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Information × Registration Number 0214U006713, 0109U000275 , R & D reports Title Study of mechanisms of structural and functional changes in nucleated cells of cord blood and erythrocytes under the effect of exo- and endocellular cryoprotectants and low temperatures. popup.stage_title Head Babijchuk Lubov Alexandrovna, Registration Date 11-02-2014 Organization Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of the Ukraine popup.description2 Objects of the study are cord blood nucleated cells, including hematopoietic stem cells, and erythrocytes of donor and cord blood. The purpose is the investigation of the mechanisms of changes in structural and functional properties of cord blood nucleated cells (CBNC), including hematopoietic stem (CD34+) cells (HSC), as well as modifications of the plasma membrane of erythrocytes of donor and cord blood, that underlie cell cryosensibility and cryostability under the influence of cryoprotectants of different effecting mechanisms and low temperatures, as well as development of new methods for cryopreservation of cord blood. Results and novelty: fundamental studies have shown that the CBNC isolation with polyglucinum following by freezing with 5 % DMSO, as well as their two-stage isolation by a centrifugation and following by freezing with 10% PEO -1500 allow to maintain up to 85% of CBNC and 95 % of HSC at the viable state. Analysis of changes in lipid membrane asymmetry, content of reactive oxygen species, the functional activity of cells in culture, as well as the study of cell ultrastructure at each stage of cryopreservation indicate the retaining of structural and functional properties of CBNC. It was found that the reduction in the absolute number and viability of the general population of CBNC occured mainly due to granulocytes, while mononuclear cells (lymphocytes, monocytes) were more cryostable. It was established that PEO and glycerol might affect lipid asymmetry and surface markers at different stages of erythrocyte preservation. Patterns of these changes depend essentially on effecting mechanisms (endo-or exocellular types) of cryoprotectants. Changes in intracellular Ca2+ levels upon application of glycerol indicate the possibility of Са2+- dependent regulatory mechanisms of erythrocytes stability under stress conditions. The results of these studies allowed developing new washing-free methods of cryopreservation of the whole cord blood and its components using non-penetrating cryoprotectant PEO -1500, as well as the efficient method for cryopreservation of CBNC with 5% DMSO. Field of use: cryobiology, cryomedicine, transfusiology, transplantation, cell therapy. Product Description popup.authors Землянських Ніна Григорівна Зубов Павло Михайлович Зубова Оксана Леонідівна Мігунова Регіна Костянтинівна Михайлова Ольга Олександрівна Невзоров В'ячеслав Павлович Рязанцев Володимир Васильович Чернявська Олена Олександрівна Щирова Валентина Анатоліївна popup.nrat_date 2020-04-02 Close