Herein, porous CuO spindle-like nanosheets were fabricated on a carbon cloth using a facile hydrothermal method, and surface morphology, microstructure, and glucose sensing performance were studied. The porous spindle-like nanosheets are constructed by nanoparticles and slit-like pores, exhibiting a hierarchical structure. When used for non-enzymatic glucose sensoring, the obtained CuO nanosheet electrode exhibits a wide linear range from 0.05 to 3.30 mM, a high sensitivity of 785.2 μA mM-1 cm-2 and a low detection limit of 0.22 μM (S/N = 3). Besides, good selectivity, stability, and reproducibility for glucose detection indicate a promising application of CuO nanosheet electrodes as non-enzymatic glucose sensors.