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Information × Registration Number 0224U000549, 0123U102365 , R & D reports Title Cryopreservation of three-dimensional cellular structures of various degrees of heterogeneity based on physical and mathematical modeling popup.stage_title Head Sukhach Oleksandr M., д.б.н. Registration Date 09-01-2024 Organization Institute of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine popup.description2 The object of study is osmotic characteristics of homogeneous three-dimensional cellular structures (spheroids) obtained from adult mesenchymal stromal cells (MSCs), L929 cells and heterogeneous three-dimensional cellular structures (spheroids and aggregates) obtained from the initial and precultured neural cells of newborn rats. The aim of this work is to determine the optimal parameters of cryopreservation (incubation time in a cryoprotective environment and cooling rate) of three-dimensional cellular structures of different degrees of heterogeneity (spheroids and aggregates) based on physical and mathematical modeling of mass transfer processes and experimental data. It was found that the formation of both homogeneous (L929 cells or MSCs) and heterogeneous (neural cells) spheroids in a droplet or on a non-adhesive surface occurs within 3-5 days of cultivation. When forming spheroids from MSCs in a droplet, the optimal cell concentration is 3×103 - 5×103 cells/droplet in a volume of 30 μL, and when forming spheroids from neural cells, the optimal concentration is 12×103 - 20×103 cells/droplet in a volume of 20 μL. According to the calculations of physical and mathematical modeling, it was determined that the optimal mode of cryopreservation of spheroids from L929 cells is cooling in the presence of 10% DMSO at a rate of 1.5-2 degrees/min to -80 °C followed by immersion in liquid nitrogen, and from human MSCs - cooling at a rate of 0.75 degrees/min in the presence of 7.5% DMSO to -40 °C followed by immersion in liquid nitrogen. It was also found that the optimal time of equilibration with DMSO for neuronal cells spheroids is: at 5 °C - (146±20) s; at 15 °C - (66±9) s; at 25 °C - (60±6) s; at 35 °C - (18±5) s. The optimal cooling rate in the presence of 10% DMSO of heterogeneous spheroids of neural cells is in the range of 0.75÷1 deg/min. Product Description popup.authors Vsevolodska Svitlana V. Kovalenko Igor F. Kovalenko Svitlana Ye. Ochenashko Olga V. Sydorenko Olga S. Trufanov Oleg V. popup.nrat_date 2024-01-09 Close