Zhang Xianjie, Sun Jiabang, Song Maomin, Li Fei, Sun Haichen
Department of General Surgery, Xuanwu Hospital, Capital University of Medical Sciences, Beijing 100053, China.
Zhonghua Wai Ke Za Zhi. 2002 Aug;40(8):618-20.
To regain bile excretory function of hepatocytes cultured in vitro.
Sandwich configuration was used to culture hepatocytes and the structure of bile canaliculi as well as the function of bile excretion was observed by immunocytochemistry and the test of FDA metabolism with a single collagen configuration as a control.
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.
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.
恢复体外培养肝细胞的胆汁排泄功能。
采用三明治结构培养肝细胞,以单层胶原结构作为对照,通过免疫细胞化学和FDA代谢试验观察胆小管结构及胆汁排泄功能。
首先,通过免疫细胞化学观察到胆小管的形成。在三明治培养的肝细胞中,观察到胆小管样结构逐渐发展为吻合网络。细胞培养24小时后,观察到胆小管的形成。随着时间推移,胆小管变得更加清晰,在120小时时建立起网络。相比之下,单层胶原结构的肝细胞几乎没有胆小管网。其次,肝细胞摄取、代谢并将化合物排泄到胆汁中的能力通过肝细胞中FDA的代谢来体现。在三明治培养的肝细胞中,96小时时显示出胆汁排泄功能,但在单层胶原系统中未观察到胆汁排泄。
培养的肝细胞在一定条件下能够恢复胆汁排泄功能。三明治培养的肝细胞可重建胆小管结构并恢复胆汁排泄功能。