Prediction of stable cutting regions is a critical requirement for high-speed milling operations. These predictions are generally made using frequency-response measurements of the tool-holder-spindle set obtained from a nonrotating spindle. However, significant changes in system dynamics occur during high-speed rotation. In this paper, a dynamic high-speed spindle-bearing system model is elaborated on the basis of rotor dynamics prediction and readjusted on the basis of experimental modal identification. The dependency of dynamic behavior on speed range is then investigated and determined with accuracy. Dedicated experiments are carried out in order to confirm model results. They show that dynamic effects due to high rotational speed and elastic deformations, such as gyroscopic coupling and spin softening, have a significant influence on spindle behavior. By integrating the modeled speed-dependent spindle transfer function in the chatter vibration stability approach of Altintas and Budak (1995, CIRPS Ann, 44(1), pp. 357–362), a new dynamic stability lobe diagram is predicted. Significant changes are observed in the stability limits constructed using the proposed approach and allow accurate prediction of cutting conditions to be established. Finally, optimization studies are performed on spindle design parameters in order to obtain a chatter vibration-free cutting operation at the desired speed and depth of cut for a given cutter.