IGF-1 gene transfected the bone marrow stem cells and its transient expressing

Objective:to transfect pIRES2-EGFP-IGF-1 into the bone marrow stem cells (MSCs) and detect its transient expression in MSCs. Methods:the target genes in the pcDNA3.1(+)-IGF-1 were subcloned into pIRES2-EGFP expressing vectors. pIRES2-EGFP-IGF-1 was transfected into MSCs by lipofectamine. After transfection of 48h,the Insulin-like growth factor I (IGF-1) expression in the transfected MSCs were detected under fluorescence microscope and by immunohistochemistry. The supernatant of the transfected MSCs cultured with L929 fibro- blasts. The effect of the recombinant IGF-1 on the L929's cell cycle was observed by flow cytometer analysis. Results:Restrictive enzyme(Nhe I,EcoR I) digestion analysis showed that recombination expression vector pIRES2-EGFP-IGF-1 has been constructed successfully. The transfected MSCs displaying green fluorescence were observed under fluorescence microscope. The immunohistochemical staining showed that positive reactant of brown-yellow could be seen in the cytoplasm of MSCs transfected by pIRES2-EGFP-IGF-1. The supernatant of the transfected MSCs can increase the L929's proliferation. Conclusion: the recombinant expression vector pIRES2-EGFP-IGF-1 is transfected into the MSCs successfully and the transfected MSCs can express IGF-1 obviously.