The ability to patterning cells is an important technique to facilitate cell-based assay and characterization. In this paper, an automated cell patterning system was developed for the fabrication of large-scale cell patterns. To resolve the challenge of the limited printable area, the cell-printing microchip and the substrate were mounted on the movable stages of the system, and large-scale cell patterns were realized through coordination between the stages. An autofocusing technique was integrated in the system to evaluate the gap between the microchip and the substrate. In order to enhance the performance of the patterning system, different experimental parameters, including the velocity of the moving stage, were examined. Yeast cells suspending in 6-aminohexanoic acid (AHA) solution were considered in this study, and a sequence of characters was successfully printed using the proposed system. The results confirm that this system offers an automatic method with high flexibility to construct large-scale cell patterns for various applications.