To investigate high cycle fatigue properties of Ti6Al4V alloys, Ti6Al4V alloy specimens were manufactured by laser powder bed fusion (LPBF) at different laser powers, and the Ti6Al4V alloys prepared at the optimal process parameters were heat treated. The surface morphology, the microstructure evolution and the static tensile and dynamic fatigue properties of heat-treated Ti6Al4V alloy specimens are investigated. The surface of as-fabricated Ti6Al4V specimens prepared at a laser power of 300 W appears smooth without apparent pores and cracks. The phases of as-fabricated Ti6Al4V specimens change from the needle-like α′ martensite to a coarsened α+β lamellar mixture upon heat treatment. The fatigue strength of heat-treated Ti6Al4V alloys reaches a value of 567.5 MPa at a stress ratio of 0.06 and 107 cycles. The propagation rate da/dN of fatigue crack and the stress intensity factor ΔK show a linear relationship in the stable propagation region.