Serum-free cryoprotectant reagent: Cell Rev™ HQ-Store, improves cell recovery rate and other intrinsic cell properties including MSC self-renewal and differentiation potentials

Background & Aim Cellular cryopreservation is a linchpin of cell therapy and manufacturing. Without proper cryoprotectant reagents, the quality of cellular properties such as basic cell viability, cell attachment, growth, differentiation, characters or even cell death could be jeopardized. The use of fetal bovine serum (FBS) in cell freezing reagents have grown to be laboratory standard practice. However, the composition of the FBS is not well defined both qualitative and quantitatively and also have batch- to – batch variations and most importantly is a loophole subjected to potential contamination of cell cultures. From the biosafety aspects, sera may contain harmful factors like mycoplasma, endotoxin, viral contaminants or prions which increase the probability of the transmission of potentially fatal diseases to human patients. Methods, Results & Conclusion To minimize the possible risks, we have developed a totally chemically defined cryoprotectant recipe to preserve the important qualities of the cells particularly mesenchymal Stem Cells (MSCs). In this study, Mesenchymal Stem Cells have been cryopreserved as adherent cell populations in widely used cryoprotectant recipe, 20% FBS 10% dimethyl sulfoxide (DMSO) and Cell Rev™ HQ-Store defined serum-free cell freezing reagents. Our serum-free cryopreservation recipe has shown a significantly improved post-thawed recovery rate of cells to 95%. Interestingly, MSCs that have been stored in our defined cryoprotectant showed more than 50% enhanced n self-renewal while retaining differentiation potentials. Most importantly, these high-quality cryopreservation remains consistent with more than 10 freeze-thaw cycles, and longer than 2 yrs extended storage. These data illustrate that cell freezing solutions affect not only cytotoxicity but also critical properties of cellular proliferation and differentiation.