IMPROVEMENT OF HUMAN ISLET FUNCTION BY ADENOVIRUS MEDIATED HO-1 GENE TRANSFER |
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| Authors: | CHEN Xiao-bo LI Yong-xiang DONG Wei-ping JIAO Yang TAN Jian-ming |
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| Institution: | CHEN Xiao-bo(Department of Urology, The First Clinical Medical College of China Three Gorges University, Yichang 443003, Hubei, China)；LI Yong-xiang(Department of Renal Transplantation and Urology, Shanghai Clinical Medical Center of Organ Transplantation, the First People's Hospital, Shanghai Jiaotong University, Shanghai 200080, China)；DONG Wei-ping(Department of Renal Transplantation and Urology, Shanghai Clinical Medical Center of Organ Transplantation, the First People's Hospital, Shanghai Jiaotong University, Shanghai 200080, China)；JIAO Yang(Department of Renal Transplantation and Urology, Shanghai Clinical Medical Center of Organ Transplantation, the First People's Hospital, Shanghai Jiaotong University, Shanghai 200080, China)；TAN Jian-ming(Department of Renal Transplantation and Urology, Shanghai Clinical Medical Center of Organ Transplantation, the First People's Hospital, Shanghai Jiaotong University, Shanghai 200080, China)； |
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| Abstract: | Objective To investigate in vitro heme oxygenase-1 gene (HO-1) delivery to human pancreatic islets by adenovirus vectors. Methods Recombinant adenovirus containing HO-1 or enhanced green fluorescent protein gene(EGFP) was generated by using the AdEasy System. The purified human pancreatic islets were infected with recombinant adenovirus vectors at various multiplicity of infection (MOI). Transduction was confirmed by fluorescence photographs and Western blot. Glucose-stimulated insulin secretion was detected by using Human insulin radioimmunoassay kits and was used to assess the function of human islets infected by recombinant adenovirus.Results Viral titers of Ad-hHO-1 and Ad-EGFP were 1.96×109 and 1.99×109 pfu/mL, respectively. Human pancreatic islets were efficiently infected by recombinant adenovirus vectors in vitro. Transfection of human islets at an MOI of 20 did not inhibit islet function. Recombinant adenovirus mediated HO-1gene transfer significantly improved the islet function of insulin release when simulated by high level glucose. Conclusion Recombinant adenovirus is efficient to deliver exogenous gene into human pancreatic islets in vitro. HO-1 gene transfection can improve human islet function. |
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