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.
Highlights
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
The results of the first successful transplantation of human cord blood hematopoietic stem/progenitor cells (HSCs), having a high proliferative and expansion potential, as well as lower immunogenicity comparing with the analogues from either adult bone marrow or mobilized peripheral blood [10, 21] were the preconditions for their use as the source of hematopoietic cells in many countries [11, 19, 20] and establishing the banks for long-term storage of preparations in a frozen state
The two-step centrifugation (NC-1) enabled to isolate more than 90% of both NCs and HSCs, if using Ficoll (NC-2) a loss of CD45+ cells ((45.2 ± 8.2)%) was observed, CD34+ cells amount was decreased ((35.9 ± 5.1)%) [14]. These changes may affect the quality of the resulting preparation, since the number of nucleated cells is the standard criterion to evaluate a cell preparation quality [18]
Summary
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. The results of the first successful transplantation of human cord blood hematopoietic stem/progenitor cells (HSCs), having a high proliferative and expansion potential, as well as lower immunogenicity comparing with the analogues from either adult bone marrow or mobilized peripheral blood [10, 21] were the preconditions for their use as the source of hematopoietic cells in many countries [11, 19, 20] and establishing the banks for long-term storage of preparations in a frozen state The functioning of these banks is based. Процедура криоконсервирования фракции ядросодержащих клеток (ЯСК), в том числе и ГСК, имеет несколько технологических этапов: выделение, обработка криопротекторными веществами и замораживание-отогрев
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