Ductular morphogenesis and functional polarization of normal human biliary epithelial cells in three-dimensional culture.
OBJECTIVE
The understanding of the physiology and function of human biliary epithelial cells (hBEC) has been improved by studies in monolayer culture systems. The aim was to develop a polarized model to elucidate the mechanisms of ductular morphogenesis and functional differentiation of hBEC.


METHODS
The morphological, phenotypic and functional properties of hBEC cultured as three-dimensional aggregates in collagen gel were assessed in medium supplemented with (or without) human hepatocyte growth factor (hHGF) and foetal bovine serum.


RESULTS
In the absence of added mitogens and serum, cells maintained as morphologically polarized aggregates, organized around a central lumen, were positive for phenotypic markers of biliary epithelium and negative for markers of other cell types. Functional markers, gamma-glutamyl-transferase, anion exchanger-2, responses to gamma interferon and forskolin induced secretion, were preserved. hHGF increased both the size and number of aggregates and induced hBEC to invade the gel and lumena forming anastomosing networks of cells.


CONCLUSIONS
Collagen gel culture in the absence of added growth factors and serum provides a model for analysis of the polarized functions of hBEC. The formation of poorly organized cords of cells in response to hHGF suggests that collagen gel culture may provide a model for the investigation of atypical ductular morphogenesis of the human biliary tract.
Ishida
Y
Y
Liver Unit, University Hospital, Edgbaston, Birmingham, UK.
Smith
S
S
Wallace
L
L
Sadamoto
T
T
Okamoto
M
M
Auth
M
M
Strazzabosco
M
M
Fabris
L
L
Medina
J
J
Prieto
J
J
Strain
A
A
Neuberger
J
J
Joplin
R
R
D000328:Adult; D000818:Animals; D001652:Bile Ducts / Q000166:cytology / Q000187:drug effects / Q000254:growth & development* / Q000502:physiology*; D002417:Cattle / Q000097:blood; D002455:Cell Division / Q000502:physiology; D016764:Cell Polarity / Q000502:physiology*; D002478:Cells, Cultured; D018920:Coculture Techniques; D003094:Collagen; D003584:Cytological Techniques; D004847:Epithelial Cells / Q000187:drug effects / Q000502:physiology; D005312:Fetal Blood; D005782:Gels; D017228:Hepatocyte Growth Factor / Q000494:pharmacology; D022781:Hepatocytes / Q000502:physiology; D006801:Humans; D010641:Phenotype; D012016:Reference Values
D005782:Gels; D017228:Hepatocyte Growth Factor; D003094:Collagen