Wood-concrete composite slab floors provide a promising solution for achieving long spans and shallow wood-based floor systems for large and tall wood buildings. In comparison with conventional wood floor systems, such long span and heavy floors have a lower fundamental natural frequency, which challenges the floor vibration controlled design. A laboratory study, including subjective evaluation and measurement of the natural frequencies and one-kN static deflections, was conducted on wood-concrete composite floors. Method of calculation of the composite bending stiffness of the wood-concrete composite floor is proposed. The design criterion for human comfort was derived from the subjective evaluation results using the calculated fundamental natural frequency and 1 kN static deflection of one meter wide strip of the composite floor. The equation to directly determine the vibration controlled spans from the stiffness and mass was derived. Limited verification was performed. Further verification is needed when more field wood-concrete composite floors become available.