Construction of Retroviral Vector Carring Insulin Gene and Its Stable Expression in Bone-marrowed Mesenchymal Stem Cells

Objective To construct retroviral vector carring human insulin gene and investigate its expression in human bone-marrowed mesenchymal stem cells(hMSCs).Methods The recombinant retroviral vector INS-pLNCX was constructed by cloning the human insulin cDNA into the retroviral vector pLNCX containing multiple cloning site.The INS-pLNCX vector was transfected into ecotropic packaging cell line PT67 by clonfectin method.Then the supernatant of PT67 was used to infect hMSCs.RT-PCR and Immunofluoresence was used to test the expression of insulin gene in hMSCS.Result The recombinant retroviral vector carring insulin gene was successfully constructed.By transfecting PT67 cell,recombinant retrovirus with infective capability was packaged and used to infect hMSCs.The infected hMSCs can stably express human insulin more than 3 weeks,but can't react to high level of glucose. Conclusion The hMSCs infected by recombinant retroviral vector carring insulin gene can provide a new way to deal with the shortage of β-cell for transplantation.