The invention relates to a micro-fluidic cell culturing chip and a preparation method thereof. A microstructure and a micro-channel are arranged on the surface of a micro-fluidic chip; two cells are respectively fixed in specific cell culturing regions; signal transmission and interaction between the cells are carried out between the cells through the micro-channel; under a microscope, the interaction between the two cells in the growing process can be intuitively studied and metabolites in the cell growing process are analyzed; a new idea and a new research technique are provided for researches on the cell biology, and the micro-fluidic cell culturing chip is mainly applied to the related field of cell biology, genetics, drug screening and the like. Through adoption of the micro-fluidic chip, co-culturing of the two cells is achieved, the operation is simple, parallel implantation and co-culturing of the two cells are realized, the usage amounts of reagents and samples are reduced, the cell implanting process is simplified, the interaction between the two cells in the growing process can be intuitively studied under the microscope, the metabolites in the cell growing process is analyzed and a brand new analysis technology is provided for researches on the cell biology and the micro-fluidic cell culturing chip provided by the invention is portable, economic, rapid, efficient and accurate.