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Simple Summary The placenta is a constantly changing organ that faces continuous tissue remodeling throughout gestation, reflected by recurrent stromal adaptation. The myofibroblasts, contractile stromal cells, are key players involved in orchestrating physiological tissue remodeling. Nevertheless, details regarding ultrastructural and functional aspects of myofibroblasts in the developed bovine placental stroma remain veiled. Here, the presence of myofibroblasts in the maternal placental stroma was confirmed by immunohistochemical and ultrastructural analysis, with the co-localization of several factors, e.g., α -smooth muscle actin, vimentin and nuclear progesterone receptor. During short-term tissue culture, the contraction of placentomal caruncle tissue slices was induced by angiotensin-II. Furthermore, a three-dimensional reconstruction demonstrates an example of a bovine placental myofibroblast. According to the present findings, it is postulated that the maternal stroma of the placenta in the cow contains a high proportion of contractile myofibroblastic cells. Herewith, a basis for further investigations regarding the functional aspects and the role of myofibroblasts in peri- and post-partum placental events in cattle was created. Abstract Myofibroblasts are contractile cells that exhibit features of both fibroblasts and smooth muscle cells. In the synepitheliochorial placenta of the cow myofibroblasts are found in the maternal stroma. However, a deeper understanding of the structure and function of the stromal myofibroblasts in the developed bovine placenta is still missing. Thus, immunohistochemical and ultrastructural analyses in bovine term placentomes, compared to non-pregnant caruncle samples, were conducted. To investigate functional aspects, contractility of placentomal caruncle slices was assessed in an in vitro contraction assay. Additionally, a three-dimensional reconstruction of a bovine placental myofibroblast was created. Immunofluorescent staining revealed a characteristic pattern, including cytoplasmic expression of α-smooth muscle actin, strong perinuclear signal for the intermediate filament vimentin and nuclear progesterone receptor staining. Ultrastructurally, stress fibers, extended cisternae of the rough endoplasmic reticulum and perinuclear intermediate filaments were observed. Moreover, in vitro stimulation with angiotensin-II, but not with prostaglandin F2α, induced contraction of placental caruncle tissue. Altogether, these results indicate that progesterone-responsive myofibroblasts represent a mesenchymal phenotype that is involved in the contractile properties of bovine placental stroma. Therefore, the present findings suggest a potential involvement of myofibroblasts in post-partum events of cattle, i.e., expulsion of fetal membranes and uterine involution.