The commonly used dimethyl sulfoxide (Me2SO) in cell banking exhibits significant side effects on both cells and the human body. Therefore

there is an urgent need for an approach to mitigate the side effects of Me2SO while with comparable efficacy. The study uses human umbilical cord mesenchymal stem cells as the research material. First

the thermal physical properties of trehalose

glucose and L-proline and their regulation of ice crystal growth were measured by a differential scanning calorimeter and a cryomicroscope. Further

cryopreservation experiments were performed to determine the optimal concentrations of each component in the cryopservation solution

and the viability and functionality of cells after cryopreservation were validated. Results show that there is no significant difference in cell viability (92.42 ± 0.28%) and recovery rate (87.80 ± 4.22%) between the use of the novel stem cell cryopreservation solution (1.25 mol/L ethylene glycol + 1.0% whey protein + 0.1 mol/L trehalose + Normosol-R) and conventional cryopreservation solution (10%Me2SO). Moreover

the novel stem cell cryopservation solution better preserves the proliferative capacity and stemness of stem cells. The proposed novel stem cell cryopservation solution addresses the "Me2SO-free" issue in cell cryopreservation

holding promising potential for clinical applications.