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Validation of the single-platform ISHAGE protocol for enumeration of CD34+ hematopoietic stem cells in umbilical cord blood in a Brazilian center
Morais, Carla Cristina Pedrosa de Lira de; Pinto, Juliana Dias Alves; Souza, Karen Wagner de; Izu, Marina; Bouzas, Luis Fernando da Silva; Braga, Flávio Henrique Paraguassú.
  • Morais, Carla Cristina Pedrosa de Lira de; Instituto Nacional de Câncer - INCA. Rio de Janeiro. BR
  • Pinto, Juliana Dias Alves; Instituto Nacional de Câncer - INCA. Rio de Janeiro. BR
  • Souza, Karen Wagner de; Instituto Nacional de Câncer - INCA. Rio de Janeiro. BR
  • Izu, Marina; Instituto Nacional de Câncer - INCA. Rio de Janeiro. BR
  • Bouzas, Luis Fernando da Silva; Instituto Nacional de Câncer - INCA. Rio de Janeiro. BR
  • Braga, Flávio Henrique Paraguassú; Instituto Nacional de Câncer - INCA. Rio de Janeiro. BR
Hematol., Transfus. Cell Ther. (Impr.) ; 44(1): 49-55, Jan.-Mar. 2022. tab, graf
Article in English | LILACS | ID: biblio-1364889
ABSTRACT
Abstract 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 AccuriTM 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 AccuriTM C6, following the protocol established for each kit, the number of CD34+ cells was recalculated, considering the cell count provided by the AccuriTM C6. Main Results In our analysis, the results showed a strong correlation between the number of CD34+ cells/μL (r2 = 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 (r2 = 0.64). Conclusion Our results showed that the AccuriTM 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.
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Full text: Available Index: LILACS (Americas) Main subject: Hematopoietic Stem Cells / Antigens, CD34 / Fetal Blood Type of study: Practice guideline Country/Region as subject: South America / Brazil Language: English Journal: Hematol., Transfus. Cell Ther. (Impr.) Journal subject: Hematologia / TransfusÆo de Sangue Year: 2022 Type: Article Affiliation country: Brazil Institution/Affiliation country: Instituto Nacional de Câncer - INCA/BR

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Full text: Available Index: LILACS (Americas) Main subject: Hematopoietic Stem Cells / Antigens, CD34 / Fetal Blood Type of study: Practice guideline Country/Region as subject: South America / Brazil Language: English Journal: Hematol., Transfus. Cell Ther. (Impr.) Journal subject: Hematologia / TransfusÆo de Sangue Year: 2022 Type: Article Affiliation country: Brazil Institution/Affiliation country: Instituto Nacional de Câncer - INCA/BR