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(American Journal of Pathology. 2000;156:2033-2043.)
© 2000 American Society for Investigative Pathology

Regular Articles

The in Vitro Differentiating Capacity of Nonparenchymal Epithelial Cells Derived from Adult Porcine Livers

Junko Kano*{dagger}, Masayuki Noguchi*, Makoto Kodama{dagger} and Takayoshi Tokiwa{dagger}{ddagger}

From the Department of Pathology,*
Institute of Basic Medical Sciences, University of Tsukuba, Tsukuba; the Department of 3D Tissue Engineering,{dagger}
National Institute for Advanced Interdisciplinary Research, Tsukuba; and the Department of Cell Physiology,{ddagger}
Kohno Clinical Medicine Research Institute, Tokyo, Japan

Specific nonparenchymal epithelial cell (NPEC) clusters derived from normal adult porcine livers demonstrate a characteristic developmental pattern in the presence of other types of nonparenchymal cells in vitro. This pattern includes scattering, colonial growth, and an emergence of duct-like structures (DLSs) in the colonies. It has been confirmed that 96% of the scattered cell clusters in these cultures develop into colonies containing DLSs. In this study, we examine the differentiation of NPEC clusters using the scattered formation as a marker of the DLS-emerged colonies. We report that the NPECs expressed albumin, {alpha}-fetoprotein, transferrin, cytokeratin (CK) 18, CK7, and c-met, but not {alpha}-1-antitrypsin (AAT), at the scattering stage. In addition, at the same stage, NPECs expressed oval-cell-related markers such as OV6, but not biliary epithelial cell (BEC) markers such as {gamma}-glutamyltransferase, CK19, and CK14. At the DLS emerging stage, hepatocyte markers, including AAT, were detectable in the cells either at the periphery of colonies or in the cells surrounded by the DLSs. On the other hand, the cells constituting DLSs expressed BEC markers, suggesting a bile duct nature of the DLSs. Furthermore, the cells in the colonies possessed an ultrastructural appearance of differentiated hepatocytes and BECs. These results suggest that certain NPECs are bipotent, and that, in culture, they mimic hepatoblast development in vivo.




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