Cell culture is the basis of cell research. In order to realize long-term dynamic culture and real-time observation of cells in a microfluidic chip, a microfluidic cell dynamic culture device was designed and fabricated. First, based on the "split" arrangement idea, a three-dimension model of the culture box, control box, and pump box were set up using the software, SolidWorks; hardware and software were designed taking the functional demands into consideration. Second, based on the Comsol software platform, the method of Joule heating was used to analyze the thermodynamic characteristics of the device's heater, namely, ITO glass; the temperature distribution was then obtained from the simulation results. Third, the entire device model was established and its stability was tested. Finally, cell culture experiments were performed. The split design can effectively reduce the volume of the incubator and achieve compatibility between the incubator and the microscopic observation system. In addition, the temperature and humidity of the device are stable, cell growth is in the "S" trend, and the cell survival rate is more than 95%. The split-body cell dynamic culture device offers a satisfactory environment condition for cell culture, in which cells can be observed in real time. As such, the device can satisfy relevant design requirements.