TY - JOUR
T1 - Ductular morphogenesis and functional polarization of normal human biliary epithelial cells in three-dimensional culture
AU - Ishida, Y
AU - Smith, S
AU - Wallace, L
AU - Sadamoto, T
AU - Okamoto, M
AU - Auth, M
AU - Strazzabosco, M
AU - Fabris, L
AU - Medina, J
AU - Prieto, J
AU - Strain, Alastair
AU - Neuberger, James
AU - Joplin, Ruth
PY - 2001/7/1
Y1 - 2001/7/1
N2 - BACKGROUND/AIMS: 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.
AB - BACKGROUND/AIMS: 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.
KW - hepatocyte growth factor
KW - collagen gel
KW - biliary epithelial cells
KW - ductular morphogenesis
UR - http://www.scopus.com/inward/record.url?scp=0034947191&partnerID=8YFLogxK
U2 - 10.1016/S0168-8278(01)00078-2
DO - 10.1016/S0168-8278(01)00078-2
M3 - Article
C2 - 11495037
VL - 35
SP - 2
EP - 9
JO - Journal of Hepatology
JF - Journal of Hepatology
IS - 1
ER -