【摘要】 目的建立大鼠肝脏干细胞的Pdx-1基因表达系。方法逆行穿刺SD大鼠乳鼠肝脏门静脉,经胶原酶Ⅳ消化并分离肝脏干细胞;免疫组织化学鉴定大鼠肝脏干细胞标志物细胞角蛋白18/19、甲胎蛋白的表达;构建pEGFP-Pdx-1表达载体并酶切鉴定,用脂质体2000介导表达载体转染大鼠肝脏干细胞后,荧光显微镜观察GFP表达,免疫组织化学鉴定Pdx-1的表达。结果肝脏门静脉逆行穿刺分离培养大鼠肝脏干细胞具有分裂增殖能力、角蛋白18/19、甲胎蛋白表达阳性;pEGFP-Pdx-1表达载体转染后的肝脏干细胞可表达绿色荧光蛋白报告基因和Pdx-1蛋白。结论成功建立了大鼠肝脏干细胞Pdx-1表达系。更多还原

【Abstract】 Objective To establish a liver stem cell line expressing pancreatic duodenal homeobox(Pdx-1)in rats.Methods Rat liver stem cells were obtained by digestion of collagenase IV and isolation after hepatic portal vein puncture.The liver stem cells were identified by using specific antibodies against cytokeratin 18 and 19 and AFP.The stem cells were transfected with pEGFP-Pdx1 expressing vector using liposome 2000,and Pdx-1 exprssion was identified by observing green fluorescence protein expression under the microscope and by using immunohistological staining.Results Rat liver stem cells isolated and cultured through hepatic portal vein puncture were capable of mitosis and proliferation,and could also express cytokeratin 18 and 19 and AFP,suggesting that they are indeed liver stem cells.Pdx-1 gene transfected cells can express green fluorescent protein reporter gene and Pdx-1 protein.Conclusion A Pdx-1 expressed rat liver stem cell line has been successfully established.更多还原