Efficient application of veterinary blood transfusion approaches for small companion animals requires readily available supply of the donor material. This can be achieved by developing of effective biobanking technologies allowing long-term storage of donor blood components via cryopreservation. Transfusion of an erythrocyte concentrate allows the successful correction of various hematological pathologies, severe bleeding, and etc. While in the past there were several approaches to cryopreserve red blood cells of dogs, to our knowledge there is virtually no data on cryopreservation of feline erythrocytes. In this paper, we performed a comprehensive parameter optimization for low temperature storage of RBCs of both species. Here, the efficiency of single-component and multicomponent cryoprotective media as well as necessary time of pre-incubation with penetrating and non-penetrating cryoprotectants prior to rapid freezing is analyzed. This study showed that glycerol was not sufficient for cryopreservation of red blood cells of the studied species under the investigated conditions. Application of 10% (v/v) ME2SO allowed for a significant reduction of canine and feline erythrocytes hemolysis after thawing. 17.5% hydroxyethyl starch demonstrated the highest cryoprotective activity for both species. It was found that dog RBCs should be incubated in cryoprotective media for 30 min at 22°C prior to freezing, while for cat RBCs 20 min is sufficient. Combination of CPAs was less effective. Presented data may be considered in further studies in veterinary transfusion and blood banking optimization.
Babesiosis is one of the most common protozoan diseases in dogs, which in the absence of the necessary veterinary care often leads to the animal death. The changes of hematological parameters in dogs during complex treatment of babesiosis and transfusion of cryopreserved erythrocytes were evaluated in the research. Erythrocytes were frozen using 20% DMSO by immersion into liquid nitrogen. Transfusion of cryopreserved erythrocytes together with antiparasitic chemotherapy has been shown to promote rapid recovery of hematological parameters of animal blood (hemoglobin, hematocrit and erythrocyte counts), as well as improves their well-being, promotes faster recovery. Cryopreserved with DMSO erythrocytes can be recommended for long-term storage of blood at cryobanks.
Cryoprotective properties of combined media of permeable (dimethyl sulfoxide) and impermeable (polyethylene glycol with m. w. 1500) cryoprotective agents during rapid cooling in liquid nitrogen of canine erythrocytes using saline and sucrose-saline media have been investigated. It was found that the use of combined solutions of cryoprotective agents based on polyethylene glycol with m.w. 1500 (15%) and dimethyl sulfoxide (2.5–10%) in saline was not quite effective for cryopreservation of canine erythrocytes. Reducing the salt concentration and adding cell-impermeable sucrose to the cryopreservation medium increase the preservation of erythrocytes after warming. The best cryoprotective properties for canine erythrocytes were demonstrated by 10% dimethyl sulfoxide based on sucrose-saline medium, with high preservation of cells after freeze-warming, mechanical and osmotic stability of warmed erythrocytes. This indicates the possibility of a long-term storage and use of cryopreserved canine erythrocytes for transfusions.
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