Abstract
Background: To develop somatic gene therapy model for diabetes mellitus, it is the most important to control it by glucose concentration. In order to develop the myoblasts that produce insulin by glucose concentration, the transfection of genes of human insulin, rat glucokinase and rat GLUT2 was conducted using C2C12, the murine myoblast cell line. Methods: pMLC-hINSmut plasmid vector to which human insulin cDNA was inserted in C2C12 cell line, pCB7/GLUT2 and pCB7/GK to which GLUT2 and glucokinase were inserted. Based on the inserted gene, C2C12/INS-GLUT2, C2C12/INS-GK and C2C12/INS-GK-GLUT2 were prepared. In each cell line, its mRNA and protein expression were measured. Also, the capability of producing insulin in low glucose (2.7 mM) and high glucose (25 mM) were compared. Results: 1. It was observed that C2C12/INS-GLUT2, C2C12/INS-GK, C2C12/INS-GK-GLUT2 cell line expressed mRNA and protein of transfected genes, respectively. 2. As for the insulin production depending on the glucose concentration in C2C12/INS, it slightly increased from 0.049 ± 0.003 μU/10 6 cells/hr to 0.197 ± 0.022 μU/10 6 cells/hr. However, in C2C12/GK-GLUT2-INS, it showed the most evident increase: from 0.251 ± 0.074 μU/10
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