In this paper, the performances of natural refrigerant R290/CO2 cascaded refrigeration system using R290 as high temperature cycle refrigerant and CO2 as low temperature cycle refrigerant were investigated experimentally. The test results show that both CO2 compressor and R290 compressor start up stably and present good running performance. The start-up power and start-up electric current of CO2 compressor are higher than that of R290 compressor. The COP (Coefficient of performance) of R290/CO2 cascaded system enhances with the increase of cooling water mass flow rate and the decrease of cooling water inlet temperature. The effect of CO2 refrigerant charge on the COP of cascaded system is obvious; however, the effect of R290 refrigerant charge on the COP of cascaded system is not obvious. Hence, the hermetic scroll type compressor is urgent to be developed and used in CO2 cycle for ensuring run safely and reliably, the value of cooling water mass flow rate and the cooling water inlet temperature must be selected reasonability for saving water resource and energy, as well as the charge amount must be determined by experiment for obtaining the optimum refrigerant charge.