Cryopreservation of Cord Blood Nucleated Cells Using Non-Penetrating Cryoprotectant PEO-1500

Lyubov A. Babijchuk, Olga A. Mykhailova, Vladimir V. Ryazantsev, Pavel M. Zubov, Regina K. Migunova

Abstract


Development of wash-free cryopreservation methods for cord blood nucleated cell fraction, including the population of hematopoietic stem/progenitor cells, is an actual task for their long-term storage. Present investigation involved the technique of cord blood nucleated cells cryopreservation, comprising the isolation of cell population by the original two-step centrifugation method, the treatment with non-penetrating cryoprotectant polyethylene oxide with molecular weight of 1500 (PEO-1500) and freezing by the two-step program. This technique enabled to preserve more than 72% CD45+ and 75% CD34+ cells with the viability indices of correspondingly 83 and 91% and higher. Herewith, an increased content of reactive oxygen species (42.7 ± 6.3%) on the background of high cell number and their viability may be considered as a cells’ physiological response to the effect of cryopreservation factors.


Probl Cryobiol Cryomed 2016; 26(1):24-34.


Keywords


hematopoietic stem/progenitor cells; nucleated cells; cord blood; cryopreservation; non-penetrating cryoprotectant; PEO-1500

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References


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DOI: https://doi.org/10.15407/cryo26.01.024

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