Characteristics of small cell colonies developing in primary cultures of adult rat hepatocytes |
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Authors: | Toshihiro Mitaka Gerald L Sattler Henry C Pitot Yohichi Mochizuki |
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Institution: | 1. Department of Pathology, Cancer Research Institute, Sapporo Medical College, Chuo-Ku, S-1, W-17, 060, Sapporo, Japan 2. McArdle Laboratory for Cancer Research, University of Wisconsin, 1400 University Avenue, 53706, Madison, WI, USA
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Abstract: | Phenotypes of the cells developing into small colonies after days of primary culture of adult rat hepatocytes in serum-free
modified Dulbecco Modified Eagles’ medium containing 10 mM nicotinamide and 10 ng/ml epidermal growth factor were analyzed
immunocytochemically, cytochemically and ultrastructurally. Albumin, cytokeratin 8 and 18 were seen by immunocytochemical
techniques in the cells of the small colonies at Day 6. Transferrin, α-antitrypsin, ceruloplasmin, and haptoglobin, proteins
secreted by mature hepatocytes, were faintly stained in these cells as was α-fetoprotein. These proteins were secreted into
the culture medium as evidenced by immunoblot analysis. γ-Glutamyltransferase, alkaline phosphatase and glucose 6-phosphatase
were not present in the cells of the small colonies as well as the surrounding hepatocytes at Day 6 of culture. In addition,
ultrastructural examinations of the cells in the small colonies indicated that these cells not only had many characteristic
mitochondria and desmosomes, but also a few small peroxisomes. Such cells, even after 20 days in culture were proliferating,
as evidenced by the intranuclear presence of the proliferating cell nuclear antigen. The potential relation of these cells
to hepatocytes which may serve as the principal reserve for replicating hepatocytes is discussed. |
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Keywords: | Hepatocytes Primary culture Colony Proliferation Cytochemistry |
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