RESEARCH IN COMPARATIVE ASPECT OF CRYOPROTECTIVE ACTION OF GLYCERIN-BASED CRYOPRESERVATIVES IN COMBINATION WITH POLIVINYL ALCOHOL OF DIFFERENT MOLECULAR WEIGHTS AT FAST AND SLOW COOLING OF HUMAN DONOR BLOOD

Hvozdiuk Y.V., Kompaniets A.M.

BIOLOGY

Scentific article

. The formation and growth of ice crystals both inside and outside the biological cell pose a serious problem in the development of methods and technologies for cryopreservation of biological objects for further use. Therefore, for several years the problem of finding and using a compound that can influence the processes of ice formation, especially the recrystallization process, is currently being actively studied in scientific institutions around the world. One of the important causes of cell death is the recrystallization of ice at the thawing stage. Inhibition of recrystallization activity is a desirable effect that can significantly improve the safety of cryopreserved material. Today the main problem is the development of effective means of cryoprotection, new cryoprotectants that will have an enhanced effect on the inhibition of ice recrystallization, which in turn will improve the cryopreservation of not only cells but also human tissues and organs. However, there are a number of important obstacles that researchers have encountered in the practice of cryopreservation: the very small number of compounds that can be obtained from natural resources, the complexity or impossibility of complete chemical synthesis of analogues; contradictory data on the effectiveness of the use of these compounds in cryopreservation of biomaterial; cytotoxicity and immunogenicity of natural antifreeze proteins. In connection with these obstacles, in recent years, researchers have turned their attention to synthetic compounds, which are also similar to natural specific antirecrystallization activity without side effects. The solution to this problem is the development of cryoprotectants, which, along with traditional cryoprotectants will use synthetic compounds – antifreeze, which has a specific antirecrystallization activity. From the list of these compounds, the attention of researchers is focused on a synthetic polymer – polyvinyl alcohol (PVA). The aim of our study was to investigate the properties of PVA of different molecular weights as an active inhibitor of ice recrystallization and the level of preservation of erythrocytes in the process of slow and rapid freezing – heating in cryoprotective media based on standard cryoprotectant glycerin of different concentrations. Materials and methods. The material of the study was erythroconcentrate obtained from human donor blood group A (II), prepared on the hemopreservative «Glugetsir». Erythroconcentrate was obtained by centrifugation of canned donor blood at 1250 g for 25 minutes. The obtained data were calculated according to the following indicators: the concentration of free hemoglobin in the supernatant and total hemoglobin of cell suspensions, determined by hemoglobin cyanide method using the kit «Hemoglobin SPL 200» (Ukraine) and spectrophotometer KFK – wave – 3–01 «ZOMZ», 540–545 nm, hematocrit was determined on a centrifuge «SM‑70», the rate of hemolysis of erythrocytes in the supernatant was calculated by the formula given in Huggins CE. (Prevention of hemolysis of large volumes of red blood cells slowly frozen and thawed in the presence of dimethylsulfoxide).Statistical processing of the results was performed using the Mann-Whitney test for small samples using the software package «Statistics». Analysis of results. As a result of a comparative study of cryoprotective media of different composition of cryoprotectants with solutions of polyvinyl alcohol in the rapid method of freezing blood cells in particular (erythrocytes) by immersing the samples in liquid nitrogen. It was found that after freezing -heating of erythrocytes in multicomponent solutions based on glycerol in combination with PVA m. m. 9 and 31 kDa, there is a tendency to increase the preservation of erythrocytes by hemolysis in comparison with the action of 20 %, 30 %, 40 % glycerol mono solutions.

erythrocytes, cryopreservation, recrystallization, polyvinyl alcohol.

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«Bulletin of problems biology and medicine» Issue 2 Part 1 (164), 2022 year, 89-99 pages, index UDK 641.528+661.188.1+678.746+547.42+612.111

10.29254/2077-4214-2022-2-1-164-89-99