Purpose:This study was conducted to analyze the air flow characteristics in a plant factory with different inlet and outlet locations using computational fluid dynamics (CFD). Methods: In this study, the flow was assumed to be a steady-state, incompressible, and three-dimensional turbulent flow. A realizable k-ε turbulent model was applied to show more reasonable results than the standard model. A CFD software was used to perform the numerical simulation. For validation of the simulation model, a prototype plant factory (5,900 mmⅹ2,800 mmⅹ2,400 mm) was constructed with two inlets (Φ250 mm) and one outlet (710 mmⅹ290 mm), located on the top side wall. For the simulation model, the average air current speed at the inlet was 5.11 m·s -1 . Five cases were simulated to predict the airflow pattern in the plant factory with different inlet and outlet locations. Results: The root mean square error of measured and simulated air current speeds was 13%. The error was attributed to the assumptions applied to mathematical modelling and to the magnitude of the air current speed measured at the inlet. However, the measured and predicted airflow distributions of the plant factory exhibited similar patterns. When the inlets were located at the center of the side wall, the average air current speed in the plant factory was increased but the spatial uniformity was lowered. In contrast, if the inlets were located on the ceiling, the average air current speed was lowered but the uniformity was improved. Conclusions: Based on the results of this study, it was concluded that the airflow pattern in the plant factory with multilayer cultivation shelves was greatly affected by the locations of the inlet and the outlet.