Understanding the molecular characteristics and metabolic processes of the mammalian endometrium is crucial for advancing biological research, particularly in veterinary obstetrics and pathology. This study established and analyzed organoids from the endometrial epithelial stem cells of five mammals with different placental types: cows (cotyledonary), dogs and cats (zonary), pigs (diffuse), and rats (discoid). The organoids from these five species were maintained for over 13 passages, successfully frozen-thawed, and confirmed by pathological analysis to retain the characteristics of the original tissues. Furthermore, integrative transcriptome analysis of the organoids and tissues from the five species highlighted key pathways, such as PI3K-Akt signaling and extracellular matrix-receptor interaction, which are crucial in cancer research. Despite the downregulation of genes associated with vascular smooth muscle contraction, the organoids exhibited significant activity of genes involved in hormone metabolism. In conclusion, our study achieves stable establishment of endometrial organoids from five mammals with different placental types and offers foundational data for organoid research. In the future, these organoids are suitable models for investigating uterine physiology, diseases, and assessing potential therapies.