The Production of Reactive Oxygen Species in Human Erythrocytes during Cryopreservation with Glycerol and Polyethylene Glycol

Abstract

Abstract—The production of reactive oxygen species in human erythrocytes during incubation in the presence of a glycerol–mannitol mixture and polyethylene glycol with a molecular weight of 1500 Da, as well as during hypothermic storage of cells cryopreserved under their protection, was studied. Analysis of the production of reactive oxygen species was carried out by flow cytometry using fluorescence indices of dichlorofluorescein. It was found that incubation in the glycerol–mannitol mixture, as well as cryopreservation and subsequent hypothermic storage, did not cause activation of the production of reactive oxygen species in erythrocytes. Polyethylene glycol caused the intensification of the production of reactive oxygen species both during incubation and storage of cryopreserved erythrocytes under hypothermia. Peculiarities of changes in the intensity of the production of reactive oxygen species in erythrocytes under the influence of cryoprotective agents and cryopreservation may have a significant impact on the stability of cells under stress and their ability to maintain functional integrity after returning to physiological conditions.