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

目的：从正常人肝脏分离的人胆管上皮细胞(BEC)与正常人肝细胞共培养，在三维培养中形成管状形态。由于分离的人肝细胞寿命短，很难长时间观察到BEG的再生。为了长期观察，我们用有功能的人肝细胞癌细胞株FLC-4代替正常人肝细胞。方法：用相差显微镜观察BEC和FLC-4以及BEC-FLC和NeIL之间有无胶原薄层的正常人肝细胞系(NHL)的形态变化。结果与结论：在BEC和FLC-4的共同培养中，没有胶原薄层的情况下，未见导管的形态发生。由于FLC4的快速增殖，在一个6孔的宏板中FLC4的数量太多。BEG与FLC或NHL之间的胶原薄层似乎对FLC 4的增殖有单独的作用。胆管形态发生与BEG、Flg4或Neil与胶原薄层共培养的形态学研究正在进行中。

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.