Glutathione Antioxidant Increases Resistance of Cord Blood Nucleated Cells During Cryopreservation with Dimethyl Sulfoxide

Authors

  • Olena Makashova Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkiv
  • Olga Mykhailova Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkiv
  • Oksana Zubova Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkiv
  • Pavlo Zubov Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkiv
  • Lubov Babijchuk Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkiv

DOI:

https://doi.org/10.15407/cryo30.01.058

Keywords:

human cord blood, nucleated cells, cryopreservation, dimethyl sulfoxide, reactive oxygen species, glutathione

Abstract

Abstract: The cryopreservation effi ciency of nucleated cells (NCs), in particular hematopoietic progenitor (HPCs) and cord blood (CB) cells was comprehensively assessed in protective mixtures, containing intracellular cryoprotectant dimethyl sulfoxide (DMSO) and antioxidant glutathione in different concentrations assessed immediately after thawing and further transfer to the in vitro conditions, simulating physiological ones. The use of 1 and 3 mM glutathione in cryoprotectant medium with 7.5 and 10% DMSO enabled preserving 85.9 and 91.2% of viable NCs and HPCs, respectively, immediately after thawing and 75–80% of viable NCs/HPCs after 1-hour incubation in the antioxidant-free Hank’s solution, which exceeded the control values. At the same time, the recovery and viability of NCs and HPCs after cryopreservation in 5% DMSO with 1mM or 3 mM glutathione were the same or even higher than those in 7.5 and 10% DMSO without glutathione. The NCs cryopreservation in the glutathione-containing solutions was esta-blished to reduce the number of DCF+ cells. The supplementation of 7.5% DMSO with 1 or 3 mM glutathione reduced the cell number with excess reactive oxygen species from 19% (in glutathione-free samples) to 11% (in glutathione-contained ones), even after transfer to the in vitro conditions, simulating physiological.

 

Probl Cryobiol Cryomed 2020; 30(1): 058–067

Author Biographies

Olena Makashova, Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkiv

Department of Cryocytology

Olga Mykhailova, Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkiv

Department of Cryocytology

Oksana Zubova, Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkiv

Department of Cryocytology

Pavlo Zubov, Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkiv

Department of Cryocytology

Lubov Babijchuk, Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkiv

Department of Cryocytology

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Published

2020-03-28

How to Cite

Makashova, O., Mykhailova, O., Zubova, O., Zubov, P., & Babijchuk, L. (2020). Glutathione Antioxidant Increases Resistance of Cord Blood Nucleated Cells During Cryopreservation with Dimethyl Sulfoxide. Problems of Cryobiology and Cryomedicine, 30(1), 58-67. https://doi.org/10.15407/cryo30.01.058

Issue

Section

Theoretical and Experimental Cryobiology