摘要：Introduction: The Wharton’s Jelly tissue possesses an attractive source for the isolation of mesenchymal stem cells (MSCs), that could be used in large scale clinical trials and in tissue engineering and regenerative medicine. However, prolonged cell culture and expansion, could affect their characteristics by causing genomic and epigenetic alterations. A possible solution to this problem would be the cryopreservation of the entire Wharton’s Jelly (WJ) tissue. The use of dimethyl sulfoxide for such a complex tissue is insufficient for its proper storage. Vitrification might be a feasible method for storage of more complex tissues. In order to find the optimum method for the storage of the WJ tissue different cryopreservation methods, including vitrification and conventional cryopreservation were applied in this study. Methods: Vitrification was performed with either the use of a commercial available solution (“CryostorTM”) or VS55 a solution that can be used in non-equilibrium approach. Conventional cryopreservation was performed using a solution that combines 10% vol/vol DMSO and Fetal Bovine Serum. The evaluation of the cryopreservation methods was accomplished by the isolation of MSCs, growth kinetics, immunophenotypic analysis, multilineage differentiation, scratch wound assay and reactive oxygen species (ROS) quantification. Results: The results clearly showed that the vitrification successfully preserved the WJ tissue at -196oC, yielding viable cells well characterized by mesenchymal stem cell properties. On the other hand, conventional cryopreservation did not achieve the same cell isolation rate when compared to vitrification methods. Conclusion: In conclusion, the vitrification is a promising method for WJ tissue banking, allowing the isolation of well-defined MSCs for future MSC-based regenerative therapies.
