Liu Chao, Schreiter Thomas, Dirsch Olaf, Gerken Guido, Oldhafer Karl J, Broelsch Christoph E, Treichel Ulrich
Department of Gastroenterology and Hepatology, University Hospital Essen, Essen, Germany.
Liver Int. 2004 Dec;24(6):669-78. doi: 10.1111/j.1478-3231.2004.0971.x.
Rodent intrahepatic bile duct may harbor bipotential liver progenitor cells. In this study, human-derived intrahepatic biliary epithelial cells (BECs) were investigated in terms of whether they have the character of liver progenitor cells.
Ten liver tissue specimens were obtained after partial hepatectomy or liver explantation. Intrahepatic BECs were isolated by density-gradient centrifugation and immunomagnetic separation using anti-human epithelial antigen and cultured in medium containing epidermal growth factor and hepatocyte growth factor. The isolated and cultured cells were characterized by immunostaining and reverse transcription polymerase chain reaction with a variety of markers for fetal hepatocytes and liver progenitor cells.
These cells had proliferated for up to 18 weeks in vitro. They continuously expressed epithelial markers (cytokeratin (CK) 8 and CK 18) as well as biliary markers (CK 7 and CK 19). Remarkably, some isolated and cultured cells also expressed markers for fetal hepatocytes and liver progenitor cells, including albumin, alpha-fetoprotein, alpha1-antitrypsin, c-kit and chromogranin-A.
Some human-derived intrahepatic BECs coexpressed markers for liver progenitor cells. This finding further supports the hypothesis that the human biliary tree may also consist of liver progenitor cells.
啮齿动物肝内胆管可能含有双潜能肝祖细胞。在本研究中,对人源性肝内胆管上皮细胞(BECs)是否具有肝祖细胞特征进行了研究。
在部分肝切除或肝脏切除术后获取10份肝组织标本。使用抗人上皮抗原通过密度梯度离心和免疫磁珠分离法分离肝内BECs,并在含有表皮生长因子和肝细胞生长因子的培养基中培养。通过免疫染色以及使用多种胎儿肝细胞和肝祖细胞标志物的逆转录聚合酶链反应对分离和培养的细胞进行鉴定。
这些细胞在体外增殖长达18周。它们持续表达上皮标志物(细胞角蛋白(CK)8和CK 18)以及胆管标志物(CK 7和CK 19)。值得注意的是,一些分离和培养的细胞还表达胎儿肝细胞和肝祖细胞的标志物,包括白蛋白、甲胎蛋白、α1-抗胰蛋白酶、c-kit和嗜铬粒蛋白A。
一些人源性肝内BECs共表达肝祖细胞标志物。这一发现进一步支持了人类胆管树也可能由肝祖细胞组成的假说。