Parenchymal cells proliferate and differentiate in an organotypic slice culture of the neonatal liver

• Published in: Anatomy and Embryology 1999-04, Vol.199 (4), p.319-327
• Main Authors: Mori, T, Iwai, M, Harada, Y, Tanaka, S, Muramatsu, A, Okanoue, T, Kashima, K, Fushiki, S
• Format: Article
• Language: English
• Subjects: Albumins - analysis; alpha-Fetoproteins - analysis; Animals; Animals, Newborn; Biomarkers - analysis; Cell Differentiation; Cell Division; Connexin 26; Connexins - analysis; Culture Techniques - methods; Enzyme-Linked Immunosorbent Assay; Gap Junction beta-1 Protein; Gestational Age; Immunohistochemistry; Keratins - analysis; Liver; Liver - cytology; Liver - embryology; Mice; Microscopy, Electron; Proliferating Cell Nuclear Antigen - analysis; Proteins; Rodents
• ISSN: 0340-2061; 1863-2653; 0340-2061
• DOI: 10.1007/s004290050231

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.