Cell patterning technology is an effective technique to understand cell behaviors by precisely controlling the position of cells revealing the major contributions of cells in basic and applied biological research in vitro. Therefore, this technology has led to the development of various micro-nano manufacturing methods. In recent years, various methods have been continuously improved and innovated based on existing methods to overcome shortcomings. We outline the latest developments in cell patterning technology, including stencil patterning, microfluidics-based on traps and droplets, and surface modification techniques such as photolithography, microcontact printing, and laser technology. In the end, we introduce the application and future prospects of cell patterning in biological research.