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Information × Registration Number 0214U008024, 0113U003764 , R & D reports Title The investigation of expresion of marker proteins epithelial stem cells and progenitor cells in oral squamous cell carcinoma. popup.stage_title Head Sydorenko Mychail V., Registration Date 03-06-2014 Organization Department of Biotechnical Problems of Diagnostics, Institute of Problems of Cryobiology and Cryomedicine of NAS of Ukraine popup.description2 Abstract. Research report: 57 pages, 12 fig., table 6., 35 sources. Object of research: cell cryoprotective agents. Objective: to study the effectiveness of a combination of different cryoprotective funds used in cryoprotective solutions to prevent cold damage cells. To investigate the influence of different concentrations of DMSO (5-25% final concentration), in combination with fetal bovine serum (FBS) at various concentrations (25-40%) and non-penetrating cryoprotective agents: glycerol, ethylene glycol, glucose. As a result, several protocols have been developed without the use of freezing vitrification method for comparing the data with those that are obtained by ultrafast freezing cells. Research methods and apparatus: cryoprotective solutions consisted of nutrient medium, serum, DMSO, glucose, glycerol, ethylene glycol in different proportions. When add the cryoprotective solution (CS) used two different protocols. Protocol 1 is the simultaneous addition to the cell suspension cryosolution and consistent reduction in temperature. Cryoprotective solution number 1 (CS №1), cooled to +4°C and slowly added to the chilled cell suspension to 200C in a ratio of 1:1. Thereafter, the cell suspension with CS №1 cooled to +4°C for 30 minutes at + 20C, and then for 30 minutes at -30C overnight, and cooled to - 800C. According to the protocol used 2 first part of cryoprotectant СS №2, cooled to +4 °C, which was added to the cell suspension at a ratio of 1:1. Samples were cooled at +4°C for 30 minutes. Then, in the second part cryovial added cryoprotectant 2:1 at - 2°C. Thereafter, the cell suspension with a CR №2 cooled for 30 minutes at -30C overnight and cooled to -800C. Results and novelty: in accordance with the results, protocol 2 was more effective than protocol 1, all the compositions of cryoprotectants. This can be explained by a gradual temperature decrease of the cell suspension with delayed addition of a penetrating cryoprotectant DMSO. It is known that at lower temperatures down to +4°C the cells metabolic processes in the cell stops. In this state, cellular organelles, and enzyme systems are less exposed to the harmful effects of DMSO, which increases the total survival cells. Of the four compositions studied cryosolution greatest protective effect in respect of cells had a solution which includes a 200mm glucose and 5% DMSO. In the future, the results will be used to determine an effective method of ultrafast cooling and preservation of cells. Keywords: cell viability, freezing, DMSO, glucose, glycerol, ethylene glycol. Product Description popup.authors Бурлака Дмитро Петрович Дем'яненко Дмитро Петрович Сидоренко Михайло Васильвич popup.nrat_date 2020-04-02 Close