scholarly journals Freezing and storage of leukodepleted erythrocyte suspensions

2017 ◽  
Vol 61 (No. 8) ◽  
pp. 443-448 ◽  
Author(s):  
DA Bala ◽  
E. Eraslan ◽  
I. Akyazi ◽  
EE Ekiz ◽  
M. Ozcan ◽  
...  

Studies on the frozen storage of human blood products have benefited veterinary transfusion medicine in recent years, but the long-term cryopreservation of canine red blood cells (RBCs) has not yet been thoroughly investigated. Further, no studies are available with respect to the frozen storage of leukocyte-depleted canine red blood cells (LD-RBCs). The objective of the current study was to investigate time-dependent effects of long-term frozen storage on leukocyte-depleted canine RBCs. Twelve healthy adult dogs meeting the criteria for blood transfusion were used in the study. Whole blood samples (450 ± 45 ml) collected from each dog were centrifuged for 5 min at 22 °C and 4200 × g in a cryogenic microcentrifuge and concentrated RBC (pRBC) suspensions were obtained. Leukocyte depletion was achieved by filtration (2.6 log<sub>10</sub>). Then, the filtrated samples were prewashed three times in 0.9% NaCl solution and were allocated into three subgroups to be evaluated at three different time points (Day 0, Month 4 and Month 6). The samples for cryopreservation were subjected to glycerolisation and then stored at –80 °C for 4- and 6-month periods. At the end of this period pRBC units were thawed by manual agitation in a water bath maintained at 36–38 °C, centrifuged and then washed in a consecutive series of 12%, 1.6% and 0.9% of NaCl + 0.2 dextrose solutions. 2,3-Diphosphoglycerate (2,3-DPG), adenosine triphosphate (ATP), supernatant haemoglobin (SupHb), sodium (Na<sup>+</sup>) and potassium (K<sup>+</sup>) levels, residual glycerol concentrations and haemograms of thawed and deglycerolised pRBC samples were evaluated together with those of Day 0. Sterility tests were performed on all samples for bacterial contamination. No statistically significant differences were noted except for Hct and SupHb levels. No bacterial contamination was noted in any of the samples on the basis of sterility tests. It was found that the described glycerolisation procedure could be a method of choice in the cryopreservation of leukocyte-depleted pRBCs (LD-pRBCs) since no negative effect was observed on the quality of the products and long-term frozen storage did not cause RBC destruction.

2016 ◽  
Vol 19 (2) ◽  
pp. 401-406 ◽  
Author(s):  
D. Aktaran Bala ◽  
M. Özcan

Abstract Human medicine studies have so far demonstrated that erythrocytes may be preserved and stored at low temperatures for decades retaining their metabolic and biochemical properties. However, detailed studies regarding this problem are not yet available in veterinary medicine. Therefore, the objective of the current study was to investigate time-dependent effects of long-term frozen storage of canine red blood cells. Twelve healthy adult dogs meeting the criteria for blood transfusion were used in the study. Whole blood samples (450 ± 45 ml) collected from each dog were centrifuged by a cryogenic microcentrifuge and packed RBC suspensions were obtained. The samples were prewashed three times in 0.9% NaCl solution and were allocated into three groups to be evaluated at three different time points (day 0 and month 4 and 6). The samples to be frozen were subjected to glycerolization and then stored at −80°C for 4 and 6-month periods. At the end of this period the packed RBC samples were thawed, centrifuged and then washed in a consecutive series of dextrose solutions. 2,3-Diphosphoglycerate (2,3-DPG), Adenosin triphosphate (ATP), supernatant hemoglobin (SupHb), sodium (Na+) and potassium (K+) levels, residual glycerol concentrations and hemograms were evaluated and compared. Sterility tests were performed on all samples for bacterial contamination. A statistically significant decrease was noted in potassium levels, which was the natural outcome of deglycerolization process. No significant change was observed in terms of other parameters due based on different time points. In conclusion, long-term frozen storage had no negative effect on the quality parameters of canine erythrocytes.


2021 ◽  
Vol 30 ◽  
pp. 096368972110360
Author(s):  
Daniel Lysak ◽  
Michaela Brychtová ◽  
Martin Leba ◽  
Miroslava Čedíková ◽  
Daniel Georgiev ◽  
...  

Cryopreserved haematopoietic progenitor cells are used to restore autologous haematopoiesis after high dose chemotherapy. Although the cells are routinely stored for a long period, concerns remain about the maximum storage time and the possible negative effect of storage on their potency. We evaluated the effect of cryopreservation on the quality of peripheral stem cell grafts stored for a short (3 months) and a long (10 years) period and we compared it to native products.The viability of CD34+ cells remained unaffected during storage, the apoptotic cells were represented up to 10% and did not differ between groups. The clonogenic activity measured by ATP production has decreased with the length of storage (ATP/cell 1.28 nM in native vs. 0.63 in long term stored products, P < 0.05). Only borderline changes without statistical significance were detected when examining mitochondrial and aldehyde dehydrogenase metabolic activity and intracellular pH, showing their good preservation during cell storage. Our experience demonstrates that cryostorage has no major negative effect on stem cell quality and potency, and therefore autologous stem cells can be stored safely for an extended period of at least 10 years. On the other hand, long term storage for 10 years and longer may lead to mild reduction of clonogenic capacity. When a sufficient dose of stem cells is infused, these changes will not have a clinical impact. However, in products stored beyond 10 years, especially when a low number of CD34+ cells is available, the quality of stem cell graft should be verified before infusion using the appropriate potency assays.


