Immunohistochemical analyses of cell-cell interactions during hepatic organoid formation from fetal mouse liver cells cultured in vitro.
Cell-cell interactions among cell types constituting the fetal liver such as hepatoblasts, stellate cells and endothelial cells lead to functional lobule development. The present study was undertaken to investigate hepatic histogenesis in the primary culture of E12.5 mouse livers, including cell-cell and cell-matrix interactions. Fetal livers were dispersed with protease treatment and cultured for 5 days. Cellular adhesion of each hepatic cell type, gene expression and extracellular matrix deposition were analyzed by immunohistochemistry and immunoblotting. Immunohistochemical analysis demonstrated that the primary culture of fetal liver cells contained at least hepatoblasts, mesenchymal cells, endothelial cells, hemopoietic cells and Kupffer cells. Although hepatoblasts, mesenchymal cells, and endothelial cells aggregated separately in the initial step, they then formed a spheroid together, adhering to the glass slide, which led to the formation of flattened hepatic organoids. Hepatoblasts more preferentially adhered to mesenchymal cells than endothelial cells. Several extracellular matrix depositions were seen in aggregates consisting of at least hepatoblasts and mesenchymal cells within 12 h, but were poor in those lacking hepatoblasts. These data show that the primary culture of fetal liver cells contains most cell types constituting fetal livers, and may be useful for studying cell-cell interactions during liver development.
Sugiyama
Yoshinori
Y
Department of Biology, Faculty of Science, Shizuoka University, Oya 836, Suruga-ku, Shizuoka City, Shizuoka, 422-8529, Japan.
Koike
Toru
T
Shiojiri
Nobuyoshi
N
D000818:Animals; D002449:Cell Aggregation; D002450:Cell Communication / Q000502:physiology*; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D047108:Embryonic Development; D005333:Fetus; D015151:Immunoblotting; D007150:Immunohistochemistry / Q000379:methods; D008099:Liver / Q000166:cytology* / Q000196:embryology*; D051379:Mice; D008809:Mice, Inbred C3H; D014746:Vimentin / Q000032:analysis
D014746:Vimentin