We applied organotypic slice culture of neonatal mouse liver tissues to maintain the parenchymal cells in ontogenesis and to investigate their proliferation and differentiation. Cultured tissue spread gradually over 3 weeks. Small basophilic cells formed several layers in the center of the cultured tissues, and a monolayer of polygonal cells was seen at the periphery. Albumin- and α-fetoprotein-immunoreactions were seen in polygonal cells, as were proliferating cell nuclear antigen-immunoreactions. Connexin 32- and 26-immunoreactions were observed in small plaques on the membrane of the polygonal cells, and electron microscopy showed gap junctional complexes. Ultrastructurally, polygonal cells had a round nucleus and abundant cytoplasmic organelles, and bile canaliculi were seen on the cytoplasmic membrane. Cytokeratin 19-immunoreactions were scattered in clusters. There were ultrastructurally bile-duct-like structures with microvilli on the inner surface of the cavity and tight junctions between their constitutent cells. Quantitative analysis of albumin-, α-fetoprotein- and cytokeratin 19- or proliferating cell nuclear antigen-immunoreactivity in parenchymal cells showed changes of their phenotypes or maintenance of their proliferation in tissue culture. Our slice-culture system enabled us to maintain and to develop parenchymal cells in the liver tissue for at least 3 weeks. The findings suggest that organotypic slice culture applied to liver tissues in ontogenesis may be a useful tool not only to maintain parenchymal cells but also to investigate their proliferation and differentiation.
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Accepted: 17 September 1998
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Mori, T., Iwai, M., Harada, Y. et al. Parenchymal cells proliferate and differentiate in an organotypic slice culture of the neonatal liver. Anat Embryol 199, 319–327 (1999). https://doi.org/10.1007/s004290050231
- Key words Tissue culture
- Structural organization