The present study was conducted in order to explore the mechanisms whereby parathyroid hormone (PTH) maintains in vitro proliferation of bone marrow mesenchymal stem cells (BMSCs). Bone marrow was isolated from Sprague Dawley (SD) rat femurs, cultured in vitro, and passaged using a cell adherent culture method. The BMSC proliferation was evaluated by the methyl thiazolyl tetrazolium (MTT) assay and the fluorescence intensity of calcium ions in BMSCs was analyzed by laser scanning confocal microscopy (LSCM). Our results show that BMSC proliferation in the experimental group treated with PTH was more significant than controls. The calcium ion fluorescence intensity in BMSCs was significantly higher for the experimental group as compared to the control group. For each group, there was significant difference in the fluorescence intensity of calcium ions in BMSCs between 7 d and 14 d. In conclusion, parathyroid hormone increased the fluorescence intensity of calcium ions in BMSCs, which might represent a key mechanism whereby BMSC proliferation is maintained.