Assessment of environmental factors for the ductular morphogenesis of isolated normal human biliary epithelial cells and the development of artificial biliary tract.

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Aims : Cocultured with normal human hepatocytes, human biiary epithelial cells(BEC) isolated from normal human liver formed ductular morphogenesis in three-dimensional culture. Because of the short life of isolated human hepatocytes, it was difficult to observe the regeneration of BEG for a long time. For the long term observation, we used the functional hepatocellular carcioma cell line FLC4 instead of normal human hepatocytes.Methods : The morphological changes in cultured BEC and FLC4 or normal human hepatic cell line(NHL) with and without the collagen thin layer, which is pipetted between BEG and FLC or NEIL, were examined daily by phase-contrast microscopy.Results and Conclusion : No ductular morphogenesis could be detected in the coculture of BEG and FLC4 without collagen thin layer. Because of the rapid proliferation of FLC4, number of FLC4 was too numerous in a 6 well macroplate. The collagen thin layer which was pipetted between BEG and FLC or NHL seemed to have an isolated effect from the proliferation of FLC4. Morphological assessment of biliary ductular morphogenesis cocultured with BEG and FLG4 or NEIL with the collagen thin layer is in progress.