabolism; D008099:Liver / Q000166:cytology* / Q000196:embryology* / Q000502:physiology; D051379:Mice / Q000196:embryology; D008854:Microscopy, Electron; D000509:alpha-Fetoproteins / Q000378:metabolism; D005723:gamma-Glutamyltransferase / Q000378:metabolism
D015415:Biomarkers; D002087:Butyrates; D000509:alpha-Fetoproteins; D020148:Butyric Acid; D007633:Keratins; D005723:gamma-Glutamyltransferase; D004121:Dimethyl Sulfoxide