With the application of Cu-Cu direct bonding technology in high-performance electronic devices, improving bonding reliability is essential for achieving high-density 3D IC integration. In this study, we innovatively applied highly (110)-oriented perpendicular nanotwinned Cu (p-ntCu) to achieve Cu-Cu direct bonding at low temperature and pressure. The anisotropic growth of p-ntCu during annealing at 200 °C resulted in the formation of ultra-large grains, which could improve the conductivity of the electrodeposited Cu film. Two Cu films were bonded at 200–250 °C with 2 MPa pressure for 1 h, and there was almost no void at the bonding interface. Anisotropic grain growth occurred within the bonding joint and grain boundary migration was observed at the bonding interface. The shear strength after bonding at 250 °C was measured as 58.3 MPa on average, indicating a relatively high quality of such Cu-Cu bonding. The p-ntCu can achieve Cu-Cu bonding and improve the conductivity by anisotropic grain growth at low temperature and pressure, which has great potential for Cu interconnect applications.