Transfusion ◽  
2017 ◽  
Vol 58 (2) ◽  
pp. 423-429 ◽  
Author(s):  
Athinoula Meli ◽  
Vicky Hancock ◽  
Heidi Doughty ◽  
Steve Smedley ◽  
Rebecca Cardigan ◽  
...  

2019 ◽  
Vol 57 (5) ◽  
pp. 617-622 ◽  
Author(s):  
Van Long Nguyen ◽  
Michael Fitzpatrick

Abstract Phosphatidylethanol (PEth) are phospholipids produced through non-oxidative ethanol metabolism. They accumulate in red blood cells and have been traditionally analysed in whole blood as potential biomarkers for moderate to long-term alcohol consumption. More recently, their analysis in dried blood spots has been gaining favour, namely, due to the ease in sampling, transport and storage conditions required. This paper aims at providing a short comparative review between analysing PEth in whole blood and dried blood spots and the potential pitfalls that researchers may face when setting up PEth testing for clinical use.


2020 ◽  
pp. 56-57
Author(s):  
V.L. Novak ◽  
B.O. Kondratsky ◽  
S.V. Primak ◽  
O.O. Tarasyuk ◽  
O.M. Tushnitsky ◽  
...  

Objective. Analysis of issues related to the safety and quality of donated blood and its components. Materials and methods. Many years of experience of hematologists, immunologists, isoserologists, morphologists, biochemists in studying the composition, morpho-functional properties of cells and blood plasma, the use of donor blood and its components in clinical practice have made it possible to reconsider method of chemotherapy “multilateral action”. Results and discussion. One of the main axioms of modern transfusion medicine: chemotherapy should be performed strictly according to the indications and those blood components that are needed to ensure the viability of the body. The development of transfusiology has proved, with few exceptions, the inexpediency and even harmfulness of the use of whole donor blood. When using blood and its components, it is necessary to clearly consider extremely important point: the use will be in a planned manner, or in special circumstances. Blood components and blood plasma preparations have unique medicinal properties and there is currently no alternative to them. Each country is recommended to switch to self-sufficiency of blood components and their derivatives, to organize their own production structures that would meet the country’s domestic needs in blood components and preparations. The main components of the ideology of component chemotherapy are: recovery of blood component deficiency is not achieved on a “drop by drop” basis; no need to completely replace the existing deficiency of a cellular or protein component. Transfusions of blood components should be treated as a responsible invasive medical procedure – an operation that can have both immediate and long-term complications and consequences. Unreasonable transfusions of whole canned blood, especially after long periods of storage, are not only ineffective, but often pose a danger. In canned blood, during storage, complex biochemical metabolic processes take place both in cells and in plasma, which ultimately reduce the quality of both the blood itself and the morpho-functional properties of its individual components. Thus, 8-day storage of erythrocytes is the threshold after which erythrocytes begin to adversely affect the patient. Morpho-functional properties of blood components are directly dependent on the shelf life and distance of transportation. During the storage of blood and erythrocyte mass, the level of ATP decreases, on which the elasticity of the erythrocyte membrane depends. The magnitude of the negative electrical charge of the surface membrane of blood cells decreases. The number of prehemolytic forms of erythrocytes and cells that are not capable of reverse transformation increases. Within 1-4 days, leukocytes die, bacteria are released, so after this period, the greatest number of complications and reactions. Leukocytes and platelets form microaggregates at an early stage of storage, which can cause microembolism and distress syndrome. Microclots are formed, which include lysed blood cells and fibrin. The number of microclots increases every day, reaching on day 21 to 100 thousand/ml, so when transfusing it is advisable to use antiplatelet filters, rather than leukocyte. The use of bed leukofilters after a long period of storage of erythromass is not advisable, because there are no leukocytes. In addition, up to 2 % of erythrocytes are lost. The pH decreases, the content of 2,3-DFG, which is responsible for oxygen transport function (decreases by 50 % on the third day), hemolysis increases (up to 200 mg% of free hemoglobin). The concentration of potassium and ammonium ions increases. The recommended threshold for erythrocyte concentrate transfusions is a hemoglobin level of less than 70 g/l in adults and most children. It is important to use fresh-frozen plasma, erythrocyte concentrate and platelets obtained from one donor. Modern blood separators technically provide such an opportunity. Conditionally acceptable number of platelets in patients with injuries is more than 50×109/l cells, and in patients with combined brain injury is 100×109/l. Platelet concentrate obtained by the manual method from 4-5 donors leads to the development of refractoriness. With increasing shelf life of platelets, the functional properties of cells deteriorate significantly. All attempts to create the so-called artificial blood in the 19th century ended at the level of scientific developments. Synthetic and bioengineered cellular components of blood, hematopoietic factors, as well as hematopoietic stem cells are considered promising in the future. One of the possible ways to solve the problem of long-term storage of blood components, especially liquid groups, is cryopreservation of individual cells (erythrocytes) at moderately low (-20; -40; -80 ºС) and ultra-low (-165-196 ºС) temperatures, followed by deglycerization (washing), the use of special solutions for resuspension. With the development of low-temperature electric refrigeration equipment in Ukraine, this has become a reality. Conclusions. Practice has shown that both the blood itself and its components can neither be produced nor extracted as minerals, it can only be shared. All of the above indicates that blood donation is and will remain the main source of cellular components of the blood for at least the next decades.


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