Freezing regimen affects preservation of canine erythrocytes when using dimethyl sulfoxide
Keywords:
cryopreservation, cryoprotectant, canine erythrocytes, dimethyl sulfoxide, freezing regimensAbstract
This study examined how dimethyl sulfoxide (DMSO) concentration and different freezing regimens influence the preservation of canine erythrocytes during cryopreservation. The degree of erythrocyte hemolysis was assessed after thawing and at each stage of cryoprotectant removal. Increasing the DMSO concentration from 7.5 to 10% reduced hemolysis immediately after freeze–thawing by more than two-fold. During cryoprotectant removal, the extent of erythrocyte damage did not differ significantly between samples frozen with 7.5 and 10% DMSO. Overall hemolysis after freeze–thawing varied markedly depending on the cooling protocol. The highest level of cell damage (73%) occurred when samples were frozen in nitrogen vapor followed by immersion in liquid nitrogen. Direct immersion of the samples into liquid nitrogen reduced hemolysis to 37%. The most effective protocol involved briefly immersing the bottom of the cryotube in liquid nitrogen prior to complete freezing, resulting in only 31% hemolysis. This approach initiates crystallization at the lower part of the cryotube, substantially reducing the risk of mechanical cell damage during ice structure formation.
Probl Cryobiol Cryomed. 2026; 36(2): 115—121
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