Objective: The objective of the study was to compare the importance of storage time and the tooth type for isolation of dental pulp stem cells (DPSCs) from extracted human teeth.Materials & Methods: 35 human teeth (premolars, first molars and third molars) were used in this study. The teeth were stored in PBS medium after extraction and divided into two groups randomly according to the time elapsed between extraction and isolation. In group one, the isolation was performed within 2 hours and in the other group it was performed 24 hours after extraction. Results: There were no significant differences between isolation time and total cell counts (p=0.483) and between isolation time and viable cells (p=0.341). Also, there were no significant differences between the first molar and the premolar related cell counts and viable cells, but both teeth groups showed significant higher viability and had higher total cell amounts than third molars after isolation. Statistically significant correlations were found between age of donors and viable cells and viability after 24 hours isolation time.Conclusion: The immediate isolation of DPSCs is not necessary after the tooth extraction. The tooth can be stored in a PBS medium at room temperature up to twenty four hours after the extraction without a significant reduction in cell viability and counts. Also, the cells obtained from younger donors might have more chance for more viability even if storage time was extended.