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[Stable expression of human basic fibroblast growth factor gene in retinal pigment epithelial cells by gene transfer
Hong Wu1, Gui-yun Wang, Xiao-guang Zhang
1Department of Ophthalmology, The Second Hospital, Jilin University, Changchun 130041,China. szgnohuw@hotmail.com
Objective:
The purpose of the present study was to explore whether gene transferred retinal pigment epithelial cells (RPE) can constantly express human basic fibroblast growth factor gene (hbFGF) and to study the feasibility of using this procedure for the treatment of retinitis pigmentosa.
Methods:
Plasmid carrying hbFGF [using green fluorescent protein (GFP) as a reporter gene] was constructed and transferred into the RPE cells by lipofectamine. The positively expressed cell clones were selected with G418 and cultured for 4 weeks. Expression of GFP gene was identified by the fluorescence microscope. Expression of hbFGF in the RPE cells was determined by in situ hybridization and immunohistochemical methods.
Results:
Reconstruction enzyme digestion demonstrated that the eukaryotic expression pcFG was constructed correctly. Expression of GFP protein in the gene transferred RPE was detected under the fluorescence microscope. Enzyme digestion and agarose gel electrophoresis analysis showed that the gene transferred RPE expressed the hbFGF gene.
Conclusions:
RPE cells are able to express hbFGF gene stably by gene transfer procedures.