体外培养肝细胞胆汁分泌功能的研究

Bile excretion of hepatocytes cultured in vitro

目的 使体外培养肝细胞获得胆汁分泌功能。 方法 利用三明治构型培养成SD大鼠肝细胞并观察其胆小管结构及其胆汁分泌功能的形成 ,并以单层胶原培养肝细胞作对照。 结果免疫细胞化学显示三明治构型肝细胞培养 2 4h后 ,胆小管结构初步形成 ,随着培养时间延长 ,胆小管结构更加清晰 ,12 0h后形成胆小管网络。而在对照组肝细胞 ,胆小管结构形成后 ,随着培养时间延长逐渐模糊 ,12 0h后胆小管结构消失 ,没有形成胆小管网络。其次 ,荧光二乙酯的肝细胞代谢显示 ,三明治构型肝细胞培养 96h后即有胆汁分泌功能 ,而在对照组 ,肝细胞没有胆汁分泌功能。 结论三明治构型培养肝细胞在体外重建体内胆小管结构 。

Objective To regain bile excretory function of hepatocytes cultured in vitro. Methods Sandwich configuration was used to culture hepatocytes and the structure of bile canaliculi as well as the function of bile excretion was observed by immnocytochemistry and the test of FDA metabolism with a single collagen configuration as a control. Results First, the formation of bile canaliculi was observed by immunocytochemistry. In sandwiched hepatocytes, the gradual development of bile canaliculi-like structures into an anastomotic network was observed. At 24 h after cell culture, the formation of bile canaliculi was observed. With time progressed, the bile canaliculi became more clear and the network was established at 120 h. In contrast, hepatocytes in single collagen configuration showed almost no network of bile canaliculi. Second, the ability for hepatocytes to internalize, metabolize and excrete compounds into bile was indicated by FDA metabolized in the hepatocytes. In sandwiched hepatocytes, the bile excretory function was shown at 96 h, but in single collagen system, no bile excretion was observed. Conclusion Cultured hepatocytes are able to regain bile excretion in a given certain condition. Sandwich-cultured hepatocytes can reestablish bile canalicular structure and regain bile excretory function.