Yamazaki Taisuke, Enosawa Shin, Kasahara Mureo, Fukuda Akinari, Sakamoto Seisuke, Shigeta Takanobu, Nakazawa Atsuko, Tokiwa Takayoshi
Department of Liver Cell Biology, Kohno Clinical Medicine Research Institute , Shinagawa-ku, Tokyo , Japan.
† Division for Advanced Medical Science, National Center for Child Health and Development , Tokyo , Japan.
Cell Med. 2012 Jun 15;3(1-3):121-126. doi: 10.3727/215517912X639441. eCollection 2012 Jan.
We sought to determine whether hepatic progenitor cells can be isolated from cirrhotic liver using epithelial cell adhesion molecule (EpCAM) or Thy-1 markers. Liver tissue with cirrhosis secondary to biliary atresia (BA) was collagenase digested, and nonparenchymal cells (NPCs) were cultivated for 24 h. Noncirrhotic NPCs derived from patients with carbamyl phosphate synthetase and ornithine transcarbamylase deficiencies were used as controls. Flow cytometric analysis demonstrated that the percentages of EpCAM- and Thy-1-positive cells were significantly higher in NPC populations derived from BA liver than in those derived from control liver. Reverse transcription polymerase chain reaction analysis revealed that EpCAM-positive sorted cells expressed EpCAM, Thy-1, albumin, and CK-19, whereas Thy-1-positive sorted cells expressed Thy-1, albumin, and CK-19. These findings indicate that EpCAM- or Thy-1-positive hepatic progenitor cells can be more efficiently isolated from BA liver than from control liver and suggest that the properties of EpCAM-positive cells are somewhat different from those of Thy-1-positive cells.
我们试图确定是否可以使用上皮细胞粘附分子(EpCAM)或Thy-1标志物从肝硬化肝脏中分离出肝祖细胞。对继发于胆道闭锁(BA)的肝硬化肝组织进行胶原酶消化,并将非实质细胞(NPCs)培养24小时。将来自氨基甲酰磷酸合成酶和鸟氨酸转氨甲酰酶缺乏症患者的非肝硬化NPCs用作对照。流式细胞术分析表明,来自BA肝脏的NPC群体中EpCAM和Thy-1阳性细胞的百分比显著高于来自对照肝脏的NPC群体。逆转录聚合酶链反应分析显示,EpCAM阳性分选细胞表达EpCAM、Thy-1、白蛋白和CK-19,而Thy-1阳性分选细胞表达Thy-1、白蛋白和CK-19。这些发现表明,与对照肝脏相比,从BA肝脏中能更有效地分离出EpCAM或Thy-1阳性肝祖细胞,并且表明EpCAM阳性细胞的特性与Thy-1阳性细胞的特性略有不同。