Ascorbic acid enhances the cold preservation period of human adipose tissue–derived mesenchymal stromal cells

INTRODUCTION: We previously developed 3% trehalose-added lactated Ringer's solution (LR-3T) and 3% trehalose- and 5% dextran-40–added lactated Ringer's solution (LR-3T-5D), which can be used to preserve adipose-derived mesenchymal stem cells (hADSCs) for 24 h at 5 and 25 °C. However, it is necessary to further extend the storage duration of cells to expand transportation zones and ensure time for quality control testing of final cell products. Therefore, we attempted to prolong the preservation duration of hADSCs by adding supplements to LR-3T-5D. We focused on ascorbic acid as an antioxidant because it is widely clinically as a nutrient. METHODS: We added the antioxidant ascorbic acid to LR-3T-5D and evaluated the viability, colony formation rate, proliferative capacity, and surface markers of hADSCs before and after preservation at 5 °C. RESULTS: Analysis of the concentration of ascorbic acid added to LR-3T-5D indicated that 1000 mg/L was the optimal concentration for maintaining the viability of hADSCs after 72 h of cold preservation. No changes were observed in the expression of specific cell surface markers or in the potential of hADSCs to differentiate into adipocytes, osteoblasts, or chondrocytes before and after cold preservation. DISCUSSION: These results suggest that cold preservation of hADSCs in LR-3T-5D supplemented with ascorbic acid helps maintain the quality of cells for use in cell therapy.