Purpose: To evaluate the cytotoxicity of temporary luting cements on bovine dental pulp-derived cells (bDPCs). Materials and methods: Four different temporary cements were tested: Rely X Temp E (3M ESPE), Ultratemp (Ultradent), GC Fuji Temp (GC), and Rely X Temp NE (3M ESPE). The materials were prepared as discs and incubated in Dulbecco's modified eagle's culture medium (DMEM) for 72 hours according to ISO 10993-5. A real-time cell analyzer was used to determine cell vitality. After seeding 200 μL of the cell suspensions into the wells of a 96-well plate, the bDPCs were cured with bioactive components released by the test materials and observed every 15 minutes for 98 hours. One-way ANOVA and Tukey-Kramer tests were used to analyze the results of the proliferation experiments. Results. All tested temporary cements showed significant decreases in the bDPCs index. Rely X Temp E, GC Fuji Temp, and Rely X Temp NE were severely toxic at both time points (24 and 72 hours) (P < .001). When the cells were exposed to media by Ultratemp, the cell viability was similar to that of the control at 24 hours (P > .05); however, the cell viability was significantly reduced at 72 hours (P < .001). Light and scanning electron microscopy examination confirmed these results. Conclusion: The cytotoxic effects of temporary cements on pulpal tissue should be evaluated when choosing cement for luting provisional restorations.