Effect of various concentrations of the cryoprotectant DMSO on safety, viability and content of reactive oxygen species in cord blood nucleated cells during cryopreservation
Keywords:
cord blood, nucleated cells, cryopreservation, reactive oxygen species
Abstract
The paper presents the results of the study of development of approaches to assess the number of cord blood nucleated cells (NC) with excessive reactive oxygen species (ROS) content with the fluorescent dye 2',7'-dihlordigidrofluorestsein diacetate by flow cytometry. Both exposure of NC for one hour with various concentrations of penetrating cryoprotectant DMSO and their cryopreservation resulted in reduced cell safety and viability that may be caused by inhibition of cell antioxidant system with the following accumulation of excessive content of ROS in cells. The results obtained showed the necessity of the addition of antioxidants into cryoprotective medium to prevent oxidative stress inside cells for improving cryopreservation techniques.
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References
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Khomenko V.I., Bychkov V.V., Bazyka D.A. State of development of hematopoietic stem cell transplantation in the Europe and world // Lik. Sprava. – 2014. – Vol. 7–8. – P. 117–121.
Kim K.M., Huh J.Y., Hong S.S., Kang M.S. Assessment of cell viability, early apoptosis, and hematopoietic potential in umbilical cord blood units after storage // Transfusion. – 2015. – Vol.55 (8). – P. 2017–2022.
Passweg J.R., Baldomero H., Peters C. et al. Hematopoietic SCT in Europe: data and trends in 2012 with special consideration of pediatric transplantation // Bone Marrow Transplant. – 2014. – Vol.49, no 6. – Р. 744–750.
Rhee S.G., Chang T.S., Jeong W., Kang D. Methods for detection and measurement of hydrogen peroxide inside and outside of cells // Mol. Cells. – 2010. – Vol.29, no 6. – Р. 539–549.
Schmid I., Krall W.J., Uittenbogaart C.H. Dead cell discrimination with 7-amino-actinomycin D in combination with dual color laser flow cytometry // Cytometry. – 1992. – Vol.13. – P. 204–208.
Schieber M., Chandel N.S. ROS function in redox signaling and oxidative stress // Curr. Biol. – 2014. – Vol.24 (10). – P. 453–462.
Shen W.J., Hsieh C.Y., Chen C.L. et al. A modified fixed staining method for the simultaneous measurement of reactive oxygen species and oxidative responses // Biochem. Biophys. Res. Commun. – 2013. – Vol.430, no 1. – P. 442–447.
Soh N. Recent advances in fluorescent probes for the detection of reactive oxygen species // Anal. Bioanal. Chem. – 2006. – Vol.386. – Р. 532–543.
Tetz L.M., Kamau P.W., Cheng A.A. et al. Troubleshooting the dichlorofluorescein assay to avoid artifacts in measurement of toxicant-stimulated cellular production of reactive oxidant species // J. Pharmacol. Toxicol. Methods. – 2013. – Vol.67, no 2. – Р. 56–60.
Yamaguchi R., Takanashi M., Ito M. et al. Plasticizer concentration in cord blood cryopreserved with DMSO // Bone Marrow Transplant. – 2014. – Vol.49 (1). – P. 157–158.
Wardman P. Fluorescent and luminescent probes for measurement of oxidative and nitrosative species in cells and tissues: progress, pitfalls, and prospects // Free Radic. Biol. Med. – 2007. – Vol.43. – Р. 995–1022.
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How to Cite
Макашова, О. Є., Бабійчук, Л. О., Зубова, О. Л., & Зубов, П. М. (1). Effect of various concentrations of the cryoprotectant DMSO on safety, viability and content of reactive oxygen species in cord blood nucleated cells during cryopreservation. The Journal of V.N.Karazin Kharkiv National University. Series «Biology», 26, 157-164. Retrieved from https://periodicals.karazin.ua/biology/article/view/6510
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CRYOBIOLOGY
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