The microenvironment and cell populations within the myometrium play crucial roles in maintaining uterine structural integrity and protecting the fetus during pregnancy. However, the specific changes occurring at the single-cell level in the human myometrium between non-pregnant (NP) and term pregnant (TP) states remain unexplored. In this study, we utilized single-cell RNA sequence (scRNA-seq) and spatial transcriptomics (ST) to construct a transcriptomic atlas of individual cells in the myometrium of NP and TP women. Integrated analysis of scRNA-seq and ST data revealed spatially distinct transcriptional characteristics and examined cell-to-cell communication patterns based on ligand-receptor interactions. We identified and categorized 87,845 high-quality individual cells into twelve populations from scRNA-seq data of 12 human myometrium tissues. Our findings demonstrated alterations in the proportions of four sub-populations of smooth muscle cells (SMCs) in TP. Moreover, an increase in monocytic cells, particularly M2 macrophages, was observed in TP myometrium samples, suggesting their involvement in the anti-inflammatory response. This study provides unprecedented single-cell resolution of NP and TP myometrium, offering new insights into myometrial remodeling during pregnancy.