Postmenopausal osteoporosis (OP) is a major concern for public health. Low-level laser therapy (LLLT) has a positive effect on the health of bone marrow mesenchymal stem cells (BMMSCs). The purpose of this study is to evaluate the influence of LLLT and oxytocin (OT) incubation-individually and in combination-on osteoporotic BMMSCs in ovariectomized rats. Twelve female rats were randomized into two groups to undergo either a sham surgery (sham group) or ovariectomy-induced osteoporosis (OVX group). MSCs harvested from the BM of healthy and OVX rats underwent culture expansion. There were five groups. In Groups one (sham-BMMSC) and two (OVX-BMMSC) the cells were held in osteogenic condition medium without any intervention. In the group three (OT), OT incubation with optimum dose was performed for 48 h (two times, 10 −12 molar). In Group four, lasertreated-OVX-BMMSCs were treated with optimum protocol of LLLT (one time, 1.2 J/cm 2 ). In Group five (laser + OT group), the OT incubation plus the laser irradiation was performed. The biostimulatory effect of LLLT is demonstrated by a significant increase in the viability of OVX-BMMSCs, cell cycle, and extracellular levels of Transforming growth factor beta (TGF-β), insulin-like growth factor-I (IGF-I), and Alkaline phosphatase (ALP) compared to control OVX-BMMSCs and/or the sham group. OT incubation and laser + OT incubation have a positive effect on OVXBMMSCs. However, LLLT is more effective statistically. We conclude that LLLT significantly improved cell viability, enhanced the osteogenic potential of the OVXBMMSCs, and increased the extracellular levels of the TGF-β, IGF-I, and ALP.