Abstract

Cryoprotectants are crucial factors in cell cryopreservation. Trehalose (Tre), a nontoxic, nonreducing, and natural disaccharide, has the potential to protect cells as a cryoprotectant. As an inducer of autophagy, Tre can influence the development of many diseases and may also have an effect on cell cryopreservation through this mechanism. In this study, human aortic endothelial cells were preserved in different cryopreservation fluids with or without dimethyl sulfoxide and Tre was added. Subsequently, the expression of the main autophagy-related genes LC3, BECN, and P62, cell death and apoptosis, and the proliferation rate were measured in different groups after cryopreservation. Our data showed that Tre can improve the expression of the autophagy-related genes LC3 and BECN and reduce the expression of P62. Dead/alive staining and flow cytometry showed that cell death and cell apoptosis were reduced during cryopreservation with Tre. In addition, the cell proliferation rate after thawing was increased in the Tre group when compared with others. These results all indicated that there might be a connection between Tre-triggered autophagy and the protective role of Tre in cell cryopreservation. Furthermore, strategies to regulate autophagy to reduce apoptosis in this process should be investigated in future research.