Instantaneous flow field and temperature field of the two-phase fluid are measured by particle image velocimetry (PIV) and steady state method during the state of onflow. A turbulent two-phase fluid model of stirred bioreactor with punched impeller is established by the computational fluid dynamics (CFD), using a rotating coordinate system and sliding mesh to describe the relative motion between impeller and baffles. The simulation and experiment results of flow and temperature field prove their warps are less than 10% and the mathematic model can well simulate the fields, which will also provide the study on optimized-design and scale-up of bioreactors with reference value.