Chabanenko O. Response of mammalian erythrocytes to posthypertonic shock and cryoprotectant removal after freezing (experimental study)

The object of the study was the phenomenon of posthypertonic shock of mammalian erythrocytes. The research was aimed to comparatively examine the effectiveness of amphiphilic compounds (chlorpromazine, trifluoperazine, sodium decyl sulfate, decyl-β,D-glucopyranoside) under posthypertonic shock of erythrocytes as well as the process of deglycerolization of cells after freezing (–196°C). Research methods were as follows: spectrophotometry, light microscopy using a thermal assembly, flow cytometry, osmometry, refractometry, as well as the method of rapid freezing of erythrocytes using cryopreservative CSRIHBT 114 based on glycerol followed by removal of the cryoprotective agent; nonparametric statistical methods. The high antihemolytic activity of amphiphilic compounds belonging to different classes of surfactants was for the first time determined both in the model of posthypertonic shock of mammalian erythrocytes and in the removal of glycerol from human erythrocytes subjected to freezing to –196°C. It was first established that the level of erythrocyte damage in posthypertonic shock depended on the concentration of glycerol and temperature. It has been shown that when using at the stage of dehydration the combined media containing glycerol and NaCl, the level of posthypertonic hemolysis of erythrocytes does not depend on total osmolality of these media, and is determined by the concentration of cryoprotectant. The developed approach of targeted correction of erythrocyte sensitivity to the action of posthypertonic shock with the use of amphiphilic compounds can be applied to increase the stability of cells under extreme conditions. Scientific results on the effectiveness of cationic, anionic and nonionic amphiphilic compounds in the deglycerolization of thawed human erythrocytes significantly expand the basis for further search for membranotropic compounds of natural origin able to exhibit antihemolytic activity at the stage of removal of glycerol from cells. The results of the thesis can be recommended to be used in the educational process when training the specialists in the field of cryobiology, membrane science, pharmacology and veterinary medicine.