Validation of the single-platform ISHAGE protocol for enumeration of CD34+ hematopoietic stem cells in umbilical cord blood in a Brazilian center.
| Autor: | Pedrosa de Lira de Morais CC; Instituto Nacional de Câncer (INCA), Rio de Janeiro, RJ, Brazil. Electronic address: clira@inca.gov.br., Dias Alves Pinto J; Instituto Nacional de Câncer (INCA), Rio de Janeiro, RJ, Brazil., Wagner de Souza K; Instituto Nacional de Câncer (INCA), Rio de Janeiro, RJ, Brazil., Izu M; Instituto Nacional de Câncer (INCA), Rio de Janeiro, RJ, Brazil., Fernando da Silva Bouzas L; Instituto Nacional de Câncer (INCA), Rio de Janeiro, RJ, Brazil., Henrique Paraguassú-Braga F; Instituto Nacional de Câncer (INCA), Rio de Janeiro, RJ, Brazil. |
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| Jazyk: | angličtina |
| Zdroj: | Hematology, transfusion and cell therapy [Hematol Transfus Cell Ther] 2022 Jan-Mar; Vol. 44 (1), pp. 49-55. Date of Electronic Publication: 2020 Dec 04. |
| DOI: | 10.1016/j.htct.2020.09.151 |
| Abstrakt: | Background: This study aims to validate the single-platform method for enumeration of CD34+ cells, by comparing the performance of two different commercial kits, as well as to evaluate the efficiency of the Accuri TM C6 cytometer in providing direct counts of absolute cell numbers. Method: We evaluated 20 samples from umbilical cord blood (UCB), comparing the two different methodologies for enumeration of CD34+ cells: single and dual-platform. For the assessment of the single-platform, Procount and SCE kits were used, both of which use fluorescent beads as a counting reference to obtain absolute CD34+ cells numbers. Moreover, after the acquisition of samples in flow cytometer Accuri TM C6, following the protocol established for each kit, the number of CD34+ cells was recalculated, considering the cell count provided by the Accuri TM C6. Main Results: In our analysis, the results showed a strong correlation between the number of CD34+ cells/μL (r 2 =0.77) when comparing the SCE kit and the current dual-platform method. On the other hand, the comparison between Procount kit and dual-platform results showed a moderate correlation for the number of CD34+/μL cells (r 2 =0.64). Conclusion: Our results showed that the Accuri TM C6 flow cytometer can be used safely, applying both the dual and single platform analysis strategy. Considering the ISHAGE protocol-based single-platform approach, as the most appropriate methodology for CD34+ cells enumeration, our results demonstrated that the SCE kit has great potential for national standardization of UCB samples analysis methodology. (Copyright © 2020 Associação Brasileira de Hematologia, Hemoterapia e Terapia Celular. Published by Elsevier España, S.L.U. All rights reserved.) |
| Databáze: | MEDLINE |
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