Evaluation of DMSO dextrose as a suitable alternative for DMSO dextran in cord blood cryopreservation

Umbilical cord blood is considered an alternative source of hematopoietic stem cells. Standard banking procedures use 50/55% DMSO in dextran 40 for cryopreservation and dextran-based solutions for thawing, however, due to the potential risk of crystallization of dextran, dextran 40 approved for clinical use has become limited or unavailable. This affects cryopreservation and thawing procedures. Carbohydrates, in particular sucrose, trehalose and glucose, have been shown to be effective in reducing cell damage during dehydration and have cryoprotective potential. We aim to study a 50/55% DMSO in 5% dextrose cryopreservation solution as an alternative to DMSO dextran.

Eighteen samples were divided into two aliquots and cryopreserved, one using standard solution and the other with DMSO dextrose experimental solution. Both aliquots were thawed and diluted with PBS or saline. Total nucleated cells counts, 7-AAD viability of CD45⁺ cells and recovery of CD34⁺ viable cells were assessed on thawed samples and compared between pair of aliquots.

No differences were observed in the total nucleated cells recovery between cryopreservation solutions, however, higher viability and CD34⁺ viable cells recoveries were observed using the experimental solution.

Results showed that DMSO dextrose cryopreservation solution had better results than the standard solution when thawed in an isotonic solution. This indicates that DMSO dextrose is probably a better alternative for direct infusion or when dextran thawing solutions are unavailable. Viability of CD45⁺ cells and recovery of CD34⁺ viable cells have positive correlation with engraftment, highlighting the relevance of the optimization of the cryopreservation and thawing process.