Abstract: Objective    To select a GLUT4HA-overexpressing C2C12 cell line with contractile activity. Methods    C2C12-GLUT4HA cell line stably overexpressing GLUT4HA was created and 23 of these clones were selected. The differentiation and contraction ability, as well as the expression level of GLUT4HA protein were detected.The clones with good myotubes ,contraction ability and high GLUT4HA protein level were divided into insulin group and its control, electrical pulse stimulation(EPS) group and its control,the amount of GLUT4HA on the cell surface was measured by enzyme-linked immunosorbent assay (ELISA) after stimulation by insulin and EPS. Results   Among the selected 23 clones, clone 17 has higher GLUT4HA protein level, good myotubes and contraction ability. In addition, GLUT4HA in clone 17 myotubes can translocate to cell surface under insulin and EPS stimulation. Conclusion     A contractile C2C12 muscle cell line stably overexpressing GLUT4HA was created. This cell line will be useful to study the mechanism of insulin- and contraction stimuli-regulated GLUT4 traffic.

Key words: muscle, skeletal, glucose transporter type 4, Insulin, muscle contraction, electrical pulse stimulation, 电脉冲刺激