2002•Zhonghua shiyan waike zazhiRequires access
Construction of human hepatocyte model transfected with hepatocyte growth factor
Yong Ping He
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Abstract
Objective To investigate hepatocyte growth factor (HGF) gene transfer and expression in human hepatocyte with LipofectAMINE in vitro.Methods HGF gene was transfected and expressed in human hepatocyte with liposome.The expression of HGF gene was detected by fluorescence microscopy,immunohistochemistry and hybridization in situ.Results After G418 selection,hepatocyte clones were obtained successfully.The positive hepatocytes of in situ hybridization of specific stained Neo probe was observed in hepatocytes transferred eukaryocyte expression vector.Under a fluorescent microscope,the expression of green fluorescent protein (GFP) was seen in hepatocytes transferred vector containing GFP gene.Positive hepatocytes of HGF immunohistochemistry stain was only observed in hepatocytes transferred HGF gene.Conclusion Hepatocyte model transferred HGF gene was successfully constructed.pEGFP HGF expression vector makes it easy to assess the expression of HGF GFP fusion protein and the protein localization on transfected hepatocytes.