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

目的：与正常人肝细胞共培养，从正常人肝中分离的人胆道上皮细胞（BEC）在三维培养中形成导管形态。由于离体人肝细胞的寿命较短，长期观察BEG的再生情况较为困难。为了长期观察，我们使用功能性肝癌细胞系FLC4代替正常的人肝细胞。方法：每天用相差显微镜观察培养的BEC和FLC4或正常人肝细胞系（NHL）在BEG和FLC或NEIL之间移液时，有无胶原薄层的形态学变化。结果与结论：无胶原薄层的BEG与FLC4共培养未见小管形态发生。由于FLC4的快速增殖，在6孔大板中FLC4的数量过多。在BEG和FLC或NHL之间移液的胶原薄层似乎对FLC4的增殖有孤立的影响。与胶原薄层共培养的BEG和FLG4或NEIL对胆管形态发生的形态学评价正在进行中。

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.