title: Parenchymal cells proliferate and differentiate in an organotypic slice culture of the neonatal liver.
abstract: 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 alpha-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 constituent cells. Quantitative analysis of albumin-, alpha-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.
authors: Third Department of Internal Medicine, Kyoto Prefectural University of Medicine, Japan. morimori@koto.kpu-m.ac.jp
authors: T
authors: T
authors: Mori
authors: M
authors: M
authors: Iwai
authors: Y
authors: Y
authors: Harada
authors: S
authors: S
authors: Tanaka
authors: A
authors: A
authors: Muramatsu
authors: T
authors: T
authors: Okanoue
authors: K
authors: K
authors: Kashima
authors: S
authors: S
authors: Fushiki
mesh_terms: D000418:Albumins / Q000032:analysis; D000818:Animals; D000831:Animals, Newborn; D015415:Biomarkers / Q000032:analysis; D002454:Cell Differentiation; D002455:Cell Division; D000072259:Connexin 26; D017630:Connexins / Q000032:analysis; D046508:Culture Techniques / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay; D005865:Gestational Age; D007150:Immunohistochemistry; D007633:Keratins / Q000032:analysis; D008099:Liver / Q000166:cytology / Q000196:embryology*; D051379:Mice; D008854:Microscopy, Electron; D018809:Proliferating Cell Nuclear Antigen / Q000032:analysis; D000509:alpha-Fetoproteins / Q000032:analysis; D000097002:Gap Junction beta-1 Protein
chemical_list: D000418:Albumins; D015415:Biomarkers; D000072259:Connexin 26; D017630:Connexins; D007633:Keratins; D018809:Proliferating Cell Nuclear Antigen; D000509:alpha-Fetoproteins; D000097002:Gap Junction beta-1 Protein