Trolox Antioxidant as a Factor in Stabilization of Human Cord Blood Nucleated Cells During Cryopreservation

Q4 Medicine

Problems of Cryobiology and Cryomedicine Pub Date : 2023-06-30 DOI:10.15407/cryo33.02.122

Pavlo Zubov, Oksana Zubova, Lyubov Babijchuk

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引用次数: 0

Abstract

The paper presents experimental data on the determination of preservation rate of human cord blood nucleated cells and their apoptosis/necrosis stages to determine the number of viable functionally active cells after cryopreservation in solutions with different concentrations of DMSO and the water-soluble analogue of vitamin E, the antioxidant trolox. Counting the number of human cord blood nucleated cells after freezing in media with the addition of trolox revealed their maximum preservation in the samples with 7.5% DMSO and 50, 70 or 200 μM of the antioxidant. Using the flow cytometry with the addition of the Annexin V FITC reagent, which specifically binds to phospholipids, and the DNA dye 7-amino-actinomycin D (7-AAD), it was established that trolox in concentrations of 50–70 μM provided an increase in the number of viable cells with intact membrane (AnnexinV─/7AAD─) by 12–16% compared to the control, which involved the use of only DMSO in the cryoprotective solution. The obtained results indicate the effectiveness of using the antioxidant trolox and the prospects of developing trolox-containing cryoprotective mixtures for freezing and long-term storage of nucleated cord blood cells, including hematopoietic progenitor cells.

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Trolox抗氧化剂在人脐带血有核细胞冷冻保存过程中的稳定作用

本文介绍了测定人脐带血有核细胞的保存率及其凋亡/坏死阶段的实验数据，以确定在不同浓度的DMSO和维生素E的水溶性类似物(抗氧化剂trolox)的溶液中冷冻保存后存活的功能活性细胞的数量。在添加了trolox的培养基中冷冻后，对脐带血有核细胞的数量进行了计数，结果表明，在7.5% DMSO和50、70、200 μM抗氧化剂的条件下，脐带血有核细胞的保存率最高。通过流式细胞术，添加特异性结合磷脂的AnnexinV FITC试剂和DNA染料7-氨基放线菌素D (7-AAD)，发现50-70 μM浓度的trolox可使具有完整膜(AnnexinV─/7AAD─)的活细胞数量比只使用DMSO的对照组增加12-16%。研究结果表明，利用抗氧化剂trolox的有效性和开发含trolox的冷冻保护混合物冷冻和长期储存有核脐带血细胞，包括造血祖细胞的前景。

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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.