Abstract: A method for predicting the cell spatial relation based on single-cell transcriptome sequencing data includes the steps of obtaining a probability matrix P of a cell-cell interaction strength matrix A based on single-cell transcriptome sequencing data; reconstructing, according to the obtained probability matrix P of the cell-cell interaction strength matrix A, a three-dimensional spatial structure in which cells interact with each other; and for each cell in the reconstructed three-dimensional spatial structure in which cells interact with each other, determining the intercellular distance threshold for each cell to interact with h cells on average to obtain an intercellular interaction network. The method requires only the single-cell transcriptome sequencing data to predict the interaction of the cells in three-dimensional space, which breaks the limitation of the existing technology that needs to obtain the spatial relationship of cells through imaging.