Rat liver stem cells gene expression of pancreatic duodenal homeobox Department of Construction and preliminary identification of.docVIP

 PAGE \* MERGEFORMAT 15 Rat liver stem cells gene expression of pancreatic duodenal homeobox Department of Construction and preliminary identification of Of: Ren Wu Chao, Li Dongmin, Wang Xuan, Gao, Han Yan, Ning Qilan, Zhang Fujun, Song Tianbao, Lu Shemin [Abstract] Objective To establish a rat liver stem cells, Pdx 1 gene expression system. Methods SD retrograde puncture the portal vein of rat liver, isolated by collagenase digestion and liver stem cells, immunohistochemical identification of rat liver stem cell marker cytokeratin 18/19, AFP expression construct pEGFP Pdx 1 expression vector and restriction enzyme digestion, using Lipofectamine 2000 mediated transfection of expression vector of rat liver stem cells, the expression of GFP fluorescence microscopy, immunohistochemistry, identification of Pdx 1 expression. Results of liver puncture the portal vein retrograde isolated rat liver stem cells have the ability to proliferate, keratin 18/19, AFP expression was positive, pEGFP Pdx 1 expression vector transfected liver stem cells expressing green fluorescent protein reporter gene and Pdx 1 protein. Conclusions The establishment of rat liver stem cell Pdx 1 expression system. [Keywords:] in rats, liver stem cells, pancreatic duodenal homeobox, expression vector 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 p