In this work, microstructure evolution of dynamic recrystallization (DRX) behavior in Cr12MoV die steel was investigated via experiments and simulations systematically. Firstly, hot compression tests were performed to obtain the true stress–strain curves. Based on the experimental results, the flow stress model was established, and Avrami equation was developed to model the kinetics of DRX. Then, the cellular automaton (CA) model was established to describe DRX behavior. In order to obtain more accurate simulation results, a microstructure enhancement, extraction and conversion program based on fingerprint image enhancement algorithm was developed to generate real initial microstructure which could be directly used in CA simulation. With real initial microstructure generation, good agreement between simulated and experimental results was achieved, indicating the high accuracy of the established CA model. Finally, the CA model was employed to investigate the hot deformation behavior of Cr12MoV die steel under multiple thermomechanical conditions. It could be found that a lower strain rate was beneficial to the occurrence of DRX. When the strain rate was beyond 1.0 s−1, the DRX fraction was very small. This work would provide a significant guidance to optimize the hot working process of Cr12MoV die steel or some other similar steels.