The Optimal Isolation and Characterization Conditions of Rabbit Dental Pulp - Derived Mesenchymal Stem Cells for Use in Regenerative Medicine

Abstract

Background and Objectives: Currently, dental pulp stem cells have gained importance in stem cell research. Our aim was to isolate mesenchymal stem cells from the rabbit dental pulp and to characterize those using different methods.Methods: Mesenchymal Stem Cells (MSCs) were obtained by enzymatic method from dental pulp of New Zealand Rabbits. MSCs that were derived from rabbit dental pulp (rabbit DPSCs) were characterized by using flow cytometry, immunofluorescence staining, quantitative Real Time PCR (qRT-PCR). For the characterization, CD29, CD44 and CD45 MSC markers were used. Adipogenic, osteogenic and chondrogenic differentiation tests were performed for multilineage differentiation. The clonogenic properties of the obtained rabbit DPSCs were demonstrated by the Colony forming unitsfibroblast (CFU-F) Test. The results were analyzed statistically.Results: Flow cytometry, immunofluorescence staining and qRT-PCR analysis showed that rabbit DPSCs were positive for CD29 and CD44 antibodies; and negative for CD45 antibody (p<0.001). Adipogenic, osteogenic and chondrogenic differentiation was demonstrated by adipo-red, alizarin red, alkaline phosphatase and safranin-O positive staining. In the CFU-F test, 127.7±13.98 colonies and 211.3±10.40 colonies were counted at 500 cells/well and 1000 cells/well, respectively (p<0.001). Conclusion:rabbit DPSCs have been successfully isolated from rabbits; and it has been proven that the cells obtained are MSCs. It has been determined that there are high potency cells according to other stem cell sources. Rabbit is a model organism frequently used in in vivo studies. Hence, rabbit dental pulp stem cells can be used in both in vitro and in vivo clinical, surgical and tissue engineering studies.