Three-dimensional culture of hepatocytes in a continuously flowing medium |
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Authors: | Kazuyoshi Takeshita William C Bowen George K Michalopoulos |
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Institution: | (1) Department of Pathology, University of Pittsburgh School of Medicine, 15261 Pittsburgh, Pennsylvania |
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Abstract: | Summary Rat hepatocytes were maintained on three-dimensional cultures on sponge discs kept in Spinner Baskets (New Brunswick Scientific
Co., New Brunswick, NJ, USA) with continuously circulating serum-free hepatocyte growth medium (HGM) containing hepatocyte
growth factor (HGF) and epidermal growth factor (EGF). Hepatocytes were embedded in polyester sponge discs with a collagen
gel at the concentration of 5 million cells/ml. Atmospheric gas containing 7% CO2 was directly bubbled into the medium. Agitation by the impeller created a continuous medium-flow through the packed hepatocytes.
Comparison between identically prepared perfused and stationery cultures showed that hepatocytes in the perfused cultures
maintain higher levels of DNA synthesis. These results demonstrate the value of perfusion systems and also show that hepatocytes
can proliferate and maintain differentiation in three-dimensional culture environments. |
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Keywords: | hepatocyte three-dimensional culture proliferation bioartificial liver hepatocyte growth medium (HGM) cytochrome P450 |
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