This study was conducted to understand the cellular proliferative effect of Photobiomodulation Therapy (PBMT) on thawed dental pulp stem cells (DPSCs) stored for 2 years. For this purpose, cells were exposed to PBMT for short period of time to evaluate the most appropriate PBMT parameter for stimulating cellular proliferation that can be used for future tissue engineering therapies. Fully characterized DPSCs were seperated into three groups according to the laser energy densities (5 J/cm² or 7 J/cm²) applied and a group was served as control in which cells did not receive any laser irradiation. The cells in laser-irradiated groups were further divided into two subgroups according to the period of application (24 h and 0 h) and exposed to Gallium– Aluminum–Arsenide diode laser irradiation. Cell viability and the proliferation rate of the cells were analyzed with the 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) assay, any PBMT related cellular cytotoxicity were determined by performing a lactate dehydrogenase assay (LDH) and statistical analysis of data were performed. The percentage of proliferation seemed to increase upon laser therapy in both different doses of irradiation (5 J/cm² and 7 J/cm²). DPSCs showed significantly higher proliferation rate upon 7 J/cm² irradiation in both 0 h and 24 h when compared to control groups. However, DPSCs irradiated with 5 J/cm² dose induced relatively lower proliferation rate when compared to 7 J/cm² dose of irradiation. According to the LDH data, PBMT exposure did not show any significant cytotoxicity at both energy densities in all different time periods. PBMT at 7 J/cm² should be an effective parameter to stimulate proliferation of long-term cryopreserved DPSCs in a short term time period. Photobiomodulation therapy may be an upcoming tool for future tissue enngineering and regenerative dentistry applications.