Selective bipotential differentiation of mouse embryonic hepatoblasts in vitro

Am J Pathol. 1997 Feb;150(2):591-602.


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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Biomarkers
  • Butyrates / pharmacology
  • Butyric Acid
  • Cell Differentiation
  • Cell Line
  • Dimethyl Sulfoxide / pharmacology
  • Embryo, Mammalian / cytology
  • Fluorescent Antibody Technique, Indirect
  • Gene Expression
  • Histocytochemistry
  • Immunohistochemistry
  • Keratins / metabolism
  • Liver / cytology*
  • Liver / embryology*
  • Liver / physiology
  • Mice / embryology
  • Microscopy, Electron
  • alpha-Fetoproteins / metabolism
  • gamma-Glutamyltransferase / metabolism


  • Biomarkers
  • Butyrates
  • alpha-Fetoproteins
  • Butyric Acid
  • Keratins
  • gamma-Glutamyltransferase
  • Dimethyl Sulfoxide