• Transfus Apher Sci · Apr 2019

    Thawing methods do not affect cell viability of CD45+ and CD34+ cells, but long-term cryopreservation of umbilical cord blood units generally decreases cell viability.

    • Jisela Dimas-González, Adán Nieto-Linares, Miriam Millán-Rocha, José Luis Salazar-Bailón, Bardo Abraham Lorenzo-Moreno, and Julieta Rojo-Medina.
    • Centro Nacional de la Transfusión Sanguínea, Av. Othón de Mendizábal 195, Zacatenco, Gustavo A. Madero, 07360, Mexico City, Mexico.. Electronic address: jisela.dimas@salud.gob.mx.
    • Transfus Apher Sci. 2019 Apr 1; 58 (2): 196-200.

    IntroductionUmbilical cord blood units (UCBUs) are collected and cryopreserved in biobanks for a future transplant. Hematopoietic stem cells and hematopoietic progenitor cells (HSC/HPC) present in UCB can be damaged due to factors such as the cryopreservation process, the thawing process, and prolonged storage time.MethodsUCBUs (n = 27) were obtained from the Biobank of the National Center of Blood Transfusion (NCBT) from Mexico. They contained three attached segments of UCBU, including 1.0-2.3 × 106 CD34+ cells prior to cryopreservation and were stored during the period from 2003 to 2015. Each UCB segment was thawed with three different methods and CD34 cells, CD45 cells, and 7-AAD were identified by flow cytometry. Furthermore, we carried out CFU assays, and trypan blue staining.ResultsViable CD45+ (vCD45+) cells, vCD34+ cells, CFU, and percentage of E-Clone were not statistically significant among three different thawing methods. The number of vCD45+ and vCD34+ cells diminished in the three thawing methods compared with the same cells prior to their cryopreservation (p < 0.0001). vCD45+ and vCD34+ cells diminished in the ≥10-year cryopreservation group (p < 0.001). In addition, percentage of recovery of vCD45+ and vCD34+ cells diminished in this same group (p =  0.013 and p < 0.0001, respectively).ConclusionThe thawing methods did not affect either cell viability (vCD45+ and vCD34+ cells) or pluripotency (CFU, percentage of E-Clone) in attached segments of UCBUs. The ≥10-year cryopreservation time in attached segments of UCBUs alters the number of vCD45+ and vCD34+ cells; however, it does not affect their pluripotency.Copyright © 2019 Elsevier Ltd. All rights reserved.

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