Effect of Cryoprotective Solutions on Metabolic Activity of Chlorococcum dissectum and Dunaliella salina Cell Cultures

Q4 Medicine

Problems of Cryobiology and Cryomedicine Pub Date : 2023-03-30 DOI:10.15407/cryo33.01.014

K. Vozovyk, N. Chernobai, N. Kadnikova, Nadiia Shevchenko

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Abstract

Development of protocols for cryopreservation of microalgae Chlorococcum dissectum Korshikov and Dunaliella salina Teodoresco requires selection of effective cryoprotectants and their concentrations. One of the stages of choosing the optimal concentrations of cryoprotectants is determining the degree of toxicity for cells at the equilibration stage. Solutions of dimethyl sulfoxide, ethylene glycol, ethanol, glycerol (5–30%) and modified PVS1 and PVS2 (50 and 75%) were used in the research. The effect of cryoprotective compounds was determined by the resazurin reduction test. The least damaging effect on the Ch. dissectum cells was made by a 10% solution of glycerol. Treatment of the samples with ethanol and dimethyl sulfoxide solutions reduced metabolic activity by 31–33%, the ethylene glycol ones did by 50%. Incubation in 75% modified PVS1, 50 and 75% PVS2 solutions reduced the metabolic activity by more than half compared to the control. Ethanol was the most toxic cryoprotectant for D. salina cells. Exposure of cells to the solutions of dimethyl sulfoxide, ethylene glycol and glycerol reduced metabolic activity by 25%. Incubation of D. salina cells in PVS did not affect the studied index.

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低温保护液对解剖绿球和盐藻细胞代谢活性的影响

制定微藻Chlorococcum dissectum Korshikov和Dunaliella salina Teodoresco的冷冻保存方案需要选择有效的冷冻保护剂及其浓度。选择冷冻保护剂的最佳浓度的阶段之一是在平衡阶段确定细胞的毒性程度。研究中使用了二甲基亚砜、乙二醇、乙醇、甘油（5–30%）和改性PVS1和PVS2（50%和75%）的溶液。通过雷萨祖林还原试验测定冷冻保护化合物的效果。对Ch.dissectum细胞的破坏作用最小的是10%的甘油溶液。用乙醇和二甲基亚砜溶液处理样品，代谢活性降低了31-33%，乙二醇溶液降低了50%。与对照相比，在75%改性PVS1、50%和75%PVS2溶液中孵育使代谢活性降低了一半以上。乙醇是盐藻细胞毒性最强的冷冻保护剂。细胞暴露在二甲基亚砜、乙二醇和甘油的溶液中，代谢活性降低了25%。盐藻细胞在PVS中孵育不影响所研究的指标。

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来源期刊

Problems of Cryobiology and Cryomedicine Medicine-Medicine (miscellaneous)

CiteScore

0.60

自引率

0.00%

发文量

期刊介绍： The Journal publishes the reviews and original papers on cryobiological and cryomedical research, in particular the elucidation of mechanisms of injuries occurring in biological objects and caused by the influence of low and ultra low temperatures; natural resistance of biologicals to cold and their recovery post effect; the development of effective methods of cryoprotection and technology of storage of biological resources under hypothermic and ultra low temperatures, application of hypothermia, cryotherapy and cryopreserved biologicals for treating various pathologies; cell and tissue based therapies and other issues of low-temperature biology and medicine, as well as development of devices and equipment for low temperature biology and medicine. The journal covers all topics related to low temperature biology, medicine and engineering. These include but are not limited to: low temperature storage of biologicals (human, animal or plant cells, tissues, and organs), including preparation for storage, thawing/warming, cell and tissue culturing etc. response of biologicals to low temperature; cold adaptation of animals and plants; utilisation of low temperature in medicine; experimental and clinical transplantation, cell and tissue based therapies; developing of cryobiological and cryomedical devices; organisation and functioning of low temperature banks etc.