Effects of Cryopreservation of intact teeth on the isolated dental pulp stem cells

Introduction: Human dental pulp stem cells (DPSCs) have been reported to be useful material for future regenerative medicine. Clinically, cryopreservation of intact teeth can successfully preserve the periodontal ligament for future autotransplantation; however, the effects of cryopreservation procedure on the properties of DPSCs are still unclear. The aim of this study was to test whether DPSCs isolated from cryopreserved teeth can express stem cell-specific markers. Methods: In this study, a novel programmable freezer coupled to a magnetic field was used to perform the cryopreservation experiments. The tested DPSCs were isolated from magnetically cryopreserved and non-cryopreserved fresh teeth with an enzyme digestion procedure. The success rate of isolation, growth curves, morphology, stem cell-specific markers, and the differentiation capacity of the isolated cells were evaluated and compared. Results: The isolation rate of dental pulp cells from magnetically cryopreserved teeth was 73%. After culture for 5 generations, there was no significant difference in cell viability between cells isolated from magnetically cryopreserved teeth and those isolated from fresh teeth. There were also no visible differences between the 2 groups of dental pulp cells in morphology, expression of stem cell markers, or osteogenic and adipogenic differentiations. Conclusions: The results suggest that cryopreserved whole teeth can be used for autotransplantation and provide a viable source of DPSCs.