Sodium Alginate as a Novel Cryoprotective Agent for Cryopreservation of Endothelial Cells in a Closed Polytetrafluoroethylene Loop.

Human umbilical vein endothelial cells (HUVECs) have wide applications in tissue engineering, drug delivery, and other fields due to their low antigenicity. Therefore, it is of great significance to effectively cryopreserve HUVECs for subsequent use (i.e., transport, long-term storage of cell banks). However, many commonly used cryoprotective agents (CPAs) are cytotoxic, so finding ideal CPAs to reduce the damage will pave the way for the application of HUVEC's cryopreservation. In this study, sodium alginate (SA) was employed as one of the main CPAs in a closed polytetrafluoroethylene (PTFE) loop used for cryopreservation with fast freezing of HUVECs. The ice crystal growth process was observed and the thermal enthalpy changes and osmolality of different solutions were tested. Moreover, the effects on cell viability and recovery were examined. The results showed that the addition of SA delayed the growth of ice crystals and decreased the number of ice crystals. Specifically, when 0.5% (w/v) SA was added to the CPAs, the cell survival increased by 10%. It is proved in this study that SA can be used as a novel CPA in combination with PTFE for the fast freezing of HUVECs, which is expected to improve the survival rate of cells and promote the exploration of protectants and cryopreservation in the future